[go: up one dir, main page]

WO2024208455A1 - Méthode de préparation d'une greffe de tissu adipeux - Google Patents

Méthode de préparation d'une greffe de tissu adipeux Download PDF

Info

Publication number
WO2024208455A1
WO2024208455A1 PCT/EP2024/051304 EP2024051304W WO2024208455A1 WO 2024208455 A1 WO2024208455 A1 WO 2024208455A1 EP 2024051304 W EP2024051304 W EP 2024051304W WO 2024208455 A1 WO2024208455 A1 WO 2024208455A1
Authority
WO
WIPO (PCT)
Prior art keywords
chamber
adipose cells
cells
adipose
ozone
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
PCT/EP2024/051304
Other languages
English (en)
Inventor
Paolo Giuseppe Gobbi Frattini
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Paolo Gobbi Frattini SRL
Original Assignee
Paolo Gobbi Frattini SRL
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Paolo Gobbi Frattini SRL filed Critical Paolo Gobbi Frattini SRL
Publication of WO2024208455A1 publication Critical patent/WO2024208455A1/fr
Anticipated expiration legal-status Critical
Pending legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A61L27/3683—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
    • A61L27/3687—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment characterised by the use of chemical agents in the treatment, e.g. specific enzymes, detergents, capping agents, crosslinkers, anticalcification agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61J—CONTAINERS SPECIALLY ADAPTED FOR MEDICAL OR PHARMACEUTICAL PURPOSES; DEVICES OR METHODS SPECIALLY ADAPTED FOR BRINGING PHARMACEUTICAL PRODUCTS INTO PARTICULAR PHYSICAL OR ADMINISTERING FORMS; DEVICES FOR ADMINISTERING FOOD OR MEDICINES ORALLY; BABY COMFORTERS; DEVICES FOR RECEIVING SPITTLE
    • A61J1/00—Containers specially adapted for medical or pharmaceutical purposes
    • A61J1/05—Containers specially adapted for medical or pharmaceutical purposes for collecting, storing or administering blood, plasma or medical fluids ; Infusion or perfusion containers
    • A61J1/10—Bag-type containers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61J—CONTAINERS SPECIALLY ADAPTED FOR MEDICAL OR PHARMACEUTICAL PURPOSES; DEVICES OR METHODS SPECIALLY ADAPTED FOR BRINGING PHARMACEUTICAL PRODUCTS INTO PARTICULAR PHYSICAL OR ADMINISTERING FORMS; DEVICES FOR ADMINISTERING FOOD OR MEDICINES ORALLY; BABY COMFORTERS; DEVICES FOR RECEIVING SPITTLE
    • A61J1/00—Containers specially adapted for medical or pharmaceutical purposes
    • A61J1/14—Details; Accessories therefor
    • A61J1/1475—Inlet or outlet ports
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61J—CONTAINERS SPECIALLY ADAPTED FOR MEDICAL OR PHARMACEUTICAL PURPOSES; DEVICES OR METHODS SPECIALLY ADAPTED FOR BRINGING PHARMACEUTICAL PRODUCTS INTO PARTICULAR PHYSICAL OR ADMINISTERING FORMS; DEVICES FOR ADMINISTERING FOOD OR MEDICINES ORALLY; BABY COMFORTERS; DEVICES FOR RECEIVING SPITTLE
    • A61J1/00—Containers specially adapted for medical or pharmaceutical purposes
    • A61J1/14—Details; Accessories therefor
    • A61J1/20—Arrangements for transferring or mixing fluids, e.g. from vial to syringe
    • A61J1/2003—Accessories used in combination with means for transfer or mixing of fluids, e.g. for activating fluid flow, separating fluids, filtering fluid or venting
    • A61J1/2048—Connecting means
    • A61J1/2058—Connecting means having multiple connecting ports
    • A61J1/2062—Connecting means having multiple connecting ports with directional valves
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61J—CONTAINERS SPECIALLY ADAPTED FOR MEDICAL OR PHARMACEUTICAL PURPOSES; DEVICES OR METHODS SPECIALLY ADAPTED FOR BRINGING PHARMACEUTICAL PRODUCTS INTO PARTICULAR PHYSICAL OR ADMINISTERING FORMS; DEVICES FOR ADMINISTERING FOOD OR MEDICINES ORALLY; BABY COMFORTERS; DEVICES FOR RECEIVING SPITTLE
    • A61J1/00—Containers specially adapted for medical or pharmaceutical purposes
    • A61J1/14—Details; Accessories therefor
    • A61J1/20—Arrangements for transferring or mixing fluids, e.g. from vial to syringe
    • A61J1/2093—Containers having several compartments for products to be mixed
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel

Definitions

  • the present invention relates to a method for the preparation of human or animal adipose tissue grafts for allotransplantation or preferably for autologous transplant.
  • the present invention relates to the treatment with ozone of the adipose tissue cells collected from the body of a human patient or an animal during the preparation of the graft.
  • graft means a suitable quantity of adipose cells extracted from the body of an individual, treated, and that are ready to be implanted in the body of the same individual (autotransplantation, autologous transplant) or of a different individual (allotransplantation).
  • adipose tissue from a point rich in adipose tissue like e.g. abdomen or thighs to a desired point like e.g. breasts or glutei is a procedure of reconstructive or aesthetic surgery known in the art. E.g., such a procedure can be used for reconstructing a breast after a surgical intervention for removing a tumour, or for a purely aesthetic intervention of additive mastoplasty.
  • adipose tissue coming from the same patient autologous transplant
  • adipose-derived stem cells in particular adipose-derived stem cells, endothelial progenitor cells, perivascular cells, macrophages, fibroblasts, lymphatic cells, which are connected to the adipose cells through the matrix of connective tissue.
  • stem cells are multipotent regenerative cells with the potential to differentiate into a plurality of other cell types performing one or more specific functions, and have the capacity of autoregeneration.
  • stem and regenerative cells are desired in the graft, in that they have a positive effect on the long-term success of grafting, e.g. promoting angiogenesis.
  • sterile solutions for rinsing unprocessed adipose cells comprise sterile water, saline, Ringer’s lactate, Ringer’s acetate, phosphate-buffered saline, crystalloid solutions, colloidal solutions, 5% glucose in water, Hartmann’s solution and the like.
  • adipose tissue grafting can lead to undesired effects like infections, necrosis, reabsorption of the graft by the body, calcifications and scarring, which derive at least in part from the lack of neovascularization of the graft and from the necrosis of the grafted tissue.
  • graft rejection is the presence of impurities in the graft, like e.g. red blood cells, white blood cells, and/or free lipids (oil).
  • impurities like e.g. red blood cells, white blood cells, and/or free lipids (oil).
  • the membranes of such cells can be damaged leaking the content of the lipid droplets inside the adipose cells. It is known that such free lipids can lead to inflammation of the surrounding tissues, and in the worst cases, to necrosis and graft rejection.
  • Body tissues are normally sterile, but once the cells have been extracted from the body, they can be contaminated by virus and/or bacteria that can be airborne or present on surfaces.
  • the removal of the rinsing liquid allows to provide a preparation containing processed cells having a reduced liquid content, which constitute a dehydrated graft that can be grafted in the desired anatomical area immediately after collection and rinsing or of a dehydrated graft that can be cryopreserved in order to implant it in a subsequent moment.
  • EP2424463B1 describes an alternative embodiment of the same invention, providing the use of a device comprising two chambers shown in the Figures 13, 14 of the document.
  • Such chambers have substantially the shape of rigid cylinders in fluid communication, and from the point of view of performing the method, said cylinders can be likened to the chambers of the flexible bags.
  • a system for separating adipose cells through centrifugation is known under the commercial name Sepax® manufactured by Biosafe SA and described in the document EPl 144026B 1.
  • a further system of separation of adipose cells is known under the commercial name Celution®, manufactured by Cytori Therapeutics and described in the document EP2380970B1.
  • a system of separation of adipose cells through fragmentation with balls is known under the commercial name Lipogems®, manufactured by Lipogems and described in the document WO2015140737A1.
  • US 2017/224869 Al discloses a method for producing a tissue product.
  • the method comprises selecting an adipose tissue, treating the tissue to remove substantially all cellular material from the tissue, suspending the tissue in a liquid to form a suspension, layering the suspension in a mould, and freezing and drying the suspension to form a porous sponge.
  • CN 106492288 A discloses injectable acellular fat matrix particles prepared from adipose tissue by a method comprising the steps of providing animal fat tissue, rinsing, disinfecting, rinsing, repeated freezing and thawing, cutting into thin slices, then soaking in water, degreasing and rinsing.
  • a method known in the art for preparing an adipose tissue graft comprises the following steps: a) Introducing a portion of unprocessed adipose tissue in a chamber of a device; b) Rinsing the unprocessed adipose tissue using a sterile liquid; c) Removing the liquid from the chamber containing said adipose cells, so obtaining a rinsed and dehydrated adipose tissue.
  • Ozone is a gas naturally present in the atmosphere; its molecule consists of three atoms of oxygen. It is a highly unstable gas tending to rapidly decompose to molecular oxygen (O2). It is a strong oxidizing agent. Ozone is soluble in water.
  • ozone has been known for a long time, but its clinical application is limited by its strong cytotoxicity at high doses.
  • the aim of the present invention is providing a method and a device for performing such method capable of improving the success of adipose cell grafting.
  • the present invention provides a method comprising the already known steps a), b), c), wherein the rinsing step b) is performed by placing said adipose cells in contact with a suitable quantity of ozone (O3).
  • Said ozone can be placed in contact with cells in the form of a liquid solution containing a suitable quantity of dissolved ozone (ozonized solution), or in the form of a mixture of oxygen/ozone gas in suitable quantities for cell fumigation.
  • the treatment with ozone is performed using a gaseous solution containing 0.1 to 8% of ozone (O3) in in a gaseous solution containing 99.9 to 92% of oxygen (O2).
  • the quantity of ozone in oxygen can vary 0.1 to 80 micrograms per millilitre.
  • the final ozonized solution contains 1 pg to 80 pg of ozone per mL of solution, equivalent to 0.1 - 8% of ozone in oxygen weight/volume.
  • the bubbling time in ultra-filtered water, or in saline solution (0,9% NaCl) or in one of the sterile liquids used for rinsing, needed to form the ozonized solution is a time comprised in 0.1 - 10 minutes, preferably 0.1-4 minutes.
  • the quantity/volume of sterile liquid to be ozonized can vary between 1 mL and 2000 mL; in case of big grafts of adipose tissues can reach 5000 mL.
  • said treatment with ozone occurs inside a device having the shape of a flexible bag shown in the Figures.
  • Said bag comprises three chambers.
  • the first chamber of said flexible bag is provided to the final user already filled with a sterile rinsing liquid chosen from the group consisting of sterile water, saline solution, Ringer’s lactate, Ringer’s acetate, phosphate-buffered saline, crystalloid solutions, colloidal solutions, 5% glucose in water solutions, Hartmann’s solutions.
  • a sterile rinsing liquid chosen from the group consisting of sterile water, saline solution, Ringer’s lactate, Ringer’s acetate, phosphate-buffered saline, crystalloid solutions, colloidal solutions, 5% glucose in water solutions, Hartmann’s solutions.
  • said flexible bag is supported by a dedicated support allowing to keep said bag in a vertical position, with its longitudinal axis alternatively parallel or perpendicular to the ground, in order to perform more efficiently the different steps of the method.
  • said support comprises a rectangular plate with a superficial face for supporting said flexible bag, an extension of one of the short sides and an extension of one of the long sides bent in the opposed direction with respect to the face supporting the flexible bag, and four hooks for connecting said flexible bag to said supporting face.
  • the separation of adipose cells from other types of cells can be performed through decantation/sedimentation, or centrifugation, or fragmentation with balls.
  • ozone is placed in contact with adipose cells using devices already on the market, which provide the use of containers different from flexible bags, generally rigid cylindrical containers.
  • EP1144026B1 Sepax
  • EP2424463B1 Puregraft
  • EP2380970B1 Celution
  • WO2015140737A1 Lipogems
  • the method of treatment with ozone according to the present invention is provided with two main advantages.
  • the first advantage is that ozone is provided with a known disinfecting activity, therefore is capable of destroying possible bacterial and/or viral contaminations present in the adipose cells, so preventing the occurrence of infections once the graft has been grafted in the body of a patient, and the potential rejection of the graft in the long-term.
  • the second advantage is that the treatment with low quantities of ozone can promote the anti-oxidant response of cells, activating adipogenesis and the stimulation of stem and regenerative cells contained in the graft of adipose tissue, so improving the probabilities of success of the grafting.
  • Stem cells treated with ozone tend to be more numerous, vital and active, increasing their efficacy when they are grafted in the patient.
  • figure 1 shows a schematic representation of a flexible bag device for preparing a cell graft according to a preferred embodiment, front view
  • figure 2 shows said flexible bag device, in a longitudinal section according to the line A- A of figure 1
  • figure 3 shows said flexible bag device, in a transversal section along the line B-B of figure 1
  • figure 4 shows schematically the possibility of connecting a bag device with a syringe containing a sterile rinsing solution or alternatively an ozone generator
  • figure 5 shows schematically the connection between an ozone generator and said flexible bag
  • figure 6 shows schematically the connection between a syringe containing an ozonized solution or alternatively ozone and said flexible bag
  • figures 7 show schematically the flexible bag device supported by a dedicated support.
  • figure 7A shows the bag vertically supported by said support, with its longitudinal axis perpendicular to the ground in a front view
  • figure 7B shows the bag vertically supported by said support with its longitudinal axis parallel to the floor in a front view
  • figure 7C shows the bag vertically supported with its longitudinal axis perpendicular to the ground in a transparent axonometric view
  • figure 7D shows the bag vertically supported by said support with its longitudinal axis parallel to the ground in a transparent axonometric view.
  • the device for preparing adipose tissue grafts that is exemplarily shown in figures 1-3 is a bag 10 made of a flexible plastic material.
  • Said bag 10 is a closed system, allowing to sterilely process unprocessed adipose cells preventing their contact with environmental air; this allows to prevent bacterial and/or viral contaminations of the graft which could lead to infections and rejection of the graft.
  • Said bag 10 when empty has a flat shape, and in a top view is provided with a substantially rectangular shape, with two short sides and two long sides.
  • the bag is manufactured by welding two equal flexible sheets of a suitable plastic material capable of being sterilized during production, and not degrading when placed in contact with ozone, according to the present invention. The material must also be capable of tolerating the temperatures needed for cryo-conserving the graft. Two welding 21, 22 divide the rectangular area of the bag in three hollow chambers 11, 12, 13.
  • Said bag 10 is provided with four holes 31, 32, 33, 34 obtained near its angles, for the connection with a support device 70, shown in figures 70.
  • the first hollow chamber 11 is provided for receiving and containing a sterile rinsing liquid (sterile water, saline, Ringer’s lactate, Ringer’s acetate, phosphate-buffered saline, crystalloid solutions, colloidal solutions, 5% glucose in water solutions, Hartmann’s solutions, ozonized solutions or the like) that is inserted through an inlet port 1 comprising a loading flexible tube 49 provided with a hermetic sealing plug 50 that can be opened through an external axial pressure and automatically closed when said pressure stops.
  • the sealing plug 50 is preferably of the kind described in EP2667839B1 and is provided with a threaded end 51.
  • the second hollow chamber 12 is provided for receiving the raw or unprocessed adipose tissue, wherein said tissue is rinsed and undergoes a treatment with a rinsing liquid and ozone (ozonized solution) according to the present invention, and successively dehydrated.
  • a rinsing liquid and ozone ozonized solution
  • said hollow chamber 12 comprises a separator filter 9 having the shape of a bag with substantially known features, which filter comprises pores of dimensions suitable for passing the impurities of the unprocessed adipose tissue, while holding adipose cells and stem and regenerative cells.
  • said filter 9 can be made of polyester or PES with porosities of 80 or 105 pm, inside a porosity field ranging 50 to 200 pm, or of Nylon.
  • Said second chamber 12 further comprises an inlet port 4 on one of its long sides, provided for sterilely introducing, preferably through a syringe, an unprocessed adipose tissue into said chamber 12, and in particular into said filter 9, and an outlet port 3 on a long side provided for sterilely extracting, preferably through a syringe, the dehydrated graft of adipose tissue after the processing of said cells occurred insides said chamber 12.
  • a tube 2 provided for connecting said first chamber 11 and said second chamber 12.
  • a known three-way valve 6 allowing to adjust the outflow of liquid from said first chamber 11 to said second chamber 12.
  • Said three-way valve 6 is provided with an inlet port 6’ that can be used even for connecting said tube 2 with a (not shown) syringe with the aim of inserting active principles of different kinds (e.g. antibiotics or inhibitors of the immune system in case of allotransplantation) inside said second chamber 12.
  • the inlet port 6’ is provided with a sealing plug of the same kind of the above- mentioned plug 50.
  • the third draining hollow chamber 13 is provided for draining impurities and liquids coming from the second chamber 12.
  • the third chamber 13 comprises an outlet port 5 allowing to drain the liquid from the third chamber 13, and eventually from said bag 10.
  • the connection between said second chamber 12 and said third draining chamber 13 occurs through a non-return valve 8 of known type (schematized only in figure 1), allowing the passage of liquids from the second chamber 12 to the third chamber 13.
  • Said valve 8 is normally closed and allows the passage of liquids in just one direction, preventing the rise of the draining liquid from said third chamber 13 to said second chamber 12.
  • the passage of the draining liquid occurs through the pressure exerted by the liquid on said valve 8. In other words, as soon as the liquid is pushed from the second chamber 12, it ends directly in the third chamber 13, and it is not possible to hold it in the second chamber 12.
  • connection between second chamber 12 and third chamber 13 is realized analogously to the connection between first chamber 11 and second chamber 12, using a tube on which there is provided a tap or a known hose clamp.
  • the ozonized solution can be kept inside said second chamber 12 for a desired time, and the closing device can be opened when the necessary contact time between ozone and cells contained in said second chamber 12 has elapsed.
  • the inlet port 4 and the outlet port 3 for cells, as well as the inlet port 1 and the outlet port 5 for liquids, are preferably provided for coupling with a syringe of standard needleless type, e.g. with a connection of Luer Lock type, well known in the art.
  • the longitudinal section of the flexible bag 10 (figure 2) allows to better observe the filter 9 inside the second chamber 12, while the transversal section (figure 3) allows to detect the relationship among chamber 12, filter 9 and tube 2.
  • the filter 9 is inside the chamber 12, and is provided with three points of access, i.e. the inlet of the tube 2, the inlet port 4 and the outlet port 3 for cells.
  • the inlet port 1 of the first chamber 11 is used alternatively for filling the first chamber 11 with a sterile rinsing solution contained in a syringe 41, or for connecting said chamber to a generator 40 of an oxygen-ozone mixture (see figure 4).
  • the first chamber 11 of the flexible bag 10 contains about 250 mL of liquid, while the chamber 13 can contain about 350 mL.
  • the flexible bag 10 can be manufactured in different dimensions, adapting the dimensions of its chambers 11, 12, 13 to operational needs, e.g. manufacturing a flexible bag suitable for preparing grafts of large quantities of adipose tissue, with ensuing increase of the dimensions of the chambers.
  • a human operator introduces a quantity of unprocessed adipose tissue (raw cells) collected from the body of a patient into the second chamber 12, in particular inside the filter 9, using a needleless syringe that is coupled to the inlet port 4.
  • the three-way valve 6 is adjusted so as not to allow the passage of substances between the chambers 11 and 12.
  • the operator introduces the sterile rinsing solution in the first chamber 11 through its specific inlet port 1.
  • the operator places the bag 10 vertically, e.g. using the support 70, shown in figures 7.
  • the three-way valve 6 is opened, so as to allow the passage due to gravity of the sterile rinsing solution from the first chamber 11 to the second chamber 12.
  • the inlet port 1 is successively connected with a generator 40 of a mixture of oxygen-ozone, bubbling the mixture oxygen-ozone in the liquid 61 for a time suitable for obtaining the desired final concentration of ozone, i.e. the ozonized solution 62 (figure 5).
  • a known quantity of oxygen-ozone mixture is taken with a syringe 41 and inserted in the first chamber 11 already containing a known quantity of sterile rinsing solution 61 (figure 6).
  • the ozonized rinsing solution by opening the three-way valve 6, the solution is channelled into the second chamber 12, and therefore ozone comes into contact with the cells held inside the filter 9.
  • a previously prepared liquid solution is inserted, containing a suitable quantity of ozone (ozonized solution), that is then channelled from the first chamber 11 to the second chamber
  • a suitable quantity of oxygen-ozone mixture is inserted, using the entry port 6’ of the second chamber 12.
  • the ozone by mixing with the liquid wherein cells are immersed, forms an ozonized solution.
  • the desired contact time is maintained, then the ozonized liquid is channelled from the second chamber 12 to the third chamber 13.
  • the flexible bag 10 is provided to the final user with the first chamber 11 already filled with a sterile rinsing solution chosen from the group consisting in sterile water, saline, Ringer’s lactate, Ringer’s acetate, phosphate-buffered saline, crystalloid solutions, colloidal solutions, 5% glucose in water solutions, Hartmann’s solutions.
  • a sterile rinsing solution chosen from the group consisting in sterile water, saline, Ringer’s lactate, Ringer’s acetate, phosphate-buffered saline, crystalloid solutions, colloidal solutions, 5% glucose in water solutions, Hartmann’s solutions.
  • the first chamber 11 in the flexible bag of standard dimensions, there are provided 250 mL of sterile rinsing solution.
  • the sterility of operation is improved and operations are simplified, in that the sterile rinsing liquid is pre-dosed, and therefore dosing the desired quantity of ozone to be inserted in the first chamber 11 for preparing the ozonized rinsing solution becomes simpler.
  • the three-way valve 6 is closed and the operator agitates and squeezes said second chamber 12, so as to delicately mix the sterile rinsing solution and the adipose cells contained in said chamber 12.
  • the separator filter 9 placed inside the second chamber 12, the action of the operator promotes the separation of impurities (white blood cells, red blood cells, where applicable free lipids) from the adipose cells and the stem and regenerative cells. This also allows to discard the excess liquids, that are drained to the third chamber 13, so obtaining a dehydrated graft, having a volume smaller than the volume of adipose tissue/cells originally collected.
  • the bag 10 is supported by a dedicated support 70, described in detail in the following, allowing to maintain its longitudinal axis perpendicular to the ground during the steps a) introduction of adipose tissue into the filter 9 and b) rinsing of the adipose tissue with the ozonized solution.
  • said support 70 is laid down on the long side of the bag 10 provided with the cell outlet port 3, so as to facilitate the outflow of all the cells from the filter 9.
  • the outflow of processed adipose cells is performed by pressing said chamber 12 with a spatula (not shown).
  • Figure 4 schematically shows an embodiment wherein a syringe 41, preferably a Luer Lock syringe, is connected to the inlet port 1 of the chamber 11 for inserting a defined quantity of sterile rinsing liquid, with the three-way valve 6 in its closed condition, so that said rinsing liquid remains inside the first chamber 11. Thereafter, the syringe 41 is removed and the same inlet port 1 is used for supplying said first chamber 11 with a defined quantity of an oxygen-ozone mixture coming from the ozone generator 40. The inlet port 1 is then closed, and this point in the chamber 11 there is provided an ozonized solution that is channelled due to gravity into the chamber 12 by manoeuvring the three-way valve 6.
  • a syringe 41 preferably a Luer Lock syringe
  • the contact between ozonized solution and adipose cells is maintained for a suitable time, and then the ozonized solution is channelled into the third chamber 13.
  • the flexible bag 10 according to figures 1-3 that is not provided with a device preventing the outflow of the ozonized solution from the second chamber 12 to the third chamber 13, the suitable contact time between ozone and cells is obtained through using a suitable quantity of rinsing liquid, that flows on the cells for the time required for obtaining the effect of ozone.
  • the removal of the processed and dehydrated adipose cells is performed, by extracting said cells, i.e. the dehydrated graft, from the outlet port 3 provided in the second chamber 12, preferably through a syringe.
  • the draining liquid flowed in the third chamber 13 is extracted from the bag 10 through the outlet port 5, which is provided with a hermetically closing plug 50 that can be opened through an external axial pressure and can close automatically when said pressure is stopped.
  • the closing plug is preferably of the type described in patent EP2667839B1.
  • Figures 7 show the bag 10 supported during its use by a three- dimensional support 70.
  • Said support 70 consists in a rectangular plate 77 made of rigid material, metal or a plastic material, provided with two perpendicular extensions 75 and
  • the two extensions 75 and 76 meet at a common edge 78.
  • Said support 70 comprises four hooks 71, 72, 73, 74 for connecting said bag 10, in particular for connecting its corresponding holes 31, 32, 33, 34 obtained in the corners of said bag 10, to said support 70.
  • said three-dimensional support 70 comprises: - a first rectangular-shaped surface 77 on which the back face of the bag 10 leans;
  • a third surface 76 having the shape of an isosceles trapezium bent to an angle of about 90° with respect to said first surface 77, that allows to said support 70 to support the bag 10 nearly vertically with respect to the ground, in a second position wherein the transversal axis B-B is perpendicular with respect to the ground.
  • a sterile pre-ozonized solution can be used, or a suitable quantity of gaseous oxygen-ozone mixture can be inserted for fumigating the cells contained in rigid chambers, which gaseous mixture combining with the solutions contained in said chambers produces an ozonized solution.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Chemical & Material Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Biomedical Technology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Transplantation (AREA)
  • Epidemiology (AREA)
  • Oral & Maxillofacial Surgery (AREA)
  • Medicinal Chemistry (AREA)
  • Botany (AREA)
  • Dermatology (AREA)
  • Fluid Mechanics (AREA)
  • Hematology (AREA)
  • Zoology (AREA)
  • Physics & Mathematics (AREA)
  • Urology & Nephrology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Materials For Medical Uses (AREA)

Abstract

L'invention concerne une méthode de préparation d'une greffe d'adipocytes déshydratés à partir d'adipocytes non traités, la méthode comprenant les étapes successives suivantes : a) introduction d'une partie de tissu adipeux non traité dans un dispositif adapté pour rincer ledit tissu adipeux ; b) rinçage dudit tissu adipeux non traité à l'aide d'un liquide de rinçage stérile ; c) retrait dudit liquide du dispositif contenant ledit tissu adipeux, de façon à obtenir une greffe d'adipocytes déshydratés ; caractérisé en ce que ledit liquide de rinçage stérile est complété par un mélange d'oxygène-ozone, de façon à obtenir une solution ozonisée liquide.
PCT/EP2024/051304 2023-04-05 2024-01-19 Méthode de préparation d'une greffe de tissu adipeux Pending WO2024208455A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
IT102023000006693 2023-04-05
IT102023000006693A IT202300006693A1 (it) 2023-04-05 2023-04-05 “Metodo per la preparazione di un innesto di tessuto adiposo”

Publications (1)

Publication Number Publication Date
WO2024208455A1 true WO2024208455A1 (fr) 2024-10-10

Family

ID=86852130

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2024/051304 Pending WO2024208455A1 (fr) 2023-04-05 2024-01-19 Méthode de préparation d'une greffe de tissu adipeux

Country Status (2)

Country Link
IT (1) IT202300006693A1 (fr)
WO (1) WO2024208455A1 (fr)

Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1144026B1 (fr) 1998-12-24 2004-07-28 Biosafe S.A. Systeme de separation sanguine convenant en particulier pour la concentration de cellules souche hematopoietiques
US20150004901A1 (en) 2013-06-28 2015-01-01 Samsung Electronics Co., Ltd. Method and apparatus for performing device-to-device communication
WO2015140737A1 (fr) 2014-03-19 2015-09-24 Lipogems International S.P.A. Dispositif et procédé de préparation de tissu adipeux pour transplantation
EP2667839B1 (fr) 2011-01-28 2016-11-09 Paolo Gobbi Frattini s.r.l. Raccord hermétique pouvant être percé sans aiguille, refermable automatiquement et de manière étanche, pour des dispositifs destinés à recueillir et à distribuer des solutions liquides à usage pharmaceutique et/ou nutritionnel
EP2424463B1 (fr) 2009-05-01 2016-12-28 Bimini Technologies LLC Systèmes, procédés et compositions permettant d'optimiser des greffons enrichis en tissus et cellules
CN106492288A (zh) 2016-10-31 2017-03-15 广州昕生医学材料有限公司 可注射的脱细胞脂肪基质微粒及其在植入剂中的应用
ITUB20154901A1 (it) * 2015-10-22 2017-04-22 Paolo Gobbi Frattini S R L Nuova sacca per la preparazione di innesti di tessuto adiposo
US20170224869A1 (en) 2016-02-08 2017-08-10 Lifecell Corporation Biologic breast implant
EP2380970B1 (fr) 2003-06-25 2017-12-20 Cytori Therapeutics, Inc. Systèmes et procédés pour séparer et concentrer des cellules régénératrices d'un tissu

Patent Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1144026B1 (fr) 1998-12-24 2004-07-28 Biosafe S.A. Systeme de separation sanguine convenant en particulier pour la concentration de cellules souche hematopoietiques
EP2380970B1 (fr) 2003-06-25 2017-12-20 Cytori Therapeutics, Inc. Systèmes et procédés pour séparer et concentrer des cellules régénératrices d'un tissu
EP2424463B1 (fr) 2009-05-01 2016-12-28 Bimini Technologies LLC Systèmes, procédés et compositions permettant d'optimiser des greffons enrichis en tissus et cellules
EP2667839B1 (fr) 2011-01-28 2016-11-09 Paolo Gobbi Frattini s.r.l. Raccord hermétique pouvant être percé sans aiguille, refermable automatiquement et de manière étanche, pour des dispositifs destinés à recueillir et à distribuer des solutions liquides à usage pharmaceutique et/ou nutritionnel
US20150004901A1 (en) 2013-06-28 2015-01-01 Samsung Electronics Co., Ltd. Method and apparatus for performing device-to-device communication
WO2015140737A1 (fr) 2014-03-19 2015-09-24 Lipogems International S.P.A. Dispositif et procédé de préparation de tissu adipeux pour transplantation
ITUB20154901A1 (it) * 2015-10-22 2017-04-22 Paolo Gobbi Frattini S R L Nuova sacca per la preparazione di innesti di tessuto adiposo
US20170224869A1 (en) 2016-02-08 2017-08-10 Lifecell Corporation Biologic breast implant
CN106492288A (zh) 2016-10-31 2017-03-15 广州昕生医学材料有限公司 可注射的脱细胞脂肪基质微粒及其在植入剂中的应用

Also Published As

Publication number Publication date
IT202300006693A1 (it) 2024-10-05

Similar Documents

Publication Publication Date Title
JP6208787B2 (ja) 脂肪吸引により抽出された小葉脂肪から移植用の組織、特に脂肪組織、を準備するための装置及び方法
CN102458302B (zh) 用于优化组织和细胞富集的移植物的系统、方法和组合物
CN108348664B (zh) 脂肪过滤装置
US20150030571A1 (en) Preparation and method for producing a preparation comprising mesenchymal stem cells
CN103402646A (zh) 用于增强从组织样品回收细胞和富含细胞的基质的方法和装置
US10006842B2 (en) Method for producing cell concentrate
JP7037510B2 (ja) 衝撃波または機械的衝撃を使用する、細胞の分離、解離、および／または脱凝集
US8454907B2 (en) Growth factor extractor
CA2661482A1 (fr) Separation de plaquettes dans des suspensions fluides
WO2024208455A1 (fr) Méthode de préparation d'une greffe de tissu adipeux
WO2012122603A1 (fr) Méthode et dispositif de traitement cellulaire
WO2020230045A1 (fr) Dispositif d'aspiration de liposuccion et de purification de graisse
CN111265550A (zh) 一种修复损伤组织的干细胞因子脂质体及其制备方法
US20250290026A1 (en) Tissue processing system and method of use thereof
WO2024116190A1 (fr) Système de traitement de tissu et sa méthode d'utilisation
WO2025186695A1 (fr) Dispositif multifonctionnel, support de dispositif, kit, utilisation de facteurs bioactifs isolés et purifiés à partir de colostrum, procédé de préparation de concentrés de plaquettes autologues enrichis
JPH05337166A (ja) 冷凍保存血液の洗浄方法
JPH05337165A (ja) 冷凍保存血液の洗浄方法およびその装置

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 24701600

Country of ref document: EP

Kind code of ref document: A1

WWE Wipo information: entry into national phase

Ref document number: 2024701600

Country of ref document: EP

NENP Non-entry into the national phase

Ref country code: DE

ENP Entry into the national phase

Ref document number: 2024701600

Country of ref document: EP

Effective date: 20251105