WO2025133708A2

WO2025133708A2 - Therapeutic use of compositions and methods for epidermal treatment

Info

Publication number: WO2025133708A2
Application number: PCT/IB2024/000779
Authority: WO
Inventors: David L STACHURA; Kristen RUEB; John AYLWORTH
Original assignee: Specbio Inc
Current assignee: Specbio Inc
Priority date: 2023-12-23
Filing date: 2024-12-23
Publication date: 2025-06-26
Anticipated expiration: 2026-06-23
Also published as: WO2025133708A3

Abstract

The disclosure relates to compositions and methods for synthesizing a composition useful for treating disorders of the skin comprising a peptide having melanin inhibition and antioxidant properties, the composition further comprising cell culture medium isolates from adipose derived mesenchymal stem cells (ADMSCs) comprising eluent compositions useful for regenerative medicine therapies of medical conditions. The disclosure further relates to methods of treatment with the compositions useful as skin therapeutics for treating skin pigmentation disorders and improving skin tone and texture.

Description

PATENT COOPERATION TREATY

INTERNATIONAL UTILITY PATENT APPLICATION

THERAPEUTIC USE OF COMPOSITIONS AND METHODS FOR EPIDERMAL TREATMENT

Field of the Invention

[0001] The inventive subject matter relates to therapeutic use of compositions and methods for treatment of various conditions of the skin comprising applying topical compositions isolated from cultivates of adipose-derived mesenchymal stem cells (ADMSCs) isolated from adipose tissue, said compositions useful for skin rejuvenation treatments, particularly facial skin rejuvenation methods and compositions. When applied to facial skin, said facial rejuvenation compositions cause minimal to no facial peeling or discomfort, reduced downtime after treatment, and low photosensitivity. The skin rejuvenation treatment applications of the inventive subject matter are useful in various skin treatment comprising reduction of fine lines, wrinkles, shrinking enlarged pores, thickening aging skin having lost plasticity, repairing sun damage, skin discoloration and hyperpigmentation, tightening loose and sagging skin, reducing stretch marks, and reducing skin dullness. The inventive subject matter further comprises compositions for culturing ADMSCs, methods of isolating and purifying secretions therefrom, and methods of use of said ADMSC culture eluent isolates, extracts, and other composition comprising secretions from ADMSCs and ADMSC growth media for use in regenerative medicine, and as therapeutics in the treatment of various skin medical conditions of patients.

Background of the Invention

[0002] The use of stem cells in the therapeutic treatment of various diseases and medical conditions has expanded with the discovery of the regenerative powers of stem cells and their products. Stem cells are undifferentiated cells that have the ability and capacity to self-renew and differentiate into different specialized cells. Totipotent stem cells can give rise to all embryonic and adult cell lineages, tissues, and cell types. Pluripotent stem cells are differentiated stem cells that can give rise to all cell types and tissues in an adult organism, having lost the ability to generate embryonic tissues. While growing, stem cells produce various proteins, hormones, cytokines, chemokines, growth factors, signaling factors, extracellular matrix proteins (ECMs) or effluents or secretions that have various specific beneficial effects on differentiated cells, for example, stimulating growth of differentiated or specialized cells, or in turn stimulate differentiated or specialized cells to produce other beneficial signal molecules or growth factors, for example cytokines and other biomolecules with immunomodulatory properties. Mesenchymal stem cells (MSCs) are multi potent stem cells having limited self-renewal and differentiation capabilities that are primarily found in bone marrow and are important for making and repairing skeletal tissues, such as cartilage, bone, and the fat found in bone marrow. As they age, MSCs predominantly convert into lipid-accumulating fat cells and can be found in adipose tissue. Adipose-derived MSCs (ADMSCs) have prominent effects in tissue regeneration given their high cell yield in adipose tissue, due to their ability to differentiate into multiple lineages and secrete growth factors, cytokines, chemokines, or other immunomodulatory biomolecules.

[0003] Though pluripotent embryonic stem cells (ESCs), isolated from a blastocyst (an implanted embryo at the sixth to eighth day of development) can give rise to more tissue types e.g. embryonic tissues, their use has severe ethical implications since isolating ESCs destroys the developing blastocyst, and moreover their limited availability hinders their use. Thus, embryonic stems cells, though very versatile, are not a good candidate for therapeutic use. MSCs, on the other hand, are more differentiated stem cells and can be sourced from adult tissues preferably bone marrow, peripheral blood, and adipose tissue, as well as from neonatal birth-associated tissues including placenta, umbilical cord, and cord blood. These neonatal birth-associated tissues as sources of stem cells are in high demand and not as easily available and pose some ethical questions as well.

[0004] Since their discovery in 1991 in bone marrow, MSCs have been used as a promising method to build tissues and generate tissue progenitor cells. MSCs also provide other benefits as immunomodulatory cells, for example modulating or regulating immune responses, such as in inflammation, by regulating various immune cell types, for example monocytes, macrophages, dendritic cells, T-cells, B-cells, and natural killer cells (NKCs). MSCs produce various signaling molecules for example cytokines, chemokines, and growth factors, with therapeutic applications in the treatment of various medical conditions. One notable example of the therapeutic use of MSC secretions is the use of extracellular vesicles with immunomodulatory functions used in the treatment of severely ill patients with severe COVID-19 complications.

[0005] Unfortunately, during in vitro culture, particularly at higher passages or cell generations, stem cells age and degenerate due to decreased telomerase activity. In addition, during long-term culture, MSCs lose their potential to differentiate and begin to exhibit morphological changes, which are undesired for therapeutic use of isolates from these cell cultures. Thus, prolonged tissue culture of MSCs is not an optimal source for bioactive stem cells or for their eluent isolates for therapeutic uses.

[0006] However, adult differentiated stem cells such as ADMSCs, given their abundant presence and availability from adipose tissue aspirates, for example from routinely performed liposuction procedures, present an abundant source of stem cells useful in the treatment of degenerative diseases or conditions due to their regenerative properties in bone, cartilage, epidermal, and other tissues.

[0007] In addition to the need for a readily available source of viable ADMSCs, there is a need for methods of cultivating ADMSCs for therapeutic uses in regenerative medicine for the treatment of various diseases or medical conditions, for example but not limited to, bone, cartilage, connective, and skin tissue conditions and diseases.

[0008] In addition, there is a need for methods and compositions for isolating, and purifying isolates of ADMSC effluent or eluent molecules and compositions for therapeutic uses in regenerative medicine for the treatment of various diseases or medical conditions, for example but not limited to inflammatory, pulmonary, bone, cartilage, connective, and skin tissue conditions and diseases, for example but not limited to pulmonary damage, joint damage, epithelial wound healing, scar reduction, trichology therapeutics, and pigmentation disorders of the skin. Thus, there continues to be a need for an effective culture medium and support matrix compositions for maximizing ADMSC propagation and growth and methods for isolating the ADMSCs effluents and secretions for therapeutic use in regenerative medicine.

[0009] In particular, there is a need for treatment of various skin conditions that benefit from the application of the rejuvenating and regenerating effects of stem-cell derived factors. Particularly, for facial skin rejuvenation, there is a need for therapeutic use of compositions and methods for treatment of various conditions of the skin comprising applying topical compositions isolated from cultivates of adipose-derived mesenchymal stem cells (ADMSCs) isolated from adipose tissue, said topical compositions useful for skin rejuvenation treatments, in particular facial skin rejuvenation treatments. When applied to facial skin, said facial rejuvenation compositions cause minimal to no facial peeling or discomfort, reduced downtime after treatment, and low photosensitivity. The skin rejuvenation treatment applications of the inventive subject matter are useful in various skin treatment comprising reduction of fine lines, wrinkles, shrinking enlarged pores, thickening aging skin having lost plasticity, repairing sun damage, skin discoloration and hyperpigmentation, tightening loose and sagging skin, reducing stretch marks, and reducing skin dullness. In addition there is a need for skin rejuvenation treatment compositions comprising components that block melanin production, and act as antioxidants to increase skin tone and texture but do not cause irritation, pain or photosensitivity, while improving the feeling of skin tightness and greatly improve skin tone and texture. The present inventive subject matter meets these and other needs as detailed below.

SUMMARY

[00010] The inventive subject matter provides therapeutic use of compositions and methods for treatment of various conditions of the skin comprising compositions isolated from cultivates of adipose-derived mesenchymal stem cells (ADMSCs) isolated from adipose tissue, said compositions useful for facial skin rejuvenation treatments. When applied to facial skin, said facial rejuvenation compositions cause minimal to no facial peeling or discomfort, reduced downtime after treatment, and low photosensitivity. The skin rejuvenation treatment applications of the inventive subject matter are useful in various skin treatment comprising reduction of fine lines, wrinkles, shrinking enlarged pores, thickening aging skin having lost plasticity, repairing sun damage, skin discoloration and hyperpigmentation, tightening loose and sagging skin, reducing stretch marks, and reducing skin dullness. In addition, the skin rejuvenation treatment compositions comprise components that block melanin production, acts as antioxidants to increase skin tone and texture but do not cause irritation, pain, or photosensitivity, while improving the feeling of skin tightness and greatly improve skin tone and texture. The compositions of the inventive subject matter comprise a penta-peptide component that blocks melanin production and functions as an antioxidant increasing skin texture and tone, while reducing irritation, pain, and photosensitivity of the treated skin. The compositions of the present inventive subject matter provide a thicker more easily applied dermal solution with none of the detractions of commercially available skin treatments presently on the market. [00011] The inventive subject matter provides methods and compositions for culturing ADMSCs and methods for isolating and purifying effluents and secretions of said ADMSCs produced during stem cell culture, said secretions useful in the treatment of various medical conditions, for example and without limitation, pulmonary damage, joint damage, epithelial wound healing, scar reduction, trichology therapeutics, and pigmentation disorders of the skin.

[00012] The compositions of the inventive subject matter comprise culture media compositions to enhance stem cell culture, specifically ADMSCs useful in the isolation and purification of ADMSC secretions produced during stem cell culture. The secretions produced during ADMSC's culture include cytokines, chemokines, ECM proteins, and growth factors having therapeutic applications in medical treatment and therapy of various medical conditions, for example and without limitation, pulmonary damage, joint damage, epithelial wound healing, scar reduction, trichology therapeutics, and pigmentation disorders of the skin.

[00013] The compositions of the inventive subject matter comprise topical therapeutic compositions useful in the treatment of wounds, including but not limited to excisional wounds, pressure sores, burns, lacerations, abrasions, skin ulcers, lesions, and chronic diabetic ulcers. The compositions of the inventive subject matter comprise topical application and/or microinjections of the compositions of the inventive subject matter for the treatment of scar tissue, including but not limited to stretch marks, fine-line scars, keloid scars, hypertrophic scars, pitted or sunken scars, and scar contractures. The compositions of the inventive subject matter further comprise intra-articular injectable therapeutic compositions to treat inflammation, osteoarthritis, cartilage damage, bone loss, and ligament damage in a variety of joints including but not limited to, elbow, shoulder, hip, and knee joints. The compositions of the inventive subject matter further comprise therapeutic compositions adapted as inhalation therapeutics used with a nebulizer or similar method of aerosolization useful for the treatment of respiratory tract damage caused by, including but not limited to, damage caused by upper or lower respiratory tract infections, bronchitis, environmental insult, and chronic respiratory tract infections including but not limited to asthma, emphysema, and chronic obstructive pulmonary disease (COPD).

[00014] The compositions of the inventive subject matter comprise solid support matrix compositions to enhance stem cell culture, specifically ADMSCs useful in the isolation and purification of ADMSC secretions produced during stem cell culture. The secretions or effluents produced during ADMSCs culture include cytokines, chemokines, ECM proteins, and growth factors having therapeutic applications in medical treatment and therapy of various medical conditions, for example and without limitation, pulmonary damage, joint damage, epithelial wound healing, scar reduction, trichology therapeutics, and pigmentation disorders of the skin.

[00015] The methods of the inventive subject matter comprise methods of synthesizing stem cell culture media elements using support matrix compositions of the inventive subject matter to synthesize a personalized regenerative medicine composition useful in the regenerative medicine therapy of a patient's degenerative medical condition, for example but not limited to inflammation, pulmonary damage, joint damage, epithelial wound healing, scar reduction, trichology therapeutics, and pigmentation disorders of the skin. BRIEF DESCRIPTION OF THE FIGURES

[00016] These and other features of the preferred embodiments of the inventive subject matter will become more apparent in the detailed description in which reference is made to the appended drawings wherein:

[00017] FIG. 1 is a diagram illustrating the method of culturing ADMSCs using the culture medium compositions of the inventive subject matter and method of isolation of effluents from different solid support culture conditions.

[00018] FIG. 2 is a diagram illustrating the method of isolating secretions or effluents from the culture of ADMSCs of the inventive subject matter.

[00019] FIG. 3 is a schematic of a method of treatment for wound healing of the present inventive subject matter.

[00020] FIG. 4 is a graph of the tyrosinase inhibition activity, preventing melanin production, of the compositions of the present inventive subject matter.

[00021] FIG. 5 is a graph of the modulated collagenase inhibition activity of the compositions of the present inventive subject matter.

[00022] FIG. 6 is a caparison image of the baseline VISIA® image (left) with a two-week post-RDS treatment VISIA® image (right) showing reduction in redness after 4 applications with FACTORFIVE™ RENEW DERMAL SOLUTION™ for a female patient (A) and a male patient (B).

[00023] FIG. 7 is a caparison image of the baseline image (left) with a two-week post-RDS treatment image (right) showing reduction appearance of fine lines after 4 applications with FACTORFIVE™ RENEW DERMAL SOLUTION™.

DETAILED DESCRIPTION

[00024] The inventive subject matter can be understood more readily by reference to the following detailed description, examples, drawings, figures, and claims, and their previous and following description. However, before the present compositions, and/or methods are disclosed and described, it is to be understood that this inventive subject matter is not limited to the specific compositions, and/or methods disclosed unless otherwise specified, as such can, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular aspects only and is not intended to be limiting.

[00025] The following description of the inventive subject matter is provided as an enabling teaching of the inventive subject matter in its best, currently known embodiments. To this end, those skilled in the relevant art will recognize and appreciate that many changes can be made to the various aspects of the inventive subject matter described herein, while still obtaining the beneficial results of the inventive subject matter. It will also be apparent that some of the desired benefits of the inventive subject matter can be obtained by selecting some of the features of the inventive subject matter without utilizing other features. Accordingly, those who work in the art will recognize that many modifications and adaptations to the inventive subject matter are possible and can even be desirable in certain circumstances and are a part of the inventive subject matter. Thus, the following description is provided as illustrative of the principles of the inventive subject matter and not in limitation thereof.

Definitions

[00026] As used throughout, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a compound" can include two or more such compounds unless the context indicates otherwise.

Ranges can be expressed herein as from "about" one particular value, and/or to "about" another particular value. When such a range is expressed, another aspect includes from the one particular value and/or to the other particular value. Similarly, when values are expressed as approximations, by use of the antecedent "about," it will be understood that the particular value forms another aspect. It will be further understood that the endpoints of each of the ranges are significant both in relation to the other endpoint, and independently of the other endpoint.

[00027] As used herein, the terms "optional" or "optionally" mean that the subsequently described event or circumstance may or may not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

[00028] As used herein, the term "adipose-derived mesenchymal stem cells (ADMSCs)" are MSCs extracted from adipose tissues, for example but not limited to, isolated from liposuction tissue aspirates.

[00029] As used herein, the term "regenerative medicine" means a treatment or therapy of a degenerative disorder or medical condition of a patient that benefits from therapeutics of stem cell extracts or secretions or effluents, for example but not limited to inflammation, pulmonary damage, joint damage, epithelial wound healing, scar reduction, trichology therapeutics, and pigmentation disorders of the skin.

[00030] As used herein, the term "regenerative medicine therapeutic" means a composition useful in the treatment or therapy of a degenerative disorder or medical condition of a patient that benefits from therapeutics of stem cell extracts or secretions or effluents, for example but not limited to wound healing, trichology therapy for example but not limited inflammation, pulmonary damage, joint damage, epithelial wound healing, scar reduction, trichology therapeutics, and pigmentation disorders of the skin.

[00031] As used herein, the term "isolation" means the process of separating a certain composition from a complex mixture by a variety of means used to separate the desired component from other undesirable components, said means are for example but not limited to, electrophoresis, gravitational or centrifugal sedimentation, immune capture, affinity columns, or other chromatographic or other separation means routinely used in the art of cell biology and cell culture.

[00032] As used herein, the term "tissue or cell culture" means the practice of growing cells derived from donor tissue in an artificial medium as routinely practiced in cell biology and cell culture laboratories.

[00033] As used herein, the term "lipoaspirates" refers to materials, compositions, and other components derived from a liposuction procedure performed on a patient or donor, said materials, compositions and other components comprising viable ADMSCs useful in the tissue and cell culture methods of the regenerative medicine methods of the present inventive subject matter.

[00034] As used herein, the term "cell culture medium" refers to a composition of optimized components for effecting the optimal growth of stem cells, for example ADMSCs, useful for the regenerative medicine therapies and therapeutic compositions of the present inventive subject matter.

[00035] As used herein, the term "cell culture matrix" refers to a composition to support the cell culture of ADMSCs comprising for example ECM and/or active compounds to promote cell growth in tissue culture media.

[00036] As used herein, the term "stem cell secretions" or "effluents" refer to compositions produced during stem cell growth during stem cell culture and released into the cell culture medium, for example but not limited to cytokines, chemokines, cell-growth factors, ECM proteins, and immunoregulatory proteins or molecules.

[00037] As used herein, the term "personalized therapeutic composition" means a therapeutic composition useful in the regenerative medicine treatment or therapy of a patient with a specific condition which the composition is specifically formulated to treat, for example a dermatologic therapeutic for wound healing, hair loss, or pigmentation disorder.

[00038] As used herein, the term "inflammation disorder" means a disease characterized by overactive inflammation response, typically associated with degenerative disorders, for example but not limited to cartilage degeneration and associated inflammation response as seen in diabetes, chronic pulmonary diseases, arthritis, rheumatoid arthritis, and other joint and cartilage degenerative disorders.

[00039] As used herein, the term "pentapeptide" refers to a five-membered peptide chain consisting of 5 amino acids selected from naturally occurring amino acids, and bioavailable modified amino acid components.

Treatment of conditions facilitated

[00040] The compositions and methods of the present inventive subject matter are used to synthesize personalized regenerative medicine therapeutics useful in the regenerative medicine treatment of medical conditions in dermatologic medical conditions, wound-healing, trichology therapy for example for hair loss, and pigmentation disorders or the skin, for example hyper-pigmentation disorders.

[00041] The compositions of the inventive subject matter comprise topical therapeutic compositions useful in the treatment of wounds, including but not limited to excisional wounds, pressure sores, burns, lacerations, abrasions, skin ulcers, lesions, and chronic diabetic ulcers. The compositions of the inventive subject matter comprise topical application and/or microinjections of the compositions of the inventive subject matter for the treatment of scar tissue, including but not limited to stretch marks, fine-line scars, keloid scars, hypertrophic scars, pitted or sunken scars, and scar contractures.

[00042] The compositions and methods of the present inventive subject matter are also used to treat conditions of the skin. The inventive subject matter provides therapeutic use of compositions and methods for treatment of various conditions of the skin comprising compositions isolated from cultivates of adipose-derived mesenchymal stem cells (ADMSCs) isolated from adipose tissue, said compositions useful for facial skin rejuvenation treatments. When applied to facial skin, said facial rejuvenation compositions cause minimal to no facial peeling or discomfort, reduced downtime after treatment, and low photosensitivity. The skin rejuvenation treatment applications of the inventive subject matter are useful in various skin treatment comprising reduction of fine lines, wrinkles, shrinking enlarged pores, thickening aging skin having lost plasticity, repairing sun damage, skin discoloration and hyperpigmentation, tightening loose and sagging skin, reducing stretch marks, and reducing skin dullness. In addition, the skin rejuvenation treatment compositions comprise components that block melanin production, and act as antioxidants to increase skin tone and texture but do not cause irritation, pain, or photosensitivity, while improving the feeling of skin tightness and greatly improve skin tone and texture. The compositions of the inventive subject matter comprise a penta-peptide component that blocks melanin production and functions as an antioxidant increasing skin texture and tone, while reducing irritation, pain, and photosensitivity of the treated skin. The compositions of the present inventive subject matter provide a thicker more easily applied dermal solution with none of the detractions of commercially available skin treatments presently on the market.

[00043] The compositions and methods of the present inventive subject matter are also used to treat conditions of the skin comprising reduction of fine lines and wrinkles, shrinking enlarged pores, thickening aging skin, repairing sun damage, skin discoloration and hyperpigmentation, tightening loose and sagging skin, reducing stretch marks, and reducing skin dullness. Said compositions comprise a composition to block melanin production and having antioxidant properties. In one embodiment said melanin-blocking antioxidant composition is a peptide. In one embodiment said peptide comprises a five-membered polypeptide, or pentapeptide. In one embodiment said pentapeptide having amino acid sequence glutamic acid, cysteine, glycine, tyrosine, and phenylalanine, or Glu- Cys-Gly-Tyr-Phe using the three-letter amino acid code, or ECGYF using the one-letter amino acid code. Said pentapeptide composition are formulated with other ingredients to provide a dermal therapeutic solution having thicker properties, making it more easily applicable to the skin to be treated with no undesirable effects of skin irritation, pain, discomfort, or photosensitivity caused by products presently on the market.

[00044] The compositions of the inventive subject matter comprise comparative advantages over formulations currently available in the marketplace. The compositions of the present inventive subject matter comprise 30% or less trichloroacetic acid (TCA) so the compositions of the present inventive subject matter can be used throughout the United States without regard to prohibitions of compositions comprising over 30% TCA. Moreover, the compositions of the present inventive subject matter comprise more kojic acid, a tyrosinase activity inhibitor, useful in the prevention of pigmentation disorders of the skin, for example brown spots. The compositions of the present inventive subject matter are pH of around 2 as more effective formulation. The compositions of the present inventive subject matter comprise a novel peptide to encourage higher antioxidant activity thereby reducing brown spots, improving extracellular matrix breakdown and restructuring, thus providing tighter skin with improved tone and texture. [00045] In one embodiment, the compositions of the inventive subject matter comprise purified water, trichloroacetic acid, glycerin, emulsifier, kojic acid, hydrogen peroxide, and disodium EDTA, and buffered with ammonium hydroxide to obtain a low pH of around 2 at room temperature. After the solution cools, an antioxidant composition is added. The resulting solution is applied to the skin with slight pressure and allowed to sit for up to 10 minutes. The application is repeated three times then washed off with water.

[00046] In one embodiment, the composition of the inventive subject matter comprises up to 37% purified water, up to 30% trichloroacetic acid, up to 6% glycerin, up to 5% emulsifier, up to 5% kojic acid, up to 1% hydrogen peroxide, up to 0.25% disodium EDTA, and q.s. of purified water to obtain the desired composition of each component buffered with 13% ammonium hydroxide to a pH of around 2 at room temperature. After the solution cools, a peptide antioxidant composition is added. The resulting solution is applied to the skin to be treated with slight pressure and allowed to sit for up to 10 minutes. The application is repeated three times then washed off with water.

[00047] In one embodiment, the composition of the inventive subject matter comprises 37% purified water, 30% trichloroacetic acid, 6% glycerin, 5% SEPIGEL emulsifier, 5% kojic acid, 1% hydrogen peroxide, 0.25% disodium EDTA, buffered with 13% ammonium hydroxide to a pH of around 2.1 at room temperature. After the solution cools, 0.3 mg/mL of ECGYF peptide composition is added. The resulting solution is applied to the skin to be treated with slight pressure and allowed to sit for up to 10 minutes. The application is repeated three times then washed off with water.

[00048] In one embodiment, the composition of the inventive subject matter comprises 37% purified water, 30% trichloroacetic acid, 6% glycerin, 5% SEPIGEL emulsifier, 5% kojic acid, 1% hydrogen peroxide, 0.25% disodium EDTA, buffered with 13% ammonium hydroxide to a pH of around 2.1 at room temperature. After the solution cools, 0.3 mg/mL of ECGYF peptide composition is added. The resulting solution is combined with a composition of ADMSC eluents and is applied to the skin to be treated with slight pressure and allowed to sit for up to 10 minutes. The application is repeated three times then washed off with water.

[00049] In one embodiment, the composition of the inventive subject matter comprises 37% purified water, 30% trichloroacetic acid, 6% glycerin, 5% SEPIGEL emulsifier, 5% kojic acid, 1% hydrogen peroxide, 0.25% disodium EDTA, buffered with 13% ammonium hydroxide to a pH of around 2.1 at room temperature. After the solution cools, 0.3 mg/mL of ECGYF peptide composition is added. The resulting solution is applied to the skin to be treated and followed with application of a composition of ADMSC eluents both applied sequentially to the skin to be treated with slight pressure and allowed to sit for up to 10 minutes. The application is repeated three times then washed off with water.

[00050] In one embodiment the therapeutic composition of the inventive subject matter is applied in two separate applications each with its own composition, RDS1 and RDS2. In one embodiment of the two-step therapeutic application, the first composition RDS1 comprises 25 to 50% purified or deionized water, 15 to 30 % trichloroacetic acid, 1 to 5% Kojic acid, 0.5 to 5% hydrogen peroxide, 1 to 10 % glycerin, 1 to 10 % SEPIGEL emulsifier, 0.1 to 1% disodium EDTA, 1 to 25% ammonium hydroxide, and 100 to 1,000 micrograms/mL Pentapeptide. In one embodiment of the two step therapeutic application, the second composition RDS2 comprises 50 to 80% purified or deionized water, 0.1 to 5% hydroxyethylcellulose, 0.01 to 1% aloe vera powder 200x, 0.01 to 1% disodium EDTA, 1 to 15% FUCOGEL 1.5P moisturizer, 1 to 10% glycerin, 0.05 to 2% sodium hyaluronate, 0.01 to 1% lecithin, 0.05 to 4% COVRACYL MV60 thickening agent, 0.5 to 10% polyolprepolymer-15, 0.1 to 2% EUXYL PE 9010 preservative, 0.05 to 5% LEXFEEL WOW emollient, 0.1 to 2% green tea extract, 0.1 to 2 % butylene glycol, 0.1 to 1% GLUADIN KERA-P LM vegetable protein lysate, 0.001 to 0.05% adenosine triphosphate, 0.001 to 0.05% sodium chondroitin sulfate, 1 to 5% DCM, 0.1 to 2% Bis (tripeptide) copper acetate solution, and 0.1 to 2% white tea extract.

Materials and Methods

[00051] The inventive subject matter relates to materials, compositions, and methods of culturing stem cells, preferably ADMSCs derived from lipoaspirates from a patient or donor obtained during a liposuction procedure. [00052] Referring to Figure 1, which is a non-limiting embodiment of the system performing the method of the present inventive subject matter, said method of culturing ADMSCs is shown using the culture medium compositions of the inventive subject matter. The compositions used in the stem cell culture medium of the method of the present inventive subject matter comprise nutrient components known to those skilled in the art of tissue or cell culture, for example without limitation, carbohydrates, amino acids, vitamins, minerals, and a pH buffer system. Said compositions known as basal medium comprise essential and non-essential amino acids, vitamins, organic and inorganic compounds, and optionally may include growth factors. Common carbon sources from carbohydrates in culture medium include glucose, fructose, sucrose, mannitol, and sorbitol. Cell and tissue culture medium compositions well known to a person of skill in the relevant art are commercially available include Dulbecco's modified Eagle medium (DMEM), Dulbecco's modified Eagle medium 2 (DMEM2), Dulbecco's modified Eagle medium F12 (DMEM F12), minimum essential medium (MEM), Roswell Park Memorial Institute (RPMI) 1640 medium, Serum-free media (SFM), Bench Stable Media, human plasma-like medium (HPLM), for example and without limitation.

[00053] Referring to Figure 1, the method of culturing stem cells of the present inventive subject matter, comprises the steps of A) thawing and culturing stem cells previously isolated from donor tissue, preferably lipoaspirates and plating on a tissue culture flask, B) said stem cells are cultured under optimal growth conditions to expand the number of cells to seed solid support culturing, including but not limited to microcarrier beads, and begin large-scale culture of up to 10 T-175 flasks with 30 mL media in each. Following large-scale culture, C) the cells are treated with trypsin to de-aggregate cells to optimize counting of cells and obtain accurate cell counts. The resulting cells D) are then cultured with different solid supports, including but not limited to microcarrier beads, and various growth conditions and media. As shown in Figure 1 step E), for example, in sample 1 the microcarrier beads are not coated with protein; in sample 2, the microcarrier beads are coated with human collagen I; in sample 3 the microcarrier beads are coated with hyaluronic acid; in sample 4, the microcarriers are coated with ECM protein mix comprising laminin, elastin, fibronectin, and glycosaminoglycans. The cells on the various microcarrier samples are grown in bioreactors at 37°C in 5% CO₂ with constant gentle agitation. At prescribed times, cells are allowed to settle to the bottom of the bioreactors and eluate is collected, and filtered as shown in Figure 2, and purified to create the dynamic culture medium extracts and compositions of the present inventive subject matter. As there are different growth conditions by the variation on the coatings on the solid support microcarrier beads, the eluents from each of the samples have different compositions of stem cell secretions, or effluents including but not limited to proteins, cytokines, chemokines, cell-growth factors, ECM proteins, and immunoregulatory proteins or molecules present in the eluate from the cell culture medium. These different stem cell secretions or effluents from the various solid support microbead conditions can be combined in different ratios to yield dynamic culture medium compositions useful for treating different ailments or therapeutic uses. All wounds are different and likely require effluents generated from different cellular conditions. For example, data indicates that ADMSC effluent isolated from cells grown in condition 1 (see Figure IE) accelerated excisional wound healing, proliferation, and migration of keratinocytes, and blastema regeneration. Excisional wounds tend to express high levels of collagen I, so higher ratios of effluent from cells grown in condition 1 and 2 are of value for their treatment, while scar treatment requires a high level of remodeling proteins expressed in the effluents before ECM repair occurs. High levels of hyaluronic acid (condition 3) stimulate more anti-inflammatory protein expression, so are useful for treating respiratory issues and joint repair. However, all wounds are slightly different, and the concentrations of effluent generated from conditions 1-4 (see Figure IE) vary for each treatment application.

[00054] In one embodiment of the present inventive subject matter, the methods for culturing stem cells comprise culturing stem cells, preferably ADMSCs, by seeding said stem cells onto microcarrier beads and growing them in spinner flasks under conditions well known to those skilled in the art.

[00055] In one embodiment of the present inventive subject matter, the methods for culturing stem cells comprise culturing stem cells, preferably ADMSCs, by seeding said stem cells onto microcarrier beads coated with a solution of human collagen I at a concentration ranging from 0.1 to 25 mg/mL, preferably ranging from 0.1 to 6 mg/mL, and growing them in spinner flasks under conditions well known to those skilled in the art.

[00056] In one embodiment of the present inventive subject matter, the methods for culturing stem cells comprise culturing stem cells, preferably ADMSCs, by seeding said stem cells onto microcarrier beads coated with a solution of hyaluronic acid at a concentration ranging from 0.1 to 10 mg/mL, preferably ranging from 0.2 to 5 mg/mL, and growing them in spinner flasks under conditions well known to those skilled in the art.

[00057] In one embodiment of the present inventive subject matter, the methods for culturing stem cells comprise culturing stem cells, preferably ADMSCs, by seeding said stem cells onto microcarrier beads coated with a solution of human-derived extracellular matrix (ECM) derived from human tissue that comprises collagen, elastin, laminin, glycosaminoglycans and other matrix proteins at a concentration ranging from 0.1 to 25 mg/mL, preferably ranging from 0.1 to 10 mg/mf, and growing them in spinner flasks under conditions well known to those skilled in the art.

[00058] In one embodiment of the present inventive subject matter, the methods for culturing stem cells comprise culturing stem cells, preferably ADMSCs, by seeding said stem cells onto microcarrier beads uncoated or coated with components comprising without limitation human collagen I, hyaluronic acid, human derived extracellular matrix (ECM) comprising collagen, elastin, laminin, glycosaminoglycans, and other matrix proteins, at a concentration ranging from 1 to 50 mg/mL. Cells are seeded at a range of lxlO⁶ to 10xl0⁶ (one to ten million) cells added to a volume ranging from 1 to 10 mL of microcarriers at a concentration of 0.01 to 0.10 g/mL beads resuspended in phosphate buffered saline (PBS) solution. After seeding, these beads with attached cells are grown in spinner flasks under conditions well known to those skilled in the art. The cells are grown in culture media comprising for example without limitation 500 mt of aMEM, 25 mt heat-inactivated human platelet lysate (h PL 4.6%), 5 ml 200 mM L-glutamine (0.93%), and 5 ml. HEPES buffer (0.93%).

[00059] Referring to Figure 2, the methods of the present inventive subject matter comprise isolation of secretions or effluents from the stem cell growth medium, comprising the step of, as shown in Figure 2A, allowing the microcarrier beads (black arrow) or other cellular support structures to settle to the bottom of the bioreactor flask. The stem cells' secreted proteins (white arrow) are suspended in the culture media. This eluent with the stem cell secretions, or effluents including but not limited to proteins, cytokines, chemokines, cell-growth factors, ECM proteins, and immunoregulatory proteins or molecules present in the eluent are gently collected. The method further comprises the step of, as shown in Figure 2B, gently filtering by gravity or suction filtering the eluent containing the cell secretions or effluents including but not limited to proteins, cytokines, chemokines, cellgrowth factors, ECM proteins, and immunoregulatory proteins or molecules (white arrow) suspended in the culture media to remove any loose microcarrier beads or other cellular support structures (black arrow) or cellular debris. The filtered eluent (as shown by the bracket) from samples 1-4 (described in Figure IF) is combined at different ratios to create the dynamic culture medium compositions of the present inventive subject matter. Modifications of these ratios allow the treatment of different ailments. For example, data indicates that ADMSC effluent isolated from cells grown in condition 1 (see Figure IE) accelerate excisional wound healing, proliferation, and migration of keratinocytes, and blastema regeneration (data not shown). Excisional wounds tend to express high levels of collagen I, so higher ratios of effluent from cells grown in condition 1 and 2 are of value for their treatment, while scar treatment requires a high level of remodeling proteins expressed in the effluents before ECM repair occurs. High levels of hyaluronic acid (condition 3) stimulate more anti-inflammatory protein expression, so are useful for treating respiratory issues and joint repair. However, all wounds are slightly different, and the concentrations of effluent generated from conditions 1-4 (see Figure IE) vary for each treatment application.

[00060] In one embodiment of the present inventive subject matter, the compositions comprising stem cell secretions or effluents comprising but not limited to proteins, cytokines, chemokines, cell-growth factors, ECM proteins, and immunoregulatory proteins or molecules, collected from the eluents of stem cell culture preferably ADMSCs as outlined above, are useful in the preparation of compositions or formulations of dynamic culture medium for various therapeutic applications, for example and without limitation, dermatological maladies, wound healing, hair growth, scar treatment, joint repair, pulmonary disease, and other skin defect or condition treatments. The concentration of components present in the dynamic culture medium is varied depending upon the therapeutic application being produced. Each culture condition outlined above is combined into a final dynamic culture medium also known as DCM formulation at ratios of components ranging from 1 to 99% so that the final concentration of the various conditions, for example conditions 1, 2, 3, and 4, as shown in Figure IE and IF, adds up to 100% of the active ingredient component of the therapeutic composition. In said therapeutic composition, the protein concentration of the dynamic culture medium or DCM ranges from 1 to 100 mg/mL depending on the therapeutic application.

[00061] Referring to Figure 3, the methods of the present inventive subject matter comprise a method of using the dynamic culture medium compositions prepared as described above to treat dermal wounds. As shown in Figure 3(A) panel A, said method comprises applying a dermatological therapeutic composition containing the dynamic culture medium enriched for secreted proteins, cytokines, chemokines, cell-growth factors, ECM proteins, and immunoregulatory proteins or molecules present in the eluate from the cell culture medium derived from ADMSCs (white arrow) to the wound to be treated, including but not limited to an excisional wound (Figure 3(A) panel A), or any breach or damage to the outer epithelial layer of the skin including scrapes, burns, pressure sores, ulcers, or ulcerations. As shown in Figure 3(A) panel B, over time the continued application of the compositions of the present inventive subject matter to the wound being treated contributes to its expedited wound healing, as shown in Figure 3(A) panel C the wound is fully healed without noticeable scar development. The application of the composition of the present invention can be varied in composition from the various growth conditions as described above to be tailored to the subject's therapeutic application optimized for a given pathology. In some embodiments of the present inventive subject matter, the topical application of the dynamic culture medium therapeutic composition can be enhanced or accelerated by microinjections or micro-needling with the compositions of the present inventive subject matter. ADMSC effluent isolated from cells grown in condition 1 (see Figure IE) accelerate excisional wound healing, proliferation, and migration of keratinocytes, and blastema regeneration (data not shown). Excisional wounds tend to express high levels of collagen I, so higher ratios of effluent from cells grown in condition 1 and 2 are of value for their treatment.

[00062] Referring to Figure 3(B) panel D, the methods of the present inventive subject matter comprise a method of using the dynamic culture medium compositions prepared as described above to treat existing scars and prevent scar formation. As shown in Figure 3(B) panel B, said method comprises applying a dermatological therapeutic composition comprising the dynamic culture medium enriched for secreted proteins, cytokines, chemokines, cell-growth factors, ECM proteins, and immunoregulatory proteins or molecules present in the eluate from the cell culture medium derived from ADMSCs (white arrow) to the scar to be treated, including but not limited to encourage scar tissue remodeling (Figure 3(B) panel B). Prior to treatment with a solution containing dynamic culture media the defect or scar to be treated (black arrow) is identified i.e. visible (Figure 3(B) panel A). As shown in Figure 3(B) panel B, a composition comprising a solution containing dynamic culture medium composition enriched for stem cell secreted proteins, cytokines, chemokines, cell-growth factors, ECM proteins, and immunoregulatory proteins or molecules present in the eluate from the cell culture medium derived from ADMSCs (white arrow) is injected into the scar or defect and surrounding area to promote remodeling of the tissue. In some embodiments of the present inventive subject matter, the application of the dynamic culture medium therapeutic composition can be enhanced or accelerated by microinjections or micro-needling with the compositions of the present inventive subject matter. In some embodiments of the present inventive subject matter, the application of the dynamic culture medium therapeutic composition is simply added topically to the scar with the compositions of the present inventive subject matter. As shown in Figure 3(B) panel C, the tissue post-treatment is fully healed without noticeable scarring or structural defects. The therapeutic application of the dynamic culture medium containing the stem cell derived secreted proteins, cytokines, chemokines, cell-growth factors, ECM proteins, and immunoregulatory proteins or molecules present in the eluate from the cell culture medium compositions of the present inventive subject matter can be applied to different scars or defects by varying the ratios of eluents from the various growth supported conditions shown in Figure IF to create an optimized therapeutic composition for a given pathology.

[00063] Scar treatment requires a high level of remodeling proteins expressed in the effluents before ECM repair occurs and requires mixing conditions 1-4 together to optimize the level of metalloproteinase and ECM proteins to break down and then rebuild ECM at the site of damage.

[00064] Referring to Figure 4, the compositions of the present invention inhibit tyrosinase activity, thereby preventing inhibition of melanin production. In a tyrosinase inhibition in vitro assay at 60 minutes' incubation, the composition of the present inventive subject matter, as shown in the gray colored bar, inhibits tyrosinase better than kojic acid, the control tyrosine inhibitor shown in the black colored bar. As shown in Figure 4, the bars represent the average of experiments (n=2), and the error bars represent the experimental standard deviation of the average.

[00065] Referring to Figure 5, the compositions of the present invention have modulated inhibition collagenase activity properties, allowing about 50% of collagenase activity. In a collagenase inhibition in vitro assay after 20-minute incubation, the compositions of the present inventive subject matter plus 0.3 mg/mL of pentapeptide composition, as shown in the dark gray colored bar has more collagenase activity than 1,10- phenathroline, the control collagenase inhibitor shown in the black colored bar, but not as much as collagenase itself as shown in the no inhibitor light gray colored bar in the middle. This modulated inhibition of collagenase is useful in the therapeutic methods of the present inventive subject matter.

[00066] The present inventive subject matter comprises compositions providing a cell culture matrix promoting the optimal growth of ADMSCs comprising human collagen I, hyaluronic acid, elastin, laminin, glycosaminoglycans, and other ECM proteins to support the growth of adult tissue derived stem cells. [00067] In addition to cell culture medium compositions, the present inventive subject matter comprises personalized regenerative medicine compositions useful in the treatment of medical conditions or disorders including, but not limited to, dermatologic medical conditions for example, wound-healing, hair loss, scars, pigmentation disorders, for example hyper-pigmentation of the skin.

[00068] The personalized regenerative medicine therapeutics of the present inventive subject matter are also useful in the treatment of inflammation disorders, joint damage, and pulmonary diseases.

Methods

[00069] In a non-limiting embodiment of the inventive subject matter, the methods of the present inventive subject matter comprise a method for synthesizing a stem cell culture medium composition for regenerative medicine, comprising the steps of providing a stem cell culture matrix composition comprising human collagen I, hyaluronic acid, elastin, laminin, glycosaminoglycans, and extracellular matrix proteins to support growth of adult tissue derived stem cells; isolating tissue from a donor; isolating stem cells from the donor tissue; culturing the stem cells derived from the donor tissue in a composition comprising said cell culture matrix composition and stem cell culture medium; and preparing said stem cell culture medium for use as a regenerative medicine therapeutic composition.

[00070] In a non-limiting embodiment of the inventive subject matter, the methods of the present inventive subject matter comprise a method for synthesizing a stem cell culture medium composition for regenerative medicine, comprising the steps of providing a stem cell culture matrix composition comprising human collagen I, hyaluronic acid, elastin, laminin, glycosaminoglycans, and extracellular matrix proteins to support growth of adult tissue derived stem cells; isolating tissue from a donor said donor tissue is adipose tissue from a lipoisolate sample; isolating adipose derived mesenchymal stem cells (ADMSCs) from the donor tissue; culturing the ADMSCs derived from the donor tissue in a composition comprising said cell culture matrix composition and stem cell culture medium; enriching said stem cell culture medium for stem cell products or secretions or effluents; and preparing said stem cell culture medium for use as a regenerative medicine therapeutic composition.

[00071] In a non-limiting embodiment of the inventive subject matter, the methods of the present inventive subject matter comprise a method for synthesizing a stem cell culture medium composition for regenerative medicine, comprising the steps of providing a stem cell culture matrix composition comprising human collagen I, hyaluronic acid, elastin, laminin, glycosaminoglycans, and extracellular matrix proteins to support growth of adult tissue derived stem cells; isolating tissue from a donor said donor tissue is adipose tissue from a liposisolate sample; isolating adipose derived mesenchymal stem cells (ADMSCs) from the donor tissue; culturing the ADMSCs derived from the donor tissue in a composition comprising said cell culture matrix composition and stem cell culture medium; enriching said stem cell culture medium for stem cell products or secretions or effluents; and preparing said stem cell culture medium for use as a regenerative medicine therapeutic composition for therapy of a patient's degenerative medical condition, for example but not limited to inflammation, pulmonary damage, joint damage, epithelial wound healing, scar reduction, trichology therapeutics, and pigmentation disorders of the skin. [00072] In a non-limiting embodiment of the inventive subject matter, the methods of the present inventive subject matter comprise a method for synthesizing a stem cell culture medium composition for regenerative medicine, comprising the steps of providing a stem cell culture matrix composition comprising human collagen I, hyaluronic acid, elastin, laminin, glycosaminoglycans, and extracellular matrix proteins to support growth of adult tissue derived stem cells; isolating tissue from a donor, performed from a lipoaspirate sample; culturing the stem cells derived from the donor tissue in a composition comprising said cell culture matrix composition and stem cell culture medium; enriching said stem cell culture medium for stem cell products or secretions or effluents; and preparing said stem cell culture medium for use as a regenerative medicine therapeutic composition.

[00073] In a non-limiting embodiment of the inventive subject matter, the methods of the present inventive subject matter comprise a method for synthesizing a stem cell culture medium composition for regenerative medicine, comprising the steps of providing a stem cell culture matrix composition comprising human collagen I, hyaluronic acid, elastin, laminin, glycosaminoglycans, and extracellular matrix proteins to support growth of adult tissue derived stem cells; isolating tissue from a donor, performed from a lipoaspirate sample; culturing the ADMSCs derived from the lipoaspirate in a composition comprising said cell culture matrix composition and stem cell culture medium; enriching said stem cell culture medium for stem cell products or secretions or effluents; and preparing said stem cell culture medium for use as a regenerative medicine therapeutic composition.

[00074] In a non-limiting embodiment of the inventive subject matter, the method of the present inventive subject matter comprise a method for synthesizing a stem cell culture medium composition for regenerative medicine, comprising the steps of providing a stem cell culture matrix composition comprising human collagen I, hyaluronic acid, elastin, laminin, glycosaminoglycans, and extracellular matrix proteins to support growth of adult tissue derived stem cells; isolating tissue from a donor, performed from a lipoaspirate sample; culturing the ADMSCs derived from the lipoaspirate in a composition comprising said cell culture matrix composition and stem cell culture medium; enriching said stem cell culture medium for stem cell products or secretions or effluents; and preparing said stem cell culture medium for use as a regenerative medicine therapeutic composition optimized for use as a personalized regenerative medicine therapeutic composition comprising optimized ratios of stem cell culture matrix to stem cell culture medium.

[00075] In a non-limiting embodiment of the inventive subject matter, the method of the present inventive subject matter comprise a method for synthesizing a stem cell culture medium composition for regenerative medicine, comprising the steps of providing a stem cell culture matrix composition comprising human collagen I, hyaluronic acid, elastin, laminin, glycosaminoglycans, and extracellular matrix proteins to support growth of adult tissue derived stem cells; isolating tissue from a donor, performed from a lipoaspirate sample; culturing the ADMSCs derived from the lipoaspirate in a composition comprising said cell culture matrix composition and stem cell culture medium; enriching said stem cell culture medium for stem cell products or secretions or effluents; and preparing said stem cell culture medium for use as a regenerative medicine therapeutic composition optimized for use as a personalized regenerative medicine therapeutic composition comprising optimized ratios of stem cell culture matrix to stem cell culture medium, said personalized regenerative medicine therapeutic composition comprising a series of compositions comprising a plurality of ratios of stem cell culture matrix to stem cell culture medium compositions.

[00076] In a non-limiting embodiment of the inventive subject matter, the method of the present inventive subject matter comprise a method for synthesizing a stem cell culture medium composition for regenerative medicine, comprising the steps of providing a stem cell culture matrix composition comprising human collagen I, hyaluronic acid, elastin, laminin, glycosaminoglycans, and extracellular matrix proteins to support growth of adult tissue derived stem cells; isolating tissue from a donor, performed from a lipoaspirate sample; culturing the ADMSCs derived from the lipoaspirate in a composition comprising said cell culture matrix composition and stem cell culture medium; enriching said stem cell culture medium for stem cell products or secretions or effluents; and preparing said stem cell culture medium for use as a regenerative medicine therapeutic composition optimized for use as a personalized regenerative medicine therapeutic composition comprising optimized ratios of stem cell culture matrix to stem cell culture medium, said personalized regenerative medicine therapeutic composition comprising a series of compositions comprising a plurality of ratios of stem cell culture matrix to stem cell culture medium compositions, where the optimized regenerative medicine therapeutic composition comprises preformulated therapeutic compositions for treating dermatological medical conditions.

[00077] In a non-limiting embodiment of the inventive subject matter, the method of the present inventive subject matter comprise a method for synthesizing a stem cell culture medium composition for regenerative medicine, comprising the steps of providing a stem cell culture matrix composition comprising human collagen I, hyaluronic acid, elastin, laminin, glycosaminoglycans, and extracellular matrix proteins to support growth of adult tissue derived stem cells; isolating tissue from a donor, performed from a lipoaspirate sample; culturing the ADMSCs derived from the lipoaspirate in a composition comprising said cell culture matrix composition and stem cell culture medium; enriching said stem cell culture medium for stem cell products or secretions or effluents; and preparing said stem cell culture medium for use as a regenerative medicine therapeutic composition optimized for use as a personalized regenerative medicine therapeutic composition comprising optimized ratios of stem cell culture matrix to stem cell culture medium, said personalized regenerative medicine therapeutic composition comprising a series of compositions comprising a plurality of ratios of stem cell culture matrix to stem cell culture medium compositions, where the optimized personalized regenerative medicine therapeutic composition comprises pre-formulated therapeutic compositions for treating inflammation, joint maladies, or pulmonary diseases.

[00078] In a non-limiting embodiment of the inventive subject matter, the methods of the present inventive subject matter comprise a method for treating conditions of the skin, thereby improving its appearance, tone, and texture. The inventive subject matter provides therapeutic use of compositions and methods for treatment of various conditions of the skin comprising compositions isolated from cultivates of adipose-derived mesenchymal stem cells (ADMSCs) isolated from adipose tissue, said compositions useful for facial skin rejuvenation treatments. When applied to facial skin, said facial rejuvenation compositions cause minimal to no facial peeling or discomfort, reduced downtime after treatment, and low photosensitivity. The skin rejuvenation treatment applications of the inventive subject matter are useful in various skin treatment comprising reduction of fine lines, wrinkles, shrinking enlarged pores, thickening aging skin having lost plasticity, repairing sun damage, skin discoloration and hyperpigmentation, tightening loose and sagging skin, reducing stretch marks, and reducing skin dullness. In addition, the skin rejuvenation treatment compositions comprise components that block melanin production, acts as antioxidants to increase skin tone and texture but do not cause irritation, pain, or photosensitivity, while improving the feeling of skin tightness and greatly improve skin tone and texture. The compositions of the inventive subject matter comprise a penta-peptide component that blocks melanin production and functions as an antioxidant increasing skin texture and tone, while reducing irritation, pain, and photosensitivity of the treated skin. The compositions of the present inventive subject matter provide a thicker more easily applied dermal solution with none of the detractions of commercially available skin treatments presently on the market.

[00079] In a non-limiting embodiment of the inventive subject matter, the methods of the present inventive subject matter comprise a method for treating conditions of the skin comprising reduction of fine lines and wrinkles, shrinking enlarged pores, thickening aging skin, repairing sun damage, skin discoloration and hyperpigmentation, tightening loose and sagging skin, reducing stretch marks, and reducing skin dullness. Said compositions comprise a composition to block melanin production and having antioxidant properties. In one embodiment said melanin-blocking antioxidant composition is a peptide. In one embodiment said peptide comprises a five-membered polypeptide, or pentapeptide. In one embodiment said pentapeptide having amino acid sequence glutamic acid, cysteine, glycine, tyrosine, and phenylalanine, or Glu-Cys-Gly-Tyr-Phe using the three- letter amino acid code, or ECGYF using the one-letter amino acid code. Said pentapeptide composition are formulated with other ingredients to provide a dermal therapeutic solution having thicker properties, making it more easily applicable to the skin to be treated with no undesirable effects of skin irritation, pain, discomfort, or photosensitivity caused by products presently on the market.

[00080] In a non-limiting embodiment of the inventive subject matter, the methods of the present inventive subject matter comprise a method for using the compositions of the inventive subject matter providing comparative advantages over formulations currently available in the marketplace. The compositions of the present inventive subject matter comprise 30% or less trichloroacetic acid (TCA) so the compositions of the present inventive subject matter can be used throughout the United States without regard to prohibitions of compositions comprising over 30% TCA. Moreover, the compositions of the present inventive subject matter comprise more kojic acid a tyrosinase activity inhibitor, useful in the prevention of pigmentation disorders of the skin, for example brown spots. The compositions of the present inventive subject matter are lower pH of around 2 as more effective formulation. The compositions of the present inventive subject matter comprise a novel peptide to encourage higher antioxidant activity thereby reducing brown spots, improving extracellular matrix breakdown and restructuring, thus providing tighter skin with improved tone and texture. [00081] In a non-limiting embodiment of the inventive subject matter, the methods of the present inventive subject matter comprise a method for using the compositions of the inventive subject matter said composition comprising purified water, TCA, glycerin, emulsifier, kojic acid, hydrogen peroxide, and disodium EDTA, and buffered with ammonium hydroxide to obtain a low pH of around 2 at room temperature. After the solution cools, an antioxidant composition is added. The resulting solution is applied to the skin with slight pressure and allowed to sit for up to 10 minutes. The application is repeated three times then washed off with water.

[00082] In a non-limiting embodiment of the inventive subject matter, the methods of the present inventive subject matter comprise a method for using the compositions of the present inventive subject matter said composition comprising up to 37% purified water, up to 30% trichloroacetic acid, up to 6% glycerin, up to 5% emulsifier, up to 5% kojic acid, up to 1% hydrogen peroxide, up to 0.25% disodium EDTA, and q.s. of purified water to obtain the desired composition of each component buffered with 13% ammonium hydroxide to a pH of around 2 at room temperature. After the solution cools, a peptide antioxidant composition is added. The resulting solution is applied to the skin to be treated with slight pressure and allowed to sit for up to 10 minutes. The application is repeated three times then washed off with water.

[00083] In a non-limiting embodiment of the inventive subject matter, the methods of the present inventive subject matter comprise a method for using the compositions of the present inventive subject matter said composition comprising 37% purified water, 30% trichloroacetic acid, 6% glycerin, 5% SEPIGEL emulsifier, 5% kojic acid, 1% hydrogen peroxide, 0.25% disodium EDTA, buffered with 13% ammonium hydroxide to a pH of around 2.1 at room temperature. After the solution cools, 0.3 mg/mL of ECGYF peptide composition is added. The resulting solution is applied to the skin to be treated with slight pressure and allowed to sit for up to 10 minutes. The application is repeated three times then washed off with water.

[00084] In a non-limiting embodiment of the inventive subject matter, the methods of the present inventive subject matter comprise a method for using the compositions of the present inventive subject matter said composition comprising 37% purified water, 30% trichloroacetic acid, 6% glycerin, 5% SEPIGEL emulsifier, 5% kojic acid, 1% hydrogen peroxide, 0.25% disodium EDTA, buffered with 13% ammonium hydroxide to a pH of around 2.1 at room temperature. After the solution cools, 0.3 mg/mL of ECGYF peptide composition is added. The resulting solution is combined with a composition of ADMSC eluents and is applied to the skin to be treated with slight pressure and allowed to sit for up to 10 minutes. The application is repeated three times then washed off with water.

[00085] In a non-limiting embodiment of the inventive subject matter, the methods of the present inventive subject matter comprise a method for using the compositions of the present inventive subject matter said composition comprising 37% purified water, 30% trichloroacetic acid, 6% glycerin, 5% SEPIGEL emulsifier, 5% kojic acid, 1% hydrogen peroxide, 0.25% disodium EDTA, buffered with 13% ammonium hydroxide to a pH of around 2.1 at room temperature. After the solution cools, 0.3 mg/mL of ECGYF peptide composition is added. The resulting solution is applied to the skin to be treated and followed with application of a composition of ADMSC eluents both applied sequentially to the skin to be treated with slight pressure and allowed to sit for up to 10 minutes. The application is repeated three times then washed off with water.

Compositions

[00086] In a non-limiting embodiment of the inventive subject matter, the compositions of the present inventive subject matter comprise a composition produced by a method for synthesizing a stem cell culture medium composition for regenerative medicine, comprising the steps of providing a stem cell culture matrix composition comprising human collagen I, hyaluronic acid, elastin, laminin, glycosaminoglycans, and extracellular matrix proteins to support growth of adult tissue derived stem cells; isolating tissue from a donor, performed from a lipoaspirate sample; culturing the ADMSCs derived from the lipoaspirate in a composition comprising said cell culture matrix composition and stem cell culture medium; enriching said stem cell culture medium for stem cell products or secretions or effluents; and preparing said stem cell culture medium for use as a regenerative medicine therapeutic composition optimized for use as a personalized regenerative medicine therapeutic composition comprising optimized ratios of stem cell culture matrix to stem cell culture medium, said personalized regenerative medicine therapeutic composition comprising a series of compositions comprising a plurality of ratios of stem cell culture matrix to stem cell culture medium compositions, where the optimized personalized regenerative medicine therapeutic composition comprises pre-formulated therapeutic compositions for treating dermatological medical conditions.

[00087] In a non-limiting embodiment of the inventive subject matter, the compositions of the present inventive subject matter comprise a composition produced by a method for synthesizing a stem cell culture medium composition for regenerative medicine, comprising the steps of providing a stem cell culture matrix composition comprising human collagen I, hyaluronic acid, elastin, laminin, glycosaminoglycans, and extracellular matrix proteins to support growth of adult tissue derived stem cells; isolating tissue from a donor, performed from a lipoaspirate sample; culturing the ADMSCs derived from the lipoaspirate in a composition comprising said cell culture matrix composition and stem cell culture medium; enriching said stem cell culture medium for stem cell products or secretions or effluents; and preparing said stem cell culture medium for use as a regenerative medicine therapeutic composition optimized for use as a personalized regenerative medicine therapeutic composition comprising optimized ratios of stem cell culture matrix to stem cell culture medium, said personalized regenerative medicine therapeutic composition comprising a series of compositions comprising a plurality of ratios of stem cell culture matrix to stem cell culture medium compositions, where the optimized personalized regenerative medicine therapeutic composition comprises pre-formulated therapeutic compositions for treating medical conditions comprising therapy of a patient's degenerative medical condition, for example but not limited to inflammation, pulmonary damage, joint damage, epithelial wound healing, scar reduction, trichology therapeutics, and pigmentation disorders of the skin.

[00088] In a non-limiting embodiment of the inventive subject matter, the compositions of the present inventive subject matter comprise compositions useful for treatment of various conditions of the skin comprising compositions isolated from cultivates of adipose-derived mesenchymal stem cells (ADMSCs) isolated from adipose tissue, said compositions useful for facial skin rejuvenation treatments. When applied to facial skin, said facial rejuvenation compositions cause minimal to no facial peeling or discomfort, reduced downtime after treatment, and low photosensitivity. The skin rejuvenation treatment applications of the inventive subject matter are useful in various skin treatment comprising reduction of fine lines, wrinkles, shrinking enlarged pores, thickening aging skin having lost plasticity, repairing sun damage, skin discoloration and hyperpigmentation, tightening loose and sagging skin, reducing stretch marks, and reducing skin dullness. In addition, the skin rejuvenation treatment compositions comprise components that block melanin production, acts as antioxidants to increase skin tone and texture but do not cause irritation, pain, or photosensitivity, while improving the feeling of skin tightness and greatly improve skin tone and texture. The compositions of the inventive subject matter comprise a penta-peptide component that blocks melanin production and functions as an antioxidant increasing skin texture and tone, while reducing irritation, pain, and photosensitivity of the treated skin. The compositions of the present inventive subject matter provide a thicker more easily applied dermal solution with none of the detractions of commercially available skin treatments presently on the market.

[00089] In a non-limiting embodiment of the inventive subject matter, the compositions of the present inventive subject matter comprise a composition useful for treating conditions of the skin comprising reduction of fine lines and wrinkles, shrinking enlarged pores, thickening aging skin, repairing sun damage, skin discoloration and hyperpigmentation, tightening loose and sagging skin, reducing stretch marks, and reducing skin dullness. Said compositions comprise a composition to block melanin production and having antioxidant properties. In one embodiment said melanin-blocking antioxidant composition is a peptide. In one embodiment said peptide comprises a five-membered polypeptide, or pentapeptide. In one embodiment said pentapeptide having amino acid sequence glutamic acid, cysteine, glycine, tyrosine, and phenylalanine, or Glu-Cys-Gly-Tyr-Phe using the three- letter amino acid code, or ECGYF using the one-letter amino acid code. Said pentapeptide composition are formulated with other ingredients to provide a dermal therapeutic solution having thicker properties, making it more easily applicable to the skin to be treated with no undesirable effects of skin irritation, pain, discomfort, or photosensitivity caused by products presently on the market.

[00090] In a non-limiting embodiment of the inventive subject matter, the compositions of the present inventive subject matter comprise compositions providing comparative advantages over formulations currently available in the marketplace. The compositions of the present inventive subject matter comprise 30% or less trichloroacetic acid (TCA) so the compositions of the present inventive subject matter can be used throughout the United States without regard to prohibitions of compositions comprising over 30% TCA. Moreover, the compositions of the present inventive subject matter comprise more kojic acid, a tyrosinase activity inhibitor, useful in the prevention of pigmentation disorders of the skin, for example brown spots. The compositions of the present inventive subject matter are lower pH of around 2 as more effective formulation. The compositions of the present inventive subject matter comprise a novel peptide to encourage higher antioxidant activity thereby reducing brown spots, improving extracellular matrix breakdown, and restructuring, thus providing tighter skin with improved tone and texture.

[00091] In a non-limiting embodiment of the inventive subject matter, the compositions of the inventive subject matter comprise purified water, tri-chloroacetic acid, glycerin, emulsifier, kojic acid, hydrogen peroxide, and disodium EDTA, and buffered with ammonium hydroxide to obtain a low pH of around 2 at room temperature, said components in therapeutically effective amounts. After the solution cools, an antioxidant composition is added. The resulting solution is applied to the skin with slight pressure and allowed to sit for up to 10 minutes. The application is repeated three times then washed off with water.

[00092] In a non-limiting embodiment of the inventive subject matter, the compositions of the present inventive subject matter comprise up to 37% purified water, up to 30% trichloroacetic acid, up to 6% glycerin, up to 5% emulsifier, up to 5% kojic acid, up to 1% hydrogen peroxide, up to 0.25% disodium EDTA, and q.s. of purified water to obtain the desired composition of each component buffered with 13% ammonium hydroxide to a pH of around 2 at room temperature. After the solution cools, a peptide antioxidant composition is added. The resulting solution is applied to the skin to be treated with slight pressure and allowed to sit for up to 10 minutes. The application is repeated three times then washed off with water.

[00093] In a non-limiting embodiment of the inventive subject matter, the compositions of the present inventive subject matter comprise 37% purified water, 30% trichloroacetic acid, 6% glycerin, 5% SEPIGEL emulsifier, 5% kojic acid, 1% hydrogen peroxide, 0.25% disodium EDTA, buffered with 13% ammonium hydroxide to a pH of around 2.1 at room temperature. After the solution cools, 0.3 mg/mL of ECGYF peptide composition is added. The resulting solution is applied to the skin to be treated with slight pressure and allowed to sit for up to 10 minutes. The application is repeated three times then washed off with water.

[00094] In a non-limiting embodiment of the inventive subject matter, the compositions of the present inventive subject matter comprise 37% purified water, 30% trichloroacetic acid, 6% glycerin, 5% SEPIGEL emulsifier, 5% kojic acid, 1% hydrogen peroxide, 0.25% disodium EDTA, buffered with 13% ammonium hydroxide to a pH of around 2.1 at room temperature. After the solution cools, 0.3 mg/mL of ECGYF peptide composition is added. The resulting solution is combined with a composition of ADMSC eluents and is applied to the skin to be treated with slight pressure and allowed to sit for up to 10 minutes. The application is repeated three times then washed off with water.

[00095] In a non-limiting embodiment of the inventive subject matter, the compositions of the present inventive subject matter comprise 37% purified water, 30% trichloroacetic acid, 6% glycerin, 5% SEPIGEL emulsifier, 5% kojic acid, 1% hydrogen peroxide, 0.25% disodium EDTA, buffered with 13% ammonium hydroxide to a pH of around 2.1 at room temperature. After the solution cools, 0.3 mg/mL of ECGYF peptide composition is added. The resulting solution is applied to the skin to be treated and followed with application of a composition of ADMSC eluents both applied sequentially to the skin to be treated with slight pressure and allowed to sit for up to 10 minutes. The application is repeated three times then washed off with water. [00096] In a non-limiting embodiment of the inventive subject matter, the therapeutic composition of the inventive subject matter is applied in two separate applications each with its own composition, RDS1 and RDS2. In one embodiment of the two-step therapeutic application, the first composition RDS1 comprises 25 to 80% purified or deionized water, 15 to 30 % trichloroacetic acid, 1 to 5% Kojic acid, 0.5 to 5% hydrogen peroxide, 1 to 10 % glycerin, 1 to 10 % SEPIGEL emulsifier, 0.1 to 1% disodium EDTA, 1 to 25% ammonium hydroxide, and 100 to 1,000 micrograms/mL pentapeptide solution. In one embodiment of the two step therapeutic application, the second composition RDS2 comprises 50 to 80% purified or deionized water, 0.1 to 5% hydroxyethylcellulose, 0.01 to 1% aloe vera powder 200x, 0.01 to 1% disodium EDTA, 1 to 15% FUCOGEL 1.5P moisturizer, 1 to 10% glycerin, 0.05 to 2% sodium hyaluronate, 0.01 to 1% lecithin, 0.05 to 4% COVRACYL MV60 thickening agent, 0.5 to 10% polyolprepolymer-15, 0.1 to 2% EUXYL PE 9010 preservative, and 0.05 to 5% LEXFEEL WOW emollient, 0.5% green tea extract, 0.54% butylene glycol, 0.01% GLUADIN KERA-P LM vegetable protein lysate, 0.003% adenosine triphosphate, 0.003% sodium chondroitin sulfate, 3% DCM, 0.15% Bis (tripeptide) copper acetate solution, and 0.1% white tea extract.

[00097] In a non-limiting embodiment of the inventive subject matter, the therapeutic composition of the inventive subject matter is applied in two separate applications each with its own composition, RDS1 and RDS2. In one embodiment of the two-step therapeutic application, the first composition RDS1 comprises about 47.81% purified or deionized water, 30 % trichloroacetic acid, 5% Kojic acid, 1.44% hydrogen peroxide, 6% glycerin, 5% SEPIGEL emulsifier, 0.25% disodium EDTA, 4.38% ammonium hydroxide, and 100 to 1,000 micrograms/mL pentapeptide solution. In a non-limiting embodiment of the inventive subject matter, the two step therapeutic application, the second composition RDS2 comprises about 79.73% purified or deionized water, 1% hydroxyethylcellulose, 0.06% aloe vera powder 200x, 0.05% disodium EDTA, 5% FUCOGEL 1.5P moisturizer, 4% glycerin, 0.1% sodium hyaluronate, 0.05% lecithin, 1% COVRACYL RH thickening agent, 2% polyolprepolymer-15, 0.8% EUXYL PE 9010 preservative, 2% LEXFEEL WOW emollient, 0.5% green tea extract, 0.54% butylene glycol, 0.01% GLUADIN KERA-P LM vegetable protein lysate, 0.003% adenosine triphosphate, 0.003% sodium chondroitin sulfate, 3% DCM, 0.15% Bis (tripeptide) copper acetate solution, and 0.1% white tea extract.

Embodiments - Compositions:

[00098] In a non-limiting embodiment of the inventive subject matter, the compositions of the present inventive subject matter comprise a composition produced by a method for synthesizing a stem cell culture medium composition for regenerative medicine, comprising the steps of providing a stem cell culture matrix composition comprising human collagen I, hyaluronic acid, elastin, laminin, glycosaminoglycans, and extracellular matrix proteins to support growth of adult tissue derived stem cells; isolating tissue from a donor, performed from a lipoaspirate sample; culturing the ADMSCs derived from the lipoaspirate in a composition comprising said cell culture matrix composition and stem cell culture medium; enriching said stem cell culture medium for stem cell products or secretions or effluents; and preparing said stem cell culture medium for use as a regenerative medicine therapeutic composition optimized for use as a personalized regenerative medicine therapeutic composition comprising optimized ratios of stem cell culture matrix to stem cell culture medium, said personalized regenerative medicine therapeutic composition comprising a series of compositions comprising a plurality of ratios of stem cell culture matrix to stem cell culture medium compositions, where the optimized personalized regenerative medicine therapeutic composition comprises pre-formulated therapeutic compositions for treating dermatological medical conditions comprising wound healing, scar healing, trichology treatments, and skin pigmentation disorders.

[00099] In a non-limiting embodiment of the inventive subject matter, the compositions of the present inventive subject matter comprise a composition produced by a method for synthesizing a stem cell culture medium composition for regenerative medicine, comprising the steps of providing a stem cell culture matrix composition comprising human collagen I, hyaluronic acid, elastin, laminin, glycosaminoglycans, and extracellular matrix proteins to support growth of adult tissue derived stem cells; isolating tissue from a donor, performed from a lipoaspirate sample; culturing the ADMSCs derived from the lipoaspirate in a composition comprising said cell culture matrix composition and stem cell culture medium; enriching said stem cell culture medium for stem cell products or secretions or effluents; and preparing said stem cell culture medium for use as a regenerative medicine therapeutic composition optimized for use as a personalized regenerative medicine therapeutic composition comprising optimized ratios of stem cell culture matrix to stem cell culture medium, said personalized regenerative medicine therapeutic composition comprising a series of compositions comprising a plurality of ratios of stem cell culture matrix to stem cell culture medium compositions, where the optimized personalized regenerative medicine therapeutic composition comprises pre-formulated therapeutic compositions for treating dermatological medical conditions comprising wound healing, scar healing, trichology treatments, and skin pigmentation disorders. Said composition further comprising purified water, tri-chloroacetic acid, glycerin, emulsifier, kojic acid, hydrogen peroxide, and disodium EDTA, and buffered with ammonium hydroxide to obtain a low pH of around 2 at room temperature, said components in therapeutically effective amounts. After the solution cools, an antioxidant composition is added. The resulting solution is applied to the skin with slight pressure and allowed to sit for up to 10 minutes. The application is repeated three times then washed off with water.

[000100] In a non-limiting embodiment of the inventive subject matter, the compositions of the present inventive subject matter comprise a composition produced by a method for synthesizing a stem cell culture medium composition for regenerative medicine, comprising the steps of providing a stem cell culture matrix composition comprising human collagen I, hyaluronic acid, elastin, laminin, glycosaminoglycans, and extracellular matrix proteins to support growth of adult tissue derived stem cells; isolating tissue from a donor, performed from a lipoaspirate sample; culturing the ADMSCs derived from the lipoaspirate in a composition comprising said cell culture matrix composition and stem cell culture medium; enriching said stem cell culture medium for stem cell products or secretions or effluents; and preparing said stem cell culture medium for use as a regenerative medicine therapeutic composition optimized for use as a personalized regenerative medicine therapeutic composition comprising optimized ratios of stem cell culture matrix to stem cell culture medium, said personalized regenerative medicine therapeutic composition comprising a series of compositions comprising a plurality of ratios of stem cell culture matrix to stem cell culture medium compositions, where the optimized personalized regenerative medicine therapeutic composition comprises pre-formulated therapeutic compositions for treating dermatological medical conditions comprising wound healing, scar healing, trichology treatments, and skin pigmentation disorders. Said composition further comprising up to 37% purified water, up to 30% trichloroacetic acid, up to 6% glycerin, up to 5% emulsifier, up to 5% kojic acid, up to 1% hydrogen peroxide, up to 0.25% disodium EDTA, and q.s. of purified water to obtain the desired composition of each component buffered with 13% ammonium hydroxide to a pH of around 2 at room temperature. After the solution cools, a peptide antioxidant composition is added. The resulting solution is applied to the skin to be treated with slight pressure and allowed to sit for up to 10 minutes. The application is repeated three times then washed off with water.

[000101] RENEW DERMAL SOLUTION™ 1 (RDS1). In a non-limiting embodiment of the inventive subject matter, the compositions of the present inventive subject matter comprise a composition produced by a method for synthesizing a stem cell culture medium composition for regenerative medicine, comprising the steps of providing a stem cell culture matrix composition comprising human collagen I, hyaluronic acid, elastin, laminin, glycosaminoglycans, and extracellular matrix proteins to support growth of adult tissue derived stem cells; isolating tissue from a donor, performed from a lipoaspirate sample; culturing the ADMSCs derived from the lipoaspirate in a composition comprising said cell culture matrix composition and stem cell culture medium; enriching said stem cell culture medium for stem cell products or secretions or effluents; and preparing said stem cell culture medium for use as a regenerative medicine therapeutic composition optimized for use as a personalized regenerative medicine therapeutic composition comprising optimized ratios of stem cell culture matrix to stem cell culture medium, said personalized regenerative medicine therapeutic composition comprising a series of compositions comprising a plurality of ratios of stem cell culture matrix to stem cell culture medium compositions, where the optimized personalized regenerative medicine therapeutic composition comprises pre-formulated therapeutic compositions for treating dermatological medical conditions comprising wound healing, scar healing, trichology treatments, and skin pigmentation disorders. Said composition further comprising about 37% purified water, about 30% trichloroacetic acid, about 6% glycerin, about 5% SEPIGEL emulsifier, about 5% kojic acid, about 1% hydrogen peroxide, about 0.25% disodium EDTA, buffered with 13% ammonium hydroxide to a pH of around 2.1 at room temperature. After the solution cools, 0.3 mg/mL of ECGYF peptide composition is added. In another embodiment of the RDS1 solution comprises 35.78 % deionized water, 29% trichloroacetic acid, 5% kojic acid, 4.33% of 30% hydrogen peroxide, 6% glycerin, 5% SEPIGEL 305, 0.25% disodium EDTA, 13.14% ammonium hydroxide from 30% ammonium hydroxide solution to adjust pH to about 2.1 at room temperature. After the solution cools, 250mg/mL ECGYF peptide is added to make up 0.12% of the RDS1 composition at an effective amount of 30 mg ECGYF peptide. The resulting solution is applied to the skin to be treated with slight pressure and allowed to sit for up to 10 minutes. The application is repeated several times preferably three times then washed off with water. This composition is referred to as the RDS1 solution comprising the ECGYF pentapeptide. [000102] RENEW DERMAL SOLUTION™ 2 (RDS2). In a non-limiting embodiment of the inventive subject matter, the compositions of the present inventive subject matter comprise a composition produced by a method for synthesizing a stem cell culture medium composition for regenerative medicine, comprising the steps of providing a stem cell culture matrix composition comprising human collagen I, hyaluronic acid, elastin, laminin, glycosaminoglycans, and extracellular matrix proteins to support growth of adult tissue derived stem cells; isolating tissue from a donor, performed from a lipoaspirate sample; culturing the ADMSCs derived from the lipoaspirate in a composition comprising said cell culture matrix composition and stem cell culture medium; enriching said stem cell culture medium for stem cell products or secretions or effluents; and preparing said stem cell culture medium for use as a regenerative medicine therapeutic composition optimized for use as a personalized regenerative medicine therapeutic composition comprising optimized ratios of stem cell culture matrix to stem cell culture medium, said personalized regenerative medicine therapeutic composition comprising a series of compositions comprising a plurality of ratios of stem cell culture matrix to stem cell culture medium compositions, where the optimized personalized regenerative medicine therapeutic composition comprises pre-formulated therapeutic compositions for treating dermatological medical conditions comprising wound healing, scar healing, trichology treatments, and skin pigmentation disorders. Said composition further comprising about 78.71% deionized water, about 0.99% hydroxyethylcellulose, about 0.06% aloe vera powder 200X, about 0.05% disodium EDTA, about 4.95% FUCOGEL 1.5P, about 3.96% glycerin, about 0.10% sodium hyaluronate, about 0.05% sunflower liquid lecithin, about 0.99% COVACRYL MV60, about 1.98% PEG-8/SMDI Copolymer, about 0.79% EUXYL PE 9010, about 1.98% LEXFEEL WOW, about 0.49% green tea extract (in glycerin), about 0.53% butylene glycol, about 0.01% GLUADIN® Kera-P LM, 0.003% adenosine triphosphate, about 0.003% sodium chondroitin sulfate, about 4% undiluted media, about 0.25% Bis (Tripeptide) copper acetate solution (SpecPed GHK2-Cu), and about 0.08% white tea extract. Said composition preferably comprising 78.714% deionized water, 0.991% hydroxyethylcellulose, 0.059% aloe vera powder 200X, 0.0495% disodium EDTA, 4.953% FUCOGEL 1.5P, 3.962% glycerin, 0.0991% sodium hyaluronate, 0.0495% sunflower liquid lecithin, 0.991% COVACRYL MV60, 1.981% PEG-8/SMDI Copolymer, 0.793% EUXYL PE 9010, 1.981% LEXFEEL WOW, 0.495% green tea extract (in glycerin), 0.535% butylene glycol, 0.0099% GLUADIN® KERA-P LM, 0.0029% adenosine triphosphate, 0.0029% sodium chondroitin sulfate, about 4% undiluted media, 0.247% Bis (Tripeptide) copper acetate solution (SpecPed GHK2-Cu), and 0.081% white tea extract.

[000103] Stem cell and stem cell component-free RDS2. In an alternate formulation of the RENEW DERMAL SOLUTION 2 (RDS2) composition, there is no stem cell culture medium and no stem cell products or secretions or effluents, said alternate stem-cell and stem cell component-free formulation comprising an optimized personalized regenerative medicine therapeutic composition comprising pre-formulated therapeutic compositions for treating dermatological medical conditions comprising wound healing, scar healing, trichology treatments, and skin pigmentation disorders. Said composition comprising about 79.31% deionized water, about 0.99% hydroxyethylcellulose, about 0.06% aloe vera powder 200X, about 0.05% disodium EDTA, about 4.99% FUCOGEL 1.5P, about 3.99% glycerin, about 0.10% sodium hyaluronate, about 0.05% lecithin, about 0.99% COVACRYL MV60, about 1.99% polyolprepolymer-15, about 0.79% EUXYL PE 9010, about 1.99% LEXFEEL WOW, about 0.49% green tea extract (in glycerin), about 0.54% butylene glycol, about 0.01% GLUADIN® KERA-P LM, about 0.003% adenosine triphosphate, about 0.003% sodium chondroitin sulfate, about 2.97% dEhO, about 0.05% ROS BIONET mix, about 0.05% KGF, about 0.05% DEFENSIN, about 0.05% EGF, about 0.25% Bis (Tripeptide) copper acetate solution (SpecPed GHK2-Cu), about 0.09% acetyl octapeptide-3, about 0.009% palmitoyl tripeptide-37 (0.005% solution, and about 0.08% white tea extract. Said composition preferably comprising 79.31% deionized water, 0.99% hydroxyethylcellulose, 0.06% aloe vera powder 200X, 0.05% disodium EDTA, 4.99% FUCOGEL 1.5P, 3.99% glycerin, 0.10% sodium hyaluronate, 0.05% lecithin, 0.99% COVACRYL MV60, 1.99% polyolprepolymer-15, 0.79% EUXYL PE 9010, 1.99% LEXFEEL WOW, 0.49% green tea extract (in glycerin), 0.54% butylene glycol, 0.01% GLUADIN® KERA-P LM, 0.003% adenosine triphosphate, 0.003% sodium chondroitin sulfate, 2.97% dhhO, 0.05% ROS BIONET mix, 0.05% KGF, 0.05% DEFENSIN, 0.05% EGF, 0.25% Bis (Tripeptide) copper acetate solution (SpecPed GHK2-Cu), 0.09% acetyl octapeptide-3, 0.009% palmitoyl tripeptide-37 (0.005% solution, and 0.08% white tea extract. ROS BIONET mix is a combination of sh-polypeptide-77, sh-polypeptide-2, Superoxide dismutase. Stock amounts of ROS BIONET are 0.6mg sh-polypeptide-77, 0.6mg sh-polypeptide-2, 1.2mg Superoxide dismutase and the amount in final product is 0.0625g. KGF is keratinocyte growth factor or sh-polypeptide-3 and the stock amount is 1.2mg and the amount in final product is 0.06g. EGF is epidermal growth factor or sH-oligopeptide-1 and the stock amount is 1.2mg and amount in final product is 0.06g. Defensin is honey bee defensin-1 or sr-honey bee defensin-1, royal jelly extract, the stock amount is 1.2mg, and the amount in final product is 0.06g .

[000104] In a non-limiting embodiment of the inventive subject matter, the compositions of the present inventive subject matter comprise a composition produced by a method for synthesizing a stem cell culture medium composition for regenerative medicine, comprising the steps of providing a stem cell culture matrix composition comprising human collagen I, hyaluronic acid, elastin, laminin, glycosaminoglycans, and extracellular matrix proteins to support growth of adult tissue derived stem cells; isolating tissue from a donor, performed from a lipoaspirate sample; culturing the ADMSCs derived from the lipoaspirate in a composition comprising said cell culture matrix composition and stem cell culture medium; enriching said stem cell culture medium for stem cell products or secretions or effluents; and preparing said stem cell culture medium for use as a regenerative medicine therapeutic composition optimized for use as a personalized regenerative medicine therapeutic composition comprising optimized ratios of stem cell culture matrix to stem cell culture medium, said personalized regenerative medicine therapeutic composition comprising a series of compositions comprising a plurality of ratios of stem cell culture matrix to stem cell culture medium compositions, where the optimized personalized regenerative medicine therapeutic composition comprises pre-formulated therapeutic compositions for treating dermatological medical conditions comprising wound healing, scar healing, trichology treatments, and skin pigmentation disorders. Said composition further comprising 37% purified water, 30% trichloroacetic acid, 6% glycerin, 5% SEPIGEL emulsifier, 5% kojic acid, 1% hydrogen peroxide, 0.25% disodium EDTA, buffered with 13% ammonium hydroxide to a pH of around 2.1 at room temperature. After the solution cools, 0.3 mg/mL of ECGYF pentapeptide composition is added. This solution is referred to as RDS1 solution. The resulting solution is applied to the skin to be treated and followed with application of a composition of ADMSC eluents, referred to as the RDS2 solution comprising ADMSC eluents, both RDS1 and RDS2 applied sequentially to the skin to be treated with slight pressure and allowed to sit for up to 10 minutes. The application is repeated three times then washed off with water.

[000105] In a non-limiting embodiment of the inventive subject matter, the compositions of the present inventive subject matter comprise a composition produced by a method for synthesizing a stem cell culture medium composition for regenerative medicine, comprising the steps of providing a stem cell culture matrix composition comprising human collagen I, hyaluronic acid, elastin, laminin, glycosaminoglycans, and extracellular matrix proteins to support growth of adult tissue derived stem cells; isolating tissue from a donor performed from a lipoaspirate sample; culturing the ADMSCs derived from the lipoaspirate in a composition comprising said cell culture matrix composition and stem cell culture medium; enriching said stem cell culture medium for stem cell products or secretions or effluents; and preparing said stem cell culture medium for use as a regenerative medicine therapeutic composition optimized for use as a personalized regenerative medicine therapeutic composition comprising optimized ratios of stem cell culture matrix to stem cell culture medium, said personalized regenerative medicine therapeutic composition comprising a series of compositions comprising a plurality of ratios of stem cell culture matrix to stem cell culture medium compositions, where the optimized personalized regenerative medicine therapeutic composition comprises pre-formulated therapeutic compositions for treating dermatological medical conditions comprising inflammation, joint disorders, and pulmonary diseases.

[000106] In a non-limiting embodiment of the inventive subject matter, the methods of the present inventive subject matter comprise a method for treating skin condition on a subject, comprising the steps of providing a skin therapeutic solution composition comprising purified water, trichloroacetic acid, glycerin, emulsifier, kojic acid, hydrogen peroxide, disodium EDTA, ammonium hydroxide buffer, human collagen 1, hyaluronic acid, elastin, laminin, glycosaminoglycans, and extracellular matrix proteins and stem cell products, secretions, and effluents eluded into cell culture medium during stem cell cultivation and growth; controlling the pH of said solution composition to about pH 2.1 at room temperature; applying in a first step said first solution composition to the skin being treated using slight pressure; allowing said solution composition to remain on the skin being treated for 10 minutes; repeating the applying of said solution composition on the skin being treated three times; washing said solution composition off the skin being treated with water; and applying in a second step a second solution comprising stem cell culture medium isolates composition comprising stem cell products, secretions, and effluents. The method further comprises wherein the first solution composition further comprises a peptide having antioxidant and melanin inhibiting functions. The method further comprises wherein the stem cell products, secretion, and effluents are adipose derived mesenchymal stem cell (ADMSC) isolates. The method further comprises wherein the peptide having antioxidant and melanin inhibiting functions is a pentapeptide. The method further comprises wherein the pentapeptide has amino acid sequence ECGYF. The method further comprises wherein the stem cell culture medium composition is applied optionally simultaneously or sequentially from the skin therapeutic composition.

[000107] In a non-limiting embodiment of the inventive subject matter, the compositions of the present inventive subject matter comprise a skin therapeutic composition comprising purified water, trichloroacetic acid, glycerin, emulsifier, kojic acid, hydrogen peroxide, disodium EDTA, and ammonium hydroxide buffer, wherein the composition has pH around 2. The composition further comprises a peptide having antioxidant and melanin inhibiting functions. The composition further comprises wherein the peptide is a pentapeptide of sequence ECGYF. The composition further comprises a stem cell culture isolate medium composition comprising stem cell products, secretions, and effluents. The composition further comprises wherein stem cell isolates are adipose derived mesenchymal stem cell isolates. The composition further comprises wherein said composition comprises up to 37% purified water, up to 30% trichloroacetic acid, up to 6% glycerin, up to 5% emulsifier, up to 5% kojic acid, up to 1% hydrogen peroxide, up to 0.25% disodium EDTA, buffered with 13% ammonium hydroxide, wherein the composition has pH around 2. The composition further comprises wherein said composition comprises 37% purified water, 30% trichloroacetic acid, 6% glycerin, 5% emulsifier, 5% kojic acid, 1% hydrogen peroxide, and 0.25% disodium EDTA, buffered with 13% ammonium hydroxide, wherein the composition has pH 2.1. The composition further comprises wherein the peptide has a concentration of 0.3 mg/mL.

[000108] In a non-limiting embodiment of the inventive subject matter, the compositions of the present inventive subject matter comprise a therapeutic composition for treating conditions of the skin, comprising two formulations applied in two steps, said therapeutic composition comprising RDS1 and RDS2 compositions where RDS1 comprises 25 to 50% purified or deionized water, 15 to 30 % trichloroacetic acid, 1 to 5% Kojic acid, 0.5 to 5% hydrogen peroxide, 1 to 10 % glycerin, 1 to 10 % SEPIGEL emulsifier, 0.1 to 1% disodium EDTA, 1 to 25% ammonium hydroxide, and 100 to 1,000 micrograms/mL pentapeptide solution; and where a second composition RDS2 comprises 50 to 80% purified or deionized water, 0.1 to 5% hydroxyethylcellulose, 0.01 to 1% aloe vera powder 200x, 0.01 to 1% disodium EDTA, 1 to 15% FUCOGEL 1.5P moisturizer, 1 to 10% glycerin, 0.05 to 2% sodium hyaluronate, 0.01 to 1% lecithin, 0.05 to 4% COVRACYL MV60 thickening agent, 0.5 to 10% polyolprepolymer-15, 0.1 to 2% EUXYL PE 9010 preservative, and 0.05 to 5% LEXFEEL WOW emollient, 0.1 to 2% green tea extract, 0.1 to 2 % butylene glycol, 0.1 to 1% GLUADIN KERA-P LM vegetable protein lysate, 0.001 to 0.05% adenosine triphosphate, 0.001 to 0.05% sodium chondroitin sulfate, 1 to 5% DCM, 0.1 to 2% Bis (tripeptide) copper acetate solution, and 0.1 to 2% white tea extract. The therapeutic composition for treating conditions of the skin, further comprising wherein said RDS1 comprises about 47.81% purified or deionized water, 30 % trichloroacetic acid, 5% kojic acid, 1.44% hydrogen peroxide, 6% glycerin, 5% SEPIGEL emulsifier, 0.25% disodium EDTA, between about 3.9% to about 4.3% ammonium hydroxide to adjust pH to about 2.1, and 100 to 1,000 micrograms/mL pentapeptide solution; and where RDS2 comprises about 79.73% purified or deionized water, 1% hydroxyethylcellulose, 0.06% aloe vera powder 200x, 0.05% disodium EDTA, 5% FUCOGEL 1.5P moisturizer, 4% glycerin, 0.1% sodium hyaluronate, 0.05% lecithin, 1% COVRACYL MV60 thickening agent, 2% polyolprepolymer-15, 0.8% EUXYL PE 9010 preservative, 2% LEXFEEL WOW emollient, 0.5% green tea extract, 0.54% butylene glycol, 0.01% GLUADIN KERA-P LM vegetable protein lysate, 0.003% adenosine triphosphate, 0.003% sodium chondroitin sulfate, 3% DCM, 0.15% Bis (tripeptide) copper acetate solution, and 0.1% white tea extract. The therapeutic composition for treating conditions of the skin, further comprising a pentapeptide having amino acid sequence ECGYF. The therapeutic composition for treating conditions of the skin, further comprising wherein said RDS1 solution further comprises human collagen 1, hyaluronic acid, elastin, laminin, glycosaminoglycans, and extracellular matrix proteins. The therapeutic composition for treating conditions of the skin, further comprising wherein said extracellular matrix proteins are stem cell products, secretions, and effluents. The therapeutic composition for treating conditions of the skin, further comprising wherein said stem cell products, secretions, and effluents are eluded into cell growth medium during adipose-derived mesenchymal stem cell (ADMSC) cultivation and growth.

EXAMPLES

[000109] EXAMPLE 1 - characterization of ADMSC condition 1 eluents

[000110] Referring to Figure 1 panel E, adipose-derived mesenchymal stem cells (ADMSCs) were cultured with conditions outlined for condition 1 (with SYNTHEMAX™ beads and no additional protein). After the eluent was collected, 50 ug of protein was digested and subjected to liquid chromatography (LC) mass spectroscopy (MS) analysis. For proteomic analysis, peptides present in the solution were compared to a database of human proteins for identification. The tables below show detectable proteins found in condition 1 ADMSC cell culture eluent compared to media with no ADMSCs present. The resulting ADMSC DCM condition 1 eluents yielded 666 different proteins identified in this analysis, and 335 proteins were overexpressed in condition 1 when compared to background. Of those 335 overexpressed proteins 18 were completely unique (not expressed in the background media at all but expressed in ADMSC DCM condition 1 growth media). As shown in Table 1 below, the 18 unique human protein eluents were measured with LC-MS analysis present in DMSC DCM condition 1 growth medium, the proteins are grouped into general biological functions, although many of these proteins gave multiple functions. Table 2 below shows that of the 335 proteins overexpressed in DCM condition 1, 317 proteins are overexpressed compared to background (media only), these proteins are also grouped into general biological functions, although many of these proteins have multiple functions. To identify proteins expressed at lower levels than detectable with GC-MS, ELISA analyses were performed. As shown in Table 3 below, ELISAs for human proteins indicated ADMSC condition 1 growth resulted in enrichment of proteins involved in angiogenesis, cell growth and differentiation, hematopoietic regulation, immune signaling, anti-inflammatory proteins, and metalloproteinase inhibitors. Table 3 below shows the ADMSC DCM condition 1 eluent proteins measured with RAYBIOTECH™ Cl and C5 ELISA assays per the manufacturer's protocol and are shown categorized by biological function, although many of these proteins have multiple functions. [000111] TABLE 1 - Unique human proteins measured with LC-MS analysis present in ADMSC DCM condition 1 eluents.

[000112] TABLE 2 - Overexpressed human protein ADMSC condition 1 eluents characterized by LC-MS.

[000113] TABLE 3 - Human proteins detected and measured with RAYBIOTECH's Cl and C5 ELISAs in ADMSC DCM condition 1 growth medium eluents.

[000114] EXAMPLE 2 - RDS1 and RDS2 in dermatological treatment method

[000115] Materials: The RDS1 and RDS2 solutions are as described above. RDS1 and RDS2 solutions are stored at 4°C and should be applied while cold and immediately stored at 4°C following application to preserve activity of components.

[000116] Dosage: approximately 2mL of RDS1 and approximately 2 g of RDS2 solution are applied per treatment to the affected area. Optionally 1 mL of RDS1 can applied to the neck area, if applicable.

[000117] Method of dermal treatment: The method comprises a first step applying RDS1 solution, neutralization of the RDS1 solution, sequentially followed by step 2 application of the RDS2 calming solution. The RDS1 and RDS2 solutions are applied by hand to the affected skin area to be treated. The area to be treated e.g. face and/or neck is cleansed thoroughly, preferably using a gentle dermal cleanser such as FACTORFIVE™ Gentle Gel Cleanser. Alcohol or degreaser agent can be used to further cleanse and disinfect the skin to be treated. The nostrils, lips, and corners of the eyes and other sensitive areas are occluded to prevent accidental application of the treatment solutions. If the area to be treated has tattoos or microbladed areas these areas are completely occluded as the procedure can lift pigments in the skin. The RDS1 solution is stored in sealed vial containers having a stopper, requiring a cannula to vent any gas in the vial. An 18-gauge needle is then used to draw 2-2.5 mL of the RDS1 solution. The RDS2 solution is provided in a pump applicator bottle. When applying to the face, the facial area can be divided into three zones i.e. forehead, cheekbone, jaw and chin, with the neck area being an optional fourth zone to be treated.

[000118] Step 1 - RDS1 Penetrating Solution Application:

[000119] Forehead Application: Step 1) Apply 1-2 drops of the RDS1 Penetrating Solution onto the middle of the forehead near the glabella. Step 2) Using firm pressure for application, apply RDS1 in a swooping motion across the forehead. Perform until the product no longer moves easily and becomes slightly tacky. Step 3) Perform spot check, examine the skin for any signs of adverse reactions or frosting. Step 4) If either are observed, neutralize the peel, and stop treatment. Step 5) Repeat 1-3 for a total of three layers. Step 6) Cover each section of the face before moving to the next.

[000120] Cheekbone Application: Step 1) Perform a second spot check. Step 2) Apply 1-2 drops of the RDS1 solution. Step 3) Using the thumbs and fingers to shape and contour the cheekbone all the way up. Step 4) Repeat 1-3 for a total of three layers. Step 5) Using a gloved finger, apply underneath the orbital rim, and around the eyelids.

[000121] Jaw & Chin Application: Step 1) Drop RDS1 along the side of the jaw and the chin. Step 2) Contour with the thumbs and forefingers under the chin, pulling upwards with firm pressure. Step 3) Move from the jawline to the mandible, up to the ear. Step 4) Repeat 1-3 for a total of three layers. Step 5) Cover each section of the face before moving to the next.

[000122] Vertical Lip Line Application: Step 1) Apply 1 droplet of RDS1 directly on to a gloved finger. Step 2) Apply to vertical lip lines staying away from the lip itself.

[000123] Neck Application (optional): Step 1) Start at the collarbone. Step 2) Apply RDS1 and pull upwards using the knuckles. Step 3) Repeat 1-2 for a total of three layers.

[000124] Neutralization: Step 1) Neutralize the peel using 4x4 gauze with cool water to wipe off any stickiness. Step 2) Using fresh gauze, dry the face fully. Step 3) Dispose of the gauze and change gloves.

[000125] Step 2 - RDS2 Calming Solution Application:

[000126] In order to calm the skin following RDS1 Penetrating Solution application, and neutralization of the RDS1 solution treatment, a sequential calming solution treatment step is performed: Step 1) Apply q.s. of RDS2 Calming Solution over the treated areas. Step 2) Optional: A separate mask, e.g. FACTORFIVE ^TM 7 Day Post Treatment Kit can optionally be applied, that evening or the next day. Place the GROWTH FACTOR FACIAL MASK™ from the 7 Day Post Treatment Kit on the face, then remove it after the designated time. Step 3) Apply SPF to the treated area.

[000127] Methods: RDS1 and RDS2 dermal treatment evaluation methodology. [000128] The treated skin of individuals was evaluated using Canfield Scientific's VISIA® Skin Analysis System per their published methodologies and a feature score assigned per the manufacturer's criteria. See

summary, VISIA® scans are performed at baseline before treatment and 2 weeks post-treatment of the 4th treatment, roughly 6 weeks from baseline to final scan. The right side, left side, and front of the face are scanned. The scores or feature counts are then averaged at each time point for an individual participant. Per Canfield's website referenced above, VISIA® score is determined by the size, area, and intensity of the skin feature. The VISIA® feature count provides a count of the number of discrete instances of the feature being evaluated, without regard to the size or intensity of each instance. The feature counts are defined as follows:

[000129] Spots: Spots are typically brown or red skin lesions including freckles, acne scars, hyper-pigmentation and vascular lesions. Spots are distinguishable by their distinct color and contrast from the background skin tone. Spots vary in size and generally have a circular shape.

[000130] Pores: Pores are the circular surface openings of sweat gland ducts. Due to shadowing, pores appear darker than the surrounding skin tone and are identified by their darker color and circular shape. The VISIA® system distinguishes pores from spots based on size; by definition, the area of a pore is much smaller than a spot.

[000131] Wrinkles: Wrinkles are furrows, folds or creases in the skin, which increase in occurrence as a result of sun exposure and are associated with decreasing skin elasticity. This skin feature has the greatest variability from image to image as it is highly dependent upon the facial expression of the client. Wrinkles are identified by their characteristic long, narrow shape.

[000132] Texture: Texture is primarily an analysis of skin smoothness. Texture measures skin color and smoothness by identifying gradations in color from the surrounding skin tone, as well as peaks and valleys on the skin surface that indicate variations in the surface texture.

[000133] Porphyrins: Porphyrins are bacterial excretions that can become lodged in pores and lead to acne. Porphyrins fluoresce in UV light and exhibit circular white spot characteristics.

[000134] UV Spots: UV spots occur when melanin coagulates below the skin surface as a result of sun damage. UV spots are generally invisible under normal lighting conditions. The selective absorption of the UV light by the epidermal melanin enhances its display and detection by VISIA®.

[000135] Red Areas: Red areas represent a potential variety of conditions, such as acne, inflammation, Rosacea or spider veins. Blood vessels and hemoglobin contained in the papillary dermis, a sub-layer of skin, give these structures their red color, which is detected by the RBX Technology in VISIA®. See

Acne spots and inflammation vary in size but are generally round in shape. Rosacea is usually larger and diffuse compared to acne, and spider veins usually are short, thin and can be interconnected in a dense network. [000136] Brown spots: Brown spots are lesions on the skin such as hyper-pigmentation, freckles, lentigines, and melasma. Brown spots occur from an excess of melanin. Melanin is produced by melanocytes in the bottom layer of the epidermis. Brown spots produce an uneven appearance to the skin and are detected in VI SI A® by RBX.

[000137] RESULTS: Dermal treatment with RDS1 and RDS2 RENEW DERMAL SOLUTION™s.

In collaboration with two dermal care practitioners, a single-arm non-randomized study investigating the effects of FACTORFIVE™ RENEW DERMAL SOLUTION™ after 4 applications was performed. FACTORFIVE™ RENEW DERMAL SOLUTION™ is a 2-component chemical peel. It is made up of RDS1 penetrating solution and RDS2 calming solution. These studies occurred over 6 weeks. The treatments were performed by a dermal care practitioner, either a Registered Nurse or an esthetician. Applications were administered every 7-10 days for a total of 4 applications. Final VISIA® scans occurred 2 weeks after the last application. A total of 10 study participants were enrolled in the study, male and female, ranging in age from 44 to 69 years old. The results are included in these data presented in Tables 4 and 5 below. VISIA® scans of participants were performed, and data were analyzed to quantify change over time following treatment. VISIA® score is determined by the size, area, and intensity of the skin feature. The VISIA® feature count provides a count of the number of discrete instances of the feature being evaluated, without regard to the size or intensity of each instance.

[000138] Table 4: VISIA® scores after 4 Applications of Renew Dermal Solution

[000139] Table 5: VISIA® feature counts after 4 applications of Renew Dermal Solution

[000140] Referring to Fig. 6 Panel A, a comparison image of baseline VISIA® image (left) with 2-week posttreatment VISIA® image (right) showing reduction in redness after 4 applications with FACTORFIVE™ RENEW DERMAL SOLUTION™ in a female patient. Referring to Fig. 6 Panel (B), a comparison image of baseline VISIA® image (left) with 2-week post-treatment photo (right) showing reduction in redness after 4 applications with FACTORFIVE™ RENEW DERMAL SOLUTION™ in a male patient. Referring to Fig. 7 , a comparison image of baseline image (left) with 2-week post-treatment image (right) showing reduction in the appearance of fine lines and wrinkles after 4 applications with FACTORFIVE™ RENEW DERMAL SOLUTION™ in a patient.

[000141] At the conclusion of the study, application of FACTORFIVE™ RENEW DERMAL SOLUTION™ improved overall skin quality as seen in Table 1 and Table 2. At 2 weeks post application 80% of participants displayed improvement in the appearance of wrinkles with an individual max of 42.3% improvement. Additionally, at 2 weeks post application 70% of participants displayed improvement in red areas with an individual max of 62.1% improvement.

[000142] Additional embodiments include any one of the embodiments described above, where one or more of its components, functionalities or structures is interchanged with, replaced by, or augmented by one or more of the components, functionalities, or structures of a different embodiment described above.

[000143] All patent and non-patent references cited herein is each hereby incorporated by reference in its entirety for all purposes.

[000144] Although several embodiments of the inventive subject matter have been disclosed in the foregoing specification, it is understood by those skilled in the art that many modifications and other embodiments of the inventive subject matter will come to mind to which the inventive subject matter pertains, having the benefit of the teaching presented in the foregoing description and associated drawings. It is thus understood that the inventive subject matter is not limited to the specific embodiments disclosed herein above, and that many modifications and other embodiments are intended to be included within the scope of the appended claims. Moreover, although specific terms are employed herein, as well as in the claims which follow, they are used only in a generic and descriptive sense, and not for the purposes of limiting the described inventive subject matter, nor the claims which follow.

Claims

What is claimed is:

1. A method for treating skin condition on a subject, comprising the steps of: providing a skin therapeutic solution composition comprising purified water, trichloroacetic acid, glycerin, emulsifier, kojic acid, hydrogen peroxide, disodium EDTA, ammonium hydroxide buffer, human collagen 1, hyaluronic acid, elastin, laminin, glycosaminoglycans, and optionally extracellular matrix proteins and stem cell products, secretions, and effluents eluded into cell culture medium during stem cell cultivation and growth, and a peptide having antioxidant and melanin inhibiting functions having amino acid sequence ECGYF; controlling the pH of said solution composition to about pH 2.1 at room temperature; applying in a first step said first solution composition to the skin being treated using slight pressure; allowing said solution composition to remain on the skin being treated for 10 minutes; repeating the applying of said solution composition on the skin being treated three times; washing said solution composition off the skin being treated with water; and applying in a second step a second solution comprising stem cell culture medium isolates composition comprising stem cell products, secretions, and effluents.

2. The method of claim 1, wherein the optional stem cell products, secretion, and effluents are adipose derived mesenchymal stem cell (ADMSC) isolates.

3. The method of claim 2, wherein the first solution composition comprises about 37% purified water, about 30% trichloroacetic acid, about 6% glycerin, about 5% SEPIGEL emulsifier, about 5% kojic acid, about 1% hydrogen peroxide, about 0.25% disodium EDTA, 13% ammonium hydroxide to buffer to a pH of around 2.1 at room temperature and an effective amount of ECGYF peptide.

4. The method of claim 3, wherein the second solution comprises about 78.71% deionized water, about 0.99% hydroxyethylcellulose, about 0.06% aloe vera powder 200X, about 0.05% disodium EDTA, about 4.95% FUCOGEL 1.5P, about 3.96% glycerin, about 0.10% sodium hyaluronate, about 0.05% sunflower liquid lecithin, about 0.99% COVACRYL MV60, about 1.98% PEG-8/SMDI Copolymer, about 0.79% EUXYL PE 9010, about 1.98% LEXFEEL WOW, about 0.49% green tea extract (in glycerin), about 0.53% butylene glycol, about 0.01% GLUADIN® Kera-P LM, 0.003% adenosine triphosphate, about 0.003% sodium chondroitin sulfate, about 4% DCM, about 0.25% GHK2-Cu, and about 0.08% white tea extract.

5. The method of claim 4 wherein: the first solution comprises 35.78 % deionized water, 29% trichloroacetic acid, 5% kojic acid, 4.33% of 30% hydrogen peroxide, 6% glycerin, 5% SEPIGEL 305, 0.25% disodium EDTA, 13.14% ammonium hydroxide to adjust pH to about 2.1 at room temperature, and 0.12% of ECGYF peptide solution of 250mg/mL; and the second solution comprises 78.714% deionized water, 0.991% hydroxyethyl-cellulose, 0.059% aloe vera powder 200X, 0.0495% disodium EDTA, 4.953% FUCOGEL 1.5P, 3.962% glycerin, 0.0991% sodium hyaluronate, 0.0495% sunflower liquid lecithin, 0.991% COVACRYL MV60, 1.981% PEG-8/SMDI Copolymer, 0.793% EUXYL PE 9010, 1.981% LEXFEEL WOW, 0.495% green tea extract (in glycerin), 0.535% butylene glycol, 0.0099% GLUADIN® Kera-P LM, 0.0029% adenosine triphosphate, 0.0029% sodium chondroitin sulfate, about 4% DCM, 0.247% GHK2-Cu, and 0.081% white tea extract.

6. The method of claim 5 wherein the optional stem cell culture medium composition is applied optionally simultaneously or sequentially from the skin therapeutic composition.

7. A skin therapeutic composition comprising purified water, trichloroacetic acid, glycerin, emulsifier, kojic acid, hydrogen peroxide, disodium EDTA, and ammonium hydroxide buffer, wherein the composition has pH around 2.

8. The composition of claim 7, further comprising a peptide having antioxidant and melanin inhibiting functions.

9. The composition of claim 8, wherein the peptide is a pentapeptide of sequence ECGYF.

10. The composition of claim 9 optionally further comprising a stem cell culture isolate medium composition comprising stem cell products, secretions, and effluents.

11. The composition of claim 10 wherein said optional stem cell isolates are adipose derived mesenchymal stem cell isolates.

12. The composition of claim 11 wherein said composition comprises up to 37% purified water, up to 30% trichloroacetic acid, up to 6% glycerin, up to 5% emulsifier, up to 5% kojic acid, up to 1% hydrogen peroxide, up to 0.25% disodium EDTA, buffered with about 1.3% ammonium hydroxide, wherein the composition has pH around 2.

13. The composition of claim 12 wherein said composition comprises 37% purified water, 30% trichloroacetic acid, 6% glycerin, 5% emulsifier, 5% kojic acid, 1% hydrogen peroxide, and 0.25% disodium EDTA, buffered with about 1.3% ammonium hydroxide, wherein the composition has pH 2.1.

14. The composition of claim 13 wherein the peptide has a concentration of 0.3 mg/mL.

15. A therapeutic composition for treating conditions of the skin, comprising two formulations applied in two steps, said therapeutic composition comprising RDS1 and RDS2 formulations wherein RDS1 comprises about 35% purified water, about 30% trichloroacetic acid, about 6% glycerin, about 5% SEPIGEL emulsifier, about 5% kojic acid, about 1% hydrogen peroxide, about 0.25% disodium EDTA, about 1.3% ammonium hydroxide to buffer to a pH of around 2.1 at room temperature, and an effective amount of ECGYF peptide; and wherein a second composition RDS2 comprises 50 to 80% purified or deionized water, 0.1 to 5% hydroxyethylcellulose, 0.01 to 1% aloe vera powder 200x, 0.01 to 1% disodium EDTA, 1 to 15% FUCOGEL 1.5P moisturizer, 1 to 10% glycerin, 0.05 to 2% sodium hyaluronate, 0.01 to 1% lecithin, 0.05 to 4% COVRACYL MV60 thickening agent, 0.5 to 10% polyolprepolymer-15, 0.1 to 2% EUXYL PE 9010 preservative, and 0.05 to 5% LEXFEEL WOW emollient, 0.1 to 2% green tea extract, 0.1 to 2 % butylene glycol, 0.1 to 1% GLUADIN KERA-P LM vegetable protein lysate, 0.001 to 0.05% adenosine triphosphate, 0.001 to 0.05% sodium chondroitin sulfate, 1 to 5% DCM, 0.1 to 2% Bis (tripeptide) copper acetate solution, and 0.1 to 2% white tea extract.

16. The therapeutic composition of claim 15 wherein,

RDS1 comprises 35.78 % deionized water, 29% trichloroacetic acid, 5% kojic acid, 4.33% of 30% hydrogen peroxide, 6% glycerin, 5% SEPIGEL 305, 0.25% disodium EDTA, 13.14% ammonium hydroxide to adjust pH to about 2.1 at room temperature, and 0.12% of ECGYF peptide solution; and; wherein RDS2 comprises 78.714% deionized water, 0.991% hydroxyethyl-cellulose, 0.059% aloe vera powder 200X, 0.0495% disodium EDTA, 4.953% FUCOGEL 1.5P, 3.962% glycerin, 0.0991% sodium hyaluronate, 0.0495% sunflower liquid lecithin, 0.991% COVACRYL MV60, 1.981% PEG-8/SMDI Copolymer, 0.793% EUXYL PE 9010, 1.981% LEXFEEL WOW, 0.495% green tea extract (in glycerin), 0.535% butylene glycol, 0.0099% GLUADIN® Kera-P LM, 0.0029% adenosine triphosphate, 0.0029% sodium chondroitin sulfate, about 4% undiluted media, 0.247% Bis GHK2-Cu, and 0.081% white tea extract.; where optionally RDS2 comprises 79.31% deionized water, 0.99% hydroxyethylcellulose, 0.06% aloe vera powder 200X, 0.05% disodium EDTA, 4.99% FUCOGEL 1.5P, 3.99% glycerin, 0.10% sodium hyaluronate, 0.05% lecithin, 0.99% COVACRYL MV60, 1.99% polyolprepolymer-15, 0.79% EUXYL PE 9010, 1.99% LEXFEEL WOW, 0.49% green tea extract (in glycerin), 0.54% butylene glycol, 0.01% GLUADIN® KERA-P LM, 0.003% adenosine triphosphate, 0.003% sodium chondroitin sulfate, 2.97% dH₂O, 0.05% ROS BIONET mix, 0.05% KGF, 0.05% DEFENSIN, 0.05% EGF, 0.25% Bis GHK2-Cu, 0.09% acetyl octapeptide-3, 0.009% palmitoyl tripeptide-37 0.005% solution, and 0.08% white tea extract.

17. The therapeutic composition of claim 16 wherein, the pentapeptide having amino acid sequence ECGYF has an effective dose of about 30 mg.

18. The therapeutic composition of claim 17, where said RDS1 solution optionally further comprises adipose derived mesenchymal stem cell growth products, secretions, and effluents.

19. The therapeutic composition of claim 18, where said optional extracellular matrix proteins are stem cell products, secretions, and effluents eluded into cell growth medium during adipose-derived mesenchymal stem cell (ADMSC) cultivation and growth.

20. Use of the therapeutic composition of claim 19 in a personalized regenerative medicine composition for treating a condition selected from dermatological medical conditions, facial rejuvenation treatment, trichology therapy to treat hair loss, and pigmentation disorders of the skin.