KR20190005981A - Skin freezing system to treat acne and skin condition - Google Patents

Abstract

Methods and systems in accordance with certain embodiments of the art include applying a substance to the skin of a human subject. The applicator is then applied to the object to cool the area of the object. After cooling the tissue, nucleation initiators are used to initiate freezing events in the tissue. The nucleation initiator may be an ice crystal that is inoculated into the skin upon contact to produce a predictable freeze event therein. The contact time between the ice crystals and the skin can be controlled to achieve the desired effect.

Description

Skin freezing system to treat acne and skin condition

Cross-reference to related application

This international application is a continuation-in-part of US Provisional Patent Application No. 62 / 334,213, filed May 10, 2016; U.S. Provisional Patent Application No. 62 / 334,317, filed May 10, 2016; U.S. Provisional Patent Application No. 62 / 334,330, filed May 10, 2016; And U.S. Provisional Patent Application No. 62 / 334,337, filed May 10, 2016, the contents of which are incorporated herein by reference; The full text of which is incorporated herein by reference.

Include by quotation of literature

The following U.S. patent applications and U. S. patents are hereby incorporated by reference in their entirety:

U.S. Patent No. 7,854,754 entitled " Cooling Apparatus for Removing Heat from Subcutaneous Lipid-Rich Cells ";

U.S. Patent No. 8,337,539 entitled " Cooling Apparatus for Removing Heat from Subcutaneous Lipid-Rich Cells ";

U.S. Patent Publication No. 2013/0158636 entitled " Cooling Device for Removing Heat from Subcutaneous Lipid-Rich Cells ";

U.S. Patent No. 8,192,474 entitled " Method of Treating Tissue ";

U.S. Patent Publication No. 2013/0066309 entitled " Tissue Treatment Method ";

U.S. Patent Publication No. 2015/0328077 entitled " Tissue Treatment Method ";

U.S. Patent No. 9,132,031 entitled " Cooling Apparatus Having Multiple Controllable Cooling Elements to Provide a Predefined Cooling Profile ";

U.S. Patent No. 9,375,345 entitled " Cooling Device Having Multiple Controllable Cooling Elements to Provide a Predefined Cooling Profile ";

U.S. Publication No. 2015/0342780 entitled " Cooling Device Having Multiple Controllable Cooling Elements to Provide a Predefined Cooling Profile ";

U.S. Patent Publication No. 2008/0077201 entitled " Cooling Device With Flexible Sensor ";

U.S. Patent Publication No. 2007/0255362 entitled " Cryoprotectant for use in therapeutic devices for improved cooling of subcutaneous lipid-rich cells ";

U.S. Patent Publication No. 2014/0005760 entitled " Cryoprotectant for use in therapeutic devices for improved cooling of subcutaneous lipid-rich cells ";

U.S. Patent Publication No. 2007/0270925 entitled " Method and Apparatus for Non-invasively Removing Heat from Subcutaneous Lipid-Rich Cells Including Coolant with Phase Transition Temperature ";

U.S. Patent Publication No. 2009/0118722 entitled " Method and Apparatus for Cooling Subcutaneous Lipid-Rich Cells or Tissues ";

U.S. Patent Publication No. 2008/0287839 entitled " Therapeutic Apparatus with Enhanced Heat Removal Method and Actuator from Subcutaneous Lipid-Rich Cells ";

U.S. Patent Publication No. 2013/0079684 entitled " Therapeutic Apparatus with Improved Heat Removal Method and Actuator from Subcutaneous Lipid-Rich Cells ";

U.S. Patent No. 8,285,390 entitled " Cooling of Subcutaneous Lipid-Rich Cells, e.g. Monitoring of Cooling of Adipose Tissue ";

U.S. Patent No. 9,408,745 entitled " Cooling Subcutaneous Lipid-Rich Cells, e.g. Monitoring of Cooling of Adipose Tissue ";

U.S. Patent Publication No. 2013/0116758 entitled " Cooling of subcutaneous lipid-rich cells, e.g. monitoring of cooling of adipose tissue ";

U.S. Patent No. 8,523,927 entitled " System for Treating Lipid-Rich Zone ";

U.S. Patent Publication No. 2014/0067025 entitled " System for Treating Lipid-Rich Zone ";

U.S. Patent Publication No. 2009/0018624 entitled " Limited Use of Disposable System Patient Protection Devices ";

U.S. Patent Publication No. 2009/0018625 entitled " Management of System Temperature to Remove Heat from Lipid-Rich Zone ";

U.S. Patent Publication No. 2009/0018626 entitled " User Interface for a System to Remove Heat from a Lipid-Rich Zone ";

U.S. Patent Publication No. 2009/0018627 entitled " A Safety System for Removing Heat from a Lipid-Rich Zone ";

U.S. Patent No. 8,275,442 entitled " Treatment Planning System and Method for Body Contouring Use ";

U.S. Patent Publication No. 2013/0158440 entitled " Treatment Planning System and Method for Body Contouring Use ";

U.S. Patent Application No. 12 / 275,002 entitled " Apparatus Having Hydrophilic Storage to Cool Subcutaneous Lipid-Rich Cells ";

U.S. Patent Application No. 12 / 275,014 entitled " Apparatus with a hydrophobic filter for removing heat from subcutaneous lipid-rich cells ";

U.S. Patent No. 8,676,338 entitled " Multiple Contour Therapy System, Method and Apparatus for Body Contouring Applications ";

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U.S. Patent Publication No. 2014/0257443 entitled " Composition for use in a system for improved cooling of subcutaneous lipid-rich tissue ";

U.S. Publication No. 2012/0239123 entitled " Apparatus, Application System and Method with Localized Thermal Zone for Removing Heat from Subcutaneous Lipid-Rich Cells ";

U. S. Patent No. 6,041, 787 entitled " Use of Freezing Aid Compounds During Cryosurgery ";

U. S. Patent No. 6,032, 675 entitled " Freeze Method for Controlled Removal of Adipose Tissue by Liposuction ";

U.S. Patent No. 9,314,368 entitled " Household Applicators and Related Devices, Systems and Methods for Non-invasive Removal of Heat from Subcutaneous Lipid-Rich Cells via Phase Change Cooling Water ";

U.S. Publication No. 2011/0238051 entitled " Household Applicators and Related Devices, Systems and Methods for Non-invasive Removal of Heat from Subcutaneous Lipid-Rich Cells Through Phase Change Cooling Water ";

U.S. Patent No. 9,545,523 entitled " Multiple-aquaculture System, Method and Apparatus for Altering Subcutaneous Lipid-Rich Tissue ";

U.S. Patent Publication No. 2014/0277302 entitled " Therapeutic System With Fluid Mixing System And Fluid-Cooled Applicator And Method Of Using It ";

U.S. Patent Publication No. 2015/0216720 entitled " Therapeutic System, Method, and Apparatus for Improving the Appearance of the Skin and Providing Other Therapies ";

U.S. Patent Publication No. 2015/0216816 entitled " Composition, Treatment System and Method for Improved Cooling of Lipid-Rich Tissue ";

U.S. Patent Publication No. 2015/0216719 entitled " Therapeutic System and Method for Treating Cellulite and Providing Other Therapies ";

U.S. Patent Application No. 14 / 662,181 entitled " Therapeutic System, Device, and Method for Cooling Targeted Tissue ";

U.S. Patent Application No. 14 / 710,407 entitled " Therapeutic System With Adjustable Gap Applicator and Method For Cooling Tissue ";

U.S. Patent Publication No. 2016/0054101 entitled " Therapeutic System, Substitute Applicator, and Method for Treating Submandibular Tissue ";

U.S. Patent Publication No. 2016/0051308 entitled " Stress Relief Coupling for Cryotherapy Devices "

U.S. Patent Publication No. 2016/0089550 entitled " Therapeutic System, Method, and Apparatus for Changing Skin Appearance ";

U.S. Patent Publication No. 2017/0007309 entitled " Therapeutic System and Method for Affecting GLAND and Other Targeted Structures ";

U.S. Patent Publication No. 2016/0317346 entitled " System and Method for Monitoring Skin and Tissue Cooling to Identify Frozen Events ";

U.S. Patent Application No. 15 / 271,121 entitled " Percutaneous Therapeutic System, Cooling Device, and Method for Cooling the Nerve ";

U.S. Patent Application No. 15 / 296,853 entitled " Vascular Therapy System, Cooling Device, and Method for Cooling Vascular Structures ";

U.S. Patent Application No. 15 / 400,885 entitled " Temperature-Dependent Adhesion Between Applicator and Skin During Tissue Cooling "; And

U.S. Provisional Patent Application No. 62 / 297,054 entitled " Cooling Cup Applicator with Contoured Head and Liner Assembly ".

Technical field

The present invention generally relates to a system for cooling tissue. In particular, some embodiments are directed to treatment systems, methods, and materials for the controlled cooling of tissue to treat acne or other diseases.

The exocrine glands found in the skin play a role in maintaining skin health, including lubrication, waterproofing, cleansing and / or cooling of the body's skin or hair follicles by discharging aqueous, oily and / or waxy materials through the skin pores or hair follicles . Excessive production and / or excess secretion of such substances by certain exocrine glands such as sebaceous glands and foot glands (e.g., sweat glands) can cause unacceptable skin disorders that have proved difficult to treat. For example, the overproduction of sebum, a waxy substance produced and secreted by sebaceous glands, is associated with the formation of cotton (e.g., blackheads, whiteheads) and other inflammatory diseases of the skin associated with acne (e.g., Inflammatory papules, pustules, nodules, etc.), which can potentially cause scarring of the skin. Overexpression of sebaceous glands associated with hair follicles is mainly found in very visible areas of the body, for example along the face, neck, upper chest, shoulders and back.

Hyperhidrosis is a condition associated with excessive perspiration caused by overproduction and secretion of sweat from sweat glands in the skin of mammals. Excessive sweating from the eccrine sweat glands distributed over almost the entire body can cause discomfort and embarrassment. For example, focal hyperhydrosis can occur in the palms, soles, face, and scalp. Apocrine sweat glands, especially apocrine glands in the armpit (i.e., axillae), have greasy-producing cells that can contribute to undesirable odors.

Treatment of these and other skin and tissue disorders is often ineffective and has non-persistent / non-persistent or undesirable side effects.

Many aspects of the invention may be better understood with reference to the following drawings. Like reference numerals designate like elements or acts. It is not necessary to make the size and the relative position of the elements in the figure necessarily proportional to each other.
1 is a schematic cross-sectional view of the skin, dermis, and subcutaneous tissue of a subject.
Figure 2 is a schematic cross-sectional view of the skin, dermis, and subcutaneous tissue of the subject of Figure 1 after treatment of sebaceous glands.
3 is a partial schematic diagram of a treatment system for non-invasively treating a targeted structure in the body of a human subject according to an embodiment of the technique.
Figure 4 is a cross-sectional view of the conduit of the treatment system of Figure 3;
5 is a flow chart illustrating a method for treating skin of a subject according to an embodiment of the technique.
Figure 6 is a plot of temperature versus time for treatment with minimal skin cooling or without skin cooling when the applicator is initially placed on the patient ' s skin to initiate a freezing event.
Figure 7 is a plot of temperature versus time for treatment with significant skin cooling.
Figure 8 is a flow chart illustrating a method for treating skin of a subject in accordance with an embodiment of the technique.
Figures 9a-9c illustrate steps of a method for preparing a treatment site in accordance with an embodiment of the disclosed technique.
Figure 10a shows a patterned hydrogel suitable for cryotherapy according to embodiments of the disclosed technology.
Figures 10b and 10c are plots of propylene glycol concentration versus length for the patterned hydrogel.
11A and 11B are side views of a hydrogel material having an ice nucleation region according to an embodiment of the technique.
Figure 12a shows an emulsifier or surfactant having a hydrophilic head and a hydrophobic tail.
Figure 12b shows the formulation captured by the emulsifier.
Figures 13a and 13b show oil-in-water emulsions and water-in-oil emulsions.
Figure 14 is a table with melting point / freezing point temperature for fat.
Figures 15a and 15b show test results performed on the skin in accordance with some embodiments of the disclosed technique.
Figure 16 shows the treatment cycle temperature profile and the temperature response measured by the temperature sensor at the applicator surface-tissue interface.
Figure 17 shows the temperature of an applicator surface that is lowered at a constant rate and then maintained at a generally constant value, in accordance with an embodiment of the technology.
18 is a plot of temperature versus time showing an exemplary treatment cycle for controlled supercooling and controlled freezing of tissue by lowering skin temperature, in accordance with an embodiment of the technique.
19A-19E are cross-sectional views of an applicator applied to a treatment site and thermal modeling.
Figures 20A-20F illustrate one method step for freezing tissue without supercooling.
Figure 21 is a plot of temperature versus time for freezing the skin multiple times according to embodiments of the disclosed technique.
Figure 22A shows a non-frozen liquid coupling medium that can act as an insulator for ice inoculation of the skin.
Figure 22b shows the coupling medium of Figure 22a with a shifted temperature profile.
23 is a plot of propylene glycol (PG) concentration in water versus water, in accordance with an embodiment of the disclosed technique.
24A-24F illustrate steps of a method for supercooling skin and then initiating a freezing event, in accordance with an embodiment of the disclosed technique.
Figure 25 is a plot of temperature for subcooling and freezing time versus tissue.
Figure 26 is a plot of temperature for a two-cycle cycle to sub-cool the time versus tissue and cause freezing.
27A-27C show the applicator and coupling media at various stages during the course.
28 shows an applicator applied to a treatment site in accordance with an embodiment of the disclosed technique.
Figure 29 is a plot of temperature versus time for a temperature profile to cause ice nucleation by activating the ice nucleating agent.
30 shows an applicator and an ice nucleating agent (INA) applied to a treatment site in accordance with an embodiment of the disclosed technique.
Figure 31 shows a plot of temperature versus time for delivering INA for nucleation.
32A-32D are IR images showing the steps of a process for freezing supercooled tissue using a hydrogel.
33A-33D are IR images showing tissue freezing using combined materials.
34A and 34B are cross-sectional views of an applicator applied to a treatment site in accordance with some embodiments of the disclosed technique.
Figure 35 is a plot of temperature versus time for inducing copper settlement using a hydrogel.
36 shows an applicator arranged to produce controlled freezing in a coupling material according to an embodiment of the disclosed technique.
Figure 37 shows an applicator having an outer nucleation element configured to initiate a freeze event at a location external to the applicator-hydrogel interface.
Figure 38 is a cross-sectional view of an applicator applied to a treatment site to provide energy-based activation.
Figure 39 is a plot of temperature versus time for supercooling skin before initiating a freezing event.
Figure 40 is a plot of temperature versus time when the epidermis is warmed by adjusting the temperature of the applicator after subcooling and before freezing.
Figure 41 shows a plot of temperature versus time for the cooling protocol and three cross-sections of the applicator, skin texture, and temperature distribution.
Figure 42 shows the temperature profile in the epidermis / dermis and the temperature distribution for one treatment protocol.
Figure 43 shows the temperature profile to the epidermis / dermis for one treatment protocol.
Figure 44 shows steps in one method of producing intradermal tissue freezing using an injectable material.
Figure 45 is a flow chart illustrating a method for preparing and freezing tissue according to aspects of the present technique.
46 is a schematic block diagram illustrating subcomponents of a computing device in accordance with an embodiment of the disclosure.

A. Overview

The present invention describes a treatment system for improving the appearance, function and health of a tissue and for performing other treatments. Some of the details set forth below are provided to describe the embodiments and methods in a manner sufficient to enable those skilled in the relevant arts to practice, make and use them. However, some of the details and advantages described below may not be necessary to practice the specific embodiments and methods of the technology. In addition, the techniques may include other embodiments and methods that are within the scope of the art but are not described in detail.

Various aspects of the technique may be used to create a cooling event (e. G., A partial freeze event, a complete freeze event, etc.) that affects a tissue, a cell, a structure, an accessory or a targeted feature To cooling the surface. The system disclosed herein may be used to treat a variety of conditions including, but not limited to, follicles (e. G., Exocrine glands, sebaceous glands, sweat glands, For example, the dermal layer, epidermal layer, subcutaneous layer, epidermis, dermis, subcutaneous sublayer (s), etc.) can be targeted. In some embodiments, the cooling event may reduce or limit the overproduction and / or excess secretion of the exocrine glands to prevent other inflammatory diseases of the skin associated with cotton and / or acne, such as inflammatory papules, pustules, Cure. For example, a cooling event allows an effective amount of heat injury to the fountain to reduce or limit the overproduction and / or excess secretion of such fountain, thereby reducing or eliminating acne or other skin conditions. The cooling event may include freezing the area of the dermis layer containing the targeted exocrine glands without affecting the non-targeted tissue. Therapeutic applicators may be configured for use along the face, neck, upper chest, shoulders, back, and other treatment areas and may target specific layers in the skin, subcutaneous tissue, specific structures, specific cells,

In certain embodiments, a method of predictably freezing the skin of a subject at a desired, predictable time includes lowering the temperature of the skin below the freezing point of the fluid in the skin to a first temperature. Contact the skin with ice crystals to inoculate the skin and create a predictable freezing event therein. The contact time of the ice crystals and the skin is controlled so that predictable freezing occurs at a desired time. In some processes, the method may be performed using an applicator applied to the skin of the subject.

In some embodiments, a method of freezing epidermal tissue once and freezing dermal tissue more than once includes applying a coupling medium to the applicator. The coupling medium has therein a medium capable of forming ice crystals. The temperature of the applicator is lowered below the freezing point of the medium to at least partially freeze the medium. The coupling medium of the applicator is disposed on the skin surface of the subject. The temperature of the applicator can be adjusted to the treatment temperature to freeze the dermis and epidermal tissue by freezing at least a portion of the medium in contact with the skin surface. In some procedures, the applicator is warmed in an amount that is sufficient to cause the dermal tissue to thaw but not enough to allow the epidermal tissue to thaw. After the dermis tissue is at least partially defrosted, the temperature of the applicator is adjusted to a second treatment temperature such that at least a portion of the defrosted dermis tissue is re-frozen and the epidermal tissue remains frozen.

In a further embodiment, the method of freezing the epidermis tissue only once and freezing the dermis tissue more than once may include at least partially freezing the dermis tissue and epidermal tissue using an applicator configured to cool the skin surface of the subject do. While the epidermis remains freeze, the applicator is warmed in an amount sufficient to cause at least a portion of the dermis tissue to thaw, and after dissolving at least a portion of the dermis tissue, the applicator is allowed to cool to refreeze at least a portion of the dermis tissue .

In one embodiment, a method for predictably freezing skin comprises supercooling the skin of the object using a coupling medium and an applicator at a first applicator temperature. The first applicator temperature is higher than the freezing point of the coupling medium. The coupling medium includes a coupling medium and a freezing point depressant that inhibits freezing of the skin. The skin is inoculated to freeze the skin without lowering the temperature of the applicator below the first applicator temperature.

In some additional embodiments, the method of treating the skin includes lowering the temperature of the skin of the subject below the freezing point of the target tissue of the skin. Cooling of the skin is monitored so that freezing does not occur therein. The amount of non-freezing cooling therapy delivered to the skin is controlled so that the target tissue reaches a predetermined first level of cooling. After the target tissue has reached the predetermined first level, the skin is frozen. The amount of freezing cooling therapy delivered to the skin is controlled so that the target tissue reaches a predetermined second level of cooling.

A system for treating a subject comprises an applicator configured to cool the skin surface of the subject when the applicator is applied to the subject, and a controller. The controller can be programmed to cause the applicator to create, maintain and / or maintain at least one ice crystal, for example, or to induce a freeze event in the skin of the object through at least one ice crystal.

At least some of the embodiments disclosed herein may be for cosmetically beneficial treatments. Thus, some therapeutic processes may be for a single purpose to change the treatment site to suit a cosmetically desirable appearance, texture, size, shape, or other desired cosmetic characteristic or characteristic. Thus, cosmetic procedures can be performed without providing any or minimal therapeutic effect. For example, some therapies may relate to goals such as reduction of acne, which does not include recovery of the subject's health, physical integrity, or physical well-being. In some embodiments, the method is used to treat acne, including skin irregularities, wrinkles, and sebaceous glands; Sweat glands to treat hyperhidrosis; Hair follicles to damage and remove hair; Or to target other targeted cells to alter the appearance or resolve the condition of the subject. Treatment can have therapeutic outcomes (whether intentionally or not), such as psychological benefits, changes in body hormone levels (due to a decrease in adipose tissue), and the like. Various aspects of the methods disclosed herein may include cosmetic treatment methods to achieve a cosmetically beneficial change in a portion of tissue within a target area. Such a cosmetic procedure may be administered by a non-medically trained human. (A) improve the appearance of the skin by tautening the skin, improving skin tone or texture, eliminating or reducing wrinkles, increasing skin smoothness and thickening the skin, and (b) Improve the appearance of cellulite, and / or (c) treat sebaceous glands, hair follicles, and / or sweat glands.

At least some embodiments of the technique include generating one or more controlled freeze events. The location and degree of freezing can be controlled to produce a therapeutic or cosmetic effect. The nucleation initiators, nucleation inhibitors, and / or therapeutic agents may be used before the freeze event, during the freeze event, and / or after the freeze event. The nucleation initiator may include, without limitation, an ice nucleating agent, an injectable material (e.g., saline, ice slurry, etc.), energy to promote ice nucleation, or other initiators that affect freezing . Nucleation inhibitors may, without limitation, include a cryoprotectant solution, a freezing temperature depressant, and / or a heating agent.

According to one aspect of the technique, the skin of the subject falls below its melting point / freezing point (" melting point "). The skin temperature is monitored to control the amount of non-freezing effect. Ice crystals contact the skin and cause freezing events in the skin. The skin may be monitored to control the amount of freezing treatment. The skin may also be monitored to detect any additional non-freezing, freezing, or thawing effects to accurately and predictably control the overall level of treatment. Skin preparation techniques can be used to enhance the absorption of substances into the skin by abrading and / or scrapping the epidermis. Examples of materials include thermal coupling gels, cryoprotectant solutions, and / or ice nuclei, which can be incorporated into or into hydrogel materials, liposomes, emulsions, nano-emulsions, nanoparticle mixtures or solutions, and / Forming agent. Nano-emulsions and nanoparticles may be preferred because their small size makes them suitable for absorption into the epidermis and dermis by traveling along the follicular opening and / or skin pore openings. The cryoprotectant may be used to increase the amount of non-cryoprotectant delivered prior to any freeze event, since the cryoprotectant may enable a significant subcooling of the skin before initiating the cryopreservation event. In one embodiment, the ice nucleating agent is used to reliably and predictably form ice crystals.

The applicator can predictably freeze the targeted tissue or structure by generating a freeze event that takes place in an expected manner. For example, an organization may initiate a freeze event at an expected time (e.g., within a certain time or within an expected time period), propagate freeze at a desired rate, achieve a desired degree of freeze, etc. Can be cooled. The treatment parameter may be selected based on the desired predictability of the freeze event. For example, the skin surface may be cooled to produce a freeze event in at least 80%, 85%, 90%, or 95% of the time in a typical patient. This provides predictable freezing. If no freeze event occurs, the skin may be warmed and cooled again to produce a freezing event.

One advantage of freezing is that, for a given amount of desired tissue damage, the process of creating freezing can take significantly less time than a process that does not involve freezing. This is because cell walls are damaged during freezing.

Damage to tissue due to freezing and cooling is dependent primarily on, for example, cooling rate, final temperature, holding time (non-freezing and / or freezing), and thawing rate. These variables can be controlled to achieve the desired cryoinjury for the target tissue.

Tissue damage of cellular scale is known to occur due to intracellular ice formation (IIF) and extracellular ice formation (EIF). Frozen injury due to IIF can be achieved by inducing irreversible damage to the tissue and by necrosis destroying the cell organelles and membranes from the inside. The freezing damage due to extracellular ice formation is mainly due to hyperosmolarity and dehydration of cells in extracellular space due to extracellular ice. This process results in direct cell death or prospective cell death (e. G., Cell apoptosis).

To achieve tissue damage, a sufficient final low temperature can be reached. Individual tissues and cells may have different susceptibility to cooling. As a result, the lethal temperature may be different between different components of the skin. Multiple cycles of the therapeutic temperature protocol should also increase efficacy.

The retention time in the frozen state improves the cryogenic tissue damage mechanism. As the ice crystals grow in size during the retention period, they will further enhance damage due to IIF and / or EIF.

Thawing is a destructive factor that promotes recrystallization (ie, recrystallization), that is, increasingly larger crystals, and rehydration of cells that cause membrane breakdown and cell death.

For skin, cooling can affect the blood microcirculation, which can lead to reversible or irreversible changes in blood vessels. During cooling, vasoconstriction of blood vessels can lead to the development of stasis and tissue ischemia in some temperature treatment protocols. During freezing, there may be damage to the endothelium of blood vessels and other cellular damage due to EIF and IIF. Vasoconstriction promotes hypoxia, a condition in which cells release vasodilating cytokines that improve intrathoracic vasodilation and reperfusion injury after thawing. Reperfusion also promotes tissue inflammation and vascular edema.

Also, partially or fully frozen tissue has higher thermal conductivity and lower specific heat than non-frozen tissue. As additional tissue is frozen, the thermal conductivity continues to increase and the specific heat continues to decrease. This change in thermal properties is due to the fact that even if the treatment temperature of the non-freezing treatment with supercooling is similar to the freezing treatment temperature, it is possible to achieve improved efficiency (for example, Improvement of the ship). Thus, using freezing, the penetration depth of cooling into the skin and surrounding tissues can be significantly faster than without freezing.

Some embodiments may be used to treat subcutaneous or subcutaneous skin or skin to treat wrinkles, fine lines, pores, warts, freckles, dark red nevi, and other vascular problems, , ≪ / RTI > subcutaneous, and subcutaneous tissue. Additionally or alternatively, the treatment may be used for the purpose of rejuvenating the skin, rebuilding the skin, solving skin pigmentation problems, interrupting pain, and the like, and administering a target such as an accessory organ, a cellular component, In order to influence the performance of the system. The accessory organs that can be treated include, without limitation, hair follicles, sebaceous glands, sweat glands, biceps, nerves, blood vessels, and the like. Cell components that can be treated include, without limitation, keratinocytes, keratinocytes, melanocytes, cebosites, fibroblasts, blood cells, collagen, elastin fibers and the like. The systems and methods disclosed herein are useful for treating the targets and conditions disclosed herein.

Reference throughout this specification to " one example, " " an example, " " one embodiment, Means that a particular feature, structure, or characteristic described in connection therewith is included in at least one embodiment of the present technique. Accordingly, the phrase " in one example, " " an example, " " an embodiment, an embodiment "does not necessarily all refer to the same example. The headings provided herein are for convenience only and are not intended to limit or interpret the scope or meaning of the description.

B. Treatment site

1 is a schematic cross-sectional view of the skin, dermis, and subcutaneous tissue of a subject. The skin 10 of the subject comprises a dermis 12 located between the epidermis 14 and the subcutaneous layer 16. [ The dermis 12 comprises a sebaceous gland 17 which produces sebum which is a waxy substance secreted to moisturize the skin and hair. Acne is a skin disease characterized by excessive sebum that can block hair follicles and / or pores. The level of sebum production may vary from person to person and may vary from person to person depending on the number and size of sebaceous glands. The sebum may flow along the healthy hair follicle 20 to moisturize the hair 23 and / or the skin 14. When the sebaceous glands 17 produce excess sebum, it can be collected and / or captured in the hair follicles. The overproduction and / or capture of sebum may result in the formation of cotton (e.g., blackheads, whiteheads) and other inflammatory diseases of the skin associated with acne (e.g., inflamed papules, pustules, You can. In some individuals, inflamed hair follicles and pores can become infected, and the condition can potentially cause scarring of the skin. The illustrated hair follicle 22 is clogged with excess sebum to form a rash or red spot. Other medical conditions associated with the hypersensitive sebaceum (17) include sebaceous glands, hyperplasia, and sebaceous adenomas. Diseases associated with irritable sebaceous glands that are non-medical but not cosmetic include oily and / or oily hairs (e.g., in the scalp).

Another skin disorder is hyperhidrosis. Hyperhidrosis is characterized by abnormal sweating due to high secretion levels of the glands 26. The eccrine sweat glands are controlled by the sympathetic nervous system and regulate body temperature. When an individual's body temperature rises, the eccrine sweat glands secrete sweat (i.e., water and other solutes) flowing through the tubules 28. Sweat evaporates from the surface of the skin to cool the body. The apocrine sweat glands (not shown) secrete the greasy-containing sweat into the hair follicle 20. The axilla (for example, axilla) and genital area often have a high concentration of apocrine sweat glands. Hyperhidrosis occurs when the sweat glands produce and secrete sweat at a level that is above the level needed to control body temperature. This condition can be general or specific to certain body parts (eg, palms, soles, forehead, scalp, face, armpits, etc.) May be localized (i. E., Local hyperhidrosis).

Figure 2 is a schematic cross-sectional view of the skin and a side view of a treatment device in the form of a thermo-applicator 104 ("applicator 104") applied to the skin for treating acne, hyperhidrosis, and other skin conditions by freezing the skin. The applicator 104 may controllably generate a predictable freeze event under treatment, to avoid over treatment, and / or undesirable side effects, such as damage to non-targeted tissue or structure. Freezing the skin to damage the tissue is difficult to control and can therefore often lead to either an incomplete treatment or an over treatment. This is because freezing the skin and underlying tissues is somewhat random and unpredictable. Water in biological tissue such as skin 10 tends to be in a liquid state for a period of time, even when its temperature is below its melting point / freezing point, a phenomenon referred to as " supercooling ". The terms " supercooling ", " supercooled ", and " supercool " refer to a state where the material is at a temperature below its freezing point / melting point but is still non- Lt; / RTI > It can be somewhat unpredictable if a freeze event will occur, if so, when the freeze event occurs during treatment, and how long the tissue will remain frozen. It is also often difficult to control freeze-thaw parameters such as freezing rate, target freezing temperature, duration of freeze events, and heating rate. These freeze-thaw parameters need to be controlled to achieve predictable therapeutic outcomes. It can be very difficult to control the freeze-thaw parameters and thus make it difficult to control the amount of treatment. If the amount of treatment is too great, undesirable side effects such as undesirable skin pigment changes may occur and, if too small, insufficient efficacy may result. This lack of control may also make it difficult to target certain tissues for treatment and minimize treatment of other specific non-targeted tissues.

The applicator 104 can accurately target tissue while minimizing or limiting impacts on non-targeted tissue. It has been found that if the ice crystals contact the skin 10 of the object in a supercooled state at a temperature below its phase transition temperature (e.g., melting / freezing temperature), the freezing event can be immediately induced in the skin . Thus, the ice crystals can be used to predictably control the onset of the freezing event. If a freeze event is triggered, it can spread rapidly through the volume of the subcooled tissue. The heat of fusion released during freezing can cause the bulk tissue to come out of its subcooled state, and the partially frozen skin can then prevent re-entry of the non-frozen tissue into the subcooled state. Also, the period from the onset of freeze events in fusion heat, subcooled tissue, to the point where the tissue is no longer in a state of further subcooling in some processes may be 1 second, 2 seconds, 3 seconds, 5 seconds, 10 seconds, or It may be any other suitable period. The time period for the supercooling may be determined by the target position, the volume of the targeted tissue, the subcooled tissue volume, the temperature profile, the tissue characteristics (e.g., the tissue moisture content), and / Composition, energy, etc.). Since the freezing propagation velocity can be strongly dependent on the supercooling temperature, the temperature of the subcooled tissue can be reduced or increased, respectively, to increase or decrease the freezing propagation velocity.

The applicator 104 may determine the amount of damage caused by the initial freeze event (e.g., by controlling the amount of supercooling that is generated prior to initiating the freeze event), the amount of damage (e.g., the temperature of the applicator The duration of the freeze event, and the thawing rate (e.g., the start of the thaw cycle, etc.). The timing of the freeze event can be precisely controlled by controlling the generation of ice crystals and when the ice crystals come into contact with the sub-cooled skin so that the frozen event can be " on command " A professional treatment method can be implemented to controllably and effectively treat a wide range of tissues while controlling and / or limiting damage to the tissue. The additives can also be used to manage freeze events at various optimal temperatures, so as to target tissue at various skin depths while controlling tissue damage, degree of damage to non-targeted tissue, and the like. By controlling when and how it freezes, the treatment process can target specific tissues without targeting other tissues, while controlling the therapeutic level of targeted tissue and its impact on non-targeted tissue.

Figure 2 shows that skin 10 after freeze-induced damage affects sebaceous glands 17 to reduce or limit sebum production. The skin 10 freezes, controllably disrupts or damages the sebaceous glands 17 or related structures, which can be an effective treatment for acne. Although the effect on the sebaceous fissile 17 is shown while the applicator 104 is applied to the skin 10, it can be seen that the sebaceous glands are reduced after treatment for a relatively long time (e. G., Days, Etc.) may be required. In Fig. 2, the sebum production levels of the two sebaceous spots 17 along the hair follicles 22 are significantly reduced to inhibit clogging, thereby minimizing, reducing or eliminating acne. The sweat glands 26 can also be targeted. For example, the applicator 104 may be a partial or complete freezing event that affects the sweat glands 26 and / or the gland tubules 28 in the area of the skin along the hands, underarms, or other locations with excessive sweat, - Freezing cooling events, or supercooling events. Other structures in the dermis or other layers of tissue can be targeted. Thus, cooling associated with controlled freezing produced by the applicator 104 can generally reduce / alleviate acne-associated inflammation and can be an important therapeutic pathway. Any and all such treatment pathways are encompassed by at least one of the embodiments of the techniques disclosed herein.

In some embodiments, the temperature-controlled surface 111 of the applicator 104 is cooled to form one or more layers (e. G., The dermal layer, e. G. Such as the epidermal layer, the subcutaneous layer, the epidermis, the dermis, and / or the underlying layer (s) of the subcutaneous layer). In order to treat acne, the skin surface of the subject may be treated with a composition that produces a temperature of -15 ° C, -10 ° C, -5 ° C, 0 ° C, 5 ° C, 10 ° C, 15 ° C, Freezing event or a freezing event in the freeze-freeze-freeze event. Local freeze events can be generated to affect the targeted structure while minimizing, limiting, or substantially preventing thermal damage to non-targeted tissue, structures, and the like. Materials used in the applicator 104 may include cryoprotectants, nucleating agents, liposomes, emulsions, hydrogels, combinations thereof, and the like. Mechanical energy (e.g., massaging), ultrasound energy, radio frequency (RF) energy, and / or freezing initiators can control freezing events, for example, by initiating, promoting, and / . In some processes, ultrasound energy is transferred to the subcooled tissue, causing freezing in the tissue. Radiofrequency energy can be used to isolate freezing to the target area and warm the tissue. The freeze initiator may be used to initiate freeze events in another material that causes freezing events in the tissue or ultimately freezing in the tissue. Examples of freeze initiators include, but are not limited to, one or more ice crystals, a cold probe, or a material that is rapidly frozen to produce a frozen event. The freezing event may include partially or completely freezing the cells, and / or the liquid or lipid in proximity to and / or within the structure to destroy, reduce, divide, modify, or otherwise affect the targeted features. The thermal damage can be managed by controlling the characteristics of the cooling event or freeze event. Such characteristics include, without limitation, the amount of cooling or freezing, the density and distribution of ice crystals, the freezing rate, and the like.

Cryotherapy can affect, without limitation, the function of the gland, the structure of the gland (e.g., the gland, the gland, etc.), the number of glands, the size of the glands, and / or the number and / or size of the cells . The freeze event may be maintained for a period of time long enough to drive the desired result. In some embodiments, in order to treat an exocrine gland, the skin of the subject may be destroyed, reduced, disrupted, modified, or affected by the cells or structure of the exocrine gland or anatomical features (e.g., ducts, pores, To generate a partial freezing event. The level of freezing can be controlled to limit unwanted tissue damage (e.g., to avoid excessive damage to the targeted tissue), such as damage to non-targeted tissue, excessive damage to the targeted tissue, and the like. The skin surface may be cooled or heated, either continuously or periodically, respectively, to increase or decrease the number and / or size of ice crystals in the target area. In one process, the tissue is exposed to a tissue volume that is at a steady-state temperature and a supercooled state, for example, about 1 second, 5 seconds, 10 seconds, 20 seconds, 30 seconds, 1 minute, Can be kept in a supercooled state for other time periods selected to reach the desired width, length, and depth of the cavity. Once the tissue is frozen, it may be used to reduce undesirable effects such as about 1 second, 5 seconds, 10 seconds, 20 seconds, 30 seconds, 1 minute, Or whilst remaining partially or fully frozen for other time periods selected to achieve the desired effect.

The applicator 104 may include one or more elements 167 for sensing cooling events, freeze events, subcooling, and the like. The thermal device 109 can be controlled based on the output from the device 167 for cooling the temperature-controlled surface 111 and ultimately cooling the patient's skin. Element 167 may include one or more temperature sensors, pressure sensors, detectors, combinations thereof, and the like. Alternatively, a separate sensor may be used to monitor the treatment site.

C. Treatment System

3 is a partial schematic diagram of a treatment system for non-invasively treating a targeted structure in the body of a human subject 101 according to an embodiment of the technique. The treatment system 100 may include an applicator 104, a connector 103, and a base unit 106. The applicator 104 may be applied to areas susceptible to acne to reduce the temperature of lipid-producing cells (e. G., Gland epithelial cells) in or near the sebaceous glands to reduce the amount of sebum secreted thereby, To reduce, to limit, or to limit. The applicator 104 may also cool sweat glands and associated structures to treat hyperhidrosis and perform other treatment procedures. The size and configuration of the applicator 104 may be selected based on the treatment site.

The connector 103 may be a cord that provides energy, fluid, and / or suction from the base unit 106 to the applicator 104. The base unit 106 may include a fluid chamber or reservoir 105 (illustrated by phantom lines) and a controller 114 carried by a housing 125 with a wheel 126. The base unit 106 may include a refrigeration unit, a cooling tower, a thermocouple chiller, a heater, or any other device capable of controlling the temperature of the cooling water in the fluid chamber 105, And / or may include an internal power supply 110 (shown in phantom). The power supply 110 may provide electrical energy (e. G., DC voltage) to operate the electrical components of the applicator 104. Urban waterworks (e.g., tap water) may be used in place of or in conjunction with fluid chamber 105. In some embodiments, the system 100 may include a pressurization device 117 that may provide suction and may include one or more pumps, valves, and / or regulators. The air pressure can be controlled by a regulator located between the pressurizing device 117 and the applicator 104. If the degree of vacuum is too low, the tissue may not be properly (or not at all) held in the applicator 104, and the applicator 104 may tend to move along the patient's skin. If the degree of vacuum is too high, undesirable patient discomfort and / or tissue damage may occur. The degree of vacuum can be selected based on the nature of the tissue and the level of comfort desired. In another embodiment, the applicator 104 does not use vacuum.

An operator may use the input / output device 118 of the controller 114 to control the operation of the treatment system 100. The input / output device 118 may display the operating status and treatment information of the applicator 104. In some embodiments, the controller 114 may be communicatively coupled to the applicator 104 to exchange data via a wired connection or a wireless or optical communication link, and may be, without limitation, a co-pending U.S. Patent No. 8,275,442 The treatment can be monitored and controlled based on one or more treatment profiles and / or patient-specific treatment plans as described in U.S. Pat. In some embodiments, the controller 114 may be inserted into the applicator 104 or other components of the system 100.

Upon receipt of the input and at the start of the treatment protocol, the controller 114 may cycle through each segment of the prescribed treatment plan. Segments can be designed for subcooling tissue, nucleating subcooled tissue, freezing tissue, thawing tissue, warming tissue, and the like. In doing so, the power supply 110 and the fluid chamber 105 provide power and cooling water to one or more of the functional components of the applicator 104, such as a thermoelectric cooler (e.g., a TEC " region & Cycle, and in some embodiments, activate features or modes such as vibration, massage, vacuum, and the like.

The controller 114 may receive a temperature reading from a temperature sensor that may be part of the applicator 104 or may be proximate to the applicator 104, patient's skin, patient protection device, and the like. While the target area of the body is being cooled or heated to the target temperature, it will be appreciated that the area of the body may, in fact, be close to, but not equal to, the target temperature due to, for example, the natural heating and cooling changes of the body. Thus, the system 100 may attempt to heat or cool the tissue to a target temperature or to provide a target heat flux, but the sensor may be capable of measuring a sufficiently close temperature or heat flux. If the target temperature or heat is not reached, the power may be increased or decreased to change the heat flux to selectively maintain the target temperature or " set-point " to affect the targeted tissue. The treatment site can be evaluated continuously or intermittently by monitoring various parameters. The skin may be continuously monitored to detect its temperature to determine if it is in a frozen state, in a non-frozen state, or in another state.

In some processes, the applicator 104 may achieve a level or amount of supercooling at a suitable temperature, for example, -15 캜, -10 캜, -5 캜, or 0 캜. After achieving a predetermined level of supercooling, the applicator 104 may automatically start the freezing event. The freeze event may be sensed and / or monitored using the applicator 104 or a separate device. The level of treatment may be controlled after initiation and / or completion of the freeze event. After one or more post freeze, the protocol is performed to thaw or otherwise thermally influence the tissue so that the treatment can be effectively tailored to specific targets and the non-targeted tissue is not treated or minimally treated to minimize treatment . For example, a freeze-thaw protocol can be used to suppress, limit, or substantially minimize permanent heat damage. In some embodiments, the post-freeze protocol may include slowly or rapidly warming the non-targeted tissue and the targeted tissue.

FIG. 4 is a cross-sectional view of a green connector 103 along line 4-4 of FIG. 3 in accordance with at least some embodiments of the technique. The connector 103 includes a body 179 (e.g., a solid or hollow body), a feed fluid line or lumen 180a ("feed fluid line 180a"), and a return fluid line or lumen 180b Return fluid line 180b "). ≪ / RTI > The body 179 may be configured (via one or more adjustable joints) to " install " in place for treatment of the object. The feed and return fluid lines 180a and 180b may comprise other materials capable of accommodating polyethylene, polyvinyl chloride, polyurethane, and / or circulating cooling water, such as water, glycols, synthetic heat transfer fluids, oils, / RTI > and / or any other suitable heat-conducting fluid. In one embodiment, each of the fluid lines 180a, 180b can be a flexible hose surrounded by a body 179. With reference now to Figures 3 and 4, the cooling water can be delivered continuously or intermittently to the applicator 104 through the supply fluid line 180a and can circulate through the applicator 104 to absorb heat . The cooling water that has absorbed the heat may flow from the applicator 104 back to the base unit 106 via the return fluid line 180b. During the warming period, the base unit 106 (FIG. 3) may heat the cooling water so that the warmed cooling water circulates through the applicator 104. The connector 103 also includes one or more wires 112 (Figure 4) to provide power to the applicator 104 and one or more steerable wires 116 to provide communication between the base unit 106 and the applicator 104. [ . ≪ / RTI > In order to provide the material, the connector 103 may include one or more tubes or lines 119 for delivering material by the applicator 104. The material may include a coupling medium, an INA, a solution (e. G., A cryoprotectant solution), and the like.

5 is a flow chart illustrating a method 140 for treating a skin of a subject according to an embodiment of the technique. Generally, the skin of the subject can be cooled below the freezing temperature of the fluid in the skin. One or more ice crystals can move into contact with the skin to create a predictable freezing event therein. The contact time between the ice crystals and the skin can be controlled to achieve the desired freezing. Details of method 140 are discussed below.

At block 142, the skin may be cooled to lower the temperature of the skin below the freezing temperature of the skin fluid. For example, the temperature of the skin may be lowered to a first temperature lower than the melting / freezing temperature of the skin fluid by 3 ° C, 5 ° C, 7 ° C, 9 ° C, 10 ° C, or 11 ° C or lower, Can be maintained. After the first period has expired, the skin temperature may be lowered to a second temperature lower than the first temperature to produce ice crystals. In another embodiment, the first temperature may be maintained at a constant temperature, for example, by varying the composition of the coupling medium to produce the ice crystals. The coupling medium may freeze and cause ice nucleation in the tissue.

In block 144 of FIG. 5, the ice crystals contact the skin of the subject and may be inoculated to the skin upon contact to produce a predictable freeze event therein. The ice crystals can be formed on the outside by the applicator. Alternatively, a catheter or other device can introduce the ice crystals into the object such that the ice crystals are physically in contact with the tissue and are initially frozen. In some processes, the formulation may be cooled and then diluted to produce one or more ice crystals therein. For example, the agent may include a cryoprotectant to protect the tissue. The concentration of the cryoprotectant in the formulation is such that the melting point / freezing point of the diluted preparation is at least equal to the temperature of the cryoprotectant so that the formation of ice crystals does not require the skin temperature to be lowered to a value below the melting / Lt; / RTI >

At block 146, the contact time between the ice crystals and the skin can be controlled. The user can maintain the applicator on the skin surface while the ice crystals are in contact with the skin surface. Upon completion of the contact period, the system may notify the object or the operator to remove the applicator from the object. The applicator can be detached from the object to stop the crystal contact. Alternatively, the applicator may be warmed to melt the ice crystals after the completion of the desired contact period. The temperature of the applicator is further controlled to set the length of the ice crystal / tissue contact and the length of the freeze event by further controlling when the temperature of the skin senses a freezing event and the temperature of the skin rises above the melting point of the ice crystal to terminate the freezing event Lt; / RTI >

In some treatments, the method 140 may include lowering the temperature of the skin of the subject to below the melting point / freezing point or temperature of the target tissue of the skin. The applicator 104 can monitor the cooling of the skin using a sensor so that freezing does not occur therein. The amount of non-freezing cooling delivered to the skin can be controlled so that the targeted tissue of the skin reaches a predetermined first level at block 142. After the targeted tissue reaches a predetermined first level, the skin is frozen (block 144). The sensor can be used to monitor and monitor freeze events. The ice crystals can be in intimate contact with the supercooled skin during the supercooling period and before the time required for the onset of the freezing event to occur without side effects. After the supercooling period has expired to produce a predetermined first level of supercooling, the ice crystal (s) may contact the skin and initiate a freezing event, and the damage associated with the initial freezing event is generally at a level or degree of supercooling It can be proportional. The freeze event can be maintained for any desired period of time, and after the freeze event, additional freeze events can further affect the tissue. The amount of freezing / cooling therapy delivered to the skin can be controlled so that it reaches a predetermined second level. In some treatments, ice crystals are used to cause skin freezing in a first-level, subcooled state. The predetermined second level may be selected to provide a therapeutically effective amount of thermal damage when combined with the first level.

A shallow skin treatment is a process in which the skin undergoes ice crystallization of the skin of a subject with a bulk temperature that is slightly below its melting point / freezing point, e.g., 0.2, 0.5, 1, 2, Or the like. The skin cooling rate may be minimal or insignificant so that relatively small tissue can be frozen when the initial freeze event is small (e.g., the fraction of initially frozen tissue will be small) when an initial freeze event occurs. Thus, early tissue damage can be located primarily in the epidermis, and in the upper layers of the dermis, and deeper layers such as dermis, fat and muscle tissue are largely unaffected. As such, treatment may be performed at sites susceptible to acne, where damage to the subcutaneous tissue can be problematic and undesirable. When a freezing event occurs, additional tissue in the skin will not enter the supercooled state because the ice crystals of the skin can suppress or prevent additional supercooling. Also, incremental cooling may result in predictable incremental freezing and, if minimal depth treatment is desired, the tissue thawing protocol may be initiated immediately or within the next freeze event.

The system 100 can also perform deeper treatments, including aggressive, deeper skin treatments, by subcooling the targeted tissue and then contacting the targeted tissue with ice crystals to induce freezing events. The subcooled tissue may include epidermal tissue, dermal tissue, subcutaneous tissue and may be stored for a period of time such as 30 seconds or 1, 2, 3, 4, 5, 7, 10, 12, 15, 20, 25 minutes, Can be cooled to about 4 ° C, 5 ° C, 6 ° C, 7 ° C, 8 ° C, 9 ° C, 10 ° C, 12 ° C, 15 ° C, 17 ° C, 20 ° C, 25 ° C, 30 ° C, . The temperature and duration of treatment may enable a controlled level of skin cooling, which is variable prior to initiating the freeze event. The overall level of treatment may also include multiple treatments, each of which individually delivers a dose that is less than the total dose of the treatment ultimately delivered. For example, after performing an initial skin subcooling and freezing event at a predetermined treatment site, the device software may be programmed to repeat the subcooling and freeze event cycles twice, optionally after the tissue re-warming / thawing step between cycles . The temperature and treatment duration for the second cycle may be the same as or different from that of the first cycle. Additional treatment cycles may be delivered. In this example, it may not be necessary to move the applicator between cycles, and the cycle may be arbitrarily separated by the tissue material on / off step. As a further alternative, further treatment at any given site may be performed later in the patient procedure after the applicator has treated other tissue sites. Another alternative is that the additional treatment may be delivered during a separate patient procedure that is performed on the same day as the first treatment, or on the next day or a few days or a week later, and the procedure may be periodically (e.g., Day, every other day, every week, every month, etc.). Any number of subsequent follow-on treatments can be performed to achieve sufficient overall level tissue treatment to produce the desired tissue response.

The method 140 may be used to perform the treatment described herein, such as the treatment discussed in connection with FIGS. 1 and 2. Frozen events can cause sebaceous gland disruption and can affect sebum production (e. G., Reduce or limit sebum production). The contact period (e.g., contact time between the skin of the subject and the ice crystals and / or between the skin of the subject and the cooling surface of the applicator) may be selected to achieve the desired thermal damage to the sebaceous glands. The systems, components, and operations disclosed herein may be mixed and matched as discussed in connection with Examples 1-4 below.

Example 1

The ice crystals can be formed along the applicator, using temperature programming. Water (e.g., a water droplet, a layer containing water, etc.) may be placed on the applicator surface (e.g., surface 111 of Figure 2) and the temperature of the applicator surface is about -20 & Deg.] C, -12 [deg.] C, or below a melting / freezing temperature of 0 [deg.] C, to another suitable temperature for producing one or more ice crystals. In some processes, the applicator may be prepared to have ice crystals on its outer surface to cause a skin freezing event to occur when the applicator contacts the skin surface. For example, one or more ice crystals of an applicator may physically contact the skin to initiate freezing events in the skin. In another process, the ice crystals may physically contact the coupling medium on the skin surface to cause a freezing event. When the coupling medium is frozen, this can cause freezing of the skin surface and subsequent freezing propagation through deeper tissue. In yet another process, the coupling medium can be absorbed into the skin, and the absorbed coupling medium can freeze and cause freezing of the skin.

The organization can be predictably slowed down or expedited as quickly as possible after the freeze event has occurred, to limit, reduce, or prevent damage and harmful side effects associated with the freeze event. After freezing has commenced, the skin may slowly or quickly warm up as soon as possible to minimize or limit damage to the epidermis. In another process, the skin is partially or completely frozen and then warmed for a predetermined period of time. According to one embodiment, the applicator 104 of FIG. 2 may, for example, use a thermoelectric element of the device 109 to warm the shallow tissue. The thermoelectric element can include a Peltier device that can operate to establish a desired temperature (or temperature profile) along the surface 111. In another embodiment, the applicator 104 has electrodes that output radio frequency energy to warm the tissue.

Absorption enhancers, cryoprotectants, INA, and coupling media may be delivered via liposomes, hydrogels, emulsions, and the like. Absorption enhancers can increase permeation and affect absorption, for example, water, INA, cryoprotectant, and the like. The skin may be warmed prior to or during exposure to the applied material to increase absorption to the epidermis while minimizing or restricting increased absorption into the dermis due to dermal-epidermal junction barriers. The nature of the tissue can be influenced by mechanically changing the skin of the subject. These properties may include absorption properties, thermal properties, and the like. For treatments that do not involve freezing, and only cooling or subcooling, it is desirable to increase absorption of the cryoprotectant to the skin to provide maximum protection against unintended freezing incidence. For treatment to include freezing, it is desirable to increase the absorption of INA and / or water to increase the likelihood that the freezing event is initiated and initiated at the desired time, and that the level of freezing injury is increased.

Figure 6 is a plot of applicator temperature versus time for treatment with minimal skin supercooling or involving skin cooling when the applicator is initially placed on a subject to initiate a freeze event according to an embodiment of the disclosed technique. The freezing event can be initiated upon placement of the frozen applicator surface on the skin. For example, an applicator surface (e.g., surface 111 shown in FIG. 2) may be cooled to a temperature of -15 占 C to form an ice crystal thereon. After raising the temperature of the applicator surface at a desired rate to a temperature suitable for placement on the object, the applicator surface can be applied to the treatment site. For example, the applicator surface may be heated to a temperature of about -4 ° C, -3 ° C, -2 ° C, -1 ° C or 0 ° C at a rate of 0.4 ° C / s, 0.5 ° C / s, or 0.6 ° C / . Skin surface, targeted tissue, etc. may be maintained at a temperature of about -3 ° C, -2 ° C, -1 ° C, 0 ° C, or 1 ° C.

The applicator may be configured to cool the skin from an initial temperature (e.g., 33 占 폚) to a lower temperature (e.g., -4 占 폚, -3 占 폚, -2 占 폚, -1 占 폚, (For example, 2 minutes, 2.5 minutes, 3 minutes, etc. 2.5 minutes are shown in Fig. 6). The applicator surface may then be lowered at a desired rate to a temperature to induce a freezing event. A frozen event (indicated by " * " in Figure 6) may occur with the applicator surface cooled at a rate of about 0.2 ° C / s, 0.25 ° C / s, 0.3 ° C / s, or any other desired rate. The applicator surface can be maintained at a temperature of about-8 C for a second treatment period (e.g., 20 seconds, 30 seconds, 40 seconds, etc.). The skin surface temperature may be slightly higher than the temperature of the applicator surface so that the temperature of the applicator surface can be selected such that the target tissue remains frozen during the desired freeze period.

After the completion of the freezing period, the applicator and skin temperature can be quickly raised to a normal temperature, e.g., room temperature or room temperature. In some procedures, the applicator may be warmed at a rate of about 1 DEG C / s, 2 DEG C / s, 2.5 DEG C / s, 3 DEG C / s, or other rate selected to thaw the frozen tissue. Figure 6 shows the temperature of the raised applicator at a rate of about 2.5 [deg.] C / s. The thawed tissue may comprise epidermal tissue, dermal tissue, subcutaneous tissue, and / or other tissue. After warming the tissue for heating, another cryotherapy procedure can be performed at the same or different sites using the same or different treatment parameters.

Example 2

The material can be applied to the skin, the applicator, or both, and can be used to create ice crystals. The material may be a coupling medium having one or more cryoprotectants and may be applied when the temperature is higher than its melting point, which may be below several degrees below 0 C, and at a temperature below the melting point / freezing point of the fluid in the skin tissue. The melting point / freezing point of the applied material may be in the therapeutic skin subcooling temperature range or other suitable temperature range. After a predetermined amount of skin supercooling has occurred, the temperature of the applied material can be lowered to its melting point or below the melting temperature to create ice crystals therein to initiate freezing events in the skin.

The cryoprotectant may include propylene glycol, glycerol, polyethylene glycol, combinations thereof, or other biocompatible agents. In some embodiments, the material is a cryoprotectant solution having a cryoprotectant mixed with water to provide the desired melting point / freezing point. The concentration of the cryoprotectant can be increased to lower the melting point / freezing point of the material. By controlling the concentration of the cryoprotectant, the properties of the material (e. G., Melting point, natural freezing point, etc.) can be controlled and thus at or below the desired temperature while suppressing or preventing the formation of ice crystals at / To enable the formation of ice crystals. The INA can be incorporated into the material to provide, for example, predictable initiation of a freezing event once the temperature of the material is lowered below the melting point / freezing point of the INA.

7 is a plot of applicator temperature versus time for treatment involving significant skin cooling in accordance with an embodiment of the disclosed technique. The applicator and skin are cooled (e.g., at -8 캜, -10 캜, -12 캜) at a desired rate (e.g., 0.5 캜 / s, 1 캜 / . The freezing event in the skin can be initiated after a supercooling period of about 3 minutes, 4 minutes, or 5 minutes at the supercooling temperature as exemplified by -10 占 폚. During this period, the skin surface and the cooled applicator surface may be at substantially the same temperature. The cryoprotectant coupling media can help limit thermal damage to non-targeted tissue and can be placed at the applicator-skin interface. In some embodiments, the cryoprotectant coupling media is propylene glycol (PG) at about 25 wt.% Or volume percent and about 75 wt.% Or volume percent water, and has a melting or freezing temperature of about -11 DEG C. The composition of the coupling medium can be adjusted to increase or decrease its melting point / freezing point. After a period of supercooling, the temperature of the applicator is further lowered to initiate a freezing event. Figure 7 shows that the freezing event is initiated in the skin with the applicator and skin surface cooled to about-10 캜 to about -18 캜. The level of freezing in tissue can be maintained while the applicator surface and the skin surface are maintained at a temperature of approximately -18 ° C for 10 seconds before rapid re-heating.

Ice crystals can be produced by diluting the pre-cooled coupling medium to raise its melting point / freezing point. The applied material may be a 25% by volume solution of PG cryoprotectant with a melting temperature of -11 캜. The tissue may be subcooled at the desired temperature (e.g., -8 C, -10 C, -12 C, etc.). After the desired amount of supercooling has occurred, the freeze event may be initiated by further lowering the temperature of the material applied to freeze the material to a temperature of about -18 占 폚 (e.g., "diving" the temperature). Instead of further lowering, or " diving " the temperature, cold water or another material is injected into the applied material at the pre-determined location to cause the cryoprotectant concentration to be increased by locally diluting the melting medium of the coupling medium to a level above the target freezing temperature , The freezing event may be initiated at a target freezing temperature (e. G., -10 C) or higher. The melting point / freezing point of the coupling medium may be close to -1 [deg.] C, -0.5 [deg.] C, or 0 [deg.] C, for example, to cause the ice crystals to form in the diluted material and initiate freezing events in the skin. The dilution may be made using 100% water, water doped with INA, or other material, thereby providing a consistent and predictable solution at a temperature of, for example, -1, -2, Freezing can be provided. Alternatively, the water and ice mixture or water, ice, and INA mixture can be injected to provide freezing at about the desired temperature (e.g., -1, -0, 5, and so on). This method is particularly advantageous when compared to treatment where the freezing event is initiated at a low temperature, e.g., -10 占 폚 or even lower (e.g., -18 占 폚 when "diving" is used to initiate a freeze event) (E.g., -8 ° C, -10 ° C, or -12 ° C), which can significantly reduce or prevent damage to non-targeted tissue, such as the epidermis, Can be used with significant skin subcooling to initiate freezing events at relatively warm temperatures, e.g., -1, -2, -3, -4, or -5 degrees Celsius.

Example 3

Energy can be used to manage the formation of ice crystals. If the aqueous coupling medium falls below their melting point / freezing point and is in a subcooled state, the ultrasound may be used to determine whether the ice crystals in the skin and / or the coupling medium are only slightly or significantly subcooled, A freezing event can be induced. Transmissive ultrasound can exclude INA, but ultrasound and INA can be used together. Ultrasound has been used to form ice crystals in aqueous coupling agents. For example, a dental clean ultrasonic probe operating at about 20 kHz and about 25 W forms ice crystals in the coupling agent. In another example, a non-dental ultrasound probe operating at about 20 kHz and 1 W forms ice crystals. Ultrasonic waves with different parameters can be selected based on the desired ice crystal formation and / or growth.

Example 4

After the tissue is in a subcooled state, a freezing event inducing or promoting component may be injected into or near the target site. The material may be a partially frozen ice or water slurry solution that produces an immediate freezing event. In some embodiments, the epidermis may be re-warmed to a temperature close to 0 ° C before the freezing event and the injection of saline ice water slurry into the dermis may initiate controlled freezing under the epidermis. A needle, catheter, or injection device may be introduced into the subject to inject the material. Figure 2 shows an optional catheter 149 that may be introduced into a subject. Once the end of the catheter 149 is positioned in the skin 10, the catheter 149 may deliver ice crystals, ice slurry, or any suitable material to the tissue. Catheter 149 may be used to initiate a freeze event at any number of treatment sites.

Various combinations of steps in embodiments 1-4 can be combined. Contact between the ice crystals and the tissue may be delayed until the desired level of skin cooling is achieved to enhance or maximize freezing damage in the dermis while limiting or minimizing side effects associated with freezing in the epidermis. A large amount of target skin is substantially subcooled and then contacted by the ice crystals to maximize freezing damage to the wrinkles while minimizing side effects. A large amount of previous supercooling can maximize the amount of tissue damage that occurs during the initial freeze event and allow the non-targeted tissue to re-warm to inhibit, restrict, or substantially prevent thermal damage to the non- can do. The epidermis may be a non-targeted tissue that can be re-heated immediately or soon after a freezing event in a targeted tissue such as the dermis. Heating can limit or minimize the amount of time the epidermis remains frozen. This is in contrast to treatment methods where little or no supercooling is used. In the case of a treatment method in which no such subcooling is used at all, treatment with little use of subcooling (using a significant subcooling and significant partial freezing during the initial freezing event since cooling is " top down " In order to obtain the same therapeutic level of treatment as in the case of the method, the epidermal tissue needs to be kept frozen longer after the onset of the freezing event, which may exacerbate the damage to the non-targeted epidermal tissue .

Deeper tissues can contact the skin immediately or as soon as the skin temperature falls below the melting point of the skin / freezing point, in order to substantially freeze significant damage and limit freeze damage to the upper skin layer. The limited surface skin freezing can be achieved with minimal damage to the dermis, fat and muscle layer, especially if the duration of the freezing event is kept relatively short. In some facial procedures, freezing injury can be limited to the skin to avoid any appreciable reduction of the underlying muscle forming the subcutaneous tissue or the supporting structure of the skin.

8 is a flow chart illustrating a method 150 for treating a skin of a subject according to an embodiment of the technique. Generally, a coupling medium may be applied to the treatment site. The treatment site can be cooled and a freezing event can be initiated to at least partially freeze the tissue. The initial step of method 150 may include coupling a heat-exchange surface of the applicator to the skin of the subject. The heat-exchange surface may be a temperature-exchange plate having an internal thermal element (e.g. a thermoelectric element, a fluid element, etc.) or an external thermal element (e.g. a heat element mounted on the back surface of a heat- May be a controlled surface (e.g., see surface 111 of FIG. 2). In some embodiments, the temperature-controlled surface may be an interface layer, a dielectric layer, or the like. Additionally or alternatively, a vacuum or suction force can be used to actively couple the skin of the patient to a temperature-controlled surface. Coupling the temperature-controlled surface to the skin of the subject can also include providing the material to the patient's skin as described in US Patent Application Publication No. 2007/0255362, herein incorporated by reference. Details of method 150 are discussed below.

At block 152, the treatment site can be prepared, for example, by mechanically, chemically, or otherwise altering the skin. The mechanical change can be measured by measuring the skin surface for a suitable length of time selected on the basis of about 30 seconds, 1 minute, 2 minutes, 3 minutes, or the desired amount of surface cleaning, permeation, and / or peeling (e.g., By intermittently or continuously brushing or scraping for the same period of time. In another embodiment, the permeability of the skin may be controlled by eliminating pores in the stratum corneum, or by producing and / or growing liquids (e.g., vacuoles of the epidermis below the stratum corneum), or by combining them. In some treatments, the adhesive strip may be applied to the skin and removed to remove the top layer of the epidermis, to clean the treatment site, to increase the permeability of the skin, or otherwise to prepare the treatment site. The uppermost layer of the epidermis is drier than the lower layer, so that when the uppermost layer is removed, the exposed lower layer has a greater moisture content, so that they are designed to freeze the tissue, particularly when the INA is used to promote freezing It is easier to freeze during the procedure. Permeability of the skin may also be increased using microdroplets, whereby a number of microscopic holes may be formed in the skin to create a path for absorption of the coupling medium. Alternatively, an ultrasound method may be used whereby ultrasound is used to stimulate micro-vibrations in the skin to increase the overall kinetic energy of the molecules that make up the coupling medium or topical formulation delivered into the skin to increase absorption . Some preferred frequencies are 20 to 40 kHz, or 1 MHz or more. Other frequencies may be used. Alternatively, increased absorption may be achieved using iontophoresis to push the topical formulation into the skin, for example, to increase absorption using an electric field. The permeation coefficient of the coupling medium to pass through the tissue (e.g., epidermal tissue) may be at least about 10%, 20%, 30%, 30%, 50% or more to achieve the desired absorption rate using one or more of the above- 40%, 50%, 60%, 70%, 80%, 90%, or 95%. Other techniques may be used to facilitate the delivery of the coupling medium or material. Different test techniques (e.g., stationary cell technology, flow diffusion techniques, etc.), algorithms, and modeling can be used to determine the permeation coefficient and can be used to determine the flow including the steady state flux .

At block 154, a coupling medium may be applied to the skin. The coupling medium may include, without limitation, water, hydrogel, cryoprotectant, emulsion, combinations thereof, and the like, prior to preparation for treatment. Applying the coupling medium may be accomplished using a liquid, gel, or sheet coupling medium, for example, using a device including a brush, spatula, spray bottle or syringe, or by hand (e.g., Hand) skin, spraying, covering, or rubbing.

The coupling medium may include one or more temperature depressants, INA, and the like. The temperature lowering agent may include, without limitation, polypropylene glycol (PPG), polyethylene glycol (PEG), propylene glycol, ethylene glycol, glycerol, dimethylsulfoxide (DMSO), or other glycols. The temperature lowering agent can also lower the freezing point of ethanol, propanol, isopropanol, butanol, and / or a solution (e.g., body fluid) to about 0 ° C to -40 ° C and more preferably to about -10 ° C to -16 ° C ≪ / RTI > other suitable alcohol compounds. Certain temperature lowering agents (e. G., PPG, PEG, etc.) may also be used to improve lubricity and to provide lubricity. Additionally or alternatively, the coupling medium may comprise one or more thickening agents, pH buffering agents, humectants, surfactants, and / or additives.

At block 156, the skin of the subject can be cooled. An applicator may be applied to the treatment site to position the applicator in thermal contact with the target tissue. The tissue may be subcooled and then frozen to limit or prevent undesired side effects. The skin surface of a human subject may be cooled to a temperature not lower than -40 ° C to avoid unwanted skin damage. The deeper targeted areas of the skin surface may still be subcooled and the shallow non-targeted tissue may be heated to come out of the subcooled state.

At block 158, the subcooled targeted site can be nucleated, for example, to cause freezing that can destroy or damage the targeted cells due to crystallization of intracellular and / or extracellular fluids. Catalysts for nucleation (e. G., Mechanical perturbation, RF energy, alternating electric field, etc.) can be provided after a protective increase in the temperature of the non-targeted skin layer. Mechanical perturbations can be vibrations, ultrasonic pulses, and / or pressure changes. The non-targeted layer of tissue may be warmed to a sufficient degree to avoid freezing upon nucleation of the targeted tissue. The treatment system disclosed herein may perform such a subcooling method using the applicators disclosed herein.

Some treatments involve freezing dermal tissue more often than adjacent epidermal tissue. At block 156, the dermis and epidermal tissue may be cooled and frozen (block 158). The skin may be warmed by an applicator (at a temperature slightly below 0 캜) in an amount sufficient to cause the dermal tissue to thaw due to internal heat, but not to further remove the epidermal tissue from the bloodstream than the dermal tissue. After thawing the dermis tissue, block 158 may be repeated, for example, by cooling the skin and re-freezing the dermis tissue while keeping the epidermis tissue freeze. The desired level of damage to dermal tissue can be achieved by repeatedly freezing and thawing the dermis layer because the main mechanism of damage during freezing is caused by ice nucleation and growth.

Some treatments include a warming / thawing (159) step after the freezing event (s), whereby the frozen and cooled tissue is manually re-powered on by the applicator. After the warming / thawing step 159, the cooling 156 and freezing 158 steps are preferably performed during the same patient treatment and optionally at any of 1, 2, 3, 4 or more times , As indicated by the arrow 153. [0060] Alternatively, the cooling 156 and freezing 158 steps may be performed during the same patient treatment, but after the applicator is moved to another treatment site and then back to the original treatment site, or after the first treatment date, Of the patient ' s treatment session. Any number of iterative sessions may be used to achieve an overall desired level of therapy. Arrows 157 and 155 show the possibility for treatment, which may optionally include repetition of skin preparation step 152 and / or repetition of coupling medium application step 154.

Figures 9a-9c illustrate steps of a method for preparing a treatment site in accordance with an embodiment of the disclosed technique. In general, the skin of a subject can be mechanically altered to promote absorption of the coupling medium. For example, a stripping element may be applied and removed from the skin surface at any number of times to remove the top of the epidermis to expose the underlying layer of tissue. The lower layer may have a relatively high moisture content and thus may be better absorbed and ingested into the epidermis by various agents, including aqueous or oily coupling media.

9A is a cross-sectional view of a stripping element 200 applied to the skin surface to cover the pores. The stripping element 200 may be peeled off from the adhesive strip (e. G., Adhesive tape) or skin to remove features from, for example, sebum, hair follicle, Lt; RTI ID = 0.0 > a < / RTI > The stripping element 200 may be a single adhesive strip (e.g., a piece of adhesive tape) that is cut to cover the entire treatment area. In another embodiment, multiple stripping elements 200 are applied to a treatment site. The adhesive properties of the stripping element can be selected based on the desired amount of mechanical change to the skin and the desired patient comfort.

Figure 9B is a cross-sectional view of a stripping element 200 that is removed from the skin and removes material 201 from the pores 203, thereby opening or closing the pore opening 202. The stripping elements may also be reapplied at any number of times to further pierce the pores 203 or otherwise prepare the treatment site.

9C is a cross-sectional view of the treatment site after the pores 203 are clean and the material 205 is applied. The material 205 may be injected into the pores 203 and absorbed by the skin. The water can be part of the material, and subsequent freezing events allow the water in the skin pores to freeze and cause further tissue damage.

The skin may be mechanically stimulated before, during, and / or after any step in the method 140 (FIG. 5) or 150 (FIG. 8) Mechanical stimulation may be used to clean / clean the area of, for example, brushing, rubbing, ultrasound application, skin scratches or treatment, or to temporarily reduce / reduce the barrier of the horny layer (i.e., the outermost layer of the epidermis of dead cells) Or stimulation or agitation by other means capable of increasing the movement (e. G., Turbulence) of the coupling medium to the skin. Without being bound by theory, it is believed that mechanical stimulation of the skin (e.g., agitation, reduction, or permeation of the stratum corneum) may enhance permeation of the coupling medium to the underlying epidermal, dermal, or other layers of tissue It is believed to be possible. In one embodiment, the skin may be mechanically stimulated for about 20 seconds to about 10 minutes. In another embodiment, the mechanical stimulation may be applied to the treatment site for about 20 seconds, about 40 seconds, about 1 minute, about 2 minutes, about 5 minutes, or about 5 minutes. In some embodiments, mechanical stimulation may be performed, for example, with a dermis agitation brush, a brush with short stiff hairs, and the like. Brushing or rubbing the skin may involve movement across the skin at the treatment site with a linear stroke, in some embodiments, in a circular or back and forth motion, or in another embodiment, to increase skin permeability to the material . The permeation rate can be increased or decreased to achieve the desired amount of absorption.

Different techniques can be used to evaluate the permeability of the skin before and / or after performing the stripping process. In one process, the cells of the treatment site can be progressively removed by applying the coupling agent to the treatment site and then applying the stripping element repeatedly or by applying a series of stripping elements. The stripping element and the treatment site may be evaluated to determine the volume of the coupling medium absorbed by the skin.

D. Materials for Treatment

The material can be used to improve thermal coupling between the skin surface and the cooling applicator, since ice crystals can be reliably created to cause on-command freeze events. In some embodiments, the material is an aqueous solution coupling agent containing a cryoprotectant. Additional materials may contain a medium and water to facilitate reliable production upon command of the ice crystals when initiation of freezing at the time of treatment is desired. Liquid water has clusters of molecules that undergo constant collisions with other molecules and clusters, which are sometimes broken and sometimes form new clusters. When water is cooled, as the temperature drops and the heat transfer of the water molecules decreases, the water molecules tend to agglomerate more strongly and the likelihood of forming a critically large cluster of molecules increases rapidly. Ice nucleation is catalyzed in the formation of a critically large cluster of molecules. As a result, the initiation of freezing or ice nucleation in a sample of water (or coupling agent) arises from nuclei with an ice-like structure. Nuclei can promote the organization of water molecules into the ice crystal lattice.

Water and aqueous coupling agents have a natural tendency to cool to a temperature well below their equilibrium freezing point before ice nucleation, i. E. They tend to undergo supercooling. There are two ways of ice nucleation in water: uniformity and non-uniformity. Nucleation is called " homogeneous " when critical nuclei are formed by spontaneous aggregation of water molecules themselves. For macroscopic amounts of water, the cluster size required for ice nucleation is often about 25 molecules. The radius of the cluster may be about three molecules. A critical radius corresponding to this size provides a temperature of -41 ° C, which is called the uniform nucleation temperature for water. Thus, the uniform nucleation temperature for water is the minimum temperature at which pure water can be cooled before freezing occurs spontaneously.

When agglomeration of water molecules is catalyzed by an external source, nucleation is referred to as " non-uniformity ". The cause of the external nucleation may be the introduction of ice crystals or other foreign materials into the subcooled sample. For example, crystallization can be caused by the physical introduction of a nucleation initiator (e. G., Seed crystals or nuclei) and a crystal structure is formed around the nucleation initiator to produce a solid.

The material may be a hydrogel, a liposome, or an emulsion, for example, an oil-in-water (O / W) emulsion, a w / o emulsion, a milk oil emulsion, or a nano-emulsion, Lt; RTI ID = 0.0 > nucleation. ≪ / RTI >

1. nucleation initiator

INA can be a material that promotes the formation of seed crystals (or initial clusters) and catalyzes the formation of uneven ice nucleation. When INA is used, water freezing occurs at a temperature higher than that required in the case of uniform nucleation, and the maximum biological ice nucleating agent is added at a temperature of -1 to -5 DEG C or higher before spontaneous water freezing otherwise occurs normally It may cause freezing at other lower temperatures. Voluntary water freezing can occur variably at -10 ° C, -15 ° C, -20 ° C, or -25 ° C or less, and the timing of spontaneous freezing is highly unpredictable. Examples of INA include biogenic-derived proteins, materials derived from gram-negative bacteria, and / or substances belonging to the genus Pseudomonas, Erwinia, or Xanthomonas do. For example, INA may be an inorganic or organic-derived material that promotes nonuniform ice nucleation. Embodiments of the present technology may include methods for producing controlled and predictable freezing of skin and subcutaneous tissue using INA.

In general, INA can promote the formation of ice crystals with water-like materials at certain temperatures, typically below a few degrees below 0 ° C. INA can be used synergistically with a thermal therapy protocol selected to control the onset and degree of freezing events during cryotherapy and can be used to promote in vivo freezing at temperatures above the natural uniform nucleation freezing point of skin tissue . Aspects of some embodiments of the present technology relate to methods for producing controlled freezing of skin and subcutaneous tissue using INA. The cooling method using INA enables the induction of ice nucleation at a specific temperature such as a temperature close to 0 ° C. Thus, INA requires freezing and variable therapeutic temperatures with the desired therapeutic / safe temperature range, and may provide an advantage to therapies that produce accurate and controllable skin and tissue damage from freeze events have.

A variety of gram-negative bacteria are known to produce INA. Among these are Pseudomonas, Erbina, or Xanthomonas among others. One of the highest levels of ice nucleating activators is the ice nucleating protein (INP) from some ice-nucleating bacteria. Protein molecules and substances located in the outer membrane of these bacteria are responsible for ice nucleation. Cells can also be lysed or otherwise produce fragments of cellular material (e.g., membranes), such as Pseudomonas syringae, in which such INA is found or captured.

One commercially available INA is SNOMAX ®, available from Snomax LLC of Englewood, Colorado, which is derived from the bacterial Pseudomonas syringae (freeze-dried protein powder). The protein acts as an ice nucleating agent to initiate a freezing process and raises the predictable freezing temperature of water to about -3 ° C. SNOMAX ® is widely used for snow removal and is safe and non-pathogenic for human use. SNOMAX ® may be a powder that exhibits 10 ¹² to 10 ¹³ ice nuclei per gram at a temperature of less than about -4 ° C. Coupling agents made with SNOMAX ® or other materials derived from the bacterial Pseudomonas syringae can have sufficient INA to produce reliable ice nucleation at the desired temperature. Bacterial / cell concentration has a direct effect on the nucleation temperature of water. INA can be used in standard powder form and can be used as an ice nucleating agent with or without the addition of water. In some embodiments, the INA can be delivered to the skin very little, for example, by a micro needle (e.g., an array of micro needle). Biocompatible INA can be delivered invasively using needles such as intradermal needles. Additionally or alternatively, the INA may also be used with non-contact cooling devices such as cooling / freeze spray.

INA can be used in cooling protocols to cause ice nucleation at temperatures of about -2 [deg.] C, -3 [deg.] C, or -4 [deg.] C. At these temperatures, the damage to the epidermal tissue may be significantly less than the damage typically caused at lower freezing temperatures. The temperature for ice nucleation can be chosen to be high enough to avoid significant skin pigmentation changes associated with frozen events.

Non-invasive applicators (e.g., the applicator 104 of FIGS. 2 and 3) may be used to control skin cooling and may include one or more temperature sensors. A temperature sensor (e.g., element 167 of FIG. 2) may be embedded along the treatment surface of the applicator and used as part of the temperature control system. The temperature control system may include one or more feedback control algorithms for controlling the applicator based on a predetermined set temperature value over one or more predetermined periods of time, And has a determined rate of change. Different feedback control algorithms can be used to treat tissue (and different temperature treatment cycles) by using different treatment temperature protocols by varying the cooling / thawing speed, pre-determining and / or predetermining the therapeutic temperature, ). The methods described herein may include using both INA to control freezing and various treatment temperature protocols and profiles.

2. Hydrogel material

Aspects of the present technology relate to methods of using hydrogel materials with freezing depressants (cryoprotectants) and / or INA to produce controlled " predictable " predictable freezing. Hydrogel materials are cross-linked polymer classes capable of absorbing large amounts of water due to their hydrophilic nature. The hydrogel material may have a suitable moisture content to control freezing, including controlling ice nucleation, ice crystallization, frozen propagation, and the like. Components of the hydrogel synthesis include monomers, initiators, and cross-linking agents. The hydrogel properties can be controlled by varying their synthesis factors, such as reaction temperature, monomer type, monomer crosslinker, ratio of crosslinker-to-monomer, monomer concentration, and type and amount of initiator. The composition of the hydrogel may be selected for particular applications by selecting the appropriate starting materials and processing techniques.

The hydrogel may be mixed with one or more freezing point depressants and manipulated to have a desired melting / freezing temperature (e.g., optimal melt temperature). Freezing agents can be inoculated into tissues. Additionally or alternatively, the hydrogel may be mixed with an INA having a set activation temperature that allows the hydrogel to be continuously frozen at a pre-determined temperature range (or a specific temperature) that differs from that associated with hydrogels without an ice nucleating agent . The combination of hydrogel, freezing point depressant, and / or INA can cause controllable freezing at desired temperatures such as -3 DEG C, -2 DEG C, -1 DEG C, or other temperatures. Temperatures close to 0 ° C can cause less damage to the epidermal tissue and are therefore very well suited to less aggressive temperature freezing protocols so that the temperature can be reduced by eliminating or minimizing significant discoloration side effects associated with freezing skin treatment, Lt; RTI ID = 0.0 > upper < / RTI >

The water contained by the hydrogel structure can be classified into four types: free water, number of gaps, number of bonds, and number of semi-bonds. The free water is located in the outermost layer and can be easily removed from the hydrogel under mild conditions. The number of spacing is not physically attached to the hydrogel network, but is physically trapped between the hydrated polymer chains. The number of bonds is directly attached to the polymer chains through hydration of the functional groups or ions. The bonded water remains as a component of the hydrogel structure and can be separated only at very high temperatures. The half-coupled number has an intermediate characteristic between the combined number and the free number. The number of free water and spacing can be removed from the hydrogel by centrifugation and mechanical compression.

The controlled freezing technique may utilize the water composition of the hydrogel. Hydrogels can be designed to have a specific freezing point or a specific freezing temperature range by having a water-monomer-crosslinker content of a specific ratio. Cryoprotectant additives such as glycols (e. G., PG) or other materials may also be used to lower their freezing point.

Hydrogels can act as initiators of predictable freezing events. As the hydrogel freezes, the hydrogel provides an " early seed " or crystalline site that is inoculated into the tissue to catalyze predictable freezing controlled at a specific temperature in the skin. In some embodiments, the predictable freezing event can be tissue freezing that occurs at least 90%, 95%, or 98% of the time that freezing is required. A predictable controlled freeze event in the hydrogel may be achieved by pre-cooling the hydrogel to a temperature below its melting point. The freeze event can be initiated by injecting a nucleation initiator (e.g., ice / water slurry) into the hydrogel to create a freeze reaching the surface of the hydrogel adjacent to the patient, which causes freezing of the skin of the subject . In another process, ultrasound or other nucleation energy can be used to generate a freezing event in the hydrogel. According to one embodiment, the additives (e. G., Cryoprotectant and / or INA) are not present on the outer surface of the hydrogel sheet or hydrogel pad and thus do not come into direct contact with the skin or other tissue being treated So that it can be segregated into a separate layer within the interior of the hydrogel. Encapsulating these materials in hydrogels eliminates the need to select proven INA materials that are tested when contacted with skin or tissue. Predictable hydrogel freezing can be enhanced by additives such as INA.

Figure 10a shows a patterned hydrogel according to an embodiment of the disclosed technique. Figures 10b and 10c are plots of PG concentration (% PG) versus length for the patterned hydrogel. Hydrogels may include water and a dispersing medium of large amounts of nucleation inhibitors (e. G., Water / PG emulsion particles or columns). Figure 10a shows a separate volume of nucleation inhibitor spaced apart in a large amount of water. In some embodiments, the hydrogel layer or sheet may contain a column of nucleation inhibiting emulsions separated by a large amount of water that is substantially free of PG. The water surrounding the column can act as an ice nucleation site. An internal enclosure (e.g., an encapsulant) can inhibit or prevent the diffusion of the water / PG emulsion. The composition of the inner enclosure can be selected based on the composition of the encapsulated material, and the pattern, number, and size of the ubiquitous nucleation inhibiting volume can be selected based on the hydrogel properties.

Figures 10b and 10c show an embodiment with propylene glycol (PG) located throughout the hydrogel, the concentration of PG being dependent on the length and width of the layer or sheet of hydrogel. The region of the hydrogel having the lowest concentration of PG has the highest melting point / freezing point and can therefore function as an ice nucleation region. An isolated freezing zone can be formed in the skin adjacent to this ice nucleation area. Other types of cryoprotectants or ingredients may replace PG. For example, PG can be replaced or compounded with PPG, PEG, DMSO, and the like.

A further embodiment is a hydrogel containing INA uniformly distributed throughout the zone where it is desired to seed the frozen propagation. In addition, the INA can be dispersed exclusively in the interior portion of a large amount of hydrogel. For example, INA may be in a hydrogel sheet to prevent INA from contacting the skin by not spreading to the surface of the sheet. The INA can seed a freeze event in the interior region of the hydrogel, and the freeze event can quickly propagate to the outer surface of the hydrogel, which eventually contacts the skin and causes a freeze event in the skin.

11A and 11B are side views of a hydrogel having an ice nucleation region. 11A shows a hydrogel material having an inner ice nucleation region between the upper and lower ice nucleation inhibition zones. The ice nucleation region can contain INA and, if present, can have a relatively low concentration of temperature rampant. In one embodiment, the ice nucleation zone may be substantially free of temperature lowering agents and may include water and ice nucleation features (e.g., INA, ice nucleating particles, etc.). The ice nucleation area may be a layer, a continuous layer, or a spot on the inner layer so that it is completely enclosed within the hydrogel.

The ice nucleation inhibition zone may have a lower melting point / freezing point than the ice nucleation zone and may contain a large amount of temperature lowering agents, for example, a large amount of PG evenly or unevenly spaced apart. The pattern, number, and composition of the freeze inhibition feature may be selected based on the desired nucleation inhibition properties.

11B shows a multilayer hydrogel material having an outer ice nucleation inhibiting layer and an inner ice nucleating layer. The outer ice nucleation inhibition layer may comprise a temperature lowering agent and the inner layer may comprise a high concentration of INA (e.g., INA mainly on a volume basis). For example, the outer ice nucleation inhibition layer may be a layer of a PG solution or a layer having a dense array of PG volumes, and the inner layer may comprise mainly or wholly water. The freezing event may be diffused through the outer anti-ice nucleation inhibiting layer initiated in the ice nucleating layer.

The hydrogel may adhere to both the patient side and the applicator side. The sticking top and bottom surfaces may help maintain contact of the subject with the skin and applicator, and in some embodiments may help to minimize or limit the migration of the hydrogel during treatment. The liner may be used to prevent contamination of the hydrogel. The side of the hydrogel in contact with the liner may stick. In some embodiments, the hydrogel may be a sheet having a uniform or variable thickness with an adhesive applied to one or more of its outer surfaces.

Hydrogels may be used in the methods discussed in connection with Figures 5, 8, and 9a-9c. For example, at block 142 of FIG. 5, the hydrogel may comprise an INA that is applicable to the skin of a subject and is capable of forming ice crystals in the presence of water. As discussed in connection with Figures 11A and 11B, the INA can be encapsulated within the polymeric structure of the hydrogel such that the INA does not come into direct contact with the skin. The hydrogel and skin may be cooled to reach a suitable cooling temperature for freezing the skin. The hydrogel may contain a freezing point depressant such that the first melting / freezing temperature of the hydrogel is lower than the second melting / freezing temperature of the fluid in the skin.

In block 144 of FIG. 5, the skin may be cooled to a temperature higher than the first freezing temperature and lower than the second freezing temperature to subcool the skin, and after a predetermined amount of subcooling has occurred, ). The temperature of the epidermis may rise above the first temperature before freezing the dermis.

With reference to the method 150 of FIG. 8, at block 154, a cross-linked polymer and a hydrogel material comprising INA can be applied. The INA can be encapsulated within the polymeric structure to prevent direct contact between the INA and the skin. In some embodiments, a sheet of hydrogel may be applied to the skin of the subject. Alternatively, the hydrogel may be injected into the skin. Other techniques may be used to apply hydrogels, which may be creams, gels, and the like. At block 156, the skin is cooled. At block 158, a freeze event may be initiated using one or more ice crystals. In another embodiment, energy is used to destroy structures containing INA to release a sufficient amount of INA to generate freeze events.

3. Liposome

Liposomal transport of a substance to a tissue can be used to deliver the substance to a specific tissue in a more effective way than just applying the substance to the skin surface only. Since the liposome is lipophilic, it can be absorbed at least into the stratum corneum and then release the substance in the liposome at a specific location or depth in the tissue of the subject. Liposomes can capture considerable " buckets " of water, thereby improving the moisture content of the skin when the liposomes are degraded and providing a more predictable freeze protection when used in conjunction with a significant amount of cryoprotectant in water in the liposome I make it. Liposome skin hydration may be more effective than direct application of water to the skin surface since the stratum corneum is normally hydrophobic.

Topically applied liposomes can enhance thermal contact between the applicator / skin and provide controlled delivery of the formulation (e.g., cryoprotectant, INA, etc.), and the liposomes are better than water or water mixed with the cryoprotectant It can penetrate the stratum corneum. In addition, liposomes can deliver different agents to different locations, allowing the agent to migrate directly to a particular targeted cell. In one embodiment, the liposome contains a cryoprotectant (e. G., Propylene glycol) and is capable of degrading to release the cryoprotectant. In another embodiment, the liposomes selectively release INA to provide controlled freezing performance through a particular tissue.

According to embodiments in which freezing is required, the substance (e.g., INA, cryoprotectant, etc.) can be incorporated into the liposome so that the liposome can controllably release the substance into the skin. In particular, liposomes can be formulated to maintain their structure upon penetration into the skin to minimize, limit, or substantially prevent release of the material. When sufficient liposomes accumulate in a particular desired tissue or layer of skin, "ordered" degradation of the liposome may be initiated, resulting in burst release of the implanted formulation. In some embodiments, the liposome may contain an INA to initiate a freeze event. Activation methods for degrading liposomes include using temperature (e.g., temperature cycling), ultrasound, or a detergent to disrupt or destroy the lipid encapsulation of the liposome. The applicator may include a heater for heating the treatment site to cause the release of the agent, or may include a transducer for delivering mechanical energy in the form of an ultrasonic wave, or may be configured to perform the method discussed with respect to Figures 5 and 8 And other components for disrupting the liposome.

The liposomes may have a composition selected based on, for example, formulation release rate, stability, and / or other desired characteristics. In some embodiments, the formulation release rate can be increased by applying energy, e. G., Ultrasound, heat, or other energy suitable for decomposing the lipid that captured the formulation. For example, the medium may comprise a first liposome for delivering the cryoprotectant to the epidermis and a second liposome for delivering the INA to the dermis. Once the first liposome is absorbed by the epidermis, they can release the cryoprotectant and protect the epidermis. After the second liposome is absorbed by the dermis through the epidermis, they release the INA into the dermis. Upon cooling the treatment site to a temperature below the melting point of the skin / freezing point, INA may cause a predictable freezing in dermal tissue. Thus, each formulation can be delivered to a particular location using liposomes. The liposome medium may be used before, during, and / or after a treatment session. In some processes, a topical medium is applied to the skin surface to deliver the cryoprotective agent prior to cooling to the shallow tissue. When the tissue is cooled and ready to be frozen, another medium (e.g., medium with INA) may be injected into deeper tissue.

4. Emulsion

An emulsion contains a droplet containing a dispersed phase, which is dispersed in a liquid medium which is a class of a dispersing system comprising two immiscible liquids and which is a continuous phase. The emulsion may be an oil-in-water (O / W) emulsion, a water-in-oil (W / O) emulsion, a oil-in-water (O / O) emulsion, or a nano-emulsion. Nano-emulsions are preferred because they can penetrate the epidermis and dermis along follicular opening and skin pore openings. Figure 12a shows an emulsifier or surfactant having a hydrophilic head and a hydrophobic tail. Figure 12b shows a formulation (e. G., Oily formulation) captured by an emulsifier to separate the formulation from water. Figures 13a and 13b show oil-in-water and water-in-oil emulsions. Referring to Figure 13a, the emulsion comprises an oil droplet which may contain the same or different agent. In single-agent emulsions, such droplets may contain the same agent. In multi-agent emulsions, different agents (e. G., Dispersed medium) may be evenly or unevenly dispersed in the dispersion medium. The dispersed medium may include a drop containing one or more cryoprotectants, INA, analgesics, agents, and the like.

Figure 14 is a table with melting point / freezing point temperature for fat. Fats can be natural only and suitable for O / W emulsions and can have relatively high melting points. For example, the fat may have a melting point / freezing point above 0 ° C and may be used in emulsions.

E. Test and treatment methods

The in vitro test of the skin using the treatment cycle shows the unpredictability of supercooled skin and the precise control freezing to the INA. In one test, a thermocouple was placed between the coupling layer (coupling medium) and the skin to detect freezing. The coupling layer of only water was tested to confirm that there was no tissue freezing without INA. The thermocouple temperature data for five tests, including two water-only tests (if no freezing has taken place) and three tests using INA (three separate freezing events have occurred), show the effect of INA and the controlled freezing To demonstrate the feasibility of the concept.

Figures 15a and 15b show the results of the test: when INA was used, the skin was continuously and predictably inoculated three times separately, while the INA was not used without the INA, the skin was not only frozen but only supercooled There was a tendency. Figure 15a shows an exemplary treatment profile designed to cause subcooling at -2 [deg.] C for 3 minutes and freezing at -5 [deg.] C. Figure 15b shows the associated temperature increase caused by three tests with INA and a freezing event and heat of fusion occurring immediately after 200 seconds, which causes the measured temperature to increase from about -1 [deg.] C to about 0 [deg.] C. A non-invasive surface cooling apparatus was used to perform the tests discussed in connection with FIGS. 15A and 15B, and INA was SNOMAX® / water solution.

Figure 16 shows an exemplary treatment cycle temperature profile and temperature response of a thermal sensor at an applicator surface-tissue interface. The target temperature (indicated by the dashed line) of the temperature-controlled surface of the applicator can be lowered at a predetermined rate and then kept constant at the preset value. One or more sensors may monitor the temperature at the contact interface. The output from the sensor can be used to precisely control the freezing run to maximize cellular changes for therapeutic purposes while restraining, limiting, or minimizing adverse therapeutic side effects. It may be desirable to control the degree of skin freezing down to epidermal-dermal layer, dermal-subcutaneous layer, or other specific depth. The degree of freezing desired can be achieved based on knowledge of the freezing point temperature of bulk tissue. In freezing events, the tissue undergoes ice nucleation and growth (pyrogenic changes). An exothermic change is when the system emits heat (e.g., from liquid to solid) around the phase change. Freezing is a pyrogenic event that releases heat for a very short period of time, and this heat release can be a reliable indicator of skin freezing. A thermal sensor may be used to sense the heat emitted by the phase change associated with the freezing event. If the tissue is in a subcooled steady state, a thermal sensor (e.g., element 167 of FIG. 2) can detect whether the temperature is stable without any significant sudden temperature change or peak at the applicator surface- have. When the subcooled tissue is frozen, the heat released is captured by the sensor as a sudden temperature increase. 15B shows the temperature increase in tests 2, 3, and 5 corresponding to the emitted heat.

Figure 17 shows that the set temperature (indicated by the dashed line) of the applicator surface was lowered at a constant rate and then maintained at a generally constant value. The temperature at the applicator-tissue interface is sensed by the thermal sensor and is represented by the solid line. After the targeted tissue is subcooled, a freeze event may be initiated. The temperature sensor can sense a temperature rise associated with the heat of fusion, and the sensed temperature rise can be identified as a phase change. Various temperature changes can be used to monitor tissue and detect phase changes. Some methods may include supercooling, and using freeze set points to intentionally treat tissue at sub-zero temperatures. The tissue temperature may be lowered at the desired time during treatment to control the onset of freezing. Thaw cycles may be included in any treatment cycle to warm the tissue at a desired rate to protect or enhance tissue damage.

The treatment cycle can be determined by selecting a supercooling parameter, a freezing parameter, and a thawing parameter. The subcooling parameter may include a cooling profile, a target temperature, and / or a time period. The cooling profile may include a cooling rate for ramp-down to reach the supercooling temperature. The target tissue can be maintained at a sub-zero target temperature during the supercooling period without phase change.

The freeze parameters may include a cooling profile to keep the tissue frozen and / or a time period during which the tissue remains frozen. The freeze parameter may be selected to increase or decrease thermal damage. After completion of the freeze time period, the cooled tissue may be warmed using a thaw cycle.

The supercooling parameters, freezing parameters, and / or thawing parameters may be obtained experimentally in vitro and / or in vivo . For example, in vivo human trials have shown that skin can be subcooled to sub-zero temperature, e.g., -20 C, without phase change. If the temperature drops sufficiently low, the tissue will freeze. Human skin tissue has been experimentally established to often freeze spontaneously at approximately -25 ° C. The temperature of spontaneous freezing depends on the tissue characteristics such as the moisture content of the tissue, the cell structure, and the like.

Figure 18 shows an exemplary treatment cycle for controlled supercooling and for controlled freezing. The temperature gradient in tissue during the cooling protocol can be estimated by biothermal heat transfer modeling, experimental testing, or a combination of both. Heat transfer modeling can be used to predict temperature gradient versus time in tissue during treatment cycles. At sub-zero temperature, other information about tissue volume, tissue depth, and tissue can be calculated. For example, a flat applicator may cool the skin and allow the targeted tissue to grow at a temperature of 0 ° C to -20 ° C, 0 ° C to -12 ° C, 0 ° C to -10 ° C, 0 ° C to -8 ° C, -20 ° C, -15 ° C, -13 ° C, -12 ° C, -11 ° C, -10 ° C, -8 ° C, -6 ° C, -5 ° C, -4 ° C, -3 ° C, 1 < 0 > C, or other suitable temperature range for cooling to a temperature equal to or lower than 0 < 0 > C. The target tissue may reach a normal steady state in which the body heat of the object counteracts a sustained heat withdrawal from the skin surface. The temperature-controlled surface of the applicator may be maintained at a suitable temperature below the desired bulk temperature of the targeted tissue during the supercooling period. To freeze the subcooled tissue, the temperature of the applicator may be further lowered, for example, to ramp down to the melting point / freezing point of the medium on the skin, the freezing point of the formulation in the skin, or the freezing point of the skin itself.

19A-19E are cross-sectional views of a cooling applicator applied to a treatment site and thermal modeling. Figure 19a shows the treatment site at the beginning of the supercooling cooling cycle where the shallow tissue begins to subcool. Figures 19b-19e show the amount of supercooled tissue that increases over time until the entire epidermis / dermis under the cooling plate is subcooled, while the subdermal layer is not subcooled (supercooled skin tissue is gray and non- The subcooled tissue is black). The subcooled tissue may be at a temperature within the range of about 0 ° C to -20 ° C. The model for cooling the skin was 0.1 mm (1.0 mm) above the subcutaneous layer of 1.0 cm (10 mm), based on a flat applicator (ramp down to 1 캜 / s and a treatment profile with a maintenance step at a target temperature of -20 캜) And skin with a dermal layer of 2 mm can be completely subcooled within 90 seconds. To freeze the entire area under the epidermal / dermal layer, skin freezing can be induced to occur at subcooling or subcooling for 90 seconds. To freeze only a portion of the underlying epidermis / dermis, skin freezing can be induced to occur 90 seconds prior (e.g., after a subcooling of 45, 60, or 75 seconds). The onset of the freeze event may be delayed for a short period of time (e. G., A few seconds) after reaching the freezing temperature due to the time required to cool the tissue to a lower set temperature and the time it takes for the ice nucleation event to occur , It may be necessary to add some additional time from the time the temperature dive is initiated to the beginning of the defrosting event, so that the tissue causes the desired amount of time to remain partially frozen. For the skin, approximately 15, 20, or 30 seconds may be the average delay time from the beginning of the temperature dive until the freezing event occurs.

When the freeze event is initiated, the entire dermis and skin under the applicator may not be completely frozen. This allows the bulk temperature of the tissue to absorb all of the heat of fusion from the freezing event to achieve 100% tissue freezing (e.g., when the heat extraction by the applicator is balanced with the warming from the dermal tissue and blood stream) As shown in FIG. As the heat of fusion is released during the freeze event, the bulk temperature of the tissue is adjusted such that the additional freezing is stopped and the skin is partially frozen if equilibrium is established (in the absence of further significant heat extraction by the applicator) Lt; RTI ID = 0.0 > 0 C. < / RTI > The temperature of the cooling plate can be adjusted to compensate for heat of fusion or other natural heating associated with the subject's body. Without being bound by theory, the skin needs to be completely frozen during the freezing event by being subcooled to a temperature of approximately -70 DEG C to completely freeze the skin, but this low supercooling temperature results in severe side effects, especially on the epidermis Which is believed to be highly undesirable.

Figures 20a-20f illustrate the steps of one method of freezing tissue without supercooling. Generally, the applicator can be applied to the treatment site after applying the coupling agent to the applicator and / or the treatment site. The applicator may freeze areas of the skin while limiting or avoiding subcooling as discussed below.

20A shows a coupling medium 209 (e.g., water, coupling gel, etc.) located along the temperature-controlled surface 211 of the applicator. A layer of coupling medium may be formed by applying water to a temperature-controlled surface 211, which may be formed at a temperature of about -20 ° C, -15 ° C, -10 ° C, or most of the coupling medium 209 Or any other suitable temperature for freezing the whole. The temperature-controlled surface 211 may then be heated to a higher temperature (e.g., -3 ° C, -2 ° C, or -1 ° C) suitable for applying the applicator to the treatment site.

Fig. 20b shows the frozen upper layer 215 and the liquid lower layer 217 of the coupling medium after the applicator has been applied to the treatment site 213. Fig. The warmed skin may contact the liquid layer 217 gel while the layer of frozen coupling medium remains freeze in contact with the temperature-controlled surface 211. The temperature-controlled surface 211 can be maintained at a temperature sufficient to hold the frozen upper layer 215 and the liquid lower layer 217. [

Fig. 20C shows a temperature-controlled surface 211 (Fig. 2C) maintained at a low temperature (e.g., -1 DEG C, -2 DEG C, -3 DEG C, etc.) for a predetermined period of time to cool the skin to its melting point / ). During this holding period, the portion of the frozen coupling medium may slightly increase in volume, thereby increasing the thickness of layer 215. The temperature of the temperature-controlled surface can be lowered or raised to increase or decrease the thickness of the frozen layer 215 while maintaining the liquid layer 217. [ In some processes, the temperature-controlled surface 211 can be maintained at a temperature in the range of about -2 ° C to about -12 ° C, about -4 ° C to about -10 ° C, or other suitable temperature range. For example, the temperature-controlled surface 211 can be maintained at a temperature of about -6 DEG C, -8 DEG C, or -10 DEG C. As the temperature is maintained at a lowered / lowered or lowered temperature, the freezing front of the coupling medium moves towards the skin until substantially all or all of the coupling medium / gel is frozen.

20D shows the entire layer of the coupling medium 209 in a frozen state. The skin surface in contact with the coupling medium 209 may be lowered to its freezing point. Due to the intimate contact of the skin with the ice crystals in the frozen coupling gel 209, the skin will gradually freeze rather than supercooled.

20E shows the frozen wire 221 and the freezing volume of the tissue. The freezing wire 221 can move deeper into the object until the desired volume of tissue is frozen.

Fig. 20f shows a temperature-controlled surface 211 maintained for two minutes at a temperature of about-8 C to produce an enlarged volume of frozen tissue. The freezing volume of the tissue can be stopped and stabilized if a steady state is established. In facial treatment, the skin can be frozen without affecting the underlying muscles and subcutaneous tissue. In some treatments, the applicator may freeze the skin without affecting the subcutaneous tissue to limit or avoid skin contour changes at the treatment site. In other treatments, the subcutaneous tissue may be frozen to inhibit, divide, or reduce subcutaneous lipid-rich cells, for example, to contour the treatment site. For example, the applicator may treat acne and may or may not profile the tissue in a single session.

The temperature of the temperature-controlled surface 211 may be increased to 18 ° C, 20 ° C, or 22 ° C at a rate of 1 ° C / s, 2 ° C / s, or 3 ° C / s. This will rapidly thaw the tissue to minimize or limit further damage to the cells. For example, the skin may be cooled at a rate of 0.25 ° C / s, held at a target temperature of -8 ° C for 2 minutes, and then thawed at a rate of 2 ° C / s. Other cooling rates, target temperatures, and thawing rates may be selected based on desired levels of freezing, thermal damage, and the like.

Targeted tissue can be frozen several more times than non-targeted tissue. A repetitive freeze-thaw cycle effectively damages or kills tissue because, besides suffering multiple cycles of harmful solution effects and mechanical ice crystal damage, cell membrane integrity after the first freeze-thaw cycle is compromised, It becomes a less effective barrier to freezing propagation in subsequent freeze-thaw cycles and the cells are much more sensitive to the formation of lethal intracellular ice in subsequent freeze-thaw cycles. In some embodiments, the targeted tissue can be frozen multiple times in a single treatment session, while non-targeted tissue, such as the epidermis, can be frozen only once. In addition, the targeted tissue can be frozen many times without overcooling any tissue. In some procedures, the dermis is repeatedly frozen to damage or destroy the target spleen without repeatedly freezing the epidermis.

21 is a plot of temperature vs. time for multiple rounds of freezing of the skin in accordance with an embodiment of the disclosed technique. After the layer of frozen coupling medium is produced at -15 占 폚, the temperature-controlled surface of the applicator is heated to -2 占 폚 to cool the skin using the technique discussed in connection with Figs. 20a-20f. The skin-applicator interface is then maintained at a temperature of about -2 [deg.] C, resulting in a freeze event (denoted by " * "). The temperature-controlled surface is then cooled to -10 DEG C and the freezing event is maintained at -10 DEG C for a period of time (e.g., 1 minute, 2 minutes, etc.). Figure 21 shows a one minute holding period. The temperature of the applicator is then raised to -1.5 DEG C for about 35 seconds. At this temperature, the epidermis in contact with the coupling medium may remain frozen. The dermis is thawed due to internal heat and, in particular, heat from the blood stream that passes through the dermis. The applicator is then re-cooled to -10 ° C or another suitable temperature for re-freezing the dermis. The second freezing event may be maintained for a period of time such as one minute, two minutes, and so on. In some cases, the dermis can be re-thawed by warming the applicator to -1.5 ° C and again frozen at -10 ° C while keeping the epidermis freeze-thawed. The thawing temperature, heating rate, cooling rate, duration of the freezing event, and re-freezing temperature can be selected based on the desired number of re-freezing and severity of thermal damage.

Using this treatment protocol, the greater the freezing rate will have a much lesser damage effect on the epidermis, since the epidermis is never thawed. In a second or subsequent freeze-thaw cycle, a much larger freezing rate can be achieved by setting the thawing temperature (e.g., -1.5 占 폚, -1 占 폚, 0.5 占 폚, 10 [deg.] C, -12 [deg.] C), which can further increase the probability of intracellular ice formation in the dermis as further described below.

Repeated freezing approaches allow complete control over most, some, or all of the variables that influence tissue viability after thawing. These variables include, without limitation, skin freezing rate, target temperature, duration of freezing, and heating rate. Skin freezing rates can not be controlled using other approaches when the skin is substantially subcooled because macroscopic freezing events occur almost instantaneously when the skin is nucleated or when ice is inoculated when the skin is in a subcooled state (Over a period of several seconds). Without being bound by theory, in the course of extracellular space ice formation induced at -2 ° C, if the tissue is then slowly cooled to -10 ° C, the intracellular water is allowed to diffuse into the extracellular space It is believed that the freezing rate is important because of the time. This helps the intracellular solute concentration to raise and lower the intracellular melting / freezing temperature, thereby reducing the probability of lethal intracellular ice formation at cooler temperatures. However, if the skin causes freezing at -10 ° C (with a larger supercooling window), there will not be enough time for cell dehydration and there will be no freezing point depression in the cell. Thus, at the cooler supercooling temperature (-10 ° C), there is greater likelihood of intracellular ice formation and associated cell damage increase. Large amounts of supercooling have been shown to correlate with greater risk of intracellular ice formation, which may sometimes be desirable, and may be undesirable in other cases, depending on which tissue is targeted and untargeted.

As discussed in connection with Figs. 20a to 20f and 21, the treatment methods disclosed herein provide for all freeze-thaw cycles without allowing significant skin cooling, which freezes excess tissue at a faster rate than the process without supercooling, Can provide complete control over thawing parameters. The pre-determined skin supercooling level can be achieved with a predetermined duration if increased tissue destruction or damage within a shorter time period and the associated effects of supercooling are therapeutically particularly interesting. Skin freezing can be induced when the temperature of the applicator, the coupling medium, and the skin is further cooled, for example, to a predetermined amount (e.g., 1, 2, 3, or 4 degrees Celsius) . For example, the applicator can be maintained at a temperature slightly cooler than the freezing point of the selected coupling medium containing the ice-nucleating agent and the freezing point depressant, ensuring that the layer of the coupling medium in contact with the applicator is frozen . By controlling the thickness of the layer of the coupling medium, the temperature gradient across the coupling medium layer can result in a temperature slightly warmer than its freezing point, so that the non-frozen coupling medium can remain in contact with the skin. In some embodiments, the coupling medium can be a hydrogel, since the hydrogel can be formulated to have an accurate thickness.

Figure 22a shows a liquid coupling medium that acts as an insulator for the binging of the skin, allowing the skin to be subcooled. To induce skin freezing, the applicator may be cooled a few more degrees to freeze the entire volume of the coupling medium. As shown in FIG. 22B, this process moves the temperature profile to the left of the freezing point of the coupling medium. When ice crystals in the coupling medium reach the skin, the supercooled skin can be frozen immediately or within a short time.

Figure 23 shows the freezing temperatures for various concentrations of PG in water. 24A-24F show that the skin is subcooled to -13 C, held at this temperature for 3 minutes, and then cooled to -15 C using a coupling medium containing 26 vol% PG with a freezing temperature of about -11.5 C RTI ID = 0.0 > a < / RTI > The concentration of the PG solution can be selected based on the desired temperature for initiating the freezing event.

Referring to Figures 24A-24F, a coupling medium may be applied to the applicator and the skin. The applicator is cooled by ramping down the temperature of the temperature-controlled surface by freezing the coupling medium, which is then heated by ramping up to -13 ° C. Keeping the temperature at -13 C will ensure that at least the layer of frozen media remains in contact with the applicator. A liquid coupling medium is applied to the skin surface. The applicator is applied to a liquid coupling medium on the skin, and then the coupling medium and skin are allowed to cool to freeze the target tissue. (E. G., A frozen layer on the applicator surface) and a thin layer of a liquid coupling medium (e. G., A freeze layer on the applicator surface) by selecting a composition of the coupling medium (e. G., Concentration of PG, glycerol, For example, a melting point / freezing point that results in the desired temperature for subcooling the skin while providing both, can be selected.

24A shows a layer of a frozen coupling medium 231 with an applicator. A paper towel-shaped carrier soaked with 26% by volume of PG / water can be placed on the temperature-controlled surface 211. If a paper towel is applied, the temperature-controlled surface 211 may be pre-cooled to freeze the coupling medium 231 quickly. In another process, the coupling medium is dispersed, sprayed, or otherwise directly applied to the temperature-controlled surface 211.

Figure 24b shows another carrier in the form of a paper towel soaked with 26 vol% PG / water applied to the skin of the subject (at room temperature). The layer of liquid coupling medium 233 (e.g., 26% PG / water) on the skin is thick enough to prevent direct contact between the frozen coupling medium 231 and the skin of the subject during deployment of the applicator . In addition, the liquid coupling medium 233 helps to improve patient comfort when placing the frozen coupling medium 231 on the coupling medium 233.

24C shows the applicator after the frozen coupling medium 231 is placed in contact with the room temperature coupling medium 233. FIG. The thickness and temperature of the liquid coupling medium 233 can be adjusted by melting only a portion of the frozen coupling medium 231 to maintain a thin layer of the frozen coupling medium 231 along the temperature- Can be selected. The control system may control the applicator to maintain the applicator surface temperature at a target temperature such as -13 DEG C below the freezing point of 26% PG (-11.5 DEG C).

The applicator can extract heat continuously or intermittently to gradually increase the volume of the frozen coupling medium during the maintenance period. 24D shows a thickened layer of the frozen coupling medium 231. Fig. The coupling medium 233 in contact with the skin may remain in a liquid state and may be close to but below the freezing temperature of -11.5 [deg.] C.

24E shows the freezing wire 221 moving toward the skin surface as the coupling medium cools. When the coupling medium in contact with the skin is frozen, it will be inoculated onto the subcooled skin and rapidly frozen over a period of a few seconds. For example, to freeze the entire coupling medium volume (i.e., coupling media 231, 233) and cause a freeze event in the skin, the applicator temperature may be about -15 ° C, -14 ° C, or -13 ° C RTI ID = 0.0 > of < / RTI >

The freezing event can be caused by a temperature that is several degrees cooler than the supercooling temperature. In one process, the skin can be cooled to a supercooling temperature of -13 DEG C, but it can cause freezing even at slightly lower temperatures such as -15 DEG C. This two-degree " dive " temperature is much smaller than that required by conventional techniques without the use of a coupling medium containing ice crystals in contact with the skin (which is approximately 10 degrees) ≪ / RTI > as a skin-bing seeding agent. For a given maximum final temperature for the applicator, a smaller dive temperature may result in a larger subcooled volume, and at tissue freezing, the frozen volume will also be larger than treatment with a larger dive temperature. After the tissue has been frozen for a desired length of time, the applicator may warm the tissue to inhibit or limit further collapse, damage, and the like. The warming and cooling cycles may be repeated any number of times in any order to thermally influence the targeted tissue.

Figure 24f shows the total volume of coupling in the tissue at the applicator-skin interface and in the underlying tissue in the frozen state. The applicator may be cooled or heated to increase or decrease the volume of the frozen tissue.

Figure 25 is a plot of temperature versus time for supercooling and freezing tissue. The skin can be cooled to -20 캜, and then freezing is initiated at -25 캜 by selecting 37 vol% PG gel with a freezing point of -19 캜. Upon freezing of the skin, the applicator can keep it at a temperature high enough to warm the epidermis quickly and allow the epidermis to remain un-frozen. For example, the epidermis may be maintained at a temperature of 1.5 [deg.] C or 2 [deg.] C. This maximizes dermal freeze exposure below, increasing dermal damage and limiting or minimizing skin-freezing exposure to reduce skin damage. Thus, warming can be used to minimize, limit, or substantially prevent thermal damage leading to poor pigmentation (skin whitening), pigmentation over-skining (skin darkening), and / or other undesirable effects.

The skin may be cooled to a temperature above the freezing point of the coupling medium to cause a freezing event. If the tissue is subcooled at -13 DEG C with a coupling medium having a slightly warmer freezing temperature (e.g., a freezing temperature of -11.5 DEG C), the skin will not be inoculated at temperatures slightly warmer than -13 DEG C. In some procedures, it may be desirable to initiate skin freezing at higher temperatures to minimize or limit damage to epidermal tissue during a freezing event. To address this need, a higher temperature melting point / freezing point can be achieved by diluting the coupling medium with a lower concentration of freezing point depressant. The melting / freezing temperature of the coupling medium may be raised to an amount sufficient to cause freezing at temperatures above the supercooling temperature. Briefly, the supercooling at time t1 can be achieved by selecting a coupling medium having a lower freezing temperature than at time t1. After subcooling, the applicator temperature ramps up to a higher temperature at time t2. A large amount of water can be delivered to the coupling medium to dilute it, confirming that the diluted coupling medium has a warmer freezing point than the temperature at t2. This will cause freezing in the diluted coupling medium to induce skin freezing more rapidly. Freezing according to a relatively warm command in a subcooled diluted coupling medium may be accomplished using, for example, energy (e.g., ultrasound), cryogenic probes (e.g., a very small cold finger probe), and / or INA . ≪ / RTI >

Figure 26 is a plot of temperature versus time for a two-cycle cycle to subcoolise tissue and then cause freezing. In general, the tissue can be sub-cooled by cycling between two temperatures (e.g., -10 ° C and -20 ° C). The freeze event is triggered at a higher temperature (e.g., -10 [deg.] C) or another suitable temperature. The freezing point of the coupling medium can be selected to ensure that the coupling medium is not frozen during the subcooling cycle. In some embodiments, the coupling medium may comprise at least 39% by volume PG having a freezing point of -20.5 [deg.] C so that the coupling medium is not frozen during the subcooling cycle at -20 [deg.] C to avoid the onset of premature skin freezing have.

At the end of the supercooling cycle, the temperature of the applicator may be raised to a higher temperature (e.g., -10 ° C) suitable for ice inoculation. A material such as cold water at 1 占 폚 may be injected through the applicator to dilute the coupling medium. The temperature and flow rate of the water may be selected so that the diluted coupling medium has a freezing point that is warmer than the predetermined value. For example, the diluted coupling medium may have a freezing point above about -10 ° C for freezing at about -10 ° C.

27A-27C show an applicator and a coupling medium. Referring now to Figure 27A, the liquid coupling medium is positioned along the temperature-controlled surface 243 of the applicator. The conduit, plate, and / or fluidic component of the applicator may be configured such that because the cooling applicator plate may be at a relatively low temperature, for example, less than -5 ° C, -10 ° C, or -12 ° C, Layer or the like to prevent unwanted freezing during operation. Additionally or alternatively, the applicator may include a diluent liquid, a cooling element (e.g., cooling water delivered by the applicator), and a thermal element (e.g., a heating element) for heating the working fluid .

Figure 27b shows an applicator and a diluted coupling medium. The diluent liquid is passed through the conduit to dilute the coupling medium. The amount of diluent liquid may be selected to achieve the desired coupling medium concentration. The coupling medium may be frozen as the applicator cools, and the temperature of the coupling medium is stabilized at a predetermined or target temperature, for example, -8 캜, -10 캜, or -12 캜. INA can be incorporated into the coupling medium or into the injected liquid to promote freezing.

Ultrasound may be used to initiate, facilitate, and / or control the freezing event. Referring now to FIG. 27A, cold water may be delivered to the ultrasonic chamber and, in some embodiments, may contact the upper surface of the liquid coupling medium. Cooling elements, cold plates, or other components of the applicator can cool water around the ultrasonic probe, which can be activated to transmit ultrasonic energy to the cooling water to cause freezing. Ultrasonic shaking (e. G., Ultrasonic shaking using a suitable frequency, power, etc.) will cause the freezing to propagate throughout the chamber and reach the coupling medium, resulting in instantaneous A freezing event can be generated.

One or more INA's may be incorporated into the coupling medium before, during, and / or after applying the coupling medium to the patient. Diluting the coupling medium to a point where the diluted melting temperature is higher than its actual temperature will cause the diluted coupling medium to freeze, which will eventually cause freezing of the skin.

Figures 28 to 31 show treatments that can entail supercooling. Supercooling cycling can cover a wide range of temperatures, and the extreme desired supercooling temperature can often be too cold for use as a freezing temperature, since this causes excessive damage to the epidermis. Coupling media that remain liquid during the supercooling cycle are often unable to inoculate because the supercooling temperature range is higher than the freezing point of the coupling medium. However, dilution of the coupling medium increases the freezing point of the coupling medium and enables freezing of the skin at a warmer applicator and skin temperature. Advantageously, the epidermal temperature can be warm enough to inhibit, restrict, or substantially prevent pigmentation degradation, hyperpigmentation, or other undesirable effects.

Dilution also allows supercooling cycling at relatively low temperatures (e.g., -10 ° C, -15 ° C, -20 ° C), and tissue can enhance or maximize the damage to the targeted tissue, The damage is frozen at relatively high temperatures (e.g., -10 ° C, -5 ° C, -4 ° C, -3 ° C, or -2 ° C) to limit or minimize damage. The targeted tissue may be tissue in the dermis and / or lower skin layer, and the non-targeted tissue may be epidermis or shallow tissue. Although enhancing or maximizing damage can be achieved by several successive treatments using different concentrations of coupling media, a single treatment can provide the desired damage to reduce treatment time and cost.

28 shows an applicator 261, a medium 262 with INA, and a coupling layer 263. INA can be placed in direct contact with the skin surface to promote predictable freezing of the skin. In some procedures, the INA is placed in direct contact with the cooling surface of the applicator. For example, media 262 may include one or more INA (e.g., SNOMAX® / water mixture). The coupling layer 263 may comprise a cellulose-derived layer and solution, for example water, and may be in direct contact with the treatment site. In some embodiments, the INA may be applied using a thin layer (paper) impregnated with an INA coupling agent, mixed with a gel-like material, delivered via a delivery device (e.g., a syringe), or sprayed onto the surface. Skilled artisans may substitute suitable coupling layer materials, chemicals, conditions, and delivery systems with other materials, chemicals, conditions, delivery systems, and the like.

Figure 29 shows a plot of temperature versus time of temperature profile for inducing ice nucleation through INA. The tissue may be subcooled to a temperature above the INA activation temperature. The temperature of the INA is then lowered to its activation temperature to initiate ice nucleation, producing a partial or complete freezing event (indicated by " * ") propagating into the skin and through the skin. The cycle can be completed by maintaining the temperature of the applicator to allow the growth of the ice crystals at the treatment site. After completing the cycle, the temperature of the applicator may be gradually raised to the desired thawing rate to warm the skin.

Figure 30 shows an applicator and INA applied to a treatment site. INA can be delivered in the coupling medium 271, on the skin surface of the subject, and / or in the skin, to predictably cause ice nucleation. The applicator may include embedded fluidics to controllably deliver the INA to the interface between the applicator and the skin of the subject. The injected INA may be at a particular therapeutic temperature or within a predetermined temperature range to inhibit ice nucleation in the interface or tissue itself, for example, in the coupling medium. In another embodiment, the INA is sprayed or otherwise delivered to the skin of the coupling medium or the subject. In some processes, the applicator cools the coupling medium and the skin of the subject. The applicator may lift the coupling medium and the INA may be sprayed onto the cooled coupling medium. After spraying the INA, the applicator can be reapplied to the treatment site to continue cooling the INA, coupling media, and tissue. Needles, rollers, and other delivery mechanisms may be used to apply one or more INAs. Other techniques may be used to provide INA implantation through the coupling medium and on or in the coupling medium and / or tissue of the subject.

Figure 31 shows a plot of temperature versus time for the delivery of INA for nucleation. The INA may be delivered to the treatment site at or below the activation temperature of the INA (indicated by the dashed line) at a particular time to initiate nucleation. As indicated by the arrows, the INA may be injected to initiate a freezing event. Thus, the activation temperature of the INA can be selected to trigger a controlled freezing event.

A variety of techniques can be used to protect targeted tissue volume and / or non-targeted tissue while affecting certain structures within, for example, epidermis, dermis, subcutaneous tissue, and the like. Targeted structures include, without limitation, hair (e.g., hair follicles), skin appendages (e.g., glands, sebaceous glands, etc.), nerves, and / or dermal components such as collagen, elastin, Blood vessels. Targeted structures can be affected while suppressing, preventing, or substantially eliminating unwanted side effects. Because accessory organs and other cells / structures can have different lethal temperatures, a multistage temperature profile can be used to target specific tissues and / or structures. In addition, preserving non-targeted tissue such as epidermis from excessive injury or damage may be beneficial, for example, in preventing pigment changes and / or scarring and promoting healing. Freezing the epidermis at a different temperature than the underlying dermis can be achieved by using the characteristic activation temperature of INA and intentionally subcooling the dermis at lower temperatures before applying INA. In some processes, the epidermis may be at a higher temperature to inhibit, restrict, or substantially prevent permanent thermal damage to the epidermis.

Some embodiments of the technique involve using a conjugated polymer containing water, a water-containing crosslinked polymer, optionally INA, and / or optionally a freezing point depressant for controlled freezing of the skin tissue. According to one preferred embodiment, the polymer may be a hydrogel for use for controlled freezing of dermal tissue. Hydrogels can be effective initiators of freezing events. As the hydrogel freezes, it may provide an initial seed or crystal site to be inoculated into the tissue to freeze the tissue, thus catalyzing predictable freezing of the skin at a certain temperature (s).

Figures 32A-32D show IR imaging of tissue freezing using supercooled tissue and hydrogel. Figure 32a shows the dermal tissue (dashed area) on both hydrogels (left and right hemisphere) and the cooling plate overlying the hydrogel. Figure 32b shows freezing at lower left side. Figure 32c shows freeze propagation along most of the left hydrogel demonstrating ice inoculation. Figure 32d shows frozen propagation through both hydrogels.

Referring now to Figure 32A, the dermal tissue in the area indicated by the dashed line contacts the two half-sheets of the hydrogel. The hydrogel sheet without PG or other freezing point depressants is located to the left of the vertical line and the hydrogel sheet with about 50% by volume PG is located to the right of the vertical line. The rectangular cooling plate is located on the hydrogel sheet. The skin surface is in direct contact with both hydrogels, which ultimately comes into direct contact with the cooling plate. In this manner, the hydrogel can be firmly held between the patient and the applicator.

Images were generated after a few minutes of tissue subcooling. The temperature of the subcooled tissue was lowered to the triggering temperature, resulting in freezing (illustrated by lighter color) in the non-PG hydrogel illustrated on the left side of Figure 32b. 32B and 32C show freeze propagation caused by using only a hydrogel. The right half of Figure 32b shows a portion of 50 vol% PG hydrogel and unfrozen adjacent skin. 32C shows freeze propagation across the hydrogel, through the tissue, and towards the hydrogel with the temperature-reducing agent. This shows that various concentrations of PG or other freezing point depressants can be included in the hydrogel to lower the melting / freezing temperature of the hydrogel to the desired value below 0 ° C. In the case of aqueous hydrogels without PG or other freezing point depressants, freezing at temperatures close to 0 ° C is inconsistent and unpredictable for controlled heterogeneous nucleation. However, the hydrogels used with INA provide the ability for controlled freezing at sub-zero temperatures, including temperatures near 0 [deg.] C in a more predictable manner (or less when used with freezing point depressants).

33A-33D illustrate a tissue freeze inoculation using combined materials, and an INA (e. G., SNOMAX < (R) >) on a supercooled hydrogel to induce freezing. Or other suitable INA derived from the bacterial Pseudomonas syringae). ≪ / RTI > Figure 33a shows the placement of the granules of the INA on the subcooled hydrogel. Figure 33 (b) shows that the INA is inoculated into the hydrogel and the freezing begins to propagate around the INA. Figure 33 (c) shows freeze propagation to tissue across the hydrogel, demonstrating the binging of the skin. Figure 33 (d) shows the frozen propagation completed. In combination, FIGS. 33A-33D illustrate the feasibility of using INA as a seed to inoculate controlled freezing in dermal tissue. The details of Figures 33a-33d are discussed below.

33A shows the non-frozen hydrogel and INA placed near the edge of the cooling plate (shown by the dashed line) while the cooling plate cools the tissue. During the initial deployment of the INA, there is no significant freezing through the tissue facing the cooling plate. INA can initiate the freezing process by acting as an ice nucleating agent and can raise the predictable freezing temperature of water to about -3 ° C. The water has a melting / freezing temperature of 0 ° C, but water tends to be subcooled, so the freezing temperature of water is often much lower than 0 ° C or -3 ° C when INA is not used. INA can initiate the freezing process by acting as an ice nucleating agent and can raise the predictable freezing temperature of water to about -3 ° C. The INA can be selected to raise the predictable freezing temperature of the water to another desired temperature. When selecting an INA, one skilled in the art will be able to select a suitable agent (e.g., an organic or an inorganic derivative) that is used at a particular desired temperature (s) and can be delivered at a particular time for a particular therapeutic purpose . Different techniques can be used to incorporate the INA into the hydrogel. For example, the INA can be sandwiched between the layers of the hydrogel such that it is totally contained and encapsulated in the hydrogel and never in contact with the skin or tissue. The encapsulant may disintegrate, destroy, or otherwise modify, releasing INA. In some embodiments, the cooling plate of the applicator may deliver the INA to the hydrogel via one or more needles, outlet ports, or other delivery means. The INA can be applied to a single site or multiple sites, or it can be mixed into a hydrogel compound. In addition, the INA can be inside a micrometer wall made of a hard or soft soluble film to avoid direct skin contact and has a disassembly controlled by passive or active means. In addition, the INA may be injected or delivered into the skin or onto the skin.

Figure 33 (b) shows the tissue after freezing has spread apart from the INA. Frozen material is shown in a lighter color compared to a darker background color representing non-frozen tissue. 33C shows freezing propagated across the hydrogel facing the cooling plate. 33D shows the frozen propagation completed to freeze all of the skin in direct contact with the hydrogel.

34A and 34B are cross-sectional views of a cooling applicator applied to a treatment site. Referring now to Figure 34A, only the hydrogel is located between the applicator and the skin of the subject. The cooling applicator may be disposed on the protective layer in the form of a thin cover layer. The protective layer may be a liner, or other component to prevent cross-contamination or degradation by the hydrogel. As the temperature-controlled surface of the cooling applicator cools, it draws heat from the skin through the hydrogel and thin cover layer, thereby eventually cooling the skin.

Figure 34B shows the hydrogel and INA located between the applicator and the skin of the subject. The INA may be a liquid, gel, cream, preformed sheet or layer located along the surface of the hydrogel. If the applicator is applied to a hydrogel, the INA may be located at the hydrogel-applicator interface. The hydrogel and / or hydrogel / INA / freezing point depressant may be selected to melt / freeze into its formulation at the specified temperature.

The hydrogels of FIGS. 34A and 34B can be used in combination with water-monomer-crosslinking agents and / or other chemicals in constituents to achieve a specific freezing point (or near temperature range) which may or may not enable skin supercooling, One or more INA, a freezing point depressant, and the like. INA may have a known activation temperature (natural freezing point) and may be a solid form (i.e., powder) or a mixed solution with the desired concentration to produce a predictable and consistent skin freeze. Such a heat-coupling material or compound can be used to provide a desired treatment, such as to enable a supercooling event, or possibly a subcooling, and to enable predictable and controlled freezing at the desired temperature and time Temperature protocols or algorithms.

35 is a plot of temperature versus time for inducing copper settlement in accordance with an embodiment of the disclosed technique. A temperature profile, protocol, and / or algorithm to cause one or more freezing enables the supercooling of tissue at the temperatures allowed by the hydrogel or hydrogel / INA. The temperature of the targeted tissue can be maintained in the supercooling temperature range during the supercooling period. The temperature of the hydrogel is then lowered to the freezing temperature to cause ice nucleation of the hydrogel or material. If undergoing a freeze event, the underlying tissue underneath may be equal to or slightly warmer than the hydrogel. In some procedures, both the hydrogel and the targeted tissue are subcooled while the temperature of the temperature-controlled surface of the applicator is generally kept constant. In another procedure, the targeted tissue can be partially frozen, while the hydrogel is subcooled. Subsequent freezing of the hydrogel may cause additional freezing of the target tissue until the desired level of freezing is achieved in the targeted tissue. Other coupling media may be used for the temperature profile shown in Fig.

Figure 36 illustrates an applicator arranged to produce controlled freezing in a coupling medium that transfers the formulation onto the surface of the coupling medium, into the coupling medium, or in another suitable position for initiating the freeze event in the coupling medium Show. The applicator may include one or more needles (e.g., micro-needle array), fluid components (e.g., conduits, pumps, valves), reservoirs (e.g. In some embodiments, the formulation is delivered out of the outlet port at the bottom of the cooling plate of the applicator.

Figure 37 shows a cooling applicator having an outer nucleation element configured to initiate a freeze event outside of the applicator-hydrogel interface. The outer nucleation element may comprise one or more energy-emitting elements capable of initiating a freezing event. In some embodiments, the outer nucleation element transfers energy (e. G., Ultrasonic energy, RF energy, etc.) to the edge region of the coupling agent to produce freezing, which includes the area of the coupling agent, And propagates directly between the applicator and the tissue site. In another embodiment, a separate nucleation mechanism may initiate nucleation, which may be a probe with nucleation elements.

38 is a cross-sectional view of a cooling applicator that provides energy-based activation. A coupling medium, a hydrogel, a hydrogel / INA mixture, or other material for producing controlled freezing may be located between the cooling applicator and the treatment site. In some embodiments, the encapsulated ice nucleating agent may be part of or located within the layer of the coupling medium. The energy can break the encapsulation and release the ice nucleating agent at the desired time. This can cause a freezing event to spread to the skin surface through the coupling medium. When ice crystals contact the skin surface, freezing can be transmitted through the skin. The operating energy may be, without limitation, mechanical energy (e.g., vibration, ultrasonic waves), electrical energy, and / or electromagnetic radiation (e.g., light).

Figure 39 shows the temperature profile for subcooling the skin before initiating the freezing event. The freeze activation can be controllably initiated to control freeze initiation, while the temperature-controlled surface of the applicator and / or the target tissue is maintained at a constant steady state temperature. It may be desirable to subcool and freeze the skin and subcutaneous tissue at a specific temperature (or temperature range) for a specified time to enable a controlled subcooling of tissue volume large enough to cause extensive freezing during tissue inoculation.

It is important to keep the epidermis cool while cooling / cooling the tissue or affecting certain structures in the dermis and subcutaneous tissues such as hair, skin, appendages, nerves, dermis components such as collagen, elastin or blood microvessels Can be advantageous. Because ancillary organs and other cells / structures may have different lethal or injury temperatures, a multistage temperature profile may be required. Furthermore, preserving the epidermis may be beneficial in preventing skin pigment changes and skin scarring. Also, preserving the epidermis may lead to more favorable healing and fewer side effects. It is possible to freeze the epidermis at a temperature different from that of the underlying dermis by using the technique described above. Specifically, the skin bulk structure can be supercooled at a low temperature and then the temperature of the epidermis can be elevated, for example, before delivery of INA or activation of nucleation. When the epidermis is frozen at a temperature of approximately -5 ° C or higher, the epidermal sensitivity is reduced. If freezing occurs under these temperatures, melanocytes and / or their melanin production in the epidermis may be excessively altered resulting in pigmentation. Thus, according to some embodiments of the disclosed technique, a temperature protocol that causes freezing of the epidermis above -5 DEG C may be used.

Figure 40 shows the temperature of the applicator warming the epidermis after subcooling and before freezing so that the applicator and / or the epidermis are at a temperature above the supercooling temperature (e.g., -6, -5, -4, Is a plot of temperature versus time when conditioned. After heating the epidermis, a freeze event is generated. For example, the temperature profile shows the activation or delivery of INA at a warmer activation temperature that is suitable for protecting tissues such as the epidermis or the upper layers of the skin. The time period for heating rate, activation temperature, and activation temperature may be selected based on the desired tissue protection and the effect on the targeted tissue. The active maintenance period can be increased or decreased to increase or decrease the protection of the epidermis.

Figure 41 shows three cross-sections of a plot of temperature versus time for a cooling protocol and an applicator and skin tissue temperature distribution. As shown in the plot, the temperature-controlled surface of the applicator is maintained at -10 ° C for 5 minutes (in the case of supercooling) and then increased to the desired rate (eg, 2 ° C / s, 2.5 ° C / s, etc.) . The subsequent freezing event is indicated by the rise in temperature after approximately time = 400 seconds. Computer modeling (COMSOL) was used to generate the results. The model was a three-dimensional bio-heat transfer model for the treatment of skin using a cooling applicator and was used to generate the plots discussed herein.

The image shows the temperature distribution in the tissue in relation to temperature profile change of the temperature-controlled surface from -10 ° C to -4 ° C. An isotherm was added at time = 380 seconds, time = 385 seconds, and time = 400 seconds (T = 0 C). The isotherm at T = 0 ° C is the boundary at which the phase change to ice crystallization (freezing of the skin) can be maximally extended (ie, the deeper tissue is warmer than 0 ° C, It will not freeze if ice nucleation occurs because it is above the temperature).

Figure 42 shows the temperature profile (logarithmic scale) at epidermis / dermis at time = 380 seconds for the applicator at -10 ° C and shows a temperature of T = 0 ° C at 2 mm depth into the skin. T = 0 ° C The depth of the isotherm is about 2 mm. Thus, if nucleation occurred at this point, freezing could be extended to about 2 mm below the skin at this point. A temperature gradient may be observed and shows a gradient of 0 DEG C at a depth of T = -8 DEG C to 2 mm at the surface of the skin.

Figure 43 is a plot of the time taken for time = 380 seconds, time = 385 seconds, and time = 400 seconds, showing the temperature of the applicator showing the temperature as the applicator stepped up from -10 占 to -4 占 폚 at 2.5 占 폚 / / Temperature profile (logarithmic scale) in the dermis. That is, the temperature profile across the depth of the skin at time = 380 seconds, time = 385 seconds, and time = 400 seconds is both before and after the applicator temperature has transitioned from -10 占 to -4 占 폚. The temperature gradient in the epidermis is above -5 ° C at time = 400 sec so that controlled freezing can be induced. The epidermis is frozen at a more optimal temperature (> -5 [deg.] C) but a better degree of skin freezing is achieved to a depth of approximately 2 mm.

Figure 44 shows a method for generating tissue freezing using an apparatus disposed within a subject. The device may be a needle that injects the material to a specific location. The inner surface of the needle may be coated to promote the transfer of the material. The outer surface of the needle may be coated with a formulation or material to treat tissue, targeted structures, and the like. Other devices may be inserted into the object to generate a freeze event.

The injected material may include, without limitation, hydrogels, hydrogel / INA, partially frozen water, ice nucleating agents, combinations thereof, and the like. An advantage of injecting a substance that produces ice crystals or ice crystals (e.g., INA) is that the frozen event takes place in a certain region. The freezing event may be initiated in the dermis or other sub-tissue layers and not in the epidermis. This limits or minimizes damage to the epidermis. Further, the epidermis may be warmed to a temperature close to or higher than its melting / freezing temperature. In some embodiments, the freezing event can be initiated in the tissue below the dermis as in subcutaneous tissue. After generating a freezing event, the same or different needles can inject additional material into the tissue. Additional materials may include cryoprotectants, liquids (e. G., Hot or saline), or other materials capable of performing the treatment.

Multiple scans can be made to generate multiple freezing events. The first material may be delivered into the tissue to produce a first freeze event and the second material may be delivered into the tissue to cause a second freezing event. For example, the first material may be suitable for completely freezing the first target area, and the second material may be suitable for generating a partial freezing event in a second target area spaced apart from the first target area have. Although the first and second target regions are at the same temperature, different degrees of severity of freezing and thermal damage can be achieved. In other treatments, the first and second target regions may be at different temperatures, and the first and second materials may be selected based on these temperatures. In this way, different types of freezing events can be generated at different locations.

44, a material having a heat-coupling or nucleating agent having a freezing point above the freezing point of the fluid in the skin tissue is used synergistically with the treatment cycle to intentionally freeze the material And subsequently cause freeze transfer into the skin at higher temperatures. The non-targeted tissue may be warmed by an applicator (e.g., through conduction), a injected warm liquid, and / or energy (e.g., RF energy). The warming cycle can be performed to warm the epidermis immediately after creating a frozen event that damages the target structure, such as the sebaceous glands. This can help prevent visible changes to the epidermis (e.g., hyperpigmentation, poor pigmentation, etc.). The injectable material can be delivered to and around the sebaceous glands, and then freezing events can be induced by temperature dives, dilution, energy, and the like.

45 is a flow chart illustrating a method 350 in accordance with aspects of the present technique. At block 352, a coupling medium may be applied to the object. At block 354, the applicator cools the tissue to a temperature suitable for the frozen event. For example, the skin surface may be lowered to a first temperature of about -2 [deg.] C to -40 [deg.] C to sub-cool the shallow tissue. In some embodiments, the first temperature may be a temperature in the range of -5 ° C to -15 ° C, -5 ° C to -20 ° C, -10 ° C to -30 ° C, or other suitable temperature range below the freezing temperature.

At block 356, the skin surface of the human subject is heated in an amount sufficient to raise the skin surface temperature to a second temperature, which may be a first temperature ratio-subcooled temperature, while the targeted area is maintained in a subcooled state do. For example, the epidermis may be heated to a temperature of at least about 0 ° C, at least about 5 ° C, at least about 10 ° C, at least about 20 ° C, at least about 30 ° C, or at least about 35 ° C. There may be a temperature gradient between the targeted tissue and the skin surface so that most of the non-targeted shallow tissue is at a non-subcooled temperature.

At block 356, the apparatus of FIG. 44 may cause nucleation in the target area so that at least a portion of the fluid and cells of the supercooled tissue are at least partially or entirely frozen. The warmed cells on the skin surface of the human subject are not frozen. Thus, cells on the surface of the skin can be protected without the use of chemical cryoprotectants. However, chemical cryoprotectants may be used to inhibit or limit hyperpigmentation or degradation of pigmentation. In some embodiments, a probe may be inserted into a subject to cause nucleation of the subcooled tissue through mechanical perturbation, ultrasound, or other suitable nucleation initiator. The freezing event can cause at least partial crystallization of a plurality of germ cells in the target region. The illustrated device of Figure 44 is arranged to generate a freezing event that causes crystallization of the cells in the sebaceous glands.

At block 358 of FIG. 45, the supercooled tissue may be used to treat acne, improve hair quality, and treat hyperhidrosis, for example, at a time of about 10 seconds, 12 seconds, 15 seconds, And other suitable time lengths sufficient for a predetermined period of time. In certain embodiments, the skin is cooled / heated to maintain the targeted tissue at least partially or totally frozen for a pre-determined time period of greater than about 10 seconds, 12 seconds, 15 seconds, or 20 seconds.

Heat may be applied to warm the epidermal cells to above freezing, but the spleen of dermis is at or near the subcooled temperature. Applying the heat may include warming a portion of most of the epidermal layer under the treatment device to a temperature greater than about 0 캜, about 5 캜, about 10 캜, about 20 캜, about 25 캜, or about 32 캜 have. The warming can be achieved before the freeze event, during the freeze event, or after the freeze event. The epidermis may be naturally heated such that body heat, warmth, or other mechanisms of the subject may avoid or limit freezing damage to these cells.

If the deeper tissue is not targeted, such tissue may be warmed using concentrated current such as concentrated ultrasound or RF energy. The applicator may include one or more electrodes, transducers, or other energy-emitting elements. For example, the applicator may cool the skin surface shown in Figure 44 to subcool the tissue comprising the dermis. The applicator may deliver RF energy or concentrated current to the underlying non-targeted subcutaneous tissue to localize the subcooling to the dermal tissue. The freezing event is then initiated in a sub-cooled dermal tissue.

The methods disclosed herein can subcool the tissue without initiating nucleation by cooling the tissue at a relatively slow rate (e.g., the temperature profile can cause slow cooling of tissue in the target area). For example, the cooling rate can be about 0.5 ° C per minute, 1 ° C, 2 ° C, 3 ° C, 4 ° C, 5 ° C, 6 ° C, 7 ° C, 8 ° C, 9 ° C, It can be fast. A preferred cooling rate is about 2 DEG C, 4 DEG C, or 6 DEG C per minute. Additionally or alternatively, the treatment device may apply a constant pressure during cooling to the subcooled temperature range to avoid pressure changes that cause accidental nucleation. In a further embodiment, the targeted tissue can be cooled while the patient is still standing (for example, without moving the treatment site), in order to mechanically disturb the supercooled tissue and avoid inadvertent crystallization.

F. Appropriate Computing Environment

46 is a schematic block diagram illustrating subcomponents of a controller-type computing device suitable for the system 100 of FIG. 3 in accordance with an embodiment of the present invention. The computing device 700 includes a processor 701, a memory 702 (e.g., SRAM, DRAM, flash, or other memory device), an input / output device 703, and / ). The computing device 700 can perform any of a wide variety of computing processing, storage, sensing, imaging, and / or other functions. The components of the computing device 700 may be received as a single unit or distributed over multiple interconnected units (e.g., via a communications network). Thus, components of computing device 700 may include local and / or remote memory storage devices and any of a wide variety of computer-readable media.

As illustrated in Figure 46, the processor 701 may include a plurality of functional modules 706, such as software modules, for execution by the processor 701. Various implementations of the source code (i.e., in a conventional programming language) may be stored in a computer-readable storage medium or may be implemented in a transmission medium as a carrier wave. The module 706 of the processor may include an input module 708, a database module 710, a process module 712, an output module 714, and optionally, a display module 716.

In operation, input module 708 accepts operator output 719 via one or more of the input / output devices described above in connection with FIG. 3 and communicates the accepted information or selection to other components for further processing. The database module 710 organizes records including patient records, treatment data sets, treatment profiles and operational records and other operator activities, and provides information to the data storage device (e.g., internal memory 702, external database, etc.) Thereby facilitating the storage and retrieval of such records from it. Any type of database cleanup can be used, including single layer file systems, tiered databases, relational databases, distributed databases, and the like.

In the illustrated example, process module 712 may generate control variables based on sensor readings 718 from sensors (e.g., sensor 167 in FIG. 2) and / or other data sources, Output module 714 may communicate the operator input to an external computing device and control variables to controller 114 (Figure 3). An output signal 720 may be used to control one or more applicators applied to the patient. In some embodiments, the output signal 720 may be an instruction to control the applicator. Display module 816 is coupled to one or more connected display devices, such as a display screen, a printer, a speaker system, etc., for processing parameters, sensor readings 818, output signals 720, input data, To transmit and transmit data. Suitable display module 716 may include a video driver that allows controller 114 to display sensor readings 718 or other treatment progress.

In various embodiments, the processor 701 may be a standard central processing unit or a secure processor. A secure processor may be a special purpose processor (e.g., a reduced instruction set processor) that can tolerate sophisticated attacks attempting to extract data or programming logic. The secure processor may not have a debug pin that allows an external debugger to monitor the execution or registration of the secure processor. In other embodiments, the system may utilize a secure field programmable gate array, smart card, or other secure device.

Memory 702 may be a combination of both standard memory, secure memory, or memory type. By using a secure processor and / or secure memory, the system can ensure that both data and instructions are highly secure and that sensitive operations such as decryption are shielded from observations. The memory 702 may be configured to cool the skin surface of the object to a temperature and to control the treatment device in response to, for example, supercooling detection, partial or complete freezing events, movement of the applicator (e.g., stripping of the applicator) May contain executable instructions. In some embodiments, the memory 702, when executed, may include a nucleation instruction word that causes the controller to instruct the applicator to change the composition of the coupling medium, scan the nucleation initiator, and the like. Additionally or alternatively, the memory 702, when executed, may include thawing instructions that allow the controller to control the applicator to heat the tissue. In some embodiments, the stored instructions can be executed to control the applicator to perform the method disclosed herein without causing undesirable effects such as significant lightning or darkening skin after more than one day of freezing event. The command may be changed based on patient information and the treatment to be performed. Other instructions and algorithms (including feedback control algorithms) may be stored and executed to perform the methods disclosed herein.

In some embodiments, the controller 114 is programmed to cause the applicator to generate or maintain at least one ice crystal and to induce a frozen event. Memory 702 may contain, for example, instructions that, when executed, cause the one or more ice crystals to contact the subject skin to cause the applicator to induce a freeze event. In one embodiment, the memory 702, when executed by the processor 701, is configured to cause the applicator to cool down the target tissue without lowering the temperature of the temperature-controlled surface below a certain level, In order to prevent the unauthorized use of the apparatus. The instructions may be used to control or communicate components of the applicator. Such components may include, without limitation, one or more thermoelectric elements, fluid elements, energy-emitting elements, and sensors. The thermoelectric element may be a Peltier element capable of selectively cooling or heating tissue. The fluidic element may be a cooling channel, conduit, or other fluidic element through which the fluid may flow to heat and / or cool the tissue. The energy-emitting device may be a radio frequency electrode, an ultrasonic electrode, or other device capable of transferring energy to control freezing, warming tissue, and the like.

Suitable computing environments and other computing devices and user interfaces are described in commonly assigned U.S. Patent No. 8,275,442 entitled " Treatment Planning System and Method for < RTI ID = 0.0 > Body Contouring < / RTI > Applications, .

G. Conclusion

Therapeutic systems, applicators and methods of treatment may be used to treat acne, hyperhidrosis, wrinkles, subcutaneous tissue, structures (e.g., epidermal, dermal, subcutaneous fat, muscle, The method and associated apparatus and system for cooling tissue in accordance with embodiments of the present invention may be used to remove one or more problems associated with prior art as discussed above and / or other problems, whether or not such problems are addressed herein At least partially. Methods of affecting the skin of a human subject include arranging an applicator of a cooling device on a subject and removing heat from the treated area to cause a pronounced decrease in subcutaneous fat tissue or to affect the appearance of the skin of the subject . Acne along the face can be treated without causing any reduction in subcutaneous fat tissue, where acne along the back can be treated while reducing subcutaneous adipose tissue. Systems, components and techniques for reducing subcutaneous adipose tissue are described in U.S. Patent No. 7,367,341 to Anderson et al., Entitled " Method and Apparatus for Selective Cleavage of Adipose Tissue by Controlled Cooling, US 2005/0251120 to Anderson et al. Entitled " METHOD AND APPARATUS FOR DETECTION AND CONTROL OF SELECTIVE FROZEN TISSUE BY TUBERCULOSIS, " and " for use in therapy devices for improved cooling of subcutaneous fat- Quot; Cryoprotective < / RTI > Agent for < RTI ID = 0.0 > Cryoprotectant < / RTI > ", the contents of which are hereby incorporated by reference in their entirety. For example, a sufficient amount of heat energy can be removed from the area, for example, to reduce wrinkles by reducing the number of visible wrinkles and / or the size of wrinkles. In another embodiment, a sufficient amount of thermal energy is removed from the treatment site to tighten the skin to the treatment site, or, in a further embodiment, to change the tissue between the skin surface of the subject's body and subcutaneous lipid-rich cells . In a further embodiment, the tissue is cooled to induce fibrosis which increases the firmness of the tissue at the treatment site. Fibrosis can be induced in the epidermis, dermis, and / or subcutaneous tissue. Vacuum applicators can stretch the skin, stress it, stress it or mechanically change it to increase damage and fibrosis to the skin, affect the fountain, control the freezing event (including initiation of freezing events), etc. .

It will be appreciated that some well-known structures or functions may not be shown or described in detail, in order to avoid unnecessarily obscuring the relevant description of the various embodiments. While some embodiments may be within the scope of the description, they may not be described in detail with reference to the figures. In addition, the features, structures, or characteristics of various embodiments may be combined in any suitable manner. The techniques disclosed herein are described in US patent application Ser. No. 61 / 943,250, filed Feb. 21, 2014, entitled " Method and Apparatus for Selective Cleavage of Adipose Tissue by Controlled Cooling " U.S. Patent No. 7,367,341 to Anderson et al., Entitled " Method and Apparatus for Detecting and Controlling the Selective Fractionation of Adipose Tissue by Controlled Cooling, " and US Patent Application Publication No. US 2005/0251120 to Anderson et al. May be used to carry out the process, the contents of which are incorporated herein by reference in their entirety. The techniques disclosed herein can be used to target tissue to tighten the skin, improve skin tone or texture, eliminate or reduce wrinkles, or increase skin smoothness, as disclosed in U.S. Patent Application No. 61 / 943,250 can do.

Unless the context expressly requires otherwise, throughout the description, the words " comprises ", " including ", and the like shall be interpreted in a generic sense as opposed to an exclusive or perfect meaning; That is, to mean "including, but not limited to". Words using singular or plural also include plural or singular, respectively. The use of the word " or " with respect to a list of two or more items encompasses all of the following interpretations of the word: any of the items of the list, all of the items of the list, and any combination of the items of the list. In the case where a convention similar to " at least one of A, B, and C, etc. " is used, this configuration is generally intended to be conventional (e.g., " systems with at least one of A, B, Includes but is not limited to systems with A alone, B alone, C alone, A and B together, A and C together, B and C together, and / or A, B and C together. In the case where a convention similar to " at least one of A, B, or C, etc. " is used, this configuration is generally intended to be customary in meaning (e. G., A system having at least one of A, B, Includes but is not limited to systems with A alone, B alone, C alone, A and B together, A and C together, B and C together, and / or A, B and C together.

Any patents, applications, and other references, including those that may be listed in the accompanying submission, are incorporated herein by reference. Aspects of the described techniques may be modified as needed to provide additional embodiments using the systems, functions, and concepts of the various references discussed above. While the above description details specific embodiments and describes the best mode contemplated, various changes may be made, regardless of how detailed the description is. Execution details can vary considerably but are encompassed by the techniques disclosed herein. The various aspects and embodiments disclosed herein are for the purpose of illustration and not of limitation, the true scope and spirit is indicated by the following claims.

Claims (140)

A method for predicting freezing of a skin of a subject at a desired predictable time,
Lowering the temperature of the skin below the freezing point of the fluid in the skin such that the skin is at a first temperature;
Contacting the skin with an ice crystal to inoculate the skin and generating a predictable frozen event therein; And
And controlling the contact time between the ice crystals and the skin such that the predictable freezing occurs at a desired time. The method of claim 1, further comprising physically contacting the skin surface with the ice crystals. The method of claim 1, further comprising introducing the ice crystals into the subject using a catheter such that the ice crystals contact the dermal layer of the skin. The method of claim 1, further comprising forming ice crystals in a cryoprotectant solution located on the skin. 5. The method of claim 4,
Maintaining the temperature of the skin at a first temperature during a first treatment period wherein the first temperature is at least 7 ° C below the freezing point of the fluid in the skin; And
Further comprising cooling the cryoprotectant solution from a temperature above the freezing point of the cryoprotectant solution to a temperature below the freezing point of the cryoprotectant solution to form the ice crystals therein after the first treatment period . The method according to claim 4, wherein, in order to raise the freezing point of the diluted concentration to a value equal to or higher than the temperature of the cryoprotectant solution so that formation of the ice crystal does not require the skin temperature to be lowered below the first temperature, Further comprising diluting the cryoprotectant concentration on the surface. 2. The method of claim 1, further comprising generating ultrasonic energy to cause the glazing to contact the skin or to the skin in contact with the skin. The method of claim 1, wherein the first temperature is at least 10 ° C below the freezing point of the fluid in the skin prior to skin inoculation. 9. The method of claim 8,
Maintaining the first temperature during the first treatment period;
Lowering the skin temperature to a second temperature lower than the first temperature after the first treatment period expires to generate the ice crystals; And
Further comprising maintaining the second temperature during a second treatment period. 10. The method of claim 9, further comprising rapidly warming the skin surface to a third temperature higher than the first or second temperature after the second treatment period has expired. 11. The method of claim 10,
Maintaining the third temperature during a third treatment period wherein the third temperature is a cooling treatment temperature within 5 [deg.] C of the freezing point causing minimal damage to the epidermal tissue; And
Further comprising the step of warming said skin surface to or above room temperature. The method of claim 1, wherein the first temperature is less than 3 ° C below the freezing point of the fluid prior to skin inoculation. The method of claim 1, further comprising maintaining the first temperature for a predetermined period of time to generate a pre-determined level of skin cooling before the ice crystals contact the skin. The method according to claim 1,
Detecting the freeze event; And
Further comprising controlling the length of time of the freezing event by controlling when the temperature of the skin rises to a temperature above the freezing point of the fluid in the skin based on the detection of the freezing event. The method of claim 1, further comprising forming the ice crystals before lowering the temperature of the skin below the freezing temperature of the fluid. 16. The method of claim 15, further comprising forming the ice crystals after lowering the temperature of the skin below the freezing temperature of the fluid. The method of claim 1, further comprising using a coupling medium in contact with the skin to help remove heat from the skin, wherein the coupling medium comprises an ice nucleating agent, a hydrogel, and / A freezing point depressant. The method according to claim 1,
Applying a coupling medium to the skin, wherein the coupling medium has a medium therein capable of forming ice crystals; And
Establishing a temperature of the applicator in thermal contact with the coupling medium to predictably freeze a portion of the medium in physical contact with the skin surface so that the frozen portion of the medium causes freezing of the skin ≪ / RTI > 19. The method of claim 18, wherein the coupling medium comprises a crosslinked polymer comprising water and wherein the coupling medium has a predetermined freezing point of greater than -40 DEG C, -25 DEG C, -20 DEG C, or -15 DEG C Having a specific ratio of water-monomer-crosslinker content. 20. The method of claim 19, wherein the crosslinked polymer is a hydrogel. 19. The method of claim 18, wherein the medium is an ice nucleating agent (INA). 22. The method of claim 21,
Biogenic proteins;
Substances derived from gram-negative co-bacteria; And / or
A substance belonging to the genus Pseudomonas, Erwinia, or Xanthomonas. 19. The method of claim 18, wherein the coupling medium comprises a hydrogel containing the ice nucleating agent of the medium so that the ice nucleating agent does not contact the skin when the coupling medium is absorbed by the skin , Way. 19. The method of claim 18, wherein the coupling medium comprises an ice nucleating agent contained in an inner layer of the hydrogel of the coupling medium or in a liposome structure of the coupling medium. The method according to claim 1,
Applying a coupling medium to the object, wherein the coupling medium has a substance therein capable of forming ice crystals;
Establishing a temperature of the cooling surface of the applicator in thermal contact with the coupling medium to at least partially freeze the material in direct contact with the cooling surface, wherein the temperature is such that the portion of the material in contact with the skin surface Liquid temperature); And
Further comprising inoculating said skin surface to lower the temperature of said cooling surface to freeze at least a portion of the material in contact with said skin surface such that said freezing event occurs predictably in said skin. 26. The method of claim 25, wherein the coupling medium comprises a hydrogel containing water. 26. The method of claim 25, wherein the coupling medium comprises a freezing point depressant and an ice nucleating agent. 26. The method of claim 25, wherein lowering the temperature of the cooling surface comprises cooling the cooling surface in an amount sufficient to cause at least partial freezing of the skin tissue without significant supercooling of the skin. 26. The method of claim 25,
Maintaining the cooling surface at a sufficiently low temperature to cool the skin tissue without freezing the skin during the first treatment period, and
At least a portion of the coupling medium in contact with the skin surface is frozen and inoculated onto the skin surface by lowering the temperature of the cooling surface after the first treatment period has expired, To be maintained during a second treatment period. 30. The method of claim 29, wherein after the second treatment period expires, the skin is rapidly heated using the applicator to rapidly thaw the skin tissue. 30. The method of claim 29, wherein after the first treatment period has expired,
Warming the applicator in an amount that is sufficient to cause the dermal tissue to thaw due to heat, but not enough to cause the epidermal tissue to thaw; And
Further comprising after the dermis tissue is thawed, the dermis tissue is re-frozen and the epidermal tissue is frozen by adjusting the temperature of the applicator to a second therapeutic temperature. The method according to claim 1,
Lowering the temperature of the cooling surface of the applicator below the freezing point of the coupling medium of the cooling surface to freeze a portion of the coupling medium; And
Further comprising applying the coupling medium of the cooling surface to the skin of the subject such that the liquid portion of the coupling medium separates the skin of the subject from the frozen portion of the coupling medium . 33. The method of claim 32, wherein after applying the coupling medium to the skin, the coupling medium is cooled through the cooling surface to propagate the freeze front through the coupling medium, To produce one or more ice crystals that cause said predictable frozen event. 33. The method of claim 32, further comprising ultrasonically freezing the medium in the coupling medium. 33. The method of claim 32, further comprising subcooling the skin of the subject through the coupling medium and the cooling surface. The epidermal tissue is frozen once and the dermis tissue is frozen more than once,
Applying a coupling medium to the applicator, wherein the coupling medium has a medium therein capable of forming ice crystals;
Lowering the temperature of the applicator below the freezing point of the medium to at least partially freeze the medium;
Disposing a coupling medium having the applicator on the skin surface of the subject;
Adjusting the temperature of the applicator to a treatment temperature for freezing at least a portion of the medium in contact with the skin surface to freeze the dermis and epidermal tissue;
Warming the applicator in an amount sufficient to cause the dermal tissue to thaw but not enough to thaw the epidermal tissue; And
The method comprising adjusting the temperature of the applicator to a second treatment temperature such that after the dermis tissue is at least partially defrosted, at least a portion of the defrosted dermis tissue is re-frozen and the epidermis remains frozen . 37. The method of claim 36, further comprising maintaining the applicator in thermal contact with the skin surface while freezing, thawing, and re-freezing the dermis tissue. 37. The method of claim 36, further comprising alternately thawing and freezing the dermis while keeping the epidermis tissue freeze. The epidermal tissue is frozen once and the dermis tissue is frozen more than once,
At least partially freezing the dermis and epidermal tissue using an applicator configured to cool the skin surface of the subject; And
While keeping the epidermal tissue freeze,
Warming the applicator in an amount sufficient to cause at least a portion of the dermal tissue to thaw,
Cooling the applicator to re-freeze at least a portion of the dermis tissue after thawing at least a portion of the dermis tissue. 40. The method of claim 39, wherein the step of warming the applicator comprises heating and defrosting the dermis tissue while raising the temperature of the temperature-controlled surface of the applicator such that the epidermal tissue remains frozen. 40. The method of claim 39, wherein cooling the applicator comprises reducing the temperature-controlled surface of the applicator and re-freezing the thawed dermal tissue. 40. The method of claim 39, further comprising at least partially freezing the dermal tissue repeatedly and then thawing the dermal tissue. 40. The method of claim 39,
Applying a first coupling medium to the cooling applicator, wherein the first coupling medium has a predetermined freezing point below 0 占 폚;
Applying the applicator to the skin surface and maintaining the temperature of the applicator such that the layer of the first coupling medium contacting the skin surface while subcooling the skin during the first treatment time is not frozen; And
Further comprising lowering the temperature of the applicator after the first treatment time expires to freeze at least a portion of the coupling medium in contact with the skin surface to produce a predictable freeze event in the skin. Way. 44. The method of claim 43, wherein the first coupling medium comprises water and a freezing point depressant. 44. The method of claim 43, wherein a second coupling medium is applied to the skin prior to applying the applicator. 40. The method of claim 39,
Detecting freezing in the skin; And
Further comprising the step of rapidly warming the applicator such that the applicator is at a temperature greater than -6 [deg.] C for a predetermined treatment time in response to detection of freezing of the skin. 47. The method of claim 46, wherein the temperature of the applicator is raised to a warm freezing temperature or a cooling therapeutic non-freezing temperature. A method for predictably freezing skin,
Supercooling the skin of a subject using a coupling medium and an applicator at a first applicator temperature wherein the first applicator temperature is higher than the freezing point of the coupling medium and wherein the coupling medium is in contact with the coupling medium and the skin Lt; / RTI > freezing point); And
And inoculating said skin to freeze said skin without lowering the temperature of said applicator below said first applicator temperature. 49. The method of claim 48, wherein the step of inoculating the skin comprises diluting at least a portion of the coupling medium to raise the freezing point of at least a portion of the coupling medium to a level equal to or higher than the first applicator temperature How to. 50. The method of claim 49, wherein the diluted coupling medium is frozen while the applicator is at the first applicator temperature. 49. The method of claim 48, wherein the step of inoculating the skin comprises applying liquid water to the skin. 49. The method of claim 48, wherein the step of inoculating the skin comprises applying at least one ice crystal to the skin. 49. The method of claim 48, further comprising inoculating said skin using ultrasound, an ice nucleating agent, and / or a small cold finger probe. 49. The method of claim 48, further comprising, prior to inoculating the skin, such that the epidermis is at a higher temperature than the dermis when the freezing event occurs and after the occurrence, so as to inhibit damage to epidermal tissue, Further comprising raising the temperature of the surface. As a method for treating skin,
Lowering the temperature of the skin of the subject below the freezing point of the target tissue of the skin;
Monitoring the cooling of the skin so that freezing does not occur therein;
Controlling the amount of non-freezing cooling therapy delivered to the skin such that the target tissue reaches a predetermined first level of cooling; And
After the target tissue has reached the predetermined first level,
The skin was frozen,
Controlling the amount of freezing and cooling therapy delivered to the skin such that the target tissue reaches a predetermined second level of cooling. 57. The method of claim 55, wherein the predetermined first level is a sub-cooled therapeutic level and the predetermined second level provides a therapeutically effective amount of thermal injury. 56. The method of claim 55, wherein the predetermined first and second levels are selected to affect sebaceous glands of the dermis without permanently damaging the epidermis. 56. The method of claim 55, wherein prior to freezing the skin, the temperature of the epidermis is raised to a value above the temperature of the dermis. 59. The method of claim 58, wherein the value is higher than -10, -9, -8, -7, -6, -5, -4, or -3 degrees Celsius. 57. The method of claim 55, wherein the predetermined first level is selected based on a target degree of initial freezing with skin depth free of the desired depth. 56. The method of claim 55, wherein immediately after freezing the skin, the temperature of the applicator in thermal contact with the surface of the epidermis is increased to a first value that is higher than the temperature of the dermis immediately prior to freezing and is sufficiently low to continue cryotherapy therapy Ascending, method. 56. The method of claim 55, further comprising lowering the temperature of the skin of the subject through the cooling surface of the applicator at an applicator temperature first value maintained until an overall level of treatment is achieved, Further comprising increasing the temperature of the cooling surface at the time of switching to the first value to be greater than the rate of change of the temperature of the applicator at the time of switching to the second value, . 56. The method of claim 55, wherein the step of freezing the skin comprises initiating a freezing event by lowering the temperature of the applicator in thermal contact with the skin surface to a predetermined amount sufficient to cause nucleation. 56. The method of claim 55, further comprising initiating the freezing by applying an ice nucleating agent to the skin. 64. A system configured to perform the method of any one of claims 1-64. A system for treating a subject,
The applicator configured to cool the skin surface of the object when the applicator is applied to the object; And
The applicator
Generating or maintaining at least one ice crystal,
To cause a freeze event in the skin of the subject through the at least one ice crystal
And a programmed controller. 67. The system of claim 66, wherein the controller, when executed, comprises an instruction to cause at least one ice crystal to contact the skin of the subject. 67. The method of claim 66, wherein the controller, when executed, causes the cooling surface of the applicator to be at a first temperature for sub-cooling the target tissue of the subject, and wherein the temperature of the cooling surface is not lowered below the first temperature And causing the skin to freeze. 69. The system of claim 68, further comprising a coupling medium having a freezing point below the first temperature. 67. The apparatus of claim 66,
Controlling the amount of non-freezing cooling therapy delivered to the skin such that the targeted tissue of the skin reaches a predetermined first level;
After the targeted tissue reaches the predetermined first level,
The skin was frozen,
Wherein the amount of freezing cooling therapy delivered to the skin is programmed to control it to reach a predetermined second level. 71. The system of claim 70, wherein the predetermined first level is a supercooled state and the predetermined second level is a frozen state. 67. The system of claim 66, wherein the applicator comprises a sensor configured to monitor the skin of the subject, wherein the controller is programmed to control the applicator based on an output from the sensor. 73. The system of claim 72, wherein the sensor comprises a temperature sensor arranged to sense the temperature of the skin of the subject. 67. The system of claim 66, wherein the controller is part of the applicator. 67. The system of claim 66, wherein the controller is communicatively coupled to the applicator. The system of any of claims 66 to 75, configured to perform the method of any one of claims 1-64. A method of treating skin of a subject,
Applying the hydrogel to the skin, wherein the hydrogel comprises an ice nucleating agent capable of forming ice crystals in the presence of water, wherein the ice nucleating agent is selected such that the ice nucleating agent is directly Encapsulated within the polymeric structure of the hydrogel so as not to contact;
Cooling the hydrogel and the skin with a cooling applicator to reach a cooling therapy temperature for the skin; And
And freezing the skin through the hydrogel. 80. The method of claim 77, wherein the step of cooling the hydrogel and the skin comprises:
Cooling the hydrogel below its freezing point;
And cooling the skin below its freezing point. 79. The method of claim 78, further comprising initiating a freezing event in the hydrogel by cooling the hydrogel to a temperature below the freezing point of the ice nucleating agent. 77. The method of claim 77, further comprising freezing the skin for a length of time sufficient to heat damage the sebaceous glands. 77. The method of claim 77, wherein the hydrogel comprises a freezing point depressant such that the first freezing point of the hydrogel is lower than a second freezing point of the fluid in the skin. 83. The method of claim 81, wherein cooling the hydrogel and the skin comprises:
Cooling the skin to a temperature higher than the first freezing point of the hydrogel and lower than the second freezing point of the skin to subcool the skin,
And after the predetermined amount of supercooling has occurred, freezing the skin. 83. The method of claim 82, wherein the skin is frozen by lowering the temperature at the skin surface to a temperature lower than the first freezing point. 83. The method of claim 82, further comprising raising the temperature of the epidermis above a first point prior to freezing the skin, wherein the temperature of the epidermis is above the first point and the skin is frozen. 83. The method of claim 82, further comprising diluting the hydrogel with water to raise the first freezing point to a freezing point above the cooling temperature. 78. The method of claim 77, wherein the hydrogel comprises water and a crosslinked polymer, wherein the coupling gel is selected such that the hydrogel has a predefined freezing point < RTI ID = 0.0 > Monomer-crosslinker content to have a specific ratio of water-monomer-crosslinker content. 78. The method of claim 77, wherein the step of freezing the skin through the hydrogel comprises contacting the skin with a frozen portion of the hydrogel to initiate freezing of the skin. 77. The method of claim 77, further comprising maintaining the hydrogel against the skin such that at least one ice crystal in the hydrogel contacts the skin to cause freezing of the skin. 77. The method of claim 77, wherein the ice nucleating agent is embedded in the polymeric structure. 80. The method of claim 77,
Derived protein;
Substances derived from gram negative co-bacteria; And / or
Pseudomonas, Erbina, and / or Xanthomonas. As a method for treating skin,
Applying a substance to the skin, wherein the substance comprises a crosslinked polymeric structure and an ice nucleating agent (INA), wherein the polymeric structure comprises water, and wherein the INA is selected from the group consisting of ice crystals And the INA is enclosed within the polymeric structure to prevent direct contact between the INA and the skin);
Cooling the material and the skin with a cooling applicator to lower the temperature of the skin; And
And freezing the skin through the material. 92. The method of claim 91,
Wherein the material is a hydrogel,
Wherein applying the substance to the skin comprises placing a sheet of the hydrogel on the surface of the skin or injecting the hydrogel into the skin. 92. The method of claim 91, wherein the material comprises an ice nucleation region and an ice nucleation inhibition region, wherein the ice nucleation region contains the INA,
Wherein applying the substance to the skin further comprises disposing the ice nucleation inhibition region between the skin surface and the ice nucleation region. 92. The method of claim 91, further comprising forming one or more ice crystals in the material to cause freezing of the skin. 92. The method of claim 91, further comprising transmitting energy to the material to cause ice crystal formation therein. 92. The method of claim 91, further comprising delivering energy to destroy the polymer structure to release a sufficient amount of the INA to produce a freezing event that causes freezing of the skin. 92. The method of claim 91, further comprising delivering a nucleating agent to the material cooled by the applicator to produce controlled freezing in the material. A method of treating skin of a subject,
Applying the hydrogel to the skin wherein the hydrogel comprises a freezing point depressant and water and the freezing point depressant is encapsulated within the polymeric structure of the hydrogel such that the freezing point depressant does not come into direct contact with the skin surface, ;
And cooling the hydrogel and the skin with a cooling applicator to reach a cooling therapy temperature for the skin. A method of treating skin of a subject,
Applying a coupling medium to the skin wherein the coupling medium comprises a freezing point depressant and a liposome containing the freezing point depressant to enhance delivery of the freezing point depressant to the skin, Emulsion, or oil-in-water emulsion); And
And cooling the coupling medium and the skin surface with an applicator to a temperature below < RTI ID = 0.0 > 0 C < / RTI > to treat the skin. 102. The method of claim 99, wherein the freezing point depressant comprises propylene glycol, glycerol, and / or polyethylene glycol. 99. The method of claim 99, wherein the coupling medium comprises the liposome, wherein the liposome and the freezing point depressant are propylene glycol liposomes. 102. The method of claim 99, wherein the coupling medium comprises the liposome, wherein the liposome contains water such that after the liposome is degraded, the freezing point depressant and / or the water is released into the skin. 102. The method of claim 99, wherein the coupling medium comprises a liposome,
Further comprising destroying the liquid encapsulation of the liposome using ultrasound energy, temperature cycling, and / or a detergent to release the freezing point depressant. 103. The method of claim 99, further comprising controlling cooling of the coupling medium and the skin surface such that the skin is not frozen. 103. The method of claim 99, further comprising controlling cooling of the coupling medium and cooling of the surface to freeze the skin. 102. The method of claim 99, wherein the coupling medium further comprises an ice nucleating agent,
Further comprising controlling cooling of the coupling medium and the skin surface to predictably freeze the skin based on nucleation caused by the ice nucleating agent. 99. The method of claim 99, wherein the coupling medium is the oil-in-water emulsion or the water-in-oil emulsion. 107. The method of claim 107, wherein the oil-in-water emulsion or the water-in-oil emulsion is a nanoemulsion. A method of treating skin of a subject,
Applying a coupling medium to the skin surface of the subject, wherein the ice nucleating agent contained in the applied coupling medium is not in direct contact with the epidermal cells for a period of time;
Applying an applicator to the skin of the subject; And
Cooling the coupling medium and the skin surface to lower the skin surface temperature below 0 ° C to cause a freeze event initiated by the ice nucleating agent to freeze the skin. 110. The method of claim 109, wherein cooling the coupling medium and the skin surface comprises removing heat from the skin after the predetermined time to start freezing the skin. 109. The method of claim 109, wherein the coupling medium comprises a liposome, an oil-in-water emulsion, a water-in-oil emulsion, a water-in-oil emulsion, and / or a nano-emulsion. 109. The method of claim 109, wherein the coupling medium comprises a freezing point depressant to lower the freezing point of the skin. 114. The method of claim 112, wherein the freezing point depressant is configured to penetrate only a shallow depth into the skin to lower the freezing point of epidermal tissue by more than the freezing point of dermal tissue. 109. The method of claim 109, further comprising the step of degrading the liposome of the coupling medium to release the ice nucleating agent to initiate the freezing event. 110. The method of claim 109, wherein cooling the coupling medium and the skin surface comprises:
Removing heat from the skin to subcool the targeted tissue in the skin;
Freezing the targeted tissue;
Freezing the targeted tissue, removing heat from the skin and / or applying heat to the skin to maintain the epidermal layer freeze while thawing and re-freezing the dermal layer. 108. The method of claim 109,
Diluting the coupling medium to raise its freezing point to produce the frozen event, and / or
Transferring energy to activate said ice nucleating agent, and / or
Further comprising lowering the temperature of the coupling medium to cause the freezing event after the fluid in the skin of the subject has cooled below its freezing point. A system for treating a subject,
An applicator configured to lower the temperature of the target area below the skin surface of the subject to lower the temperature of the target tissue from a natural body temperature to a temperature for freezing the target tissue;
A coupling medium comprising a freezing point depressant, and a liposome, oil-in-water emulsion, water-in-oil emulsion, or oil-in-water emulsion containing said freezing point depressant to enhance delivery of said freezing point depressant to said skin; And
And wherein the applicator comprises a controller programmed to cool the coupling medium and the skin surface to a temperature below 0 ° C to freeze the skin for a period of time. 117. The system of claim 117, configured to perform the method of any one of claims 77 to 116. A method of treating a subject,
Mechanically changing the skin of the subject to promote absorption of the coupling medium;
Applying the coupling medium to the mechanically altered skin such that the coupling medium is absorbed into the skin;
Cooling said mechanically altered skin to subcool the targeted tissue; And
Using ice crystals to initiate a freeze event in said skin to freeze said subcooled targeted tissue. 120. The method of claim 119, wherein the freezing of the subcooled targeted tissue causes damage to cells contributing to acne. The method of claim 119, wherein mechanically changing the skin of the subject comprises:
Applying an adhesive strip to said skin surface;
And removing the adhesive strip from the skin surface to increase the permeability of the skin to the coupling medium. 120. The method of claim 119, wherein mechanically changing the skin of the subject comprises mechanically stimulating the skin by brushing the skin surface. 124. The method of claim 122, further comprising brushing the skin surface for at least 2 minutes. 120. The method of claim 119, wherein the step of mechanically changing the skin of the subject causes at least mild exfoliation of the stratum corneum. 120. The method of claim 119, wherein the coupling medium comprises a cryoprotectant and / or an ice nucleating agent. 120. The method of claim 119, further comprising mechanically altering the skin of the subject to increase the coefficient of transmission of the coupling medium through the skin's skin layer by at least 10%. A method of treating a targeted tissue of a subject,
Increasing the permeability of the skin of the subject at the treatment site to promote absorption of the coupling medium to the skin of the subject;
Applying the coupling medium to the skin of the subject having increased permeability;
Applying an applicator to the treatment site; And
Using the applicator to remove heat from the treatment site to generate a freeze event in the skin of the subject for a predetermined length of time. 127. The method of claim 127, further comprising increasing the permeability coefficient of the coupling medium by at least 10% for skin absorption. 127. The method of claim 127,
Applying a stripping element to the treatment site; And
Further comprising removing the stripping element from the treatment site to increase the permeability of the skin of the subject. 127. The method of claim 127, further comprising mechanically stimulating the skin of the subject to increase the coefficient of transmission of the coupling medium to the skin by at least 10%. As a method of treating acne,
Stimulating the skin of the subject to increase the permeability coefficient of the epidermis of the subject to the coupling medium;
Applying a coupling medium to the stimulated skin; And
Removing heat from the skin of the subject, cooling the applied coupling medium and creating a freezing event in the skin to affect tissue associated with acne. 132. The method of claim 131,
Applying an adhesive strip to the skin; And
Further comprising removing the adhesive strip from the skin to irritate the skin. 133. The method of claim 131, further comprising brushing the skin surface to mechanically irritate the skin. 132. The method of claim 131, further comprising cooling the skin of the subject to maintain the freezing event for a length of time sufficient to destroy cells contributing to acne. 132. The method of claim 131, wherein the coupling medium comprises a liposome and / or an emulsion. 132. The method of claim 131,
Applying an applicator to the skin surface; And
Further comprising using the applicator to remove heat from the skin of the subject to produce the freeze event. A method of treating a subject,
Applying an adhesive strip to the skin surface;
Removing the adhesive strip from the skin surface to increase the permeability of the skin to the coupling medium;
Applying the coupling medium to the skin such that the coupling medium is absorbed into the skin; And
Cooling the skin to subcool the targeted tissue. 143. The method of claim 137, further comprising initiating a freeze event in the skin using an ice crystal to freeze the supercooled targeted tissue. A method of treating a subject,
Brushing the skin surface to increase the permeability of the skin to the coupling medium;
Applying the coupling medium to the skin such that the coupling medium is absorbed into the skin; And
Cooling the skin to subcool the targeted tissue. A system configured to perform the method of any one of claims 119 to 139.

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