Methods and compositions for skin repair, rejuvenation, and comfort.

Abstract

A composition of rhPDGF-BB for the regeneration, rejuvenation, strengthening, comfort or aesthetic usefulness of soft tissue in the field of cosmetics and cosmeceuticals, comprising rhPDGF-BB combined with a carrier selected from the group consisting of different types of collagen, gelatin, alginates, polysaccharides, glycosaminoglycans, fibrin, hyaluronic acid and derivatives or combinations thereof, wherein damage to skin or soft tissue is treated by application of the disclosed composition.

Description

Technical Field

[0001] Priority Claim This application claims priority to U.S. Provisional Patent Application No. 63 / 523,584, filed on June 27, 2023, and U.S. Provisional Patent Application No. 63 / 522,989, filed on June 23, 2023, the entire contents of which are incorporated herein by reference.

[0002] The present invention relates to methods and compositions for using recombinant human platelet-derived growth factor BB (rhPDGF-BB) for the repair, rejuvenation, enhancement, and regeneration of skin and soft tissues, and for improving its comfort and aesthetics.

[0003] Description of Electronically Submitted Text File The content of the text file electronically submitted with this specification is incorporated herein by reference in its entirety: a computer-readable copy of the sequence listing named 093544-00002-01CIP1-CON_SL, recording date: 2024-05-01, file size 2,353 bytes.

Background Art

[0004] There is a well-known demand for cosmetic treatments to repair and restore skin damage. Aging and exposure to environmental factors cause wrinkles, sagging, and the development of grooves and creases in the skin. For example, the normal use of facial and neck muscles, such as by frowning or squinting, can also form grooves or creases in the facial and neck areas over time. The damage caused by these various factors results in an aesthetically displeasing cosmetic appearance. Aesthetic treatments such as micro-needling, micro-coaring, scraping, lasers, dermal fillers, and other injections are aimed at reversing skin damage, but there remains a great need for minimally invasive treatments that can effectively repair the skin with minimal pain or no pain at all and with a rapid recovery time.

[0005] rhPDGF-BB is known in the art for its use in bone regeneration and strengthening for both orthopedic and periodontal indications, and more recently, novel methods and compositions have been discovered for treating wounds such as ulcers and burns. The cellular and molecular mechanisms involved in bone regeneration and wound healing are highly complex, fundamentally different, and often directly opposed. Different processes involve fundamentally different extracellular matrix formation, calcification vs. non-calcification, different cellular signaling, and different cell types. Furthermore, the repair environment is very different, including vascular distribution for blood supply, cellular protein production involved in the repair process, different innervation, and different biomechanics (i.e., skin elasticity vs. bone stiffness). Therefore, PDGF compositions, dosage ranges, and treatment protocols known to treat bone repair in the body had to be adapted for treating wounds.

[0006] These known uses of PDGF delivered to bone or damaged skin within the body are generally expected to be ineffective and may even cause deterioration of aesthetics. For example, PDGF methods and compositions known in the art may stimulate osteoblast proliferation and bone formation, leading to skin calcification, or fibroblast growth and collagen production, which may result in better wound healing, but may also cause hypertrophic scarring, leading to severe complications and poor appearance. Furthermore, PDGF stimulates angiogenesis, i.e., the formation of new blood vessels, and an increase in facial blood vessels can be a serious cosmetic side effect of using PDGF in cosmetic procedures. Therefore, current methods, including delivery systems and therapeutic compositions of PDGF for treating bone and tissue within the body, would not be used by those skilled in the art to promote skin repair and rejuvenation.

[0007] However, given recent developments in using PDGF to treat different types of wounds, based on changes in composition and treatment regimens, there is a demand for effective use of PDGF to improve, repair, regenerate, or rejuvenate cosmetic damage to the dermis or subcutaneous layers of the skin in a patient-friendly and consistent manner. [Overview of the Initiative]

[0008] This disclosure describes compositions of rhPDGF-BB, and corresponding methods for improving skin repair and appearance, as well as aiding skin regeneration and rejuvenation, by using the disclosed compositions. These embodiments leverage the properties of rhPDGF-BB to treat tissue destruction such as that caused by skin and other connective tissue injuries, injuries or trauma, and disease processes, as well as defects in healthy skin integrity, such as those resulting from aging. The disclosed compositions of rhPDGF-BB may be used to improve tissue integrity and strengthening, for aesthetic utility in cosmetics and cosmeceuticals, or to improve comfort and reduce pain, inflammation, redness, and swelling resulting from skin and connective tissue destruction.

[0009] In one embodiment, an rhPDGF-BB-based composition for treating damaged or diseased skin and connective tissue comprises a composition of rhPDGF-BB and a biocompatible carrier selected from the group consisting of different types of collagen, gelatin, alginates, polysaccharides, glycosaminoglycans, fibrin, and their derivatives or combinations. Preferably, the composition comprises rhPDGF-BB and hyaluronic acid as a carrier. In another embodiment, an rhPDGF-BB-based composition for treating damaged or diseased skin and connective tissue comprises a composition of rhPDGF-BB alone (i.e., without a carrier). Those skilled in the art will understand that the disclosed compositions may include any combination of the above-described components and are not limited to the compositions described above.

[0010] This disclosure also describes a method for treating skin and connective tissue by applying any of the disclosed compositions. In one embodiment, the present invention provides a cosmetic method for treating a skin lesion of a subject, wherein the skin lesion does not involve tendons, ligaments or bones, and the method comprises (i) preparing a composition comprising, or essentially comprising, sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution and a sterile biocompatible carrier, wherein the biocompatible carrier is hyaluronic acid, collagen, gelatin, alginate, polysaccharide, glycosaminoglycan, fibrin, derivatives thereof or combination thereof, and (ii) applying the composition on or into the skin lesion, wherein the carrier delivers bioactive rhPDGF on or into the skin lesion to result in regeneration, rejuvenation or repair of the skin lesion. In a further embodiment, the present invention provides a method for reducing or improving pain or discomfort caused by skin destruction of a subject, wherein the destruction does not involve tendons, ligaments, or bones, and the method comprises (i) preparing a composition comprising or essentially comprising sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution, and optionally further comprising a sterile biocompatible carrier which is hyaluronic acid, collagen, gelatin, alginate, polysaccharide, glycosaminoglycan, fibrin, derivatives thereof, or combinations thereof, and (ii) applying the composition on or into the skin destruction to reduce pain or discomfort caused by the skin destruction.

[0011] In some embodiments, the composition includes hyaluronic acid as a carrier. In exemplary embodiments, the method comprises the steps of: mixing 1 to 3 milliliters (mL) of hyaluronic acid (a linear polysaccharide also known as hyaluronan or hyaluronate) with 0.5 to 0.3 mL of rhPDGF-BB solution in sodium acetate buffer having a pH between 5.5 and 6.5 to form a composition; and applying the composition to the surface of damaged or diseased skin or connective tissue. The composition may be applied to intact, undamaged, but aged skin or connective tissue, or to skin or connective tissue damaged by accidental or intentional damage to the dermis or subcutaneous layer of the skin. In some embodiments, skin damage is caused by aging, inflammation, radiation damage, ultraviolet damage, or a combination thereof. In some embodiments, skin damage is caused by surgical incision. In some embodiments, skin damage is scarring. In some embodiments, skin destruction is caused by aesthetic procedures described herein, such as microneedling, microcoring, dermaplaning, dermal ablation, dermal filling, or laser resurfacing. In aesthetic procedures, the compositions disclosed herein may be applied to the skin or other destruction, either alone or before, during, and / or after other aesthetic procedures, as further described herein. For example, the compositions may be applied topically to skin destruction, or before, during, and / or after microneedling, microcoring, dermal ablation, dermaplaning, dermal filling, or laser resurfacing procedures.

[0012] The above treatment may be repeated regularly for up to approximately six times, for example, weekly, every few weeks, monthly, or every few months, followed by a healing period of varying lengths depending on clinical requirements, after which it may be repeated.

[0013] The embodiments of the present invention provided in this summary are for illustrative purposes only and are intended to provide an overview of the selected embodiments disclosed herein. This summary is illustrative and selective and does not limit the scope of any claim, nor does it provide the full scope of the embodiments of the present invention disclosed or contemplated herein, nor should it be construed as limiting or restricting the scope of the present disclosure or any claimed embodiments of the invention. [Brief explanation of the drawing]

[0014] Figures 1A–1D show histology of injured skin biopsies treated with either PDGF example composition 1 or standard treatment, stained with hematoxylin and eosin. Dotted lines indicate the boundary between the epidermis and dermis, and circles highlight stained inflammatory cells. [Figure 1A] This is a micrograph of stained cells from a biopsy of injured skin treated with Example Composition 1, taken 21 days after the injury. [Figure 1B] This is a micrograph of stained cells from a biopsy of injured skin, taken 21 days after the injury and treated according to standard procedures. [Figure 1C] This is a micrograph of stained cells from a biopsy of injured skin treated with Example Composition 1, taken 42 days after the injury. [Figure 1D] This is a micrograph of stained cells from a biopsy of injured skin, taken 21 days after the injury and treated according to standard procedures.

[0015] Figures 2A and 2B show histology of injured skin biopsies treated with either PDGF example composition 1 or standard treatment, stained with Mallory's trichrome. [Figure 2A] This is a micrograph of stained cells from a biopsy of injured skin treated with Example Composition 1, taken 42 days after the injury. [Figure 2B] This is a micrograph of stained cells from a biopsy of injured skin, taken 42 days after the injury and treated according to standard procedures.

[0016] [Figure 3]These are photographic images of patients treated with PDGF Example Composition 1 prior to micro-needling. These images were taken immediately after treatment, 20 minutes after treatment, and 1 hour after treatment.

[0017] [Figure 4] These are photographic images of patients treated with PDGF Example Composition 1 prior to micro-needling. These images were taken immediately after treatment, 1 hour after treatment, and 2 hours after treatment.

[0018] [Figure 5] These are photographic images of patients taken 1 hour after treatment with PDGF Example Composition 1 prior to micro-needling, or 1 hour after treatment with micro-needling using a previous standard treatment.

[0019] [Figure 6] These are photographic images of melasma patients treated with PDGF Example Composition 1 prior to micro-needling. These images were taken immediately after treatment and 5 days after treatment.

[0020] [Figure 7A] These are photographic images of patients who underwent facelift surgery, with sutured incisions made on both sides of the face, treated with PDGF Example Composition 1 on one side and a control treatment of topical Aquaphor on the other side. [Figure 7B] These are photographic images of patients who underwent facelift surgery, with sutured incisions made on both sides of the face, treated with PDGF Example Composition 1 on one side and a control treatment of topical Aquaphor on the other side. [Figure 7C] These are photographic images of patients who underwent facelift surgery, with sutured incisions made on both sides of the face, treated with PDGF Example Composition 1 on one side and a control treatment of topical Aquaphor on the other side. [Figure 7D] These are photographic images of patients who underwent facelift surgery, with sutured incisions made on both sides of the face, treated with PDGF Example Composition 1 on one side and a control treatment of topical Aquaphor on the other side. [Figure 7E] These are photographic images of a patient who underwent a facelift, in which sutured incisions were made on both sides of the face; one side was treated with PDGF example composition 1, and the other side was treated with topical Aquaphor as a control.

[0021] [Figure 8A] These are photographic images of wounds caused by facelift bandages two weeks post-operatively, treated with a PDGF composition.

[0022] [Figure 8B] These are photographic images of wounds caused by facelift bandages 2 weeks post-surgery, treated with a PDGF composition 6 weeks later.

[0023] [Figure 9] These are photographic images of patients treated with PDGF example composition 1 after broadband laser and Moxi® laser treatment. These images were taken immediately after treatment and 4 hours after treatment.

[0024] [Figure 10] These are photographic images of patients treated with a 300 μg PDGF solution after Morpheus 8 treatment. These images were taken immediately before, immediately after, 3 days after, and 30 days after treatment.

[0025] Figures 11A-11B and 12A-12B show photographic images of a patient with wounds on the cheek and scalp after the Mohs procedure, where the cheek wound was sutured and treated with PDGF example composition 1, and the scalp wound was treated with skin substitute. [Figure 11A] This is a photograph of the cheek wound immediately after the Mohs procedure. [Figure 11B] This is a photographic image of a sutured cheek wound after Mohs surgery, treated with PDGF example composition 1, 10 days later. [Figure 12A] This is a photograph of the scalp wound immediately after the Mohs procedure. [Figure 12A]This is a photograph of a scalp wound treated with a skin substitute 10 days after a Mohs procedure. [Modes for carrying out the invention]

[0026] The following detailed description is provided to enable those skilled in the art to construct and use the present invention. For illustrative purposes, specific details are given to provide a complete understanding of the present invention. However, it will be apparent to those skilled in the art that these specific details are not necessary to carry out the present invention. Descriptions of specific uses are provided only as representative examples. Various modifications to preferred embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be applied to other embodiments and uses without departing from the scope of the present invention. The present invention is not intended to be limited to the embodiments shown, but rather to provide the broadest possible scope consistent with the principles and features disclosed herein.

[0027] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those commonly understood by those skilled in the art to which this disclosure belongs. Methods and materials are described herein for use in this disclosure; other suitable methods and materials known in the art may also be used. Materials, methods, and examples are illustrative and not intended to limit. All publications, patent applications, patents, sequences, database entries, and other references referenced herein are incorporated by reference in their entirety. In case of any conflict, this specification, including definitions, shall prevail.

[0028] Certain terms and phrases are used throughout this disclosure and have the following meanings in the context of ongoing disclosures.

[0029] As used herein, the article "a" means one or plural unless otherwise specified.

[0030] The meanings of the abbreviations are as follows: As is clear from their usage, "C" means Celsius or degrees Celsius, "μL" or "uL" or "ul" means microliter, "mL" means milliliter, "L" means liter, "mm" means millimeter, "nm" means nanometer, "mM" means millimolar concentration, "μM" or "uM" means micromolar concentration, "M" means molar concentration, "mmol" means millimoles, "μmol" or "uMol" means micromoles, "g" means grams, "μg" or "ug" means micrograms, "ng" means nanograms, "%w / v" means weight / volume percent, and "wt%" means weight percent.

[0031] As used in the claims and specification of this specification, the terms “comprising,” “contain,” “containing,” “including,” and “having” shall be considered to indicate an open group which may include other elements not specified. The terms “a,” “an,” and the singular form of the word should be interpreted as including the plural form of the same word, and such terms mean that one or more things are offered. The terms “one” or “single” may be used to indicate that one and only one thing is intended. Similarly, if a specific number of things is intended, other specific integer values ​​such as “2” may be used. The terms “preferably,” “preferred,” “prefer,” “optionally,” “may,” and similar terms are used to indicate that the item, condition, or process mentioned is an optional (not essential) feature of the invention.

[0032] As used herein, the term "consisting essentially of" refers to an element required of a given embodiment. This term allows for the presence of additional elements that do not substantially affect the fundamental and novel or functional characteristics of that embodiment of the invention.

[0033] Some embodiments of this specification assume numerical ranges. Where a numerical range is provided, unless otherwise indicated, the range includes its endpoints. Unless otherwise indicated, a numerical range includes all values ​​and subranges within it, as if it were explicitly written out.

[0034] Some values ​​in this specification are modified by the term “about.” In some cases, the term “about” in relation to a reference number may include a range of values ​​plus or minus 10% from that value. For example, the term “about 10” may include amounts from 9 to 11. In other embodiments, the term “about” in relation to a reference number may include a range of values ​​plus or minus 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% from that value.

[0035] composition As described herein, recombinant human platelet-derived growth factor BB (rhPDGF-BB) has been observed to effectively accelerate the repair or rejuvenation of skin or connective tissue, such as improving the appearance of skin or tissue, or improving the quality or integrity of tissues such as skin, by inducing an immediate influx of regenerating cells through a process called chemotaxis. Furthermore, rhPDGF-BB also stimulates the proliferation or growth of regenerating cells through a process called mitosis induction. In a third mechanism, rhPDGF-BB enhances the tissue environment for healthy tissue growth or rejuvenation or regeneration by inducing new capillaries and blood vessels to grow and stabilize. Therefore, compositions containing rhPDGF-BB are useful in treating tissue destruction such as that caused by skin and other connective tissue injuries, injuries or trauma, and disease processes, as well as defects in healthy skin integrity, such as those resulting from aging. Furthermore, the rhPDGF-BB composition is useful for aesthetic purposes in cosmetics and cosmeceuticals, including improving the appearance of the skin, by improving tissue integrity and strengthening, or by improving comfort and reducing pain, inflammation, redness, and swelling caused by skin and connective tissue damage.

[0036] In one embodiment, the present invention provides a composition comprising recombinant human platelet-derived growth factor BB (rhPDGF-BB). In some embodiments, the composition comprises sterile rhPDGF-BB in a physiological solution. In some embodiments, the composition is useful for promoting skin regeneration, rejuvenation, and repair. For example, the composition may be useful for treating skin damage in a subject. In some embodiments, the composition is useful for preventing or reducing pain or discomfort caused by skin damage in a subject. In some embodiments, the composition is useful for improving pain or discomfort caused by skin damage in a subject. In some embodiments, the composition may be useful for preventing or reducing redness, swelling, and / or inflammation caused by skin damage. Where used throughout this disclosure, the term “composition” may be used interchangeably with the terms “therapeutic composition” or “cosmetic composition.”

[0037] In some embodiments, the composition (e.g., therapeutic or cosmetic) comprises rhPDGF-BB and a biocompatible carrier. In some embodiments, the composition comprises sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution and a sterile biocompatible carrier. A “composition” of rhPDGF-BB may be used for application to intact or damaged skin or connective tissue and may comprise a therapeutic or cosmetic composition of rhPDGF-BB and a carrier. Compositions comprising rhPDGF-BB and a carrier are particularly useful for promoting skin regeneration, rejuvenation, and repair, for example, in subjects with skin damage. As used herein, the term “carrier” is intended to broadly refer to any biocompatible material that can function as a delivery vehicle for PDGF, while the term “matrix” is intended to refer to a carrier that acts as a substrate for cell adhesion and / or vascular endoproliferation and / or provides means for capturing PDGF within its structure (e.g., through interconnected pores), thereby enabling ongoing, delayed, or long-term delivery of PDGF as the disruption heals. Biocompatible carriers may be sterile and / or porous. Preferably, biocompatible carriers are sterile. In some embodiments, carriers are non-porous. The term “non-porous” means that the carrier does not contain interconnected pores within its structure that can capture PDGF.

[0038] In some embodiments, the composition (e.g., therapeutic or cosmetic) contains only rhPDGF-BB in a buffer without a carrier. As described herein, such compositions may be useful for treating skin damage or for promoting the repair, regeneration, and rejuvenation of skin damage in a subject. As described herein, such compositions may be useful for preventing, reducing, or improving pain or discomfort at a site of skin damage in a subject. Such compositions may also be useful for preventing or reducing swelling, redness, and / or inflammation at a site of skin damage in a subject. Such compositions may also be useful for preventing or reducing scarring at a site of skin damage in a subject. In some embodiments, when the composition contains PDGF without a carrier, the composition may be combined with other aesthetic compositions such as dermal fillers. For example, the composition may be combined with a filler containing cross-linked hyaluronic acid (e.g., Juvederm®). For example, the composition may be combined with a filler containing calcium hydroxyapatite (e.g., Radiesse®). For example, the composition may be combined with a filler containing poly-L-lactic acid (e.g., Sculptra®). When using a PDGF-only composition, the concentration of PDGF applied may be higher than in a composition containing a carrier. This is because the carrier assists in delivery, reducing the amount of PDGF required.

[0039] The composition is prepared using sterile rhPDGF-BB, with or without containing a carrier. In some embodiments, sterile PDGF comprises a pre-formulated sterile PDGF physiological solution, while in other embodiments, sterile PDGF comprises a lyophilized sterile powder containing PDGF that is reconstituted with sterile water or a buffer. The buffer used to reconstitute the lyophilized rhPDGF-BB or to prepare the PDGF physiological solution may include, but are not limited to, water, saline, carbonate, phosphate (e.g., phosphate-buffered saline), histidine, acetate (e.g., sodium acetate), acetic acid, and HCl, as well as organic buffers such as lysine, Tris buffer (e.g., tris(hydroxymethyl)aminoethane), N-2-hydroxyethylpiperazine'-N'-2-ethanesulfonic acid (HEPES), and 3-(N-morpholino)propanesulfonic acid (MOPS). Preferably, the buffer is sterile. The buffer can be selected based on its biocompatibility with PDGF and its ability to prevent undesirable protein modifications. The buffer can also be selected based on its suitability for the intended application. In some embodiments, sodium acetate buffer is used. In some embodiments, sodium chloride buffer is used. In some embodiments, phosphate-buffered saline is used. In some embodiments, phosphate-buffered saline and water are used. In some embodiments, sodium chloride buffer or sodium acetate buffer is used. In some embodiments, both sodium acetate and sodium chloride are used as buffers. In some embodiments, the buffer contains sodium acetate and water. In some embodiments, the buffer contains sodium chloride and water. The buffer can be used at different molar concentrations, e.g., about 0.1 mM to about 500 mM, about 0.1 mM to about 100 mM, about 1 mM to about 50 mM, about 5 mM to about 40 mM, about 10 mM to about 30 mM, or about 15 mM to about 25 mM, or about 100 mM to about 200 mM, or any molar concentration within these ranges. In some embodiments, the acetate buffer is used at a molar concentration of about 20 mM. In some embodiments, the buffer contains about 20 mM sodium acetate.In some embodiments, the buffer solution contains about 20 mM sodium acetate and water. In some embodiments, the buffer solution contains about 0.90% w / v (0.15 M) sodium chloride or phosphate-buffered saline. In some embodiments, the buffer solution contains about 0.15 M sodium chloride and water. In some embodiments, the buffer solution contains about 20 mM sodium acetate, about 0.15 M sodium chloride and water. Therefore, the compositions of the present invention contain a PDGF physiological solution containing PDGF in the buffer solution described herein. In some embodiments, the composition contains about 20 mM sodium acetate (in addition to rhPDGF-BB and optionally a carrier) and optionally water. In some embodiments, the composition contains about 0.15 M sodium chloride (in addition to rhPDGF-BB and optionally a carrier) and optionally water. In some embodiments, the composition contains about 20 mM sodium acetate (in addition to rhPDGF-BB and optionally a carrier), about 0.15 M sodium chloride, and optionally water. Preferably, the buffer solution is about 20 mM sodium acetate and water. Buffers or physiological solutions may contain pH values ​​of 6.0 ± 0.5 (i.e., 5.5 to 6.5).

[0040] As described herein, compositions may be prepared using sterile PDGF, sterile buffer, and / or sterile biocompatible carriers. In some embodiments, the final composition is sterile. This is particularly important when the composition is applied to surgical incisions or other skin lesions penetrating the skin surface (as described herein).

[0041] The carrier may include hyaluronic acid, various types of collagen, alginates, fibrin, gelatin, glycosaminoglycans, polysaccharides, derivatives thereof, or combinations thereof. In some embodiments, the carrier includes collagen. In some embodiments, the carrier includes alginates. In some embodiments, the carrier includes fibrin. In some embodiments, the carrier includes gelatin. In some embodiments, the carrier includes glycosaminoglycans. For example, the carrier may include hyaluronic acid. In some embodiments, the carrier includes polysaccharides. In some embodiments, the carrier includes a combination of polysaccharides and alginates. In some embodiments, the carrier includes a combination of polysaccharides and collagen. In some embodiments, the carrier includes hyaluronic acid but does not include other carrier components described herein. For example, the carrier may include or consist of hyaluronic acid but not alginates, fibrin, gelatin, glycosaminoglycans, or other polysaccharides. For example, the carrier may include or consist of hyaluronic acid but not collagen. For example, the carrier may contain or consist of hyaluronic acid and not contain fibrin. For example, the carrier may contain or consist of hyaluronic acid and not contain any other polysaccharides.

[0042] The "collagen" in the rhPDGF-BB-based composition not only provides a matrix or scaffold for intracellular proliferation but also absorbs proteases that are detrimental to the integrity or healing of skin or connective tissue. The collagen component may be animal-derived collagen. It may contain at least 90% type I collagen, at least 10% type III collagen, hydrolyzed collagen, monomeric collagen, or cross-linked collagen. It may be freeze-dried collagen or gel-like collagen.

[0043] The compositions described herein have been shown to be particularly effective when hyaluronic acid (HA) is used as a carrier. Hyaluronic acid is useful as a carrier because it helps enhance skin hydration, elasticity, and repair and / or maintenance of the extracutaneous tissue (ECM). The inventors have also observed that HA allows for better penetration and absorption of the compositions into the target skin than other carriers such as collagen. In particular, fluid HA helps to deliver the compositions directly to the skin after aesthetic procedures (such as microneedling, microcoring, or laser treatment) or to channels / pores formed in the skin. Any reference to “hyaluronic acid” as used herein includes hyaluronic acid or its physiologically acceptable salts, e.g., sodium salts, potassium salts, zinc salts, and silver salts, or derivatives or mixtures thereof. Hyaluronic acid may be present in an amount of about 0.1% to about 2.5% by weight of the whole composition, for example, about 0.1% to about 2.5% by weight, about 0.1% to about 1.0% by weight, about 0.2% to about 1.0% by weight, about 0.3% to about 1.0% by weight, about 0.4% to about 1.0% by weight, about 0.5% to about 1.5% by weight, about 0.5% to about 1.0% by weight, about 0.5% to about 0.8% by weight, or about 0.6% to 0.8% by weight. Hyaluronic acid may be present in an amount of about 0.5% to about 1.0% by weight of the whole composition, for example, about 0.5% to about 0.6% by weight, about 0.7% by weight, about 0.8% by weight, about 0.9% by weight, or about 1.0% by weight. Preferably, hyaluronic acid is present in an amount of about 0.75% by weight of the whole composition. Hyaluronic acid is available in a molecular weight range of 4,000 Daltons to 8,000,000 Daltons. In the compositions disclosed herein, the hyaluronic acid has a molecular weight between approximately 100,000 Da and 1,000,000 Da. For example, the molecular weight may be between approximately 125,000 Da and 1,000,000 Da. For example, the molecular weight may be between approximately 150,000 Da and 900,000 Da. For example, the molecular weight may be between approximately 200,000 Da and 850,000 Da. The composition may contain hyaluronic acid that is a blend of at least two molecular weights.For example, the carrier may include a blend of (i) hyaluronic acid having a weight between approximately 100,000 Da and 300,000 Da and (ii) hyaluronic acid having a weight between approximately 700,000 Da and 950,000 Da. For example, the carrier may include a blend of (i) hyaluronic acid having a weight between approximately 125,000 Da and 275,000 Da and (ii) hyaluronic acid having a weight between approximately 775,000 Da and 925,000 Da. For example, the carrier may include a blend of (i) hyaluronic acid having a weight between approximately 150,000 Da and 250,000 Da and (ii) hyaluronic acid having a weight between approximately 800,000 Da and 900,000 Da. For example, the carrier may include a blend of (i) hyaluronic acid having a weight between approximately 100,000 Da and 150,000 Da and (ii) hyaluronic acid having a weight between approximately 750,000 Da and 900,000 Da. For example, the carrier may include a blend of (i) hyaluronic acid having a weight between approximately 175,000 Da and 225,000 Da and (ii) hyaluronic acid having a weight between approximately 825,000 Da and 875,000 Da. For example, the carrier may include a blend of (i) hyaluronic acid having a weight between approximately 190,000 Da and 210,000 Da and (ii) hyaluronic acid having a weight between approximately 840,000 Da and 860,000 Da. For example, the carrier may include a blend of (i) hyaluronic acid having a weight of about 200,000 Da and (ii) hyaluronic acid having a weight of about 850,000 Da. In some embodiments, the two molecular weights are blended in a ratio between 30:70 and 70:30. In some embodiments, the two molecular weights are blended in a ratio between 35:65 and 65:35. In some embodiments, the two molecular weights are blended in a ratio between 40:60 and 60:40. In some embodiments, the two molecular weights are blended in a ratio of 50:50. Preferably, the hyaluronic acid used to prepare the composition is formulated as a serum or gel. In some embodiments, the hyaluronic acid is a serum. In some embodiments, the hyaluronic acid is a gel. Thus, the resulting composition prepared using a serum or gel carrier and a PDGF solution has a formulation (e.g., liquid or fluid) suitable for both topical and injectable administration.

[0044] Furthermore, the PDGF-BB used in the (therapeutic or cosmetic) compositions of the present invention may originate from any source, such as a natural source, a synthetic source, or a recombinant source, and may be intact or cleaved. According to one embodiment, PDGF is produced by recombinant DNA technology. When PDGF is produced by recombinant DNA technology, a DNA sequence encoding a single monomer (e.g., PDGF B chain) is inserted into cultured cells for the expression of the B chain monomer. The monomer is then extracted and isolated from the cell culture and refolded to form a biologically active homodimer (e.g., PDGF-BB), which may be further processed for further purification. According to one embodiment, the cultured cells are mammalian cells or eukaryotic cells such as yeast cells, or prokaryotic cells such as Escherichia coli (E. coli) cells. The rhPDGF-BB produced through these recombinant technologies can be purified according to the techniques outlined, for example, in International Publication No. 2005 / 077973. In some embodiments, substantially all of the rhPDGF-BB contained in the compositions described herein is intact and cleaved. For example, the rhPDGF-BB contained in the composition may be at least about 1%, or at least about 5%, or at least about 10%, or at least about 15%, or at least about 20%, or at least about 25%, or at least about 30%, or at least about 35%, or at least about 40%, or at least about 45%, or at least about 50%, or at least about 55%, or at least about 60%, or at least about 65%, or at least about 70%, or at least about 75%, or at least about 80%, or at least about 85%, or at least about 90%, or at least about 95%, or at least about 97% by weight of uncut rhPDGF-BB. In some embodiments, the PDGF-BB used in the (therapeutic or cosmetic) composition of the present invention is cut.

[0045] In some embodiments, the rhPDGF-BB contained in the composition of the present invention is an rhPDGF-BB that contains, or is essentially derived from, an amino acid sequence having at least about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or about 100% identity with SEQ ID NO: 1 provided below: [ka]

[0046] In some embodiments, the rhPDGF-BB contained in the composition of the present invention contains, or is essentially composed of, an amino acid sequence having at least about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or about 100% identity with SEQ ID NO: 1, or at least about 80%, or at least about 85%, or at least about 90%, or at least about 95%, or at least about 97% by weight of uncut rhPDGF-BB. The term "identity" refers to the degree to which two amino acid sequences are identical (residue-wise) across a comparison window. Identity can be determined by comparing the positions in each sequence that can be aligned for comparison purposes. Standard homology software such as BLAST can be used. Molecules are identical at positions if equivalent positions in the sequences being compared are occupied by the same amino acids. The percentage of identity refers to a function of the number of identical amino acids at positions shared by the sequences being compared.

[0047] As used herein, the terms “promoting skin regeneration, rejuvenation and / or repair” or “leading to regeneration, rejuvenation and / or repair of a skin disruption” refer to a process in which the condition of the skin in question is improved and / or a skin disruption is repaired. In some embodiments, the skin in question may have an improved aesthetic appearance. The skin in question may be intact or damaged. In some embodiments, the skin in question may have skin disruption that affects its appearance. In some embodiments, promoting or leading to regeneration, rejuvenation and / or repair includes reducing wrinkles, sagging and / or grooves in the skin in question. In some embodiments, promoting or leading to regeneration, rejuvenation and / or repair includes reducing the size of pores in the skin in question. In some embodiments, promoting or leading to regeneration, rejuvenation and / or repair includes increasing tissue volume and / or tissue firmness. In some embodiments, promoting or resulting in regeneration, rejuvenation, and / or repair includes increasing skin elasticity. In some embodiments, promoting or resulting in regeneration, rejuvenation, and / or repair includes reducing sunspots or irregular pigmentation on the skin in question. In some embodiments, promoting or resulting in regeneration, rejuvenation, and / or repair includes reducing sunburn. In some embodiments, promoting or resulting in regeneration, rejuvenation, and / or repair includes reducing rough skin texture. In some embodiments, promoting or resulting in regeneration, rejuvenation, and / or repair includes improving skin texture. In some embodiments, promoting or resulting in regeneration, rejuvenation, and / or repair includes reducing skin dryness. In some embodiments, promoting or resulting in regeneration, rejuvenation, and / or repair includes improving skin function.In some embodiments, promoting or resulting in regeneration, rejuvenation, and / or repair includes stimulating cell growth. In some embodiments, promoting or resulting in regeneration, rejuvenation, and / or repair includes stimulating angiogenesis. In some embodiments, promoting or resulting in regeneration, rejuvenation, and / or repair includes reducing, preventing, or improving pain or discomfort experienced by the subject at, for example, a site of skin damage. Those skilled in the art can easily determine whether a treatment promotes or results in skin regeneration, rejuvenation, and repair by using standard methods available in the art. For example, by looking for a reduction in wrinkles, grooves, and sagging of the subject's skin; a reduction in redness or other discoloration, smoother skin, increased tissue volume, reduced redness, reduced swelling, reduced pain or discomfort, reduced pigmentation, reduced scarring, or reduced pore size.

[0048] As used herein, the terms “disruption” or “skin disruption” refer to a disorder or defect of skin or connective tissue that may result from a genetic disorder (a vesicular condition); an intentional or accidental acute injury such as a surgical defect or a traumatic defect; and a surgical intervention such as abdominoplasty and other types of surgical tissue flaps. In some embodiments, skin disruption is an intentional surgical defect. For example, skin disruption may be a surgical incision (i.e., a sutured incision, an adhesive incision, a stapled incision, or a surgical scar). For example, skin disruption may be an incision from a cosmetic or non-cosmetic surgery. For example, an incision may be from a facelift. For example, an incision may be from abdominoplasty. For example, an incision may be from breast augmentation or breast reduction. For example, an incision may be from surgery to remove skin cancer or lesions (such as Mohs microsurgery). An incision may be an open wound or flap that is in the process of being closed (e.g., by sutures, adhesive or staples). An incision may be a closed incision (such as an incision closed by sutures, adhesive or staples). In some embodiments, skin disruption may be caused by an aesthetic procedure described herein, such as microneedling, microcoring, dermaplaning, dermal ablation, dermal filling or laser resurfacing. Furthermore, “disruption” or “skin disruption” may be the result of aging, inflammation, radiation damage, ultraviolet (e.g., sun) damage, or other damage to the skin or connective tissue. In some embodiments, skin disruption is caused by aging, inflammation, radiation damage, ultraviolet damage, or a combination thereof. In some embodiments, skin disruption is caused by aging. Skin disruption caused by aging may include wrinkles, sagging, and grooves. Skin damage caused by inflammation, radiation damage, and / or UV damage may include sunburn, sunspots, irregular pigmentation, rough texture, and dryness.In some embodiments, skin disturbances may include one or more of the following: wrinkles, sagging, grooves, sunburn, sunspots, irregular pigmentation, rough texture, and dryness. Aging and UV damage can also result in loss of skin elasticity, as well as significant weight loss. In some embodiments, skin disturbances may also be skin laxity. Skin laxity includes sagging, wrinkles, and drooping of the skin. Aging and UV damage can also result in an increased appearance of large skin pores. In some embodiments, skin disturbances may be large skin pores. In some embodiments, skin disturbances may be scars. Scars may be surgical scars. Surgical scars may be scars from breast reconstruction, breast augmentation, or breast reduction, such as the bottom of a linear incision scar of the breast. Surgical scars may be from abdominal wall reconstruction. Scars may be acne scars. In some embodiments, skin disturbances may be irregular pigmentation, such as hyperpigmentation. In some embodiments, skin disturbances may be melasma. Skin lesions can be present on any part of the subject's body. In some embodiments, skin lesions are located on the subject's head, face, scalp, chest, neck, arms, hands, and / or legs. In some embodiments, skin lesions are present on the subject's face, scalp, and / or neck. Skin lesions can affect the aesthetic appearance of the skin. The lesions may be in relation to intact or damaged skin or tissue. In one example, the treatable area is at least about 1 cm² in area. 2 And 5cm 2 Less than 5-15cm 2 Between, or 15cm 2 This may include burns or chemical burns that exceed a certain area and cover a large portion of the body's surface area.

[0049] In the methods and compositions disclosed herein (for example, for treating skin damage to a subject, for example, for improving the aesthetics of the skin, or for improving pain or discomfort caused by skin damage to a subject), the skin damage does not involve tendons, ligaments, or bones. For example, the skin damage may be located in the epidermis and dermis, or in the epidermis, dermis, and subcutaneous tissue. In some embodiments, the skin damage does not extend below the subcutaneous tissue. In some embodiments, the skin damage extends only to the dermal or subcutaneous layer of the skin. In some embodiments, the skin damage does not extend below the subcutaneous tissue. As described above, the cellular and molecular mechanisms involved in bone regeneration and wound healing are very complex, fundamentally different, and often directly opposed. Different processes involve fundamentally different extracellular matrix formation, calcification vs. non-calcification, different cellular signaling, and different cell types. Furthermore, the repair environment is very different, including vascular distribution for blood supply, cellular protein production involved in the repair process, different innervation, and different biomechanics (i.e., skin elasticity vs. bone stiffness). Therefore, PDGF compositions, dosage ranges, and treatment protocols known to treat bone repair within the body would not be used by those skilled in the art to treat skin damage. Similarly, known methods involved in the repair of deep tissue wounds (including deep partial-thickness or full-thickness wounds) typically involve the step of performing wound debridement to remove necrotic or infected tissue, followed by the application of a wound dressing to reduce infection. Such steps are not required in the methods and uses disclosed herein. Therefore, the methods and uses disclosed herein (in particular those performed to improve the aesthetics of the skin) do not include any step of performing skin debridement to remove necrotic or infected tissue. In some embodiments, the methods and uses disclosed herein also do not include any step of applying a wound dressing to the treatment site.

[0050] As used herein, the terms “preventing or reducing pain or discomfort” or “improving pain or discomfort” refer to the reduction or elimination of pain or discomfort experienced by a subject as a result of skin damage, for example, at the site of an incision following a cosmetic or surgical intervention described herein, such as an incision made by a facelift. Other skin damage known to cause pain or discomfort includes skin damage caused by aging, inflammation, radiation damage, ultraviolet damage, or a combination thereof, such as sunspots, sunburn, and areas of dry or rough skin. The application of cosmetic procedures such as microneedling, microcoring, dermal ablation, dermaplaning, dermal fillers, or laser resurfacing may also cause pain or discomfort. The compositions described herein may also be applied in combination with these procedures (described herein) to reduce / prevent or improve pain or discomfort caused by the procedures. A person skilled in the art can determine whether a treatment improves pain or discomfort by monitoring the level of pain or discomfort in the subject over time after the treatment.

[0051] As used herein, the term “preventing or reducing redness, swelling and / or inflammation” refers to a reduction in redness, swelling and / or inflammation experienced by the subject as a result of skin destruction, for example, at the site of an incision following a cosmetic or surgical intervention described herein, such as an incision made by a facelift. In further examples, redness, swelling and / or inflammation may be prevented or reduced in scars such as surgical scars or acne scars. The application of cosmetic procedures such as microneedling, microcoring, dermal ablation, dermaplaning, dermal fillers or laser resurfacing may also cause redness, swelling and / or inflammation of the subject's skin. The compositions described herein may also be applied in combination with these procedures (either before, during, and / or after, as described herein) to prevent or reduce redness, swelling and / or inflammation caused by the procedures. A person skilled in the art can determine whether the treatment reduces redness, swelling, and / or inflammation by monitoring the appearance of the skin over time after the treatment.

[0052] The concentration of PDGF in embodiments of the present invention can be determined by using an enzyme-linked immunoassay as described in U.S. Patent Nos. 6,221,625, 5,747,273, and 5,290,708, which are incorporated herein by reference, or by using any other assay known in the art for determining PDGF concentration. A (therapeutic or cosmetic) composition disclosed herein, comprising rhPDGF-BB and a carrier or rhPDGF-BB in a buffer, may contain rhPDGF-BB in amounts between about 0.001 mg / mL and about 5 mg / mL, or between about 0.1 mg / mL and about 1 mg / mL, or between about 0.2 mg / mL and about 0.4 mg / mL. The rhPDGF-BB in a (therapeutic or cosmetic) composition disclosed herein may contain rhPDGF-BB in amounts of about 0.001 mg / mL, or about 0.01 mg / mL, or about 0.3 mg / mL, or about 0.5 mg / mL, or about 1.0 mg / mL. The rhPDGF-BB in the compositions disclosed herein may contain at least about 0.001 mg / mL, or at least about 0.01 mg / mL, or at least about 0.3 mg / mL, or at least about 0.5 mg / mL, or at least about 1.0 mg / mL of rhPDGF-BB. In some embodiments, the (therapeutic or cosmetic) composition may contain rhPDGF-BB in amounts of about 0.001 mg / mL, or about 0.01 mg / mL, or about 0.02 mg / mL, or about 0.03 mg / mL, or about 0.04 mg / mL, or about 0.05 mg / mL, or about 0.06 mg / mL, or about 0.07 mg / mL, or about 0.08 mg / mL, or about 0.09 mg / mL, or about 0.1 mg / mL, or It contains at a concentration of approximately 0.2 mg / mL, or approximately 0.25 mg / mL, or approximately 0.3 mg / mL, or approximately 0.35 mg / mL, or approximately 0.4 mg / mL, or approximately 0.5 mg / mL, or approximately 0.6 mg / mL, or approximately 0.7 mg / mL, or approximately 0.8 mg / mL, or approximately 0.9 mg / mL, or approximately 1 mg / mL, or approximately 2 mg / mL, or approximately 3 mg / mL, or approximately 4 mg / mL, or approximately 5 mg / mL.In some embodiments, the composition contains rhPDGF-BB in an amount of at least about 0.001 mg / mL, or at least about 0.01 mg / mL, or at least about 0.02 mg / mL, or at least about 0.03 mg / mL, or at least about 0.04 mg / mL, or at least about 0.05 mg / mL, or at least about 0.06 mg / mL, or at least about 0.07 mg / mL, or at least about 0.08 mg / mL, or at least about 0.09 mg / mL, or at least about 0.1 mg / mL, or at least about 0.2 mg / mL, The composition contains rhPDGF-BB at a concentration of at least about 0.25 mg / mL, or at least about 0.3 mg / mL, or at least about 0.35 mg / mL, or at least about 0.4 mg / mL, or at least about 0.5 mg / mL, or at least about 0.6 mg / mL, or at least about 0.7 mg / mL, or at least about 0.8 mg / mL, or at least about 0.9 mg / mL, or at least about 1 mg / mL, or at least about 2 mg / mL, or at least about 3 mg / mL, or at least about 4 mg / mL, or at least about 5 mg / mL. In some embodiments, the composition contains rhPDGF-BB at a concentration between about 0.04 mg / mL and 0.08 mg / mL. For example, the composition may contain rhPDGF-BB at a concentration between about 0.05 mg / mL and 0.08 mg / mL. For example, the composition may contain rhPDGF-BB at a concentration between about 0.06 mg / mL and 0.08 mg / mL. For example, the composition may contain rhPDGF-BB at a concentration between approximately 0.07 mg / mL and 0.08 mg / mL. For example, the composition may contain rhPDGF-BB at a concentration of approximately 0.075 mg / mL. For example, the final concentration of rhPDGF-BB may be between approximately 0.04 mg of rhPDGF-BB per mL of composition and 0.08 mg of rhPDGF-BB per mL of composition. For example, the final concentration of rhPDGF-BB may be between approximately 0.05 mg of rhPDGF-BB per mL of composition and 0.08 mg of rhPDGF-BB per mL of composition. For example, the final concentration of rhPDGF-BB may be between approximately 0.06 mg of rhPDGF-BB per mL of composition and 0.08 mg of rhPDGF-BB per mL of composition.For example, the final concentration of rhPDGF-BB may be between approximately 0.07 mg of rhPDGF-BB per mL of composition and 0.08 mg of rhPDGF-BB per mL of composition. For example, the final concentration of rhPDGF-BB may be approximately 0.075 mg of rhPDGF-BB per mL of composition. If the composition does not contain a carrier molecule (i.e., PDGF alone in buffer), the concentration of PDGF in the composition may be higher because it is not diluted by the carrier molecule. For example, a composition containing only PDGF may contain rhPDGF-BB at a concentration between approximately 0.01 and 1.0 mg / mL. The composition may contain rhPDGF-BB at a concentration between approximately 0.1 mg / mL and 0.5 mg / mL. The composition may contain rhPDGF-BB at a concentration between approximately 0.2 mg / mL and 0.4 mg / mL. The composition may contain rhPDGF-BB at a concentration of approximately 0.3 mg / mL. For example, the final concentration of rhPDGF-BB may be between approximately 0.01 mg of rhPDGF-BB per mL of composition and 1.0 mg of rhPDGF-BB per mL of composition. For example, the final concentration of rhPDGF-BB may be between approximately 0.1 mg of rhPDGF-BB per mL of composition and 0.5 mg of rhPDGF-BB per mL of composition. For example, the final concentration of rhPDGF-BB may be between approximately 0.2 mg of rhPDGF-BB per mL of composition and 0.4 mg of rhPDGF-BB per mL of composition. For example, the concentration of rhPDGF-BB may be approximately 0.3 mg of rhPDGF-BB per mL of composition.

[0053] In some embodiments, the total amount of rhPDGF-BB contained in the (therapeutic or cosmetic) composition is less than approximately 1000 μg / mL, or less than approximately 900 μg / mL, or less than approximately 800 μg / mL, or less than approximately 700 μg / mL, or less than approximately 600 μg / mL, or less than approximately 500 μg / mL, or less than approximately 400 μg / mL, or less than approximately 300 μg / mL, or less than approximately 200 μg / mL, or less than approximately 100 μg / mL, or 90 μg / The amount is less than mL, or less than approximately 80 μg / mL, or less than approximately 70 μg / mL, or less than approximately 60 μg / mL, or less than approximately 50 μg / mL, or less than approximately 25 μg / mL, or less than approximately 10 μg / mL, or less than approximately 9 μg / mL, or less than approximately 8 μg / mL, or less than approximately 7 μg / mL, or less than approximately 6 μg / mL, or less than approximately 5 μg / mL, or less than approximately 2.5 μg / mL, or less than approximately 1 μg / mL, or less than approximately 0.1 μg / mL. In some embodiments, the composition contains between approximately 1 μg of rhPDGF-BB and 10 μg of rhPDGF-BB per mL. In some embodiments, the composition contains between approximately 1 μg of rhPDGF-BB and 9 μg of rhPDGF-BB per mL. In some embodiments, the composition contains between approximately 1 μg of rhPDGF-BB and 8 μg of rhPDGF-BB per mL. In some embodiments, the composition contains between approximately 1 μg and 7 μg of rhPDGF-BB per mL. In some embodiments, the composition contains between approximately 1 μg and 6 μg of rhPDGF-BB per mL. In some embodiments, the composition contains between approximately 1 μg and 5 μg of rhPDGF-BB per mL. In some embodiments, the composition contains between approximately 1 μg and 80 μg of rhPDGF-BB per mL. In some embodiments, the composition contains between approximately 1 μg and 100 μg of rhPDGF-BB per mL. In some embodiments, the composition contains between approximately 0.1 μg and 1,000 μg of rhPDGF-BB per mL.In some embodiments, the composition contains between approximately 10 μg and 80 μg of rhPDGF-BB per mL. In some embodiments, the composition contains between approximately 20 μg and 80 μg of rhPDGF-BB per mL. In some embodiments, the composition contains between approximately 30 μg and 80 μg of rhPDGF-BB per mL. In some embodiments, the composition contains between approximately 40 μg and 80 μg of rhPDGF-BB per mL. In some embodiments, the composition contains between approximately 40 μg and 50 μg of rhPDGF-BB per mL. In some embodiments, the composition contains between approximately 70 μg and 80 μg of rhPDGF-BB per mL. In some embodiments, the composition contains approximately 75 μg of rhPDGF-BB per mL. For example, the final concentration of rhPDGF-BB may be between approximately 40 μg of rhPDGF-BB per mL of composition and 80 μg of rhPDGF-BB per mL of composition. For example, the final concentration of rhPDGF-BB may be between approximately 50 μg of rhPDGF-BB per mL of composition and 80 μg of rhPDGF-BB per mL of composition. For example, the final concentration of rhPDGF-BB may be between approximately 60 μg of rhPDGF-BB per mL of composition and 80 μg of rhPDGF-BB per mL of composition. For example, the final concentration of rhPDGF-BB may be between approximately 70 μg of rhPDGF-BB per mL of composition and 80 μg of rhPDGF-BB per mL of composition. For example, the final concentration of rhPDGF-BB may be between approximately 75 μg of rhPDGF-BB per mL of composition. If the final composition does not contain a carrier molecule (i.e., PDGF alone in physiological solution), the final concentration of PDGF in the composition may be higher because it is not diluted by the carrier molecule. For example, a PDGF-only composition may contain between approximately 1 μg of rhPDGF-BB per mL and 1,000 μg of rhPDGF-BB per mL. For example, a PDGF-only composition may contain between approximately 300 μg of rhPDGF-BB per mL and 900 μg of rhPDGF-BB per mL.For example, a PDGF-only composition may contain between approximately 400 μg of rhPDGF-BB and 800 μg of rhPDGF-BB per mL. For example, a PDGF-only composition may contain between approximately 500 μg of rhPDGF-BB and 700 μg of rhPDGF-BB per mL. For example, a PDGF-only composition may contain between approximately 100 μg of rhPDGF-BB and 500 μg of rhPDGF-BB per mL. For example, a PDGF-only composition may contain between approximately 200 μg of rhPDGF-BB and 400 μg of rhPDGF-BB per mL. For example, a PDGF-only composition may contain approximately 300 μg of rhPDGF-BB per mL. For example, the final concentration of rhPDGF-BB may be between approximately 1 μg of rhPDGF-BB per mL of composition and 1,000 μg of rhPDGF-BB per mL of composition. For example, the final concentration of rhPDGF-BB may be between approximately 300 μg of rhPDGF-BB per mL of composition and 900 μg of rhPDGF-BB per mL of composition. For example, the final concentration of rhPDGF-BB may be between approximately 400 μg of rhPDGF-BB per mL of composition and 800 μg of rhPDGF-BB per mL of composition. For example, the final concentration of rhPDGF-BB may be between approximately 500 μg of rhPDGF-BB per mL of composition and 700 μg of rhPDGF-BB per mL of composition. For example, the final concentration of rhPDGF-BB may be between approximately 100 μg of rhPDGF-BB per mL of composition and 500 μg of rhPDGF-BB per mL of composition. For example, the final concentration of rhPDGF-BB may be between approximately 200 μg of rhPDGF-BB per mL of composition and 800 μg of rhPDGF-BB per mL of composition. For example, the final concentration of rhPDGF-BB may be between approximately 200 μg of rhPDGF-BB per mL of composition and 400 μg of rhPDGF-BB per mL of composition. For example, the concentration of rhPDGF-BB may be approximately 300 μg of rhPDGF-BB per mL of composition.

[0054] Various amounts of rhPDGF-BB can be used in the (therapeutic or cosmetic) compositions of the present invention. In some embodiments, the total amount of rhPDGF-BB contained in the (therapeutic or cosmetic) composition is less than about 50 mg, or less than about 25 mg, or less than about 10 mg, or less than about 5 mg, or less than about 2.5 mg, or less than about 1 mg, or less than about 0.1 mg, or less than about 0.09 mg, or less than about 0.08 mg, or less than about 0.07 mg, or less than about 0.06 mg, or less than about 0.05 mg, or less than about 0.04 mg, or less than about 0.03 mg, or less than about 0.02 mg, or less than about 0.01 mg, or less than about 0.001 mg. In some embodiments, the total amount of rhPDGF-BB contained in the (therapeutic or cosmetic) composition is about 50 mg, or about 25 mg, or about 10 mg, or about 1.0 mg, or about 0.5 mg, or about 0.1 mg, or about 0.09 mg, or about 0.08 mg, or about 0.07 mg, or about 0.06 mg, or about 0.05 mg, or about 0.04 mg, or about 0.03 mg, or about 0.02 mg, or about 0.01 mg, or about 0.001 mg. In some embodiments, the total amount of rhPDGF-BB contained in the composition is at least about 50 mg, or at least about 25 mg, or at least about 10 mg, or at least about 1.0 mg, or at least about 0.5 mg, or at least about 0.1 mg, or at least about 0.09 mg, or at least about 0.08 mg, or at least about 0.07 mg, or at least about 0.06 mg, or at least about 0.05 mg, or at least about 0.04 mg, or at least about 0.03 mg, or at least about 0.02 mg, or at least about 0.01 mg, or at least about 0.001 mg.

[0055] The concentration of PDGF in the embodiments of the present invention is less than about 10 mg / g, or less than about 5 mg / g, or less than about 1 mg / g, or less than about 0.5 mg / g, or less than about 0.1 mg / g, or less than about 0.09 mg / g, or less than about 0.08 mg / g, or less than about 0.07 mg / g, or less than about 0.06 mg / g, or less than about 0.05 mg / g, or less than about 0.04 mg / g, or less than about 0.03 mg / g, or less than about 0.02 mg / g, or less than about 0.01 mg / g, or less than about 0.001 mg / g. In some embodiments, the concentration of PDGF in the composition of the present invention is between approximately 0.001 mg / g and approximately 1 mg / g, or between approximately 0.05 mg / g and approximately 5 mg / g, or between approximately 0.05 mg / g and approximately 15 mg / g, or between approximately 0.01 mg / g and approximately 1 mg / g, or between approximately 0.25 mg / g and approximately 0.5 mg / g. In some embodiments, the concentration of PDGF in the composition of the present invention is at least about 10 mg / g, or at least about 5 mg / g, or at least about 1 mg / g, or at least about 0.5 mg / g, or at least about 0.1 mg / g, or at least about 0.09 mg / g, or at least about 0.08 mg / g, or at least about 0.07 mg / g, or at least about 0.06 mg / g, or at least about 0.05 mg / g, or at least about 0.04 mg / g, or at least about 0.03 mg / g, or at least about 0.02 mg / g, or at least about 0.01 mg / g, or at least about 0.001 mg / g.

[0056] The ratio of rhPDGF-BB solution to the support is such that the support is 1 cm 3 Approximately 0.025 mL of solution per 1 cm of support 3 The ratio of rhPDGF-BB to the support is between approximately 5 mL of solution per unit, or 1 cm of support. 3 Approximately 1.0 μg of rhPDGF-BB per 1 cm² of carrier 3 Therapeutic or cosmetic compositions comprising an rhPDGF-BB solution and a carrier, in which the amount is approximately 750 μg of rhPDGF-BB per unit, are also provided herein.

[0057] In some embodiments, when the composition contains rhPDGF-BB solution in a carrier such as hyaluronic acid, the ratio of rhPDGF-BB solution to the carrier is 1:2. In some embodiments, when the composition contains rhPDGF-BB solution in a carrier such as hyaluronic acid, the ratio of rhPDGF-BB solution to the carrier is 1:1. In some embodiments, the ratio of rhPDGF-BB solution to the carrier is 1:3. In some embodiments, the ratio of rhPDGF-BB solution to the carrier is 1:4. In some embodiments, the ratio of rhPDGF-BB solution to the carrier is 1:5. In some embodiments, the ratio of rhPDGF-BB solution to the carrier is 1:6. In some embodiments, the ratio of rhPDGF-BB solution to the carrier is 1:7. In some embodiments, the ratio of rhPDGF-BB solution to the carrier is 1:8. In some embodiments, when the composition contains rhPDGF-BB solution in a carrier such as hyaluronic acid, the ratio of rhPDGF-BB solution to the carrier is 1:9. In some embodiments, the ratio of rhPDGF-BB solution to the carrier is 1:10. In some embodiments, when the composition contains rhPDGF-BB solution in a carrier such as hyaluronic acid, the ratio of rhPDGF-BB solution to the carrier is 2:1. In some embodiments, the ratio of rhPDGF-BB solution to the carrier is 3:1. In some embodiments, the ratio of rhPDGF-BB solution to the carrier is 4:1. In some embodiments, the ratio of rhPDGF-BB solution to the carrier is 5:1. In some embodiments, the ratio of rhPDGF-BB solution to the carrier is 6:1. In some embodiments, the ratio of rhPDGF-BB solution to the carrier is 7:1. In some embodiments, the ratio of rhPDGF-BB solution to the carrier is 8:1.

[0058] The inventors have observed that the compositions described herein require only a single PDGF growth factor to be effective in the methods and uses described herein (for example, in promoting skin repair, rejuvenation and regeneration (e.g., reducing wrinkles, grooves, sagging, pigmentation, sunspots, rough texture or dryness), reducing, preventing or improving pain or discomfort caused by skin damage, and preventing or reducing redness, swelling or inflammation caused by skin damage). Other growth factors such as TGF and EGF are not required in the compositions to achieve these effects. As a result, the compositions of the present invention, which require only a single growth factor, are easy to prepare and avoid the unnecessary use of multiple growth factors. As described herein, PDGF can achieve effective skin repair and regeneration by (i) chemotaxis, which causes an immediate influx of regenerating cells, including stem cells, fibroblasts, and smooth muscle, that promote the repair or rejuvenation of skin or connective tissue and improve the quality or integrity of tissues such as skin; (ii) mitosis induction, which promotes the proliferation or growth of regenerating cells, including stem cells; (iii) angiogenesis, which promotes the growth of new capillaries and blood vessels, which strengthens the tissue environment for the growth, rejuvenation, or regeneration of healthy tissue; (iv) production of collagen, elastin, glycosaminoglycans, and hyaluronic acid (all of which are important for maintaining healthy tissue); and (v) anti-apoptotic effects to improve cell survival. Other growth factors have different mechanisms of action and do not achieve the same broad range of regenerative and restorative effects as PDGF. For example, transforming growth factors (e.g., TGF) are known to stimulate inflammatory cells and result in scar tissue formation by high-density collagen. Fibroblast growth factors (e.g., FGF) are known to induce abnormal angiogenesis. Therefore, these growth factors are not considered suitable for aesthetic skin repair and regeneration. VEGF stimulates angiogenesis but does not promote other regenerative processes achieved with PDGF. Similarly, connective tissue growth factor (CTGF) promotes connective tissue growth (like PDGF) but does not promote other regenerative processes achieved with PDGF.Therefore, VEGF and CTGF are not as useful as PDGF in promoting skin repair, regeneration, and recovery. Accordingly, in some embodiments, the composition does not contain any additional growth factors (i.e., no growth factors other than recombinant human PDGF-BB (rhPDGF-BB)). In some embodiments, the composition does not contain any additional growth factors, such as any transforming growth factor (e.g., TGF-beta), fibroblast growth factor (e.g., FGF), hepatocyte growth factor (e.g., HGF), vascular endothelial growth factor (e.g., VEGF), connective tissue growth factor (e.g., CTGF), insulin-like growth factor (e.g., IGF), or epidermal growth factor (e.g., EGF). IGF and EGF have been reported to have some useful effects for wound healing. In some embodiments, the composition does not contain VEGF, CTGF, and / or TGF (e.g., TGF-beta). In particular, the composition does not contain TGF-beta. In particular, the composition does not contain VEGF. In particular, the composition does not contain CTGF.

[0059] The inventors have further observed that certain additional components increase the risk of potential adverse interactions, as well as the cost of preparing the composition. Furthermore, certain additional components may also reduce the bioavailability of PDGF. For example, skincare compositions containing alcohol are thought to have a drying effect on the skin. Other components such as carboxymethylcellulose, guar gum, or cellulose may stimulate inflammation. In some embodiments, the composition does not contain one or more of alcohol, carboxymethylcellulose, guar gum, and cellulose. In some embodiments, the composition does not contain any alcohol. In some embodiments, the composition does not contain carboxymethylcellulose. In some embodiments, the composition does not contain guar gum. In some embodiments, the composition does not contain cellulose.

[0060] However, other additional components may be included in the composition to further assist in promoting skin repair, regeneration, and recovery. In some embodiments, the compositions described herein further include additional components such as topical anesthetics, including but not limited to carbokaines, lidocaine, epinephrine, or combinations thereof.

[0061] The present invention also provides a kit comprising the compositions described herein. In some embodiments, the compositions are contained in a pre-formulated syringe. In some embodiments, the present invention provides two compositions for use in the methods and uses described herein. One composition comprises a carrier such as hyaluronic acid. The other composition comprises rhPDGF-BB. These two compositions may be contained in a kit. In further embodiments, the present invention also provides a kit comprising both compositions. Each composition may be contained in a pre-formulated syringe.

[0062] The compositions may be prepared in different formulations depending on their intended use. For example, the compositions described herein may be formulated as creams, gels, serums, balms, sun creams, after-sun creams, foundations, tint creams, tint sun creams, solutions, suspensions, emulsions, ointments, foams, pastes, lotions, powders, soaps, surfactant-containing cleansing oils, or sprays. Preferably, the compositions are formulated as serums or gels.

[0063] Method and use of composition The inventors have observed that compositions of rhPDGF-BB (therapeutic or cosmetic) containing or not containing hyaluronic acid or other carriers disclosed herein stimulate skin cell growth, angiogenesis, or collagen deposition, enhance tissue volume, thereby improving skin firmness and tightening. This improvement may exceed the improvement observed in the skin by using hyaluronic acid alone. Furthermore, the compositions also stimulate angiogenesis, thereby further improving skin health, texture, fullness, or color. In addition, the compositions reduce discomfort or pain or promote healing after microneedling or microcoring procedures, going beyond the comfort or healing obtained with hyaluronic acid alone. The compositions also reduce any pain, swelling, redness, inflammation, or scarring experienced by the patient undergoing the procedure, provide a cooling, tingling, and soothing effect on the skin, and can be applied to the skin before, during, and / or after aesthetic procedures such as microneedling and microcoring to promote repair, pain reduction, and healing. Therefore, the disclosed compositions and methods improve human function, texture, comfort, and / or appearance or aesthetic outcomes. These improved outcomes enhance patient satisfaction.

[0064] As a result, the treatment procedures described herein result in more satisfied patients immediately after treatment of damaged, affected, or deficient skin or connective tissue, with medical benefits lasting for several weeks to several months. The compositions and methods disclosed herein heal cosmetic damage to the dermis or subcutaneous layers of the skin in a patient-friendly and consistent manner. As discussed herein, these results are unexpected based on the use of PDGF known in the art.

[0065] In further embodiments, the present invention provides a method for treating a subject using the compositions described herein. In some embodiments, the method may include cosmetically treating a subject using the compositions described herein. In further embodiments, the present invention provides the compositions described herein for use in a method for treating a subject. In further embodiments, the present invention provides the use of the compositions described herein in the manufacture of a pharmaceutical for treating a subject. All methods and uses described herein may be carried out using any of the compositions described herein.

[0066] As used herein, the term “subject” refers to any living organism, such as humans and animals, that requires treatment, generally vertebrates. This term includes mammals such as primates, domestic animals (e.g., cattle, horses, pigs, sheep, goats), companion animals (e.g., cats, dogs), and laboratory animals (e.g., mice, rabbits, rats). In some embodiments, the subject is human. In some embodiments, the subject has the skin lesions described herein.

[0067] In a further embodiment, the present invention provides a method for promoting the regeneration, rejuvenation, or repair of target skin, comprising the steps of: (1) preparing a composition comprising, or essentially comprising, sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution and a sterile biocompatible carrier, wherein the biocompatible carrier is hyaluronic acid, collagen, gelatin, alginate, polysaccharide, glycosaminoglycan, fibrin, derivatives thereof, or combinations thereof; and (2) applying the composition to a region of target skin, wherein the carrier delivers bioactive rhPDGF onto or into the target skin to regenerate, rejuvenate, or repair the target skin. This method may be a cosmetic treatment method for promoting the regeneration, rejuvenation, or repair of target skin.

[0068] In a further embodiment, the present invention provides a composition for use in promoting the regeneration, rejuvenation, or repair of target skin, comprising, or essentially comprising, sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution and a sterile biocompatible carrier, wherein the biocompatible carrier is hyaluronic acid, collagen, gelatin, alginate, polysaccharide, glycosaminoglycan, fibrin, derivatives thereof, or combinations thereof, applied to a region of target skin, wherein the carrier delivers bioactive rhPDGF onto or into the target skin to regenerate, rejuvenate, or repair the target skin.

[0069] In a further embodiment, the present invention provides a use of a composition in the manufacture of a pharmaceutical for promoting the regeneration, rejuvenation, or repair of target skin, comprising, or essentially comprising, sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution and a sterile biocompatible carrier, wherein the biocompatible carrier is hyaluronic acid, collagen, gelatin, alginate, polysaccharide, glycosaminoglycan, fibrin, derivatives thereof, or combinations thereof, wherein the carrier delivers bioactive rhPDGF onto or into target skin to regenerate, rejuvenate, or repair the target skin.

[0070] In a further embodiment, the present invention provides a method for promoting the regeneration, rejuvenation, or repair of a target skin, comprising the steps of (1) preparing a composition comprising or essentially comprising sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution, and (2) applying the composition to a target area of ​​skin. This method may be a cosmetic treatment method for promoting the regeneration, rejuvenation, or repair of a target skin.

[0071] The area of ​​skin to which the composition is applied may have skin defects as defined herein. For example, skin defects may result from aging, inflammation, radiation damage, ultraviolet damage, or a combination thereof (e.g., wrinkles, sagging, grooves, skin laxity, sunburn, solar lentigines, irregular pigmentation, rough texture, dryness, large pores, etc.). For example, skin defects may be incisions from cosmetic surgery, such as a facelift. For example, skin defects may be scars as described herein (e.g., surgical scars or acne scars). For example, skin defects may be hyperpigmentation or melasma. For example, skin defects may result from percutaneous aesthetic procedures such as microneedling, microcoring, dermal ablation, or dermal filling. The area of ​​skin to which the composition is applied may be scheduled for percutaneous aesthetic procedures that introduce skin defects (e.g., microneedling, microcoring, dermal ablation, or dermal filling). In this context, the composition is administered in preparation for treatment to assist in the regeneration, rejuvenation, or repair of the skin. In some embodiments, the skin disruption does not involve tendons, ligaments, or bones. In some embodiments, the skin extends only to the dermis or subcutaneous layer of the skin in question. Methods and uses for promoting the regeneration, rejuvenation, or repair of the skin in question may be carried out using any of the compositions described herein. For example, the method and use may be carried out using a composition comprising rhPDGF-BB and a carrier containing hyaluronic acid, as described herein. For example, the concentration of rhPDGF-BB may be between about 40 μg of rhPDGF-BB per mL of composition and 80 μg of rhPDGF-BB per mL of composition, for example, about 75 μg of rhPDGF-BB per mL of composition. For example, hyaluronic acid may be present at a concentration of about 0.75% by weight of the composition. In some embodiments, the method and use are carried out using a composition comprising rhPDGF-BB in a physiological solution without a carrier. In these embodiments, the concentration of rhPDGF-BB may be between approximately 100 μg of rhPDGF-BB per mL of composition and 500 μg of rhPDGF-BB per mL of composition, for example, approximately 300 μg of rhPDGF-BB per mL of composition.Alternatively, in these embodiments, the concentration of rhPDGF-BB may be between approximately 100 μg of rhPDGF-BB per mL of composition and 1,000 μg of rhPDGF-BB per mL of composition.

[0072] In some embodiments, the present invention provides a method (e.g., a cosmetic method) for promoting the repair of skin incisions resulting from a surgical procedure (e.g., a cosmetic surgery procedure such as a facelift), comprising: (1) preparing a composition comprising, or essentially comprising, sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution and a sterile biocompatible carrier comprising hyaluronic acid; and (2) applying the composition (e.g., topically) to or into the incision, thereby allowing the carrier to deliver bioactive rhPDGF to the or into the incision to repair the skin of the subject. Promoting the repair of surgical skin incisions may include promoting the repair of the skin of the subject and / or reducing scarring of the skin of the subject.

[0073] In a further embodiment, the present invention provides a method for treating a target skin lesion, comprising the steps of (1) preparing a composition comprising, or essentially comprising, sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution and a sterile biocompatible carrier, wherein the biocompatible carrier is hyaluronic acid, collagen, gelatin, alginate, polysaccharide, glycosaminoglycan, fibrin, derivatives thereof, or a combination thereof; and (2) applying the composition to or into the lesion, wherein the carrier delivers bioactive rhPDGF to or into the lesion, resulting in regeneration, rejuvenation, or repair of the lesion. This method may be a cosmetic method for treating skin lesions to regenerate, rejuvenate, or repair the lesion.

[0074] In a further embodiment, the present invention provides a method for treating a skin lesion in a target area, wherein the lesion does not involve tendons, ligaments, or bones, and the method comprises (1) preparing a composition comprising, or essentially comprising, sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution and a sterile biocompatible carrier, wherein the biocompatible carrier is hyaluronic acid, collagen, gelatin, alginate, polysaccharide, glycosaminoglycan, fibrin, derivatives thereof, or combinations thereof; and (2) applying the composition to or into the lesion, wherein the carrier delivers bioactive rhPDGF to or into the lesion to result in regeneration, rejuvenation, or repair of the lesion. The method may be a cosmetic method for treating skin lesions to regenerate, rejuvenate, or repair the lesion.

[0075] In further embodiments, the present invention provides a composition for use in a method of treating a target skin lesion, comprising, or essentially comprising, sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution and a sterile biocompatible carrier, wherein the biocompatible carrier is hyaluronic acid, collagen, gelatin, alginate, polysaccharide, glycosaminoglycan, fibrin, derivatives thereof, or combinations thereof, which is applied on or into the skin lesion, the carrier delivering bioactive rhPDGF on or into the skin lesion to regenerate, rejuvenate, or repair the lesion. In some embodiments, the skin lesion does not involve tendons, ligaments, or bones.

[0076] In further embodiments, the present invention provides the use of a composition in the manufacture of a pharmaceutical for use in a method of treating skin destruction of a subject, comprising, or essentially comprising, sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution and a sterile biocompatible carrier, wherein the biocompatible carrier is hyaluronic acid, collagen, gelatin, alginate, polysaccharide, glycosaminoglycan, fibrin, derivatives thereof, or combinations thereof, wherein the carrier delivers bioactive rhPDGF on or into the skin destruction to regenerate, rejuvenate, or repair the destruction. In some embodiments, the skin destruction does not involve tendons, ligaments, or bones.

[0077] In further embodiments, the present invention provides a method for treating a skin lesion of a subject, comprising the steps of (1) preparing a composition comprising, or essentially comprising, sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution, and (2) applying the composition to or into the lesion to bring about regeneration, rejuvenation, or repair of the lesion. The method may be a cosmetic method for treating a skin lesion to regenerate, rejuvenate, or repair the lesion. In some embodiments, the composition does not contain the carrier described herein. The methods and uses described herein with carrier-free compositions may be administered by injection as described herein. In some embodiments, the skin lesion does not involve tendons, ligaments, or bones.

[0078] In further embodiments, the present invention provides compositions comprising, or essentially comprising, sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution for use in a method of treating skin destruction of a subject, which are applied on or in the skin destruction to regenerate, rejuvenate, or repair the destruction. In some embodiments, the skin destruction does not involve tendons, ligaments, or bones.

[0079] In further embodiments, the present invention provides the use of a composition comprising, or essentially comprising, sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution in the manufacture of a pharmaceutical for use in a method of treating a target skin lesion, wherein the composition is applied on or in the skin lesion to regenerate, rejuvenate, or repair the lesion. In some embodiments, the skin lesion does not involve tendons, ligaments, or bones.

[0080] The methods and uses for treating skin damage described herein may be applied to any skin damage described herein. For example, skin damage may result from aging, inflammation, radiation damage, ultraviolet damage, or a combination thereof (e.g., wrinkles, sagging, grooves, skin laxity, sunburn, sunspots, irregular pigmentation, rough texture, dryness, large pores). For example, skin damage may be caused by surgical incisions, such as in cosmetic surgery, such as a facelift. For example, skin damage may be scars, such as surgical scars or acne scars, as described herein. For example, skin damage may result from penetrating skin aesthetic procedures such as microneedling, microcoring, dermal ablation, dermaplaning, or laser treatment. The area of ​​skin to which the composition is applied may be scheduled for a penetrating skin aesthetic procedure that introduces skin damage (e.g., microneedling, microcoring, dermal ablation, dermaplaning, or laser treatment). In this situation, the composition is applied in preparation for the procedure to assist in skin regeneration, rejuvenation, or repair. In some embodiments, the skin disruption does not involve tendons, ligaments, or bones. In some embodiments, the skin disruption extends only to the dermis or subcutaneous layer of the skin in question. Methods and uses for treating the skin disruption in question may be carried out using any of the compositions described herein. For example, methods and uses may be carried out using a composition comprising rhPDGF-BB and a hyaluronic acid-containing carrier. For example, rhPDGF-BB may have a concentration between about 40 μg of rhPDGF-BB per mL of composition and 80 μg of rhPDGF-BB per mL of composition, for example, about 75 μg of rhPDGF-BB per mL of composition. For example, the composition may contain the hyaluronic acid carrier described herein, which may be present at a concentration of about 0.75% by weight of the composition. In some embodiments, the composition may contain PDGF in a physiological solution without the carrier described herein. In these embodiments, the rhPDGF-BB may have a concentration between approximately 100 μg of rhPDGF-BB per mL of composition and 500 μg of rhPDGF-BB per mL of composition, for example, approximately 300 μg of rhPDGF-BB per mL of composition.Alternatively, in these embodiments, the rhPDGF-BB may have a concentration between approximately 100 μg of rhPDGF-BB per mL of composition and 1,000 μg of rhPDGF-BB per mL of composition.

[0081] In some embodiments, the present invention provides a method (e.g., a cosmetic method) for treating skin damage caused by surgical incisions (e.g., cosmetic surgery such as a facelift), comprising: (1) preparing a composition comprising, or essentially comprising, sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution and a sterile biocompatible carrier comprising hyaluronic acid; and (2) applying the composition on or into the incision (e.g., topically) so that the carrier delivers bioactive rhPDGF on or into the incision to regenerate, rejuvenate, or repair the skin damage.

[0082] In some embodiments, the present invention provides a method (e.g., a cosmetic method) for treating skin defects (such as wrinkles, sagging, grooves, skin laxity, sunburn, sunspots, irregular pigmentation, rough texture, dryness, enlarged pores, etc.) caused by aging, inflammation, radiation damage, ultraviolet damage, or a combination thereof, comprising (1) preparing a composition comprising, or essentially comprising, sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution and a sterile biocompatible carrier comprising hyaluronic acid; and (2) applying the composition (e.g., topically) to the skin defects, wherein the carrier delivers bioactive rhPDGF to the defects or into the defects to regenerate, rejuvenate, or repair the skin defects. In some embodiments, the method may be performed before, during, and / or after cosmetic procedures such as microneedling, microcoring, dermal ablation, dermaplaning, dermal filling, or laser resurfacing procedures as described herein.

[0083] In some embodiments, the present invention provides a method (e.g., a cosmetic method) for treating skin defects (such as wrinkles, sagging, grooves, skin laxity, sunburn, sunspots, irregular pigmentation, rough texture, dryness, and enlarged pores) caused by aging, inflammation, radiation damage, ultraviolet damage, or a combination thereof, comprising (1) preparing a composition comprising or essentially comprising sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution, and (2) applying the composition to the skin defects (e.g., topically or by injection) to regenerate, rejuvenate, or repair the skin defects. In some embodiments, the method may be performed before, during, and / or after cosmetic procedures such as microneedling, microcoring, dermal ablation, dermaplaning, dermal filling, or laser resurfacing procedures as described herein. For example, a PDGF-only composition may be added to a dermal filler applied to the skin of a subject by injection. For example, a PDGF-only composition may be applied before, during, or after an aesthetic procedure such as microneedling or laser treatment (i.e., ablative or non-ablative).

[0084] In some embodiments, the present invention provides a method for treating a target scar (e.g., a cosmetic method) comprising: (1) preparing a composition comprising, or essentially comprising, sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution and a sterile biocompatible carrier comprising hyaluronic acid; and (2) applying the composition to the scar (e.g., topically or by injection) so that the carrier delivers bioactive rhPDGF to the scar or into the scar to regenerate, rejuvenate, or repair the target skin. In some embodiments, the method reduces redness, swelling, and inflammation of the scar site. In some embodiments, the method reduces the visibility of the scar. In some embodiments, the method improves the aesthetic appearance of the scar. In some embodiments, the method may be performed before, during, and / or after cosmetic procedures such as microneedling, microcoring, dermal ablation, dermaplaning, dermal filling, or laser treatment as described herein. The scar may be a surgical scar or an acne scar.

[0085] Methods and uses for treating skin defects may include reducing wrinkles, sagging and / or grooves in the skin of the subject. Methods and uses for treating skin defects may include reducing enlarged pores in the skin of the subject. Methods and uses for treating skin defects may include increasing tissue volume and / or tissue firmness. Methods and uses for treating skin defects may include increasing skin elasticity. Methods and uses for treating skin defects may include reducing skin laxity. Methods and uses for treating skin defects may include reducing sunspots or irregular pigmentation in the skin of the subject. Methods and uses for treating skin defects may include reducing sunburn in the skin of the subject. Methods and uses for treating skin defects may include reducing rough skin texture. Methods and uses for treating skin defects may include improving skin texture. Methods and uses for treating skin defects may include reducing skin dryness. Methods and uses for treating skin defects may include improving skin function. Methods and uses for treating skin damage may include stimulating cell growth to promote regeneration, rejuvenation, or repair of the skin in question. Methods and uses for treating skin damage may include stimulating angiogenesis to promote regeneration, rejuvenation, or repair of the skin in question. Methods and uses for treating skin damage may include preventing or reducing scarring of the skin in question. Methods and uses for treating skin damage may include improving the aesthetic appearance of the skin.

[0086] In further embodiments, the present invention provides a method for preventing or reducing pain or discomfort caused by skin damage to a subject, comprising the steps of (1) preparing a composition comprising or essentially comprising sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution, and (2) applying the composition to an area of ​​skin of a subject that is affected or will be affected by skin damage to prevent or reduce pain or discomfort caused by the damage. In some embodiments, the skin damage does not involve tendons, ligaments, or bones. In some embodiments, the composition further comprises a carrier as described herein, for example, hyaluronic acid.

[0087] In further embodiments, the present invention provides compositions comprising, or essentially comprising, sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution for use in a method of preventing or reducing pain or discomfort caused by skin damage to a subject, which are applied to an area of ​​skin affected or likely to be affected by skin damage to prevent or reduce pain or discomfort caused by the damage. In some embodiments, the skin damage does not involve tendons, ligaments, or bones. In some embodiments, the composition further comprises a carrier as described herein, for example, hyaluronic acid.

[0088] In further embodiments, the present invention provides the use of a composition comprising, or essentially comprising, sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution in the manufacture of a pharmaceutical for use in a method of preventing or reducing pain or discomfort caused by skin destruction of a subject, wherein the composition is applied to an area of ​​skin of a subject that is affected or will be affected by skin destruction to prevent or reduce pain or discomfort caused by the destruction. In some embodiments, the skin destruction does not involve tendons, ligaments, or bones. In some embodiments, the composition further comprises a carrier as described herein, for example, hyaluronic acid.

[0089] In further embodiments, the present invention provides a method for improving pain or discomfort caused by skin damage to a target, comprising the steps of (1) preparing a composition comprising or essentially comprising sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution, and (2) applying the composition to or into the skin damage to reduce the pain or discomfort caused by the damage. In some embodiments, the composition further comprises a carrier as described herein, for example, hyaluronic acid.

[0090] In further embodiments, the present invention provides a method for improving pain or discomfort caused by skin destruction of a subject, wherein the destruction does not involve tendons, ligaments, or bones, and the method comprises (1) preparing a composition comprising or essentially comprising sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution, and (2) applying the composition on or into the skin destruction to reduce pain or discomfort caused by the destruction. In some embodiments, the composition further comprises a carrier as described herein, for example, hyaluronic acid.

[0091] In further embodiments, the present invention provides compositions comprising, or essentially comprising, sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution for use in a method of improving pain or discomfort caused by skin destruction of a subject, which are applied on or in skin destruction to prevent or reduce pain or discomfort caused by the destruction. In some embodiments, the skin destruction does not involve tendons, ligaments, or bones. In some embodiments, the composition further comprises a carrier as described herein, for example, hyaluronic acid.

[0092] In further embodiments, the present invention provides the use of a composition comprising, or essentially comprising, sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution in the manufacture of a pharmaceutical for use in a method of improving pain or discomfort caused by skin destruction of a subject, wherein the composition is applied on or in skin destruction to prevent or reduce pain or discomfort caused by the destruction. In some embodiments, the skin destruction does not involve tendons, ligaments, or bones. In some embodiments, the composition further comprises a carrier as described herein, for example, hyaluronic acid.

[0093] Methods and uses for preventing, reducing, or improving pain or discomfort caused by skin damage may be carried out using any of the compositions described herein. For example, the methods and uses may be carried out using a composition comprising rhPDGF-BB in a physiological solution without a carrier, as described herein. The physiological solution may comprise the buffer described herein. In these embodiments, the rhPDGF-BB may have a concentration between about 100 μg of rhPDGF-BB per mL of composition and 1000 μg of rhPDGF-BB per mL of composition, for example, about 500 μg of rhPDGF-BB per mL of composition. Alternatively, the methods and uses may be carried out using a composition comprising rhPDGF-BB and a carrier, preferably hyaluronic acid, as described herein. In these embodiments, the rhPDGF-BB may have a concentration between about 40 μg of rhPDGF-BB per mL of composition and 80 μg of rhPDGF-BB per mL of composition, for example, about 75 μg of rhPDGF-BB per mL of composition. For example, hyaluronic acid may be present in the composition at a concentration of approximately 0.75% by weight.

[0094] The methods and uses described herein for preventing, reducing or improving pain or discomfort may be applied to any skin lesions described herein. For example, the methods and uses may be performed to reduce or improve pain or discomfort caused by surgical incisions. For example, the incision may be due to surgery, such as an incision in a facelift. In some embodiments, the present invention provides a method (e.g., a cosmetic method) for reducing (or improving) pain or discomfort caused by a skin incision resulting from a surgery (e.g., a cosmetic surgery such as a facelift), comprising: (1) preparing a composition comprising, or essentially comprising, recombinant human PDGF-BB (rhPDGF-BB) and a biocompatible carrier comprising hyaluronic acid; and (2) applying the composition on or into the incision (e.g., topically) so that the carrier delivers bioactive rhPDGF on or into the incision to reduce pain or discomfort caused by the incision.

[0095] The methods and uses may be implemented to prevent, reduce or improve pain or discomfort caused by aesthetic procedures such as microneedling, microcoring, dermal ablation, dermaplaning, dermal filling, or laser treatment. For example, the methods and uses may be implemented to prevent, reduce or improve pain or discomfort caused by a microneedling procedure. In some embodiments, the present invention provides a method (e.g., a cosmetic method) for preventing or reducing (or improving) pain or discomfort caused by a microneedling procedure performed on skin of interest, comprising: (1) preparing a composition comprising, or essentially comprising, recombinant human PDGF-BB (rhPDGF-BB) and a biocompatible carrier comprising hyaluronic acid; and (2) applying the composition (e.g., topically) onto the skin of interest before, during and / or after a microneedling procedure is performed (on the same area of ​​skin of interest), thereby allowing the carrier to deliver bioactive rhPDGF onto the skin of interest to prevent or reduce (or improve) pain or discomfort caused by the procedure. The same method can also be used to prevent or reduce (or improve) pain or discomfort caused by other skin-penetrating aesthetic procedures such as microcoring, dermal ablation, dermaplaning, dermal filling, or laser treatments.

[0096] The methods and uses may be implemented to reduce or improve pain or discomfort caused by skin damage resulting from aging, inflammation, radiation damage, ultraviolet damage, or a combination thereof. As described herein, skin damage resulting from aging, inflammation, radiation damage, and / or ultraviolet damage includes rough skin, dry or itchy skin, sunburn (or other thermal skin damage), and several immunological conditions. In some embodiments, the present invention provides a method (e.g., a cosmetic method) for reducing (or improving) pain or discomfort caused by skin damage resulting from aging, inflammation, radiation damage, ultraviolet damage, or a combination thereof, comprising: (1) preparing a composition comprising, or essentially comprising, recombinant human PDGF-BB (rhPDGF-BB) and a biocompatible carrier comprising hyaluronic acid; and (2) applying the composition (e.g., topically) over skin damage, wherein the carrier delivers bioactive rhPDGF over the skin damage to reduce pain or discomfort caused by the damage. In some embodiments, the present invention provides a method (e.g., a cosmetic method) for reducing (or improving) pain or discomfort caused by skin damage (such as rough skin, dry skin and / or itchy skin, or sunburn) resulting from aging, inflammation, radiation damage, ultraviolet damage, or a combination thereof, comprising: (1) preparing a composition comprising, or essentially comprising, sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution and a sterile biocompatible carrier comprising hyaluronic acid; and (2) applying the composition (e.g., topically) onto the skin of the subject before, during and / or after a microneedling procedure (on the same area of ​​the skin of the subject), thereby allowing the carrier to deliver bioactive rhPDGF over the skin damage to reduce pain or discomfort caused by the damage. In some embodiments, the method and use may include reducing (or improving) pain or discomfort by reducing roughness of the skin of the subject. In some embodiments, the method and use may include reducing (or improving) pain or discomfort by reducing dryness of the skin of the subject.In some embodiments, the method and use may include reducing (or improving) pain or discomfort by reducing itching of the skin in question. In some embodiments, the method and use may include reducing (or improving) pain or discomfort caused by sunburn or other thermal skin injury. In some embodiments, as described herein, the composition may be formulated as an after-sun treatment and applied topically to the skin in question.

[0097] In further embodiments, the present invention provides a method for preventing or reducing redness, swelling, and / or inflammation caused by skin disruption of a subject, comprising the steps of (1) preparing a composition comprising or essentially comprising sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution, and (2) applying the composition to an area of ​​skin of a subject affected or likely to be affected by skin disruption to prevent or reduce redness, swelling, and / or inflammation caused by skin disruption. In some embodiments, the skin disruption does not involve tendons, ligaments, or bones. The method may be a cosmetic treatment method for preventing or reducing redness, swelling, and / or inflammation caused by skin disruption. In some embodiments, the composition further comprises a carrier as described herein, for example, hyaluronic acid.

[0098] In further embodiments, the present invention provides compositions comprising, or essentially comprising, sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution for use in a method of preventing or reducing redness, swelling and / or inflammation caused by skin disruption of a subject, which are applied to an area of ​​skin affected or likely to be affected by skin disruption to prevent or reduce redness, swelling and / or inflammation caused by the disruption. In some embodiments, the skin disruption does not involve tendons, ligaments or bones. In some embodiments, the composition further comprises a carrier as described herein, for example, hyaluronic acid.

[0099] In further embodiments, the present invention provides the use of a composition comprising, or essentially comprising, sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution in the manufacture of a pharmaceutical for use in a method of preventing or reducing redness, swelling and / or inflammation caused by skin disruption of a subject, wherein the composition is applied to an area of ​​skin of a subject that is affected or will be affected by skin disruption to prevent or reduce redness, swelling and / or inflammation caused by the disruption. In some embodiments, the skin disruption does not involve tendons, ligaments or bones. In some embodiments, the composition further comprises a carrier as described herein, for example, hyaluronic acid.

[0100] Methods and uses for preventing or reducing redness, swelling, and / or inflammation caused by skin damage may be carried out using any of the compositions described herein. For example, the methods and uses may be carried out using a composition comprising rhPDGF-BB in a physiological solution without a carrier, as described herein. The physiological solution may comprise the buffer described herein. In these embodiments, the rhPDGF-BB may have a concentration between about 100 μg of rhPDGF-BB per mL of composition and 1000 μg of rhPDGF-BB per mL of composition, for example, about 500 μg of rhPDGF-BB per mL of composition. Alternatively, the methods and uses may be carried out using a composition comprising rhPDGF-BB and a carrier, preferably hyaluronic acid, as described herein. For example, the rhPDGF-BB may have a concentration between about 40 μg of rhPDGF-BB per mL of composition and 80 μg of rhPDGF-BB per mL of composition, for example, about 75 μg of rhPDGF-BB per mL of composition. For example, hyaluronic acid may be present in the composition at a concentration of approximately 0.75% by weight.

[0101] The methods and uses described herein for preventing or reducing redness, swelling, and / or inflammation may be applied to any skin lesions described herein. The methods and uses may be performed to prevent or reduce redness, swelling, and / or inflammation caused by surgical incisions. For example, the incision may result from a cosmetic surgery, such as a facelift. In some embodiments, the present invention provides a method (e.g., a cosmetic method) for reducing redness, swelling, or inflammation caused by a skin incision resulting from a surgery of interest (e.g., a cosmetic surgery such as a facelift), comprising: (1) preparing a composition comprising, or essentially comprising, sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution and a sterile biocompatible carrier comprising hyaluronic acid; and (2) applying the composition (e.g., topically) to or into the incision, wherein the carrier delivers bioactive rhPDGF to or into the incision to reduce redness, swelling, or inflammation caused by the incision.

[0102] The methods and uses may be carried out to prevent or reduce redness, swelling, and / or inflammation of a scar. For example, the scar may be a surgical scar or an acne scar, as described herein. In some embodiments, the present invention provides a method (e.g., a cosmetic method) for reducing redness, swelling, or inflammation of a scar of interest, comprising: (1) preparing a composition comprising, or essentially comprising, sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution and a sterile biocompatible carrier comprising hyaluronic acid; and (2) applying the composition on or into the scar (e.g., topically or by injection) so that the carrier delivers bioactive rhPDGF on or into the incision to reduce redness, swelling, or inflammation at the scar site. In some embodiments, the method reduces the visibility of the scar. In some embodiments, the method improves the aesthetic appearance of the scar.

[0103] The method and use may be performed to prevent or reduce redness, swelling, and / or inflammation caused by skin-penetrating aesthetic procedures such as microneedling, microcoring, dermal ablation, dermaplaning, dermal filling, or laser treatment. For example, the method and use may be performed to prevent or reduce redness, swelling, and / or inflammation caused by microneedling procedures. In some embodiments, the present invention provides a method (e.g., a cosmetic method) for reducing redness, swelling, or inflammation caused by a microneedling procedure performed on a target skin, comprising: (1) preparing a composition comprising, or essentially comprising, sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution and a sterile biocompatible carrier comprising hyaluronic acid; and (2) applying the composition (e.g., topically) onto the target skin before, during, and / or after a microneedling procedure (on the same area of ​​the target skin), thereby allowing the carrier to deliver bioactive rhPDGF onto the target skin to reduce redness, swelling, or inflammation caused by the procedure. The same method can also be used to prevent or reduce redness, swelling, and / or inflammation caused by other percutaneous aesthetic procedures such as microcoring, dermal ablation, dermaplaning, dermal filling, or laser treatment.

[0104] All methods and uses described herein may be carried out using the compositions described herein. In some embodiments, the compositions may contain rhPDGF-BB (as described herein) in a physiological solution without a carrier. In these embodiments, the compositions preferably contain rhPDGF-BB between about 100 μg / mL and 500 μg / mL, for example, about 300 μg / mL of rhPDGF-BB. In these embodiments, the rhPDGF-BB may have a concentration between about 100 μg of rhPDGF-BB per mL of composition and 500 μg of rhPDGF-BB per mL of composition, for example, about 300 μg of rhPDGF-BB per mL of composition. Alternatively, in these embodiments, the rhPDGF-BB may have a concentration between about 100 μg of rhPDGF-BB per mL of composition and 1,000 μg of rhPDGF-BB per mL of composition. In some embodiments, the composition may comprise rhPDGF-BB in a physiological solution (as described herein) and a carrier (as described herein), preferably hyaluronic acid. For example, the composition may comprise rhPDGF-BB between about 40 μg / mL and 80 μg / mL, for example, about 75 μg / mL of rhPDGF-BB. For example, the rhPDGF-BB may have a concentration between about 40 μg of rhPDGF-BB per mL of composition and 80 μg of rhPDGF-BB per mL of composition, for example, about 75 μg of rhPDGF-BB per mL of composition. In some embodiments, the composition may comprise (i) hyaluronic acid at a concentration between about 0.1% by weight and 1.5% by weight, (ii) rhPDGF-BB at a concentration between about 40 μg / mL and 80 μg / mL, (iii) sodium acetate at a concentration between about 10 mM and 30 mM (for example, about 20 mM), and (iv) water. For example, the composition may comprise (i) hyaluronic acid at a concentration between approximately 0.5% by weight and 1% by weight, (ii) rhPDGF-BB at a concentration between approximately 40 μg / mL and 80 μg / mL, (iii) sodium acetate at a concentration between approximately 10 mM and 30 mM (e.g., 20 mM), and (iv) water. For example, the composition may comprise (i) hyaluronic acid at a concentration of approximately 0.75% by weight, (ii) rhPDGF-BB at a concentration of approximately 75 μg / mL, (iii) sodium acetate at approximately 20 mM, and (iv) water.Preferably, the composition does not contain any additional growth factors (i.e., no growth factors other than recombinant human PDGF-BB (rhPDGF-BB)). In some embodiments, the composition does not contain any additional growth factors, such as any transforming growth factor (e.g., TGF-beta), fibroblast growth factor (e.g., FGF), vascular endothelial growth factor (e.g., VEGF), connective tissue growth factor (e.g., CTGF), hepatocyte growth factor (e.g., HGF), insulin-like growth factor (e.g., IGF), or epidermal growth factor (e.g., EGF). In particular, the composition does not contain TGF-beta, VEGF, and / or CTGF.

[0105] Water may be present in an amount of about 0.1% to about 99% by weight of the whole composition, for example, at least about 10% by weight, or at least about 20% by weight, or at least about 30% by weight, or at least about 40% by weight, or at least about 50% by weight, or at least about 60% by weight, or at least about 70% by weight, or at least about 80% by weight, or at least about 85% by weight, or at least about 90% by weight, or at least about 91% by weight, or at least about 92% by weight, or at least about 93% by weight, or at least about 94% by weight, or at least about 95% by weight, or at least about 96% by weight, or at least about 97% by weight, or at least about 98% by weight, or at least about 99% by weight.

[0106] All methods and uses described herein may be applied to skin disturbances described herein. For example, skin disturbances may result from aging, inflammation, radiation damage, ultraviolet damage, or a combination thereof (e.g., wrinkles, sagging, grooves, skin laxity, sunburn, sunspots, irregular pigmentation, rough texture, dryness, large pores, etc.). For example, skin disturbances may be incisions from cosmetic surgery, such as facelifts, abdominal reshaping, or surgical procedures such as breast reconstruction, breast augmentation, or breast reduction. For example, skin disturbances may be scars, such as surgical scars or acne scars. For example, skin disturbances may result from penetrating skin aesthetic procedures such as microneedling, microcoring, dermal ablation, dermaplaning, dermal filling, or laser treatment. The area of ​​skin to which the composition is applied may be scheduled for a penetrating skin aesthetic procedure that introduces skin disturbance (e.g., microneedling, microcoring, dermal ablation, dermaplaning, or laser treatment). In this context, the composition is applied in preparation for treatment to assist in the regeneration, rejuvenation, or repair of the skin. In some embodiments, the skin disruption does not involve tendons, ligaments, or bones. In some embodiments, the skin disruption extends only to the dermis or subcutaneous layer of the skin in question. In some embodiments, the skin disruption is located within the epidermis and dermis, or within the epidermis, dermis, and subcutaneous tissue. In some embodiments, the skin disruption does not extend below the subcutaneous tissue. In some embodiments, the skin disruption does not extend below the subcutaneous tissue.

[0107] The methods and uses described herein may be cosmetic treatment methods performed to (i) improve the appearance or aesthetics of the skin in question, (ii) improve the function of the skin in question, (iii) stimulate cell growth (e.g., to promote skin regeneration, rejuvenation, or repair), (iv) reduce pain and / or discomfort of skin lesions, (v) stimulate angiogenesis (e.g., to promote skin regeneration, rejuvenation, or repair), (vi) improve the texture of the skin in question, (vii) increase the volume and / or firmness of the skin in question, (viii) prevent or reduce scarring of the skin in question, and / or (ix) reduce redness, swelling, and / or inflammation of skin lesions. In some embodiments, the composition is applied to the skin lesion to improve the appearance or aesthetics of the skin in question. In some embodiments, the composition is applied to the skin lesion to improve the function of the skin in question. In some embodiments, the composition is applied to the skin lesion to stimulate cell growth (e.g., to promote skin regeneration, rejuvenation, or repair). In some embodiments, the composition is applied to a skin lesion to reduce pain and / or discomfort of the lesion. In some embodiments, the composition is applied to a skin lesion to stimulate angiogenesis (for example, to promote skin regeneration, rejuvenation, or repair). In some embodiments, the composition is applied to a skin lesion to improve the texture of the skin in question. In some embodiments, the composition is applied to a skin lesion to enhance the volume and / or firmness of the skin in question. For example, the composition may reduce wrinkles, grooves, and / or sagging. For example, the composition may reduce sunspots or irregular pigmentation of the skin in question. For example, the composition may reduce sunburn. For example, the composition may increase the elasticity of the skin in question. For example, the composition may reduce sagging of the skin in question. For example, the composition may reduce rough skin texture. For example, the composition may reduce dry skin. For example, the composition may improve itchy skin. In some embodiments, the composition is applied to a skin lesion to prevent or reduce scarring of the skin in question.

[0108] In some embodiments, the methods and uses described herein further include a step of preparing the skin or other skin lesion site before applying the composition. In some embodiments, the skin or skin lesion is anesthetized with an anesthetic such as lidocaine and disinfected with an antiseptic such as alcohol. In some embodiments, the skin or lesion is washed with soap before applying the composition.

[0109] The methods and uses described herein may further include an initial step of forming a composition. In some embodiments, the method or use includes forming a composition by adding sterile recombinant human PDGF-BB (rhPDGF-BB) to a physiological solution. For example, the methods and uses described herein may include an initial step of forming a composition comprising or essentially comprising sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution. In some embodiments, the method or use includes forming a composition by combining (i) sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution and (ii) a sterile biocompatible carrier, as described herein. For example, the methods and uses described may include an initial step of forming a composition comprising (i) sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution and (ii) a sterile biocompatible carrier which is hyaluronic acid, collagen, gelatin, alginate, polysaccharide, glycosaminoglycan, fibrin, derivatives thereof or combinations thereof.

[0110] In some embodiments, the method for treating a target skin lesion as described herein comprises (i) forming a composition comprising, or essentially comprising, sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution and a sterile biocompatible carrier, wherein the biocompatible carrier is hyaluronic acid, collagen, gelatin, alginate, polysaccharide, glycosaminoglycan, fibrin, derivatives thereof, or a combination thereof; and (ii) applying the composition to or into the skin lesion, wherein the carrier delivers bioactive rhPDGF to or into the skin lesion to result in regeneration, rejuvenation, or repair of the skin lesion.

[0111] In some embodiments, the methods described herein for improving or reducing / preventing pain or discomfort caused by skin damage include (i) forming a composition comprising or essentially comprising sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution, and (ii) applying the composition on or into skin damage to result in a reduction of pain or discomfort caused by skin damage.

[0112] In some embodiments, the methods described herein for improving or reducing / preventing pain or discomfort caused by skin damage include (i) forming a composition comprising, or essentially comprising, sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution and a sterile biocompatible carrier, wherein the biocompatible carrier is hyaluronic acid, collagen, gelatin, alginate, polysaccharide, glycosaminoglycan, fibrin, derivatives thereof, or a combination thereof; and (ii) applying the composition on or into skin damage to reduce pain or discomfort caused by skin damage.

[0113] The “physiologic solution” described herein for forming the compositions of the present invention may include the buffers described herein. In some embodiments, the physiological solution contains sodium acetate. In some embodiments, the physiological solution contains sodium chloride. In some embodiments, the physiological solution contains sodium acetate and sodium chloride. In some embodiments, the physiological solution contains sodium acetate, sodium chloride and water. In some embodiments, the physiological solution contains about 20 mM sodium acetate and optionally water. In some embodiments, the physiological solution contains about 0.15 M sodium chloride and optionally water. In some embodiments, the physiological solution contains about 20 mM sodium acetate, about 0.15 M sodium chloride and optionally water. In some embodiments, the physiological solution contains about 20 mM sodium acetate, water and optionally about 0.15 M sodium chloride. The physiological solution may have a pH value of 6.0 + / - 0.5 (i.e., 5.5 to 6.5).

[0114] The sterile rhPDGF-BB solutions described herein for forming the compositions of the present invention may contain rhPDGF-BB at concentrations of at least 40 μg / mL, 50 μg / mL, 60 μg / mL, 70 μg / mL, 80 μg / mL, 90 μg / mL, 100 μg / mL, 200 μg / mL, 300 μg / mL, 400 μg / mL, 500 μg / mL, 600 μg / mL, 700 μg / mL, 800 μg / mL, 900 μg / mL, and 1,000 μg / mL. The solutions may contain rhPDGF-BB at concentrations between approximately 100 μg / mL and 500 μg / mL. The solutions may contain rhPDGF-BB at concentrations between approximately 300 μg / mL and 1,000 μg / mL. The solution may contain rhPDGF-BB at a concentration between approximately 200 μg / mL and 400 μg / mL. The solution may contain rhPDGF-BB at a concentration of approximately 300 μg / mL.

[0115] In some embodiments, forming a composition may involve combining or mixing an rhPDGF-BB solution (e.g., a sterile physiological solution of rhPDGF-BB) with a carrier described herein, such as a hyaluronic acid gel or serum described herein. In some embodiments, the rhPDGF-BB solution and the carrier (e.g., hyaluronic acid serum / gel) are mixed in a 1:2 ratio. In some embodiments, the rhPDGF-BB solution and the carrier (e.g., hyaluronic acid serum / gel) are mixed in a 1:3 ratio. In some embodiments, the rhPDGF-BB solution and the carrier (e.g., hyaluronic acid serum / gel) are mixed in a 1:4 ratio. In some embodiments, the rhPDGF-BB solution and the carrier (e.g., hyaluronic acid serum / gel) are mixed in a 1:5 ratio. In some embodiments, the rhPDGF-BB solution and the carrier (e.g., hyaluronic acid serum / gel) are mixed in a 1:6 ratio. In some embodiments, the rhPDGF-BB solution and the carrier (e.g., hyaluronic acid serum / gel) are mixed in a ratio of 1:7. In some embodiments, the rhPDGF-BB solution and the carrier (e.g., hyaluronic acid serum / gel) are mixed in a ratio of 1:8. In some embodiments, the rhPDGF-BB solution and the carrier (e.g., hyaluronic acid serum / gel) are mixed in a ratio of 1:8. In some embodiments, the rhPDGF-BB solution and the carrier (e.g., hyaluronic acid serum / gel) are mixed in a ratio of 1:10. In some embodiments, the HA gel or serum described herein for forming a composition may be an HA serum or gel containing about 0.5% to 2%, for example, about 0.75% to about 1.75%, about 0.75% to about 1.5%, or about 1% to about 1.25% of HA. In some embodiments, the HA gel or serum described herein for forming a composition may be an HA serum or gel containing about 1% of HA. In some embodiments, the HA gel or serum described herein for forming a composition may be an HA serum or gel containing about 1.25% HA.

[0116] In some embodiments, forming a composition may involve combining or mixing an rhPDGF-BB solution (e.g., a sterile physiological solution of rhPDGF-BB) with a carrier described herein, such as a hyaluronic acid gel or serum described herein. In some embodiments, the rhPDGF-BB solution and the carrier (e.g., hyaluronic acid serum / gel) are mixed in a 1:1 ratio. In some embodiments, the rhPDGF-BB solution and the carrier (e.g., hyaluronic acid serum / gel) are mixed in a 2:1 ratio. In some embodiments, the rhPDGF-BB solution and the carrier (e.g., hyaluronic acid serum / gel) are mixed in a 3:1 ratio. In some embodiments, the rhPDGF-BB solution and the carrier (e.g., hyaluronic acid serum / gel) are mixed in a 4:1 ratio. In some embodiments, the rhPDGF-BB solution and the carrier (e.g., hyaluronic acid serum / gel) are mixed in a 5:1 ratio. In some embodiments, the rhPDGF-BB solution and the carrier (e.g., hyaluronic acid serum / gel) are mixed in a 6:1 ratio. In some embodiments, the rhPDGF-BB solution and the carrier (e.g., hyaluronic acid serum / gel) are mixed in a 7:1 ratio. In some embodiments, the rhPDGF-BB solution and the carrier (e.g., hyaluronic acid serum / gel) are mixed in an 8:1 ratio.

[0117] In some embodiments, the method may further include the step of mixing the combination of rhPDGF-BB and the carrier with sodium chloride and / or water. For example, the method may include the steps of (i) combining a solution of rhPDGF-BB (e.g., rhPDGF-BB in a buffer as described herein, e.g., sodium acetate buffer) with a carrier (e.g., hyaluronic acid as a serum or gel), and (ii) mixing rhPDGF-BB and the carrier with sodium chloride and / or water. For example, the method may include the steps of (i) combining a solution of rhPDGF-BB in 20 mM sodium acetate buffer with a hyaluronic acid serum or gel (as described herein), and (ii) mixing rhPDGF-BB and hyaluronic acid with sodium chloride and / or water. The buffer and carrier may be as described herein. The final concentration of rhPDGF-BB in the composition is as described herein. For example, the composition may contain between approximately 40 μg and 80 μg of PDGF per mL, for example, approximately 75 μg of PDGF per mL.

[0118] In exemplary embodiments, 0.5 mL of a 300 ug / mL rhPDGF-BB solution and 1.5 mL of hyaluronic acid (HA) gel or serum (which itself is a blend of (i) hyaluronic acid having a weight between approximately 100,000 Da and 150,000 Da and (ii) hyaluronic acid having a weight between approximately 750,000 Da and 900,000 Da) are thoroughly mixed to form an rhPDGF-BB-based therapeutic composition. The resulting mixture contains 150 μg of PDGF in 2 mL of gel, i.e., 75 μg of PDGF per mL, and can be applied to skin damage following microneedling or other aesthetic procedures. In some embodiments, sodium chloride and / or water are added to the rhPDGF-BB therapeutic composition before applying the composition to the target skin. In further exemplary embodiments, 0.5 mL of a 300 ug / mL rhPDGF-BB solution is mixed with 3 mL of hyaluronic acid (HA) gel or serum (which itself is a blend of (i) hyaluronic acid having a weight between 100,000 Da and 150,000 Da and (ii) hyaluronic acid having a weight between 750,000 Da and 900,000 Da) to form the rhPDGF-BB composition. The resulting mixture contains 150 μg of PDGF in 3.5 mL of composition, i.e., 42.86 μg of PDGF per mL, and can be applied to skin damage after microneedling or other aesthetic procedures. In some embodiments, sodium chloride and / or water are added to the rhPDGF-BB therapeutic composition before applying the composition to the target skin.

[0119] The methods and uses described herein may involve administering (i.e., injecting) the composition through one or more needles. For example, the composition may be administered (i.e., injected) into or on a lesion through one or more needles. For example, the composition may be administered (i.e., injected) into or on the skin of a target through one or more needles. The needles may be single-use needles. For example, the composition may be injected into a site of skin lesion using a single-use needle. Suitable skin lesions for injection of the composition include scars such as surgical scars or acne scars. Skin lesions caused by aging, inflammation, radiation damage, ultraviolet damage, or a combination thereof (such as wrinkles, sagging, grooves, skin laxity, sunburn, solar lentigines, irregular pigmentation, rough texture, dryness, large pores, etc.) may also be suitable for injection. The compositions described herein may be mixed with other aesthetic compositions and injected into the skin or skin lesions. For example, the composition may be mixed with dermal fillers and injected into the skin or skin lesions. Exemplary dermal fillers include calcium hydroxyapatite fillers (e.g., Radiese®) (calcified dermal fillers for nasolabial folds), which are injected or placed along the mandibular line and / or cheekbones to provide contouring. Other exemplary fillers include poly-L-lactic acid-containing fillers (e.g., Sculptra®) and cross-linked hyaluronic acid-containing fillers (e.g., Juvederm®). In some embodiments, one or more needles are contained within a microchannel microinjector (e.g., an Aquagold® device). The microchannel microinjector device includes multiple hollow needles containing the composition and delivering it to the skin or skin lesions of the subject. In an exemplary microinjector, a reservoir is located at the top of the device that can be used to deliver the composition to the hollow needles for application to the skin of the subject. The composition can be injected into the skin or skin lesions in any area of ​​the subject's body. For example, the composition can be injected into the skin of the subject's head, face, scalp, chest, neck, arms, hands, and / or legs. For example, the composition may be injected into the skin of the face, neck, and / or scalp of the subject.

[0120] In some embodiments of the present invention, a needle or other penetrating device is hollow and delivers a therapeutic or cosmetic composition containing rhPDGF-BB, with or without a carrier, to destruction during treatment. The needle may be a microneedle used in the microneedling treatment described above, or the needle of a single injection-ready syringe. In some embodiments, the therapeutic or cosmetic composition is manufactured and pre-formulated in a syringe that can be used by the user in the treatment method disclosed herein, without preparation, mixing, or filling before or during syringe treatment. The syringe may contain a needle or may be used without a needle to deliver the therapeutic composition to destruction. In some embodiments, the needle penetration depth is between about 0.1 mm and 5 mm, generally between about 1.0 mm and 2.0 mm. In other embodiments, the needle is between about 0.5 mm and 5 inches. In other embodiments, the needle is of the length and gauge required to deliver the therapeutic composition to destruction.

[0121] The methods and uses described herein may include topical application of the composition. For example, the composition may be applied topically to the skin of the subject. For example, the composition may be applied topically to a skin lesion. In some embodiments, the composition may be applied topically to a skin incision. In further examples, the composition may be applied topically to a skin incision resulting from surgery. The surgery may be a cosmetic surgery such as a facelift. The composition may be applied topically during surgery. For example, the composition may be applied to an open surgical wound or flap when closing the incision (e.g., using staples, adhesives or sutures). The composition may be applied topically to an incision after surgery, for example, immediately after the incision has been closed. The composition may be applied during and / or after surgery. For example, the composition may be applied when closing the incision (e.g., using staples, adhesives or sutures) and immediately after the incision has been closed. The incision may be re-treated daily or periodically. For example, the incision may be re-treated using the re-treatment timing schedule described herein. For example, the treatment of an incision may be repeated once or twice a day for 2, 3, 4, 5, 6, 7, 8, 9, or 10 days. The duration of retreatment may be adjusted according to the severity of the incision. In the case of deeper wounds / incisions (e.g., after Mohs surgery), the composition may be reapplied once or twice a day for 7 to 10 days post-surgery. In the case of more superficial incisions, the composition may be reapplied once or twice a day for 4 to 5 days post-surgery. In some embodiments, the composition may be applied topically to skin having damage caused by aging, inflammation, radiation damage, ultraviolet damage, or a combination thereof. In some embodiments, the composition is applied to the surface of damaged skin or connective tissue. In some embodiments, the therapeutic or cosmetic composition may be applied to body parts such as, but not limited to, the face, scalp, and / or neck (e.g., for repair or rejuvenation). The composition may be applied topically to skin or skin damage in any area of ​​the body in question. For example, the composition may be applied topically to the head, face, scalp, back, chest, abdomen, neck, arms, hands, and / or legs of the subject.In some embodiments, the composition is applied before, during, and / or after aesthetic procedures (such as microneedling, microcoring, dermal ablation, dermaplaning, dermal filling, or laser skin penetration as described herein) to repair and rejuvenate skin or other damage. In other embodiments, the composition may be applied to these or other body parts for aesthetic utility. However, those skilled in the art will understand that the presented embodiments are not limited to the uses described above and can be implemented in any suitable use relating to the medical industry. Topical application typically involves applying about 0.5 ml to 1 ml of the composition.

[0122] Furthermore, the application of a therapeutic or cosmetic composition may involve penetration of the skin (e.g., intact skin) by mechanical or chemical intervention to assist in the delivery of the therapeutic or cosmetic composition to or into the skin of the target. For example, in the methods and uses described herein, the composition may be applied topically to the skin or skin lesion of the target, and mechanical or chemical means may be used to penetrate the skin and assist in the delivery of the applied composition into the skin or skin lesion. In some embodiments of the present invention, the mechanical or chemical intervention may include energy-based devices (e.g., those used to perform microneedling, microcoring, and laser resurfacing), excipients, and / or decomposition to assist in the delivery of the therapeutic or cosmetic composition across the stratum corneum to or into the skin of the target and to enhance clinical efficacy. Penetration may allow for the delivery of the composition into the dermis and / or epidermis. Such delivery methods for rhPDGF-BB-based compositions are not intended by methods known in the art for treating bone and tissue in the body. In some embodiments, skin penetration is achieved by performing microneedling, microcoring, dermabrasion, dermaplaning, or laser skin resurfacing using mechanical devices, lasers, or scraping tools, and by applying therapeutic or cosmetic compositions to the skin (e.g., face, neck, and / or scalp) before, during, or after treatment using such tools. Lasers are also frequently used for aesthetic purposes and include ablative and non-ablative lasers. Non-ablative laser treatments, such as fractional erbium (Er) lasers, intense pulsed light lasers, alexandrite lasers, and neodymium (Nd) lasers, are commonly used for treating fine lines, mild wrinkles, and newly acquired hyperpigmentation abnormalities, as well as to improve skin firmness and texture. Non-ablative lasers produce a skin rejuvenating effect by heating the target tissue while leaving the epidermis intact.Ablative laser treatments, such as carbon dioxide lasers, pulsed dye lasers (PDLs), and erbium (Er) lasers, are commonly used to treat deep lines and wrinkles, acne scars, severe discoloration (brown or red spots), hyperpigmentation disorders, skin texture issues, and scarring. Ablative lasers can create channels in the skin somewhat similar to mechanical microneedling and can be used to remove the outer epidermal layer from the entire skin while heating water molecules in the dermis to induce a healing response. In some embodiments, the laser penetrating the skin or skin destruction is an ablative laser. In some embodiments, the laser penetrating the skin or skin destruction is a carbon dioxide laser, a pulsed dye laser (PDL), or an erbium laser (e.g., an erbium-doped yttrium aluminum garnet (Er:YAG) laser). Non-ablative laser treatments may also be used in combination with the treatment methods and uses described herein. In some embodiments, the composition may be applied to the target skin or skin lesion before, during, and / or after a laser resurfacing procedure using a non-ablative laser. In some embodiments, the non-ablative laser procedure includes broadband light (BBL) therapy or intense pulsed light (IPL) therapy, which uses short bursts of light that penetrate the skin and work to rejuvenate it. In some embodiments, the device that penetrates the skin or skin lesion is a microneedle pen. In some embodiments, the device that penetrates the skin or skin lesion is a microcoring device. Dermaplaning (or dermablading) is a procedure that involves scraping away the upper epidermal layer of the skin, exfoliating the skin, and removing dead skin cells and fine facial hairs. In some embodiments, the scraping tool that penetrates the skin or skin lesion is a dermaplaning scalpel, such as a dermaplaning tool or skin harvester. In some embodiments, the device that penetrates the skin or skin lesion is a microchannel microinjector (e.g., Aquagold®) that penetrates the skin using a hollow needle.When this procedure is used, the composition is contained within a hollow needle of a microinjector and can be applied to the skin or skin lesions via the needle. This differs from procedures that introduce channels or lesions into the epidermis or dermis of the target skin (such as microneedling, microcoring, or laser penetration) to facilitate the delivery of the locally applied composition to the target skin. In some embodiments, several tools are used to penetrate the skin and assist in the delivery of the composition. For example, both lasers and microneedling may be applied to the target skin or skin lesions, and the composition may be applied before, during, and / or after the aesthetic procedure is performed. For example, the skin can be penetrated using a device that delivers radiofrequency (RF) energy through microneedles (e.g., Morpheus 8).

[0123] The methods and uses described herein may include applying the composition before, during, and / or after an aesthetic treatment (e.g., a skin-penetrating aesthetic treatment) is performed on the skin in question. In some embodiments, the composition may be applied before the aesthetic treatment (e.g., a skin-penetrating aesthetic treatment) is performed on the skin in question. In some embodiments, the composition may be applied during the aesthetic treatment (e.g., a skin-penetrating aesthetic treatment) is performed on the skin in question. In some embodiments, the composition may be applied after the aesthetic treatment (e.g., a skin-penetrating aesthetic treatment) is performed on the skin in question. In some embodiments, the composition may be applied before and during the aesthetic treatment (e.g., a skin-penetrating aesthetic treatment) is performed on the skin in question. In some embodiments, the composition may be applied before and after the aesthetic treatment (e.g., a skin-penetrating aesthetic treatment) is performed on the skin in question. In some embodiments, the composition may be applied before, during, and after the aesthetic treatment (e.g., a skin-penetrating aesthetic treatment) is performed on the skin in question. As used herein, the term “skin-penetrating aesthetic treatment” refers to a treatment that penetrates the skin using mechanical devices (e.g., needles such as microneedles, microcoring needles, or microchannel microinjectors), scraping tools (e.g., for dermal ablation or dermaplaning), or lasers (ablative or non-ablative lasers). In some embodiments, compositions may be applied topically before, during, and / or after microneedling, microcoring, microchannel microinjection, dermabrasion, dermaplaning, dermal filling, or laser skin resurfacing. When a composition is applied “before” an aesthetic treatment, this involves topically applying the composition before the aesthetic treatment is performed. In some embodiments, this involves applying the composition at least 5 minutes, 15 minutes, 20 minutes, 25 minutes, 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 12 hours, 24 hours, 2 days, 3 days, 4 days, 5 days, or 6 days before the aesthetic treatment is performed.For example, the composition may be applied immediately before the procedure (e.g., 5-10 minutes prior). When the composition is applied "between" the aesthetic procedure, this involves topical application of the composition throughout the procedure (e.g., in a single treatment appointment between skin penetration procedures). For example, an aesthetic procedure (e.g., microneedling, microcoring, or laser penetration) may be performed by dividing the skin in question into various sub-regions (or "quarters"), each sub-region may be subjected to the aesthetic procedure (e.g., using vertical, horizontal, and oblique movements across the same area of ​​skin or lesion being treated). This procedure may be repeated up to three times on the same sub-region in a single treatment appointment. The composition may be reapplied (e.g., topically) between each repetition of the procedure on the sub-region (or quarter) of the skin in question. When the composition is applied "after" the aesthetic procedure, this involves topical application of the composition after the aesthetic procedure has been performed. In some embodiments, this involves applying the composition within 5 minutes, 10 minutes, 15 minutes, 20 minutes, 25 minutes, 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 12 hours, or 24 hours after the aesthetic treatment is performed. In some embodiments, the composition may be applied up to 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, or 14 days after the aesthetic treatment is performed. For example, the composition may be applied immediately after the aesthetic treatment, for example, within 5 to 10 minutes. The composition may be reapplied to the target skin periodically according to the retreatment timing schedule described herein. For example, the target skin may be retreated by applying the composition once or twice a day for up to 14 days. For example, the target skin can be re-treated by applying the composition two to three times a day for up to 14 days. In some embodiments, the composition is applied before, during, and / or after microneedling (e.g., topically). In some embodiments, the composition is applied before microneedling (e.g., topically).In some embodiments, the composition is applied (e.g., topically) during microneedling (i.e., between repeated penetration steps of the microneedling procedure). In some embodiments, the composition is applied (e.g., topically) after microneedling. In some embodiments, the composition is applied (e.g., topically) before, during, and / or after a laser resurfacing procedure. In some embodiments, the composition is applied (e.g., topically) before a laser resurfacing procedure. In some embodiments, the composition is applied (e.g., topically) during a laser resurfacing procedure. In some embodiments, the composition is applied (e.g., topically) after a laser resurfacing procedure. These aesthetic procedures help penetrate the target skin and deliver PDGF into or onto the skin surface. When the composition is applied before, during, and / or after an aesthetic procedure, the composition may help promote skin regeneration, rejuvenation, or repair. When the composition is applied before, during, and / or after an aesthetic procedure, it may help prevent or reduce pain or discomfort, or prevent or reduce redness, swelling, and inflammation.

[0124] Any skin degeneration described herein can be adequately treated using the compositions of the present invention, applied before, during, and / or after aesthetic treatments. For example, scars (such as surgical scars or acne scars) can be treated using the methods and uses described herein, involving the application of the compositions before, during, and / or after aesthetic treatments such as microneedling, microcoring, dermal ablation, dermaplaning, or laser treatments. For example, scars may be treated using the compositions described herein, applied before a microneedling treatment is performed. For example, scars may be treated using the compositions described herein, applied after a laser resurfacing treatment is performed. Skin damage caused by aging, inflammation, radiation damage, ultraviolet damage, or a combination thereof (such as wrinkles, sagging, grooves, skin laxity, sunburn, solar lentigines, sunburn, irregular pigmentation, rough texture, dryness, and large pores) may also be treated using the methods and uses described herein, which involve applying the compositions described herein before, during, and / or after aesthetic treatments such as microneedling, microcoring, dermal ablation, dermaplaning, or laser treatments. For example, skin damage caused by aging (such as wrinkles, sagging, grooves, skin laxity, rough texture, dryness, and large pores) may be treated using the compositions described herein, which are applied before a microneedling treatment is performed. For example, skin damage caused by aging (such as wrinkles, sagging, grooves, skin laxity, rough texture, dryness, and large pores) may be treated using the compositions described herein, which are applied after a microneedling treatment is performed. For example, skin damage caused by aging (such as wrinkles, sagging, grooves, skin laxity, rough texture, dryness, and enlarged pores) can be treated using the compositions described herein, which are applied before the laser resurfacing procedure is performed. For example, skin damage caused by aging (such as wrinkles, sagging, grooves, skin laxity, rough texture, dryness, and enlarged pores) can be treated using the compositions described herein, which are applied after the laser resurfacing procedure is performed.For example, skin damage caused by hyperpigmentation or melasma may be treated with a composition described herein, applied before a microneedling procedure is performed. For example, skin damage caused by hyperpigmentation or melasma may be treated with a composition described herein, applied after a microneedling procedure is performed. For example, skin damage caused by hyperpigmentation or melasma may be treated with a composition applied before a laser resurfacing procedure is performed. For example, skin damage caused by hyperpigmentation or melasma may be treated with a composition applied after a laser resurfacing procedure is performed.

[0125] In some embodiments, the present invention provides a method (e.g., a cosmetic method) for treating skin defects (wrinkles, sagging, grooves, sunburn, sunspots, irregular pigmentation, rough texture, and dryness) caused by aging, inflammation, radiation damage, ultraviolet damage, or a combination thereof, comprising: (1) preparing a composition comprising, or essentially comprising, sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution and a sterile biocompatible carrier comprising hyaluronic acid; and (2) applying the composition (e.g., topically) to the skin defects before, during, and / or after a penetrating aesthetic treatment (e.g., microneedling, microcoring, dermal ablation, or dermal filling) is performed (e.g., on the same area of ​​skin of the subject), thereby allowing the carrier to deliver bioactive rhPDGF to the defects to regenerate, rejuvenate, or repair the skin of the subject. Advantageously, the penetrating aesthetic treatment penetrates the skin of the subject, facilitating the delivery of the composition to the skin defects. In some embodiments, the skin destruction does not involve tendons, ligaments, or bones. In some embodiments, the skin destruction extends only to the dermis or subcutaneous layer of the skin in question.

[0126] In some embodiments, the present invention provides a method (e.g., a cosmetic method) for treating skin defects (wrinkles, sagging, grooves, sunburn, sunspots, irregular pigmentation, rough texture, and dryness) caused by aging, inflammation, radiation damage, ultraviolet damage, or a combination thereof, comprising: (1) preparing a composition comprising, or essentially comprising, sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution and a sterile biocompatible carrier comprising hyaluronic acid; and (2) applying the composition (e.g., topically) to the skin defects before, during and / or after a microneedling procedure (e.g., on the same area of ​​skin of the subject), so that the carrier delivers bioactive rhPDGF to the defects to regenerate, rejuvenate, or repair the skin of the subject. Advantageously, the microneedling procedure penetrates the skin of the subject, facilitating the delivery of the composition to the skin defects. In some embodiments, the skin defects do not involve tendons, ligaments, or bones. In some embodiments, the skin destruction extends only to the dermis or subcutaneous layer of the skin in question.

[0127] In some embodiments, the present invention provides a method (e.g., a cosmetic method) for treating skin defects (wrinkles, sagging, grooves, sunburn, sunspots, irregular pigmentation, rough texture, and dryness) caused by aging, inflammation, radiation damage, ultraviolet damage, or a combination thereof, comprising: (1) preparing a composition comprising, or essentially comprising, sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution and a sterile biocompatible carrier comprising hyaluronic acid; and (2) applying the composition (e.g., topically) to the skin defects after a microneedling procedure has been performed (e.g., on the same area of ​​the skin of the subject), so that the carrier delivers bioactive rhPDGF to the defects, thereby regenerating, rejuvenating, or repairing the skin of the subject. Advantageously, the microneedling procedure penetrates the skin of the subject, facilitating the delivery of the composition to the skin defects. In some embodiments, the skin defects do not involve tendons, ligaments, or bones. In some embodiments, the skin destruction extends only to the dermis or subcutaneous layer of the skin in question.

[0128] As described herein, the application of the composition may include penetration of the skin (e.g., intact skin) by mechanical or chemical intervention to aid in the destruction or delivery of the composition onto or into the skin of the target. Furthermore, the application of the therapeutic composition may include performing microneedling, microcoring, or dermabrasion using mechanical devices, lasers, or scraping tools, and applying the therapeutic composition to the skin of the face, neck, and / or scalp before, during, and / or after the use of such tools. In some embodiments, the penetration depth is between about 0.2 mm and 0.5 mm, between about 0.2 mm and 3 mm, or between about 0.5 mm and 3 mm, depending on the thickness of the skin. For example, the penetration depth may be between about 0.2 mm and 0.5 mm. For example, the penetration depth may be between about 0.5 mm and 1.0 mm. For example, the penetration depth may be between about 1.0 mm and 2.0 mm. In some embodiments, the penetration depth of the needle or laser is between approximately 0.2 mm and 0.5 mm, or between approximately 0.5 mm and 3 mm, depending on the thickness of the skin, and is generally between approximately 0.5 mm and 1.0 mm or 1.0 mm and 2.0 mm. For example, the penetration depth of the needle or laser may be between approximately 0.2 mm and 0.5 mm. For example, the penetration depth of the needle or laser may be between approximately 0.2 mm and 1.0 mm. For example, the penetration depth of the needle or laser may be between approximately 0.5 mm and 1.0 mm. For example, the penetration depth of the needle or laser may be between approximately 1.0 mm and 2.0 mm. Some procedures described herein penetrate the skin more deeply. For example, the Morpheus8 laser procedure typically penetrates between 4 mm and 8 mm. Therefore, in some embodiments, the penetration depth of the needle or laser may be between 4 mm and 8 mm. In some embodiments of the methods and uses disclosed herein, the action of skin penetration by microneedling or microcoring delivers a composition (e.g., a composition of rhPDGF-BB and hyaluronic acid or other carrier) into the subepithelial dermal tissue of the skin of the body part being treated. In exemplary embodiments, the hyaluronic acid is a serum or gel. In some embodiments, the hyaluronic acid is applied to the disruption prior to the penetration of disruption by microneedling, microcoring or other treatment.In some embodiments, hyaluronic acid is a serum or gel. In some embodiments, hyaluronic acid has a pH value of 5.0 to 7.0. Hyaluronic acid may have the weight % described herein.

[0129] In some embodiments, microneedling, laser, or other penetration of the skin is repeated up to six times. For example, skin penetration may be repeated up to two, three, four, five, or six times. In some embodiments of the present invention, microneedling is repeated three times, such as with vertical, horizontal, and diagonal movements across the same area of ​​skin or other destruction being treated. Penetration of the same area of ​​skin may also be repeated with movements in the same direction. In another embodiment of the present invention, the area of ​​skin being treated is divided into sub-regions, and the initial and repeated penetrations of the skin are performed in each sub-region. In one embodiment, the sub-region is a 1.5 square inch area of ​​skin, but any size of sub-region is intended herein. In some embodiments, the composition may be locally reapplied between each repeated penetration.

[0130] In some embodiments, the (therapeutic or cosmetic) composition remains on the skin or other damaged areas for at least 24 hours after application. In some embodiments, the (therapeutic or cosmetic) composition remains on the skin for up to 48 hours. After a minimum of 24 hours, the user can wash the treatment area with water and reapply the composition or hyaluronic acid.

[0131] As used herein, “user” may include a dermatologist, plastic surgeon, physician, esthetician, person receiving treatment, or other person qualified to perform the relevant method for treating a person’s lesion as disclosed herein. Thus, in some embodiments of the invention, a patient (or subject) may complete the method and treat and / or re-treat their lesion. In another example, there may be two or more different users of the method throughout the course of treatment and re-treatment described herein. For example, a patient (or subject) may complete the treatment in the office of a dermatologist or esthetician and receive re-treatment in the comfort of their home.

[0132] In some embodiments of the present invention described herein, therapeutic or cosmetic compositions are added during the manufacturing process of known compositions to compositions known in the art for repairing and rejuvenating skin and other damage in aesthetic treatments, such as skin and body contouring fillers, and the resulting compositions are applied to skin damage before, during, or after aesthetic treatments. Unlike most proteins that are inactivated during heat sterilization techniques, rhPDGF-BB has heat-resistant properties that allow it to withstand the thermal effects of moist heat sterilization better than other proteins. These heat-resistant properties make it possible to add rhPDGF-BB to existing manufacturing processes for use in aesthetic compositions and treatment methods.

[0133] The methods and uses described herein may further include a monitoring step in which the effectiveness of the treatment is monitored during the treatment period, and the treatment step of the method ("step 2") may be repeated as necessary. For example, a method for promoting skin regeneration, rejuvenation or repair described herein may further include a step of monitoring the regeneration, rejuvenation or repair of the skin in question during the treatment period, and a step of re-treating the destruction by repeating step (2) of the method. For example, a method for treating skin destruction described herein may further include a step of monitoring the regeneration, rejuvenation or repair of the skin destruction during the treatment period, and a step of re-treating the destruction by repeating step (2) of the method. For example, a method for preventing or reducing pain or discomfort caused by skin destruction or a method for improving pain or discomfort caused by skin destruction described herein may further include a step of monitoring pain or discomfort during the treatment period, and a step of re-treating the destruction by repeating step (2) of the method. For example, a method for preventing or reducing redness, swelling and / or inflammation caused by skin damage as described herein may further include the steps of monitoring redness, swelling and / or inflammation during the treatment period and repeating step (2) of the method to re-treat the damage.

[0134] For effective results, the above treatment may be repeated up to six times, approximately weekly, monthly, semi-annually, or annually. In some embodiments of the present invention, the composition is re-administered once or twice per day for up to 14 days. For example, the composition may be re-administered once or twice per day for up to 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 days. In some embodiments, the composition may be re-administered two to three times per day for up to 14 days (for example, up to 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 days). In some embodiments of the present invention, the composition is re-administered two to three times per day for up to 14 days.

[0135] In some embodiments of the present invention, a timing schedule is provided for periodic retreatment of a lesion, i.e., for periodic reapplication of a (therapeutic or cosmetic) composition to the lesion. The actual number and frequency of retreatments (i.e., treatment intervals) should be determined based on several factors, including the severity of the lesion, the degree to which the natural lesion healing environment is impaired, the patient's age, and the duration of the lesion. For more severe lesions or lesions accompanied by a more impaired healing environment, the number and frequency of retreatments should be increased. Furthermore, the prescribed number and / or frequency of treatments can be adjusted during the treatment period based on the healing rate of the lesion, i.e., the number and / or frequency of retreatments can be increased if the healing of the lesion is slower, or decreased if the healing of the lesion is faster.

[0136] In some embodiments of the present invention, the retreatment frequency is at least about once every six weeks, such as at least about 2 days, at least about 3 days, at least about 4 days, at least about 5 days, at least about 6 days, at least about 7 days, at least about 8 days, at least about 9 days, at least about 10 days, at least about 11 days, at least about 12 days, at least about 13 days, at least about 14 days, or at least about 15 days, or a combination thereof. In some embodiments of the present invention, the retreatment frequency is at least about six weeks, at most within at least about 2 days, at least about 3 days, at least about 4 days, at least about 5 days, at least about 6 days, at least about 7 days, at least about 8 days, at least about 9 days, at least about 10 days, at least about 11 days, at least about 12 days, at least about 13 days, at least about 14 days, or at least about 15 days, or a combination thereof. In some embodiments, the retreatment frequency is once every 2 to 42 days, or once every 3 to 42 days, or once every 2 to 28 days, or once every 3 to 28 days, or once every 2 to 7 days, or once every 2 to 3 days, or once every 3 to 7 days, or once every 4 to 21 days, or once every 7 to 28 days, or once every 7 to 21 days, or once every 7 to 14 days, or once every 10 to 15 days, or once every 12 to 14 days. In some embodiments, the retreatment frequency is once every 2 days, once every 3 days, once every 4 days, once every 5 days, once every 6 days, once every 7 days, once every 8 days, once every 9 days, once every 10 days, once every 12 days, once every 14 days, once every 15 days, once every 21 days, once every 28 days, once every 30 days, once every 35 days, once every 42 days, or a combination thereof. In another aspect of the present invention, the retreatment frequency for the number of days listed above is more than once per day, for example, once or twice per day or two or three times per day.

[0137] In some embodiments, the retreatment frequency is substantially the same throughout the treatment period, and the retreatment frequency is at least once every two days, at least once every three days, at least once every four days, at least once every five days, at least once every six days, at least once every seven days, at least once every eight days, at least once every nine days, at least once every ten days, at least once every eleven days, at least once every twelve days, at least once every thirteen days, at least once every fourteen days, or at least once every fifteen days, and so on, up to at least once per year.

[0138] In some embodiments, the destruction is retreated at least once, at least twice, at least three times, at least four times, or at least five times over the treatment period, or at least six times over the treatment period. According to another aspect of the present invention, the destruction is retreated between 0 and 6 times, between 0 and 7 times, or between 0 and 8 times over the treatment period. According to another aspect of the present invention, the destruction is treated between 1 and 8 times, between 2 and 7 times, or between 3 and 6 times over the treatment period. According to another aspect of the present invention, the destruction is retreated 1, 2, 3, 4, 5, 6, 7, 8, 10, or 20 times over the treatment period. According to another aspect of the present invention, the destruction is retreated between 0 and 46 times, between 1 and 46 times, between 0 and 20 times, between 1 and 20 times, between 0 and 27 times, or between 1 and 27 times.

[0139] In some embodiments, the cumulative total amount of rhPDGF-BB administered for destructive action during the treatment period is preferably greater than 0 mg, but less than about 100 mg, less than 50 mg, or less than about 25 mg, or less than 20 mg, or less than about 15 mg, or less than about 10 mg, or less than about 5 mg, or less than about 4 mg, or less than about 3 mg, or less than 2 mg, or less than 1 mg of rhPDGF-BB. In some embodiments, the cumulative total amount of rhPDGF-BB administered for destructive action during the treatment period is preferably between about 0.1 mg and about 50 mg, or between about 0.5 mg and about 25 mg, or between about 1 mg and about 10 mg, or between about 2.5 mg and about 8 mg, or between about 3 mg and about 7 mg, or between about 4 mg and about 6 mg. In some embodiments, the cumulative total amount of rhPDGF-BB administered for destructive action during the treatment period is between approximately 0.05 mg and approximately 0.1 mg, for example between approximately 0.06 mg and approximately 0.1 mg, between approximately 0.07 mg and approximately 0.09 mg, or between approximately 0.07 mg and approximately 0.08 mg. In some embodiments, the cumulative total amount of rhPDGF-BB administered for destructive action during the treatment period is between approximately 0.1 mg and approximately 1 mg, for example between approximately 0.2 mg and approximately 1 mg, between approximately 0.3 mg and approximately 1 mg, between approximately 0.4 mg and approximately 1 mg, between approximately 0.5 mg and approximately 1 mg, between approximately 0.6 mg and approximately 1 mg, between approximately 0.7 mg and approximately 1 mg, between approximately 0.8 mg and approximately 1 mg, or between approximately 0.9 mg and approximately 1 mg. In some embodiments, the cumulative total amount of rhPDGF-BB administered for destructive action during the treatment period is approximately 0.2 mg, 0.3 mg, 0.4 mg, 0.5 mg, 0.6 mg, 0.7 mg, 0.8 mg, 0.9 mg, 1 mg, 1.1 mg, 1.2 mg, 1.3 mg, 1.4 mg, 1.5 mg, 1.6 mg, 1.7 mg, 1.8 mg, 1.9 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, or 100 mg.

[0140] Various retreatments may involve the same or different doses of rhPDGF-BB with respect to the exact amount of rhPDGF-BB applied to the disruption (i.e., the “absolute dose”) or the amount of rhPDGF-BB applied per square centimeter (cm²) of disruption area (i.e., the “area dose”). In some embodiments, each treatment involves applying an absolute dose of rhPDGF-BB between approximately 10 μg and approximately 50 mg, or between approximately 10 μg and approximately 25 mg, or between approximately 10 μg and approximately 20 mg, or between approximately 10 μg and approximately 15 mg, or between approximately 10 μg and approximately 10 mg, or between approximately 10 μg and approximately 5 mg of rhPDGF-BB, or between approximately 10 μg and approximately 1 mg. In some embodiments, each treatment is administered in an area dose between approximately 10 μg and 1.0 mg of PDGF per cm², or between approximately 10 μg and 0.5 mg of PDGF per cm², or between approximately 10 μg and 0.25 mg of PDGF per cm², or between approximately 10 μg and 0.1 mg of PDGF per cm², or between approximately 10 μg and 0.05 mg of PDGF per cm². In some embodiments, each treatment with rhPDGF-BB is preferably between approximately 10 μg and 1000 μg of PDGF per cm², or between approximately 0.01 mg and 50 mg of PDGF per cm², or between approximately 0.05 mg and 25 mg of PDGF / cm², or between approximately 0.1 mg and 10 mg of PDGF per cm², or between approximately 0.2 mg and 2 mg of PDGF per cm². In some embodiments, each treatment involves administering an area dose of at least about 10 μg of rhPDGF per 1 cm² of destroyed surface area, or at least about 25 μg of rhPDGF per 1 cm² of destroyed surface area, or at least about 50 μg of rhPDGF per 1 cm² of destroyed surface area, or at least about 100 μg of rhPDGF per 1 cm² of destroyed surface area, or at least about 250 μg of rhPDGF per 1 cm² of destroyed surface area, or at least about 500 μg of rhPDGF per 1 cm² of destroyed surface area, or at least 1,000 μg of rhPDGF per 1 cm² of destroyed surface area.In some embodiments, each treatment involves a dose of rhPDGF ranging from approximately 100 μg per cm² of fractured surface area to approximately 1,000 μg per cm² of fractured surface area, or from approximately 300 μg per cm² of fractured surface area to approximately 9,000 μg per cm² of fractured surface area, or from approximately 400 μg per cm² of fractured surface area to approximately 8,000 μg per cm² of fractured surface area, or from approximately 10 μg per cm² of fractured surface area to approximately 500 μg per cm² of fractured surface area. The area dose is administered between F, or between approximately 10 μg of rhPDGF per 1 cm² of destroyed surface area and approximately 100 μg of rhPDGF per 1 cm² of destroyed surface area, or between approximately 15 μg of rhPDGF per 1 cm² of destroyed surface area and approximately 375 μg of rhPDGF per 1 cm² of destroyed surface area, or between approximately 30 μg of rhPDGF per 1 cm² of destroyed surface area and approximately 190 μg of rhPDGF per 1 cm² of destroyed surface area, or between approximately 30 μg of rhPDGF per 1 cm² of destroyed surface area and approximately 300 μg of rhPDGF per 1 cm² of destroyed surface area. In some embodiments, each treatment is administered in an area dose between approximately 0.1 μg of rhPDGF per 1 cm² of destroyed surface area and approximately 10 μg of rhPDGF per 1 cm² of destroyed surface area. In some embodiments, each treatment involves administering an area dose between approximately 0.2 μg of rhPDGF per 1 cm² of destroyed surface area and approximately 10 μg of rhPDGF per 1 cm² of destroyed surface area. In some embodiments, each treatment involves administering an area dose between approximately 0.2 μg of rhPDGF per 1 cm² of destroyed surface area and approximately 1 μg of rhPDGF per 1 cm² of destroyed surface area. In some embodiments, each treatment involves administering an area dose between approximately 0.5 μg of rhPDGF per 1 cm² of destroyed surface area and approximately 1 μg of rhPDGF per 1 cm² of destroyed surface area. In some embodiments, each treatment involves administering an area dose between approximately 0.5 μg of rhPDGF per 1 cm² of destroyed surface area and approximately 10 μg of rhPDGF per 1 cm² of destroyed surface area. In some embodiments, each treatment involves administering an area dose between approximately 1 μg of rhPDGF per 1 cm² of surface area and approximately 10 μg of rhPDGF per 1 cm² of surface area.In some embodiments, each treatment involves administering an area dose between approximately 1 μg of rhPDGF per 1 cm² of destroyed surface area and approximately 5 μg of rhPDGF per 1 cm² of destroyed surface area. In some embodiments, each treatment involves administering an area dose between approximately 2 μg of rhPDGF per 1 cm² of destroyed surface area and approximately 5 μg of rhPDGF per 1 cm² of destroyed surface area. In some embodiments, each treatment involves administering an area dose between approximately 10 μg of rhPDGF per 1 cm² of destroyed surface area and approximately 100 μg of rhPDGF per 1 cm² of destroyed surface area. In some embodiments, each treatment involves administering an area dose between approximately 100 μg of rhPDGF per 1 cm² of destroyed surface area and approximately 1,000 μg of rhPDGF per 1 cm² of destroyed surface area. In some embodiments, each treatment involves administering an area dose between approximately 300 μg of rhPDGF per 1 cm² of damaged surface area and approximately 1,000 μg of rhPDGF per 1 cm² of damaged surface area.

[0141] In some embodiments, initial treatment with a composition according to the present invention may be the most important treatment. PDGF promotes the destructive healing process through its effects on cell proliferation (mitosis induction) and directional cell movement (chemotaxis) as well as re-angiogenesis (generation of new blood vessels). PDGF can also improve cell survival by stimulating anti-apoptotic pathways. Many cells have been shown to have receptors (binding sites) for PDGF, including connective tissue cells (skin, bone, cartilage, tendons, and ligaments), vascular cells, and nerve cells. PDGF recruits and activates cells involved in mitosis induction, such as stem cells, fibroblasts, and smooth muscle cells. Cells having receptors for PDGF respond by migrating toward the site of destruction (where PDGF is present at high levels as a result of application of a therapeutic composition according to the present invention) and then proliferating after binding to PDGF. Since PDGF receptors are rapidly degraded after activation, cell proliferation is controlled and limited by the presence of locally available PDGF, as well as by cell-cell interactions that allow cells to progress from the proliferative phase of destructive healing to the proliferative phase of matrix deposition, which ultimately leads to complete healing. As a result, a definitive bolus of rhPDGF-BB must be administered during the initial treatment to ensure that the patient's natural destruction and healing process is properly activated. Therefore, in some embodiments, the initial treatment is at least 0.1 μg of PDGF to a maximum of 500 μg of PDGF per 1 cm² of fracture surface area, at least 10 μg of PDGF per 1 cm² of fracture surface area to a maximum of 5000 μg of PDGF per 1 cm² of fracture surface area, or at least 20 μg of PDGF per 1 cm² of fracture surface area to a maximum of 1000 μg of PDGF per 1 cm² of fracture surface area, or at least 30 μg of PDGF per 1 cm² of fracture surface area to a maximum of 600 μg of PDGF per 1 cm² of fracture surface area, or at least 40 μg of PDGF per 1 cm² of fracture surface area The treatment involves applying a composition containing an area dose of PDGF such that the area dose is at most 400 μg of PDGF per 1 cm² of broken surface area, or at least 50 μg of PDGF per 1 cm² of broken surface area up to a maximum of 350 μg of PDGF per 1 cm² of broken surface area, or at least 60 μg of PDGF per 1 cm² of broken surface area up to a maximum of 300 μg of PDGF per 1 cm² of broken surface area, or at least 200 μg of PDGF per 1 cm² of broken surface area up to a maximum of 2000 μg of PDGF per 1 cm² of broken surface area. In some embodiments, the initial treatment involves at least 0.1 μg of PDGF per 1 cm² of broken surface area, at least 0.2 μg of PDGF per 1 cm² of broken surface area, at least 0.4 μg of PDGF per 1 cm² of broken surface area, at least 0.6 μg of PDGF per 1 cm² of broken surface area, and at least 0.8 μg of PDGF, at least 1 μg of PDGF per 1 cm² of fracture surface area, at least 2 μg of PDGF per 1 cm² of fracture surface area, at least 3 μg of PDGF per 1 cm² of fracture surface area, at least 4 μg of PDGF per 1 cm² of fracture surface area, at least 5 μg of PDGF per 1 cm² of fracture surface area, at least 6 μg of PDGF per 1 cm² of fracture surface area, at least 7 μg of PDGF per 1 cm² of fracture surface area, at least 8 μg of PDGF per 1 cm² of fracture surface area, per 1 cm² of fracture surface area At least 9 μg of PDGF, at least 10 μg of PDGF per 1 cm² of fracture surface area, or at least 20 μg of PDGF per 1 cm² of fracture surface area, or at least 25 μg of PDGF per 1 cm² of fracture surface area, or at least 30 μg of PDGF per 1 cm² of fracture surface area, or at least 40 μg of PDGF per 1 cm² of fracture surface area, or at least 50 μg of PDGF per 1 cm² of fracture surface area, or at least 60 μg of PDGF per 1 cm² of fracture surface area, or fracture surface area At least 70 μg of PDGF per 1 cm², or at least 80 μg of PDGF per 1 cm² of fracture surface area, or at least 90 μg of PDGF per 1 cm² of fracture surface area, or at least 100 μg of PDGF per 1 cm² of fracture surface area, or at least 250 μg of PDGF per 1 cm² of fracture surface area, or at least 500 μg of PDGF per 1 cm² of fracture surface area, or at least 600 μg of PDGF per 1 cm² of fracture surface area, or at least 600 The treatment involves administering a therapeutic composition containing a surface dose of μg of PDGF, or at least 700 μg of PDGF per 1 cm² of disrupted surface area, or at least 800 μg of PDGF per 1 cm² of disrupted surface area, or at least 900 μg of PDGF per 1 cm² of disrupted surface area, or at least 1,000 μg of PDGF per 1 cm² of disrupted surface area, or at least 2,000 μg of PDGF per 1 cm² of disrupted surface area, or at least 5,000 μg of PDGF per 1 cm² of disrupted surface area.

[0142] In some embodiments, each treatment applied involves a PDGF solution concentration ranging from approximately 4 μl per 1 cm³ of carrier (which may be hyaluronic acid gel or serum) to approximately 40 mL per 1 cm³ of carrier, or from approximately 0.1 mL per 1 cm³ of carrier to approximately 30 mL per 1 cm³ of carrier, or from approximately 0.2 mL per 1 cm³ of carrier to approximately 20 mL per 1 cm³ of carrier, or from approximately 0.1 mL per 1 cm³ of carrier to approximately 1 cm³ of carrier The amount of PDGF solution is between approximately 10 mL per cm³ of carrier, or between approximately 0.25 mL of PDGF solution per cm³ of carrier and approximately 5 mL of PDGF solution per cm³ of carrier, or between approximately 0.25 mL of PDGF solution per cm³ of carrier and approximately 2.5 mL of PDGF solution per cm³ of carrier, or between approximately 0.1 mL of PDGF solution per cm³ of carrier and approximately 1 mL of PDGF solution per cm³ of carrier, or between approximately 0.5 mL of PDGF solution per cm³ of carrier and approximately 1.5 mL of PDGF solution per cm³ of carrier. In a particular embodiment, the PDGF solution contains approximately 0.3 mg / mL of rhPDGF-BB.

[0143] In some embodiments, each treatment is between approximately 0.01 μg of PDGF per 1 cm³ of carrier and approximately 5 mg of PDGF per 1 cm³ of carrier, or between approximately 1.2 μg of PDGF per 1 cm³ of carrier and approximately 12 mg of PDGF per 1 cm³ of carrier, or between approximately 30 μg of PDGF per 1 cm³ of carrier and approximately 9 mg of PDGF per 1 cm³ of carrier, or between approximately 60 μg of PDGF per 1 cm³ of carrier and approximately 6 mg of PDGF per 1 cm³ of carrier, or between approximately 75 μg of PDGF per 1 cm³ of carrier and approximately 3 mg of PDGF per 1 cm³ of carrier. Alternatively, administer a dose of PDGF between approximately 75 μg and 1.5 mg per cm³ of carrier, or between approximately 75 μg and 750 μg per cm³ of carrier, or between approximately 120 μg and 600 μg per cm³ of carrier, or between approximately 150 μg and 450 μg per cm³ of carrier, or between approximately 75 μg and 225 μg per cm³ of carrier.

[0144] In some embodiments, the absolute initial PDGF treatment dose may be greater than the subsequent retreatment dose. The absolute initial PDGF treatment dose may be about 10%, 20%, 30%, 40%, or 50%, or up to about 300%, higher than each of the subsequent retreatment PDGF doses.

[0145] In some embodiments, the methods for treating acute skin destruction described herein may be similar to the methods for treating chronic wounds described in U.S. Patent No. 10,071,182. However, the compositions are applied to acute destruction more frequently but with fewer applications (e.g., once or twice or two or three times a day for up to 14 days). The methods for treating acute skin or connective tissue destruction may also be similar to those used after microneedling or other aesthetic procedures.

[0146] In some embodiments described herein, the compositions of the present invention used in the methods and uses described herein further include additional components such as local anesthetics. In some embodiments of the methods and uses described herein, a carrier such as hyaluronic acid further includes the above-mentioned components (one or more local anesthetics) and is applied to the skin or other lesion separately from the composition before, during, and / or after the administration procedure.

[0147] example Embodiments of the compositions, methods, uses, and systems of the present invention are described in more detail in the following non-limiting examples.

[0148] Example 1: Preparation of rhPDGF-BB composition in a hyaluronic acid carrier Example 1 describes a method for treating skin damage by applying a therapeutic composition comprising an rhPDGF-BB solution and hyaluronic acid, according to one embodiment. The method described below uses a composition of rhPDGF-BB solution containing hyaluronic acid. However, those skilled in the art will understand that any of the rhPDGF-BB-based compositions described herein may be used in conjunction with the method described in Example 1.

[0149] In this exemplary method, the first step may include dispensing a certain amount (e.g., 3 mL) of hyaluronic acid into a small dish. In one example, the hyaluronic acid may, but is not limited to, be in the form of a hyaluronic acid serum.

[0150] The next step may involve dispensing a certain amount (e.g., 0.5 mL) of rhPDGF-BB solution into a dish containing hyaluronic acid serum. In one embodiment, the rhPDGF solution may consist of 300 ug / mL of rhPDGF-BB in 20 mM sodium acetate buffer having a pH of 6.0 + / - 0.5 (i.e., 5.5 to 6.5). Thus, the dish contains the rhPDGF-BB solution and hyaluronic acid serum in a ratio of 1:6.

[0151] Furthermore, in this exemplary embodiment, the next step may include thoroughly mixing the dispensed solution with hyaluronic acid to form an rhPDGF-BB-based therapeutic composition having a uniform consistency. The concentration of rhPDGF-BB in this exemplary therapeutic composition is 42.85 ug / mL. In alternative embodiments, the composition may comprise the rhPDGF-BB composition, hyaluronic acid, and alginate. In yet another alternative embodiment, the composition may comprise the rhPDGF-BB composition and collagen or gelatin, the collagen may be soluble or insoluble. In yet another alternative embodiment, rhPDGF-BB may be a powder, such as one produced by freeze-drying or lyophilization, and the rhPDGF powder may be added to a carrier such as hyaluronic acid serum or collagen, or collagen, gelatin, or fibrin, or other types of solutions.

[0152] Exemplary Composition 1: An exemplary composition is prepared by mixing 0.5 mL of 300 ug / mL rhPDGF-BB solution (300 ug / mL rhPDGF-BB in 20 mM sodium acetate buffer) with 1.5 mL of 1% hyaluronic acid (HA) gel or serum. The HA gel or serum is a 50:50 blend of (i) hyaluronic acid having a weight between 100,000 Da and 150,000 Da and (ii) hyaluronic acid having a weight between 750,000 Da and 900,000 Da. The resulting mixture contains 150 ug of PDGF in 2 mL of gel and has a final concentration of 75 ug of PDGF per mL. The rhPDGF-BB solution and hyaluronic acid serum are present in a 1:3 ratio. The resulting mixture contains 0.75 wt% hyaluronic acid. Water may be added to the rhPDGF-BB therapeutic composition before administration, or it may be present in the rhPDGF-BB solution, HA gel, or serum, or both.

[0153] Exemplary Composition 2: A further exemplary composition is prepared by mixing 0.5 mL of 300 ug / mL of rhPDGF-BB solution (300 ug / mL of rhPDGF-BB in 20 mM sodium acetate buffer) with 3 mL of 1% hyaluronic acid (HA) gel or serum (which itself is a blend of (i) hyaluronic acid having a weight between 100,000 Da and 150,000 Da and (ii) hyaluronic acid having a weight between 750,000 Da and 900,000 Da). The resulting mixture contains 150 ug of PDGF in 3.5 mL of composition and has a final concentration of 42.86 μg of PDGF per mL. The rhPDGF-BB solution and hyaluronic acid serum are present in a 1:6 ratio. The resulting mixture contains 0.86 wt% hyaluronic acid. Water may be added to the rhPDGF-BB therapeutic composition before administration, or it may be present in the rhPDGF-BB solution, HA gel, or serum, or both.

[0154] Example 2: Investigation of the effects of PDGF applied to damaged skin. Histological analysis (hematoxylin and eosin staining) was performed on injured skin, specifically surgical wounds, treated with the PDGF composition described as "Example Composition 1" in Example 1, or on control biopsy specimens treated according to standard treatment, which involves applying Aquaphor® to injured skin and covering it with a wound dressing. Hematoxylin stains cell nuclei purple / blue, and eosin stains the extracellular matrix and cytoplasm pink. Skin biopsies were performed 21 days after injury (Figures 1A and 1B) and 42 days after injury (Figures 1C-1D, 2A and 2B). Clusters of inflammatory cells are indicated by white circles in Figures 1A-1D. Microscopic analysis of stained cells taken from skin wounds treated with the PDGF composition showed fewer inflammatory cells compared to cells from skin wounds treated with standard treatment (Figure 1B) (Figure 1A). 42 days after injury, biopsy specimens of skin wounds treated with PDGF compositions had more collagen compared to controls (Figure 1D and Figure 2B) (Figure 1C and Figure 2A). Furthermore, the collagen was more organized in PDGF-treated cells, and microscopic images showed that the tissue had improved blood supply and skin perfusion compared to controls (Figure 2B) (Figure 2A).

[0155] Example 3: Local treatment during microneedling Two patients underwent microneedling procedures on their faces and necks to treat age-related skin damage. Immediately after the microneedling procedures, the patients' skin was red and swollen at the penetration sites.

[0156] Prior to the microneedling procedure, approximately 1 mL of the PDGF composition (see "Example Composition 1" in Example 1) was applied topically to and around the microneedling site. During the procedure, the composition was "microneedled in" into the skin during quarter-section needle penetration.

[0157] Patient 1 had photographs taken immediately after the microneedling procedure, as well as 20 minutes and 1 hour after the procedure. By 20 minutes, redness and swelling had significantly decreased, and within 1 hour, the patient's skin had returned to a normal color (Figure 3). Patient 2 had photographs taken immediately after the microneedling procedure, as well as 1 hour and 2 hours after the procedure. By 1 hour, only a small amount of redness and swelling remained on the patient's skin, and by 2 hours, the patient's skin was no longer swollen and had returned to a normal color (Figure 4). Patient 2 compared the effect of using the PDGF composition immediately after the microneedling procedure with the previous standard treatment. Photographs taken 1 hour after microneedling with the previous standard treatment showed that the patient's skin was still very red and swollen. In comparison, photographs taken 1 hour after performing the microneedling procedure and treating the patient's skin with the PDGF composition showed a significant reduction in redness and swelling of the patient's skin (Figure 5).

[0158] Example 4: Local treatment of melasma patients undergoing microneedling Patients with melasma were treated with microneedling on the face and neck. Prior to the microneedling procedure, approximately 1 mL of the PDGF composition (see "Example Composition 1" in Example 1) was applied topically to and around the microneedling site. During the procedure, the composition was "microneedled in" into the skin during quarter-section needle penetration. Immediately after the microneedling procedure, the patient's skin was red and swollen. Photographs were taken immediately after the microneedling procedure and 5 days later. By 5 days post-microneedling, the patient's skin was no longer red or swollen, and there was an improvement in the aesthetic appearance of the skin (Figure 6). This result was highly unexpected from a single microneedling session, as hyperpigmentation in melasma is typically difficult to treat.

[0159] Example 5: Local treatment of skin damage resulting from facelift surgery The patient underwent a facelift. Incisions were made on both sides of the patient's face, in front of the ears and along the hairline, and closed with sutures. On one side of the patient's face, approximately 0.5 mL of the PDGF composition (see "Example Composition 1" in Example 1) was applied topically during incision closure and immediately after skin suturing. The composition was reapplied twice daily for 4–5 days post-surgery. On the other side of the patient's face, the incision was treated with standard treatment of applying 0.5 mL of Aquaphor® twice daily for 5 days.

[0160] Patient photographs were taken at 1 week (Figures 7A-7B), 2 weeks (Figure 7C), 3 weeks (Figure 7D), and 8 weeks (Figure 7E) postoperatively. At 1 week postoperatively, there was a significant difference in incision healing. Incisions treated with the PDGF composition were less red, and the skin healed better than incisions treated with the control standard treatment (Figures 7A and 7B). In Figure 7B, purpura around the incision treated with the PDGF composition was also reduced. Figure 7C shows the incision at 2 weeks postoperatively, where the incision treated with the PDGF composition continued to show improved healing compared to the control. By 3 weeks postoperatively (Figure 7D), the incision treated with the PDGF composition was completely healed with no redness or swelling. By 8 weeks postoperatively (Figure 7E), the incision treated with the PDGF composition was shown to be virtually scar-free, while the incision treated with the standard treatment still showed signs of scarring and redness.

[0161] A neck wound caused by a facelift bandage was identified during a follow-up visit two weeks post-surgery (Figure 8A). A PDGF composition was applied topically to the wound, followed by re-application twice daily for approximately 5–7 days. Six weeks after the initial application, the wound closed with minimal scarring (Figure 8B).

[0162] Example 6: Local treatment after laser treatment To improve the aesthetic appearance of the skin, the patient was treated with a broadband laser and level 3 Moxi®. During the procedure, the outer layer of the epidermis was removed by the laser. Immediately after the procedure, approximately 1 mL of a PDGF composition (see "Example Composition 1" in Example 1) was applied topically to and around the laser-treated area. Photographs were taken immediately after the procedure and 4 hours later (Figure 9). Immediately after the procedure, the patient's face was red and inflamed. After 4 hours, swelling and redness had decreased.

[0163] The second patient was treated with a Morpheus 8 laser. During the procedure, the outer layer of the facial epidermis was removed with the laser. After the procedure, approximately 1 mL of PDGF solution (300 μg / mL) containing only PDGF was applied topically to the skin on and around the laser-treated area. Photographs were taken immediately before, immediately after, 3 days after, and 30 days after the procedure (Figure 10). Before the procedure, the patient had visible wrinkles on the cheeks and around the eyes. After the procedure, the patient's skin was red and swollen. 3 days after the procedure, the redness of the skin was gone, and the wrinkles had decreased. 30 days later, the facial skin appeared smoother and firmer, and the wrinkles had further decreased.

[0164] Example 7: Local treatment of skin damage after Mohs surgery. The patient's scalp and cheek skin lesions were excised using the Mohs procedure (Figures 11A and 12A). The open cheek wound was sutured, and a PDGF composition (see "Example Composition 1" in Example 1) was applied topically 1-2 times per day for 10 days. After 10 days, the cheek suture closure was healed, showing minimal redness and scarring (Figure 11B). The open scalp surgical wound was treated with skin substitute. After 10 days, the scalp wound did not dry, and epithelialization of the open wound progressed well, although the wound healing process was considerably slower than that of the cheek wound (Figure 12B).

[0165] The present invention is described with reference to various specific preferred embodiments and techniques. However, it should be understood that many variations and modifications can be made while remaining within the spirit and scope of the invention. It will be apparent to those skilled in the art that methods, devices, device elements, materials, procedures, and techniques other than those specifically described herein can be applied to the implementation of the invention as broadly disclosed herein without relying on excessive experimentation. All functional equivalents known in the art of the methods, devices, device elements, materials, procedures, and techniques described herein are intended to be encompassed by the invention. Whenever a scope is disclosed, all sub-scopes and individual values ​​are intended to be encompassed. The present invention is not limited by the disclosed embodiments, but includes those shown in the drawings or illustrated herein, which are given as examples rather than limitations. Furthermore, it should be understood that various embodiments of networks, devices, and / or modules described herein contain optional features that can be applied individually or together with any other embodiments shown or intended herein to be mixed and adapted with the features of such networks, devices, and / or modules.

[0166] Although the present invention has been described in terms of a limited number of embodiments, those skilled in the art who are interested in this disclosure will understand that other embodiments can be devised that do not depart from the scope of the invention disclosed herein.

[0167] The embodiments, modifications, and sequences described herein should provide indicators of the usefulness and versatility of the invention. Other embodiments that do not provide all of the features and advantages described herein may also be utilized without departing from the spirit and scope of the invention. Such modifications and variations are considered to be within the scope of the invention.

[0168] Where the methods and processes described herein describe certain events occurring in a particular order, those skilled in the art will recognize that the order of certain steps may be modified, and that such modifications constitute variations of the present invention. Furthermore, certain steps may be performed sequentially, or simultaneously in parallel processes, if possible.

Claims

1. A cosmetic method for treating skin damage in a target area, wherein the skin damage does not involve tendons, ligaments, or bones, and the method is (i) A step of preparing a composition comprising sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution and a sterile biocompatible carrier, wherein the biocompatible carrier is hyaluronic acid, collagen, gelatin, alginate, polysaccharide, glycosaminoglycan, fibrin, derivatives thereof, or a combination thereof. (ii) A step of applying the composition on or into the skin lesion, wherein the carrier delivers bioactive rhPDGF on or into the skin lesion to bring about regeneration, rejuvenation or repair of the skin lesion. Methods that include...

2. The method according to claim 1, wherein the composition does not contain additional growth factors, and optionally the composition does not contain transforming growth factors (e.g., TGF beta), vascular endothelial growth factor (e.g., VEGF), or connective tissue growth factor (e.g., CTGF).

3. The method according to claim 1 or 2, wherein the skin damage is caused by aging, inflammation, radiation damage, ultraviolet damage, or a combination thereof, and optionally the skin damage includes one or more of wrinkles, sagging, grooves, skin laxity, sunburn, sunspots, irregular pigmentation, rough texture, dryness, and large pores.

4. The method according to any one of claims 1 to 3, wherein the skin destruction is caused by an incision made by a surgical procedure such as a facelift, abdominal wall reconstruction, or breast reconstruction, breast augmentation, or breast reduction.

5. The method according to any one of claims 1 to 4, wherein the skin destruction is a scar such as a surgical scar or an acne scar.

6. The method according to any one of claims 1 to 5, wherein the skin damage is hyperpigmentation or irregular pigmentation such as melasma.

7. The method according to any one of claims 1 to 6, wherein the skin destruction extends only to the dermis or subcutaneous layer of the target skin.

8. The method according to any one of claims 1 to 7, further comprising the steps of monitoring the regeneration, rejuvenation, or repair of the destruction during the treatment period, and repeating step (2) to re-treat the skin destruction.

9. The method according to claim 8, wherein the skin damage is retreated 1 to 6 times.

10. The method according to any one of claims 1 to 9, wherein the skin destruction is treated once or twice a day for a maximum of 14 days.

11. The method according to any one of claims 1 to 10, further comprising the step of (i) forming the composition by combining sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution with (ii) a sterile biocompatible carrier which is hyaluronic acid, collagen, gelatin, alginate, polysaccharide, glycosaminoglycan, fibrin, derivatives thereof or combinations thereof.

12. The method according to any one of claims 1 to 11, wherein the sterile rhPDGF-BB constitutes less than about 90 μg per gram of the composition.

13. The method according to claim 12, wherein the sterile rhPDGF-BB constitutes less than about 80 μg per gram of the composition.

14. The method according to claim 12, wherein the sterile rhPDGF-BB constitutes less than about 70 μg per gram of the composition.

15. The method according to claim 12, wherein the sterile rhPDGF-BB constitutes less than about 60 μg per gram of the composition.

16. The method according to claim 12, wherein the sterile rhPDGF-BB constitutes less than about 50 μg per gram of the composition.

17. The method according to claim 12, wherein the sterile rhPDGF-BB constitutes less than approximately 25 μg per gram of the composition.

18. The method according to claim 12, wherein the sterile rhPDGF-BB constitutes less than about 10 μg per gram of the composition.

19. The method according to claim 12, wherein the sterile rhPDGF-BB constitutes less than about 1.0 μg per gram of the composition.

20. The method according to any one of claims 1 to 11, wherein the sterile rhPDGF-BB has a final concentration of less than about 90 μg of rhPDGF-BB per 1 mL of the composition.

21. The method according to claim 20, wherein the sterile rhPDGF-BB has a final concentration of less than approximately 80 μg of rhPDGF-BB per 1 mL of the composition.

22. The method according to claim 20, wherein the sterile rhPDGF-BB has a final concentration of less than approximately 70 μg of rhPDGF-BB per 1 mL of the composition.

23. The method according to claim 20, wherein the sterile rhPDGF-BB has a final concentration of less than approximately 60 μg of rhPDGF-BB per 1 mL of the composition.

24. The method according to claim 20, wherein the sterile rhPDGF-BB has a final concentration of less than approximately 50 μg of rhPDGF-BB per 1 mL of the composition.

25. The method according to claim 20, wherein the sterile rhPDGF-BB has a final concentration of less than approximately 25 μg of rhPDGF-BB per 1 mL of the composition.

26. The method according to claim 20, wherein the sterile rhPDGF-BB has a final concentration of less than approximately 10 μg of rhPDGF-BB per 1 mL of the composition.

27. The method according to claim 20, wherein the sterile rhPDGF-BB has a final concentration of less than approximately 1.0 μg of rhPDGF-BB per 1 mL of the composition.

28. The method according to claim 20, wherein the sterile rhPDGF-BB has a final concentration between approximately 1.0 μg of rhPDGF-BB per 1 mL of the composition and 80 μg of rhPDGF-BB per 1 mL of the composition, and optionally, the sterile rhPDGF-BB has a final concentration between approximately 40 μg of rhPDGF-BB per 1 mL of the composition and 80 μg of rhPDGF-BB per 1 mL of the composition, for example, approximately 75 μg of rhPDGF-BB per 1 mL of the composition.

29. The method according to any one of claims 1 to 28, wherein the composition is applied topically to the skin lesion.

30. The method according to any one of claims 1 to 29, wherein the composition is applied to the skin lesion before, during and / or after an aesthetic procedure is performed that penetrates the target skin or skin lesion using a mechanical device, laser or scraping tool, for example, before, during and / or after a microneedling, microcoring, dermal ablation, dermaplaning, dermal filling or laser skin resurfacing procedure is performed.

31. The method according to claim 30, wherein the composition is applied to the skin lesion before a microneedling, microcoring, dermal ablation, dermaplaning, dermal filling, or laser skin resurfacing procedure is performed, for example, before a microneedling procedure is performed.

32. The method according to claim 30, wherein the composition is applied to the skin after a microneedling, microcoring, dermal ablation, dermaplaning, dermal filling, or laser skin resurfacing procedure has been performed, for example, after a microneedling procedure.

33. The method according to any one of claims 30 to 32, wherein the machine, laser, or scraping tool penetrates the skin fracture to a depth of about 0.5 mm to 3 mm.

34. The method according to claim 33, wherein the machine, laser, or scraping tool penetrates the skin to a depth of about 1 mm to 2 mm.

35. The method according to claim 34, wherein the machine, laser, or scraping tool penetrates the skin fracture to a depth of about 0.5 mm to 1.5 mm.

36. The method according to any one of claims 30 to 35, wherein the penetration of the skin is repeated, and the composition is optionally reapplied between the repeated penetrations.

37. The method according to claim 36, wherein the penetration of the skin is repeated up to four times.

38. The method according to any one of claims 1 to 37, wherein the composition further comprises a carbokaine, lidocaine, epinephrine, or a combination thereof.

39. The method according to any one of claims 1 to 38, wherein the biocompatible carrier is collagen.

40. The method according to any one of claims 1 to 38, wherein the biocompatible carrier is an alginate.

41. The method according to any one of claims 1 to 38, wherein the biocompatible carrier is fibrin.

42. The method according to any one of claims 1 to 38, wherein the biocompatible carrier is gelatin.

43. The method according to any one of claims 1 to 38, wherein the biocompatible carrier is a glycosaminoglycan.

44. The method according to any one of claims 1 to 38, wherein the biocompatible carrier is a combination of a polysaccharide and an alginate.

45. The method according to any one of claims 1 to 38, wherein the biocompatible carrier is a combination of polysaccharide and collagen.

46. The method according to any one of claims 1 to 38, wherein the biocompatible carrier is a combination of collagen and polysaccharide.

47. The method according to any one of claims 1 to 38, wherein the biocompatible carrier is a polysaccharide.

48. The method according to claim 47, wherein the polysaccharide is hyaluronic acid.

49. The method according to claim 47, wherein the hyaluronic acid is a serum or gel.

50. The method according to claim 48 or 49, wherein the composition comprises rhPDGF-BB and hyaluronic acid in a ratio of 1:3, 1:4, 1:5, or 1:

6.

51. The method according to claim 49 or 50, wherein the hyaluronic acid is a 50:50 blend of two molecular weights: (i) hyaluronic acid having a weight between about 100,000 Da and 300,000 Da and (ii) hyaluronic acid having a weight between about 700,000 Da and 950,000 Da, and optionally, the hyaluronic acid is a 50:50 blend of (i) hyaluronic acid having a weight between about 100,000 Da and 150,000 Da and (ii) hyaluronic acid having a weight between about 750,000 Da and 900,000 Da.

52. The composition is Approximately 0.5% to 1% by weight of hyaluronic acid, rhPDGF-BB with a final concentration between approximately 40 μg / mL and 80 μg / mL, A sodium acetate solution of approximately 10 mM to approximately 30 mM, preferably approximately 20 mM sodium acetate, Water and The method according to any one of claims 1 to 51, comprising or comprising.

53. The method according to any one of claims 1 to 52, wherein the composition is applied to the skin lesion to improve the appearance or aesthetics of the skin in question.

54. The method according to any one of claims 1 to 53, wherein the composition is applied to the skin lesion to improve the function of the target skin.

55. The method according to any one of claims 1 to 54, wherein the composition is applied to the skin fracture to reduce the pain and / or discomfort of the fracture.

56. The method according to any one of claims 1 to 55, wherein the composition is applied to the skin lesion to stimulate cell growth.

57. The method according to any one of claims 1 to 56, wherein the composition is applied to the skin lesion to stimulate angiogenesis.

58. The method according to any one of claims 1 to 57, wherein the composition is applied to the skin lesion to improve the texture of the skin, for example, to reduce wrinkles, grooves and / or sagging.

59. The method according to any one of claims 1 to 58, wherein the composition is applied to the skin lesion, for example, to increase tissue volume, to reduce wrinkles, grooves and / or sagging.

60. The method according to any one of claims 1 to 59, wherein the composition is applied to the skin damage to prevent or reduce scarring of the skin of the target.

61. The method according to any one of claims 1 to 60, wherein the composition is applied to the skin lesion to prevent or reduce redness, swelling, and / or inflammation at a site where an aesthetic procedure such as surgical incision, scarring, or microneedling, microcoring, dermal ablation, dermaplaning, dermal filling, or laser skin resurfacing has been or will be performed.

62. The method according to any one of claims 1 to 61, wherein the subject is a human.

63. The method according to any one of claims 1 to 62, wherein the composition is applied in or on the skin lesion via one or more needles, and optionally the composition is mixed with a dermal filler and applied by injection in or on the skin lesion.

64. A method for improving pain or discomfort caused by skin damage to a target, wherein the damage does not involve tendons, ligaments, or bones, and the method is: (1) A step of preparing a composition containing sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution, (2) A step of applying the composition on or into the skin lesion to reduce the pain or discomfort caused by the skin lesion. A method that includes this.

65. The method according to claim 64, wherein the composition does not contain additional growth factors, and optionally the composition does not contain transforming growth factors (e.g., TGF beta), vascular endothelial growth factor (e.g., VEGF), or connective tissue growth factor (e.g., CTGF).

66. The method according to claim 642 or 65, wherein the composition further comprises a sterile biocompatible carrier, the biocompatible carrier being hyaluronic acid, collagen, gelatin, alginate, polysaccharide, glycosaminoglycan, fibrin, derivatives thereof, or combinations thereof.

67. The method according to claim 66, wherein the biocompatible carrier is hyaluronic acid.

68. The method according to claim 67, wherein the hyaluronic acid is a serum or gel.

69. The method according to claim 67 or 68, wherein the composition comprises rhPDGF-BB and hyaluronic acid in a ratio of 1:3, 1:4, 1:5, or 1:

6.

70. The method according to any one of claims 67 to 69, wherein the hyaluronic acid is a 50:50 blend of two molecular weights: (i) hyaluronic acid having a weight between about 100,000 Da and 300,000 Da and (ii) hyaluronic acid having a weight between about 700,000 Da and 950,000 Da, and optionally, the hyaluronic acid is a 50:50 blend of (i) hyaluronic acid having a weight between about 100,000 Da and 150,000 Da and (ii) hyaluronic acid having a weight between about 750,000 Da and 900,000 Da.

71. The method according to any one of claims 64 to 70, wherein the skin damage is caused by aging, inflammation, radiation damage, ultraviolet damage, or a combination thereof, and optionally the skin damage includes rough skin, dry skin or itchy skin that causes discomfort, or the skin damage is sunburn.

72. The method according to any one of claims 64 to 70, wherein the skin destruction is caused by an incision resulting from a surgical procedure such as an incision after a facelift, or by a scar such as a surgical scar.

73. The method according to any one of claims 64 to 70, wherein the skin destruction is caused by a skin-penetrating aesthetic procedure such as microneedling, microcoring, dermal ablation, dermaplaning, dermal filling, or laser skin resurfacing.

74. The method according to any one of claims 64 to 73, wherein the skin destruction extends only to the dermis or subcutaneous layer of the target skin.

75. The method according to any one of claims 64 to 74, further comprising the steps of monitoring the pain or discomfort of the skin damage during the treatment period and repeating step (2) to re-treat the skin damage.

76. The method according to claim 75, wherein the skin damage is retreated 1 to 6 times.

77. The method according to any one of claims 64 to 76, wherein the skin destruction is treated once or twice a day for a maximum of 14 days.

78. The method according to any one of claims 64 to 77, further comprising the step of forming the composition by adding sterile recombinant human PDGF-BB (rhPDGF-BB) to a physiological solution, and optionally further comprising the step of combining the sterile recombinant human PDGF-BB (rhPDGF-BB) in a physiological solution with a sterile biocompatible carrier which is hyaluronic acid, collagen, gelatin, alginate, polysaccharide, glycosaminoglycan, fibrin, derivatives thereof, or a combination thereof.

79. The method according to any one of claims 65 to 78, wherein the composition comprises an rhPDGF-BB solution containing less than about 90 μg / g of rhPDGF-BB.

80. The method according to claim 79, wherein the composition comprises an rhPDGF-BB solution containing less than about 80 μg / g of rhPDGF-BB.

81. The method according to claim 79, wherein the composition comprises an rhPDGF-BB solution containing less than about 70 μg / g of rhPDGF-BB.

82. The method according to claim 79, wherein the composition comprises an rhPDGF-BB solution containing less than 60 μg / g of rhPDGF-BB.

83. The method according to claim 79, wherein the composition comprises an rhPDGF-BB solution containing less than about 50 μg / g of rhPDGF-BB.

84. The method according to claim 79, wherein the composition comprises an rhPDGF-BB solution containing less than about 25 μg / g of rhPDGF-BB.

85. The method according to claim 79, wherein the composition comprises an rhPDGF-BB solution containing less than about 10 μg / g of rhPDGF-BB.

86. The method according to claim 79, wherein the composition comprises an rhPDGF-BB solution containing less than 1.0 μg / g of rhPDGF-BB.

87. The method according to any one of claims 64 to 86, wherein the composition comprises an rhPDGF-BB solution containing less than about 90 μg / mL of rhPDGF-BB.

88. The method according to claim 87, wherein the composition comprises an rhPDGF-BB solution containing less than 80 μg / mL of rhPDGF-BB.

89. The method according to claim 87, wherein the composition comprises an rhPDGF-BB solution containing less than 70 μg / mL of rhPDGF-BB.

90. The method according to claim 87, wherein the composition comprises an rhPDGF-BB solution containing less than 60 μg / mL of rhPDGF-BB.

91. The method according to claim 87, wherein the composition comprises an rhPDGF-BB solution containing less than 50 μg / mL of rhPDGF-BB.

92. The method according to claim 87, wherein the composition comprises an rhPDGF-BB solution containing less than 25 μg / mL of rhPDGF-BB.

93. The method according to claim 87, wherein the composition comprises an rhPDGF-BB solution containing less than 10 μg / mL of rhPDGF-BB.

94. The method according to claim 87, wherein the composition comprises an rhPDGF-BB solution containing less than 1.0 μg / mL of rhPDGF-BB.

95. The method according to any one of claims 64 to 78, wherein the sterile rhPDGF-BB has a final concentration between approximately 200 μg of rhPDGF-BB per 1 mL of the composition and 400 μg of rhPDGF-BB per 1 mL of the composition, and optionally, the sterile rhPDGF-BB has a final concentration of approximately 300 μg of rhPDGF-BB per 1 mL of the composition.

96. The method according to any one of claims 66 to 78, wherein the sterile rhPDGF-BB has a final concentration between approximately 1.0 μg of rhPDGF-BB per 1 mL of the composition and 80 μg of rhPDGF-BB per 1 mL of the composition, and optionally, the sterile rhPDGF-BB has a final concentration between approximately 40 μg of rhPDGF-BB per 1 mL of the composition and 80 μg of rhPDGF-BB per 1 mL of the composition.

97. The composition is Approximately 0.5% to 1% by weight of hyaluronic acid, rhPDGF-BB with a final concentration between approximately 40 μg / mL and 80 μg / mL, A sodium acetate solution of approximately 10 mM to approximately 30 mM, preferably approximately 20 mM sodium acetate, Water and The method according to any one of claims 64 to 96, comprising or comprising.

98. The composition is rhPDGF-BB with a final concentration between approximately 200 μg / mL and 400 μg / mL, A sodium acetate solution of approximately 10 mM to approximately 30 mM, preferably approximately 20 mM sodium acetate, Water and The method according to any one of claims 64 to 65, 71 to 77 and 79 to 96, comprising or including.

99. The method according to any one of claims 64 to 98, wherein the composition is applied topically to the skin lesion.

100. The method according to any one of claims 64 to 99, wherein the composition is applied to the skin lesion, for example, before, during, and / or after an aesthetic procedure is performed that penetrates the target skin or skin lesion using a mechanical device, laser, or scraping tool, or before, during, and / or after a procedure such as microneedling, microcoring, dermal ablation, dermaplaning, dermal filling, or laser skin resurfacing is performed.

101. The method according to claim 100, wherein the composition is applied to the skin lesion before a microneedling, microcoring, dermal ablation, dermaplaning, dermal filling, or laser skin resurfacing procedure is performed, for example, before a microneedling procedure is performed.

102. The method according to claim 100, wherein the composition is applied to the skin after a microneedling, microcoring, dermal ablation, dermaplaning, dermal filling, or laser skin resurfacing procedure has been performed, for example, after a microneedling procedure.

103. The method according to any one of claims 100 to 102, wherein the machine, laser, or scraping tool penetrates the skin fracture to a depth of about 0.5 mm to 3 mm.

104. The method according to claim 103, wherein the machine, laser, or scraping tool penetrates the skin to a depth of about 1 mm to 2 mm.

105. The method according to claim 103, wherein the machine, laser, or scraping tool penetrates the skin fracture to a depth of about 0.5 mm to 1.5 mm.

106. The method according to any one of claims 100 to 105, wherein the penetration of the skin is repeated, and the composition is optionally reapplied between the repeated penetrations.

107. The method according to claim 106, wherein the penetration of the skin is repeated up to four times.

108. The method according to any one of claims 64 to 107, wherein the composition is applied to the skin lesion to improve the appearance or aesthetics of the skin in question.

109. The method according to any one of claims 64 to 108, wherein the composition is applied to the skin lesion to improve the function of the skin of the subject.

110. The method according to any one of claims 64 to 109, wherein the composition is applied to the skin lesion to stimulate cell growth.

111. The method according to any one of claims 64 to 110, wherein the composition is applied to the skin lesion to stimulate angiogenesis.

112. The method according to any one of claims 64 to 111, wherein the composition is applied to the skin lesion to improve the texture of the skin, for example, to reduce wrinkles, grooves and / or sagging.

113. The method according to any one of claims 64 to 112, wherein the composition is applied to the skin lesion, for example, to increase tissue volume, to reduce wrinkles, grooves and / or sagging.

114. The method according to any one of claims 64 to 113, wherein the composition is applied to the skin damage to prevent or reduce scarring of the skin of the subject.

115. The method according to any one of claims 64 to 114, wherein the composition is applied to the skin lesion to prevent or reduce redness, swelling, and / or inflammation at a site where an aesthetic procedure such as surgical incision, scarring, or microneedling, microcoring, dermal ablation, dermaplaning, dermal filling, or laser skin resurfacing has been performed or is to be performed.

116. The method according to any one of claims 64 to 115, wherein the composition is applied to the skin damage in order to regenerate, rejuvenate, or repair the skin damage.

117. The method according to any one of claims 64 to 116, wherein the subject is a human.

118. The method according to any one of claims 64 to 117, wherein the composition is administered in or on the skin lesion via one or more needles, and optionally the composition is mixed with a dermal filler and administered by injection in or on the skin lesion.

119. The method according to claim 64, wherein the composition comprises an rhPDGF-BB solution containing less than 1,000 μg / mL of rhPDGF-BB.

120. The method according to claim 64, wherein the composition comprises an rhPDGF-BB solution containing less than 900 μg / mL of rhPDGF-BB.

121. The method according to claim 64, wherein the composition comprises an rhPDGF-BB solution containing less than 800 μg / mL of rhPDGF-BB.

122. The method according to claim 64, wherein the composition comprises an rhPDGF-BB solution containing less than 700 μg / mL of rhPDGF-BB.

123. The method according to claim 64, wherein the composition comprises an rhPDGF-BB solution containing less than 600 μg / mL of rhPDGF-BB.

124. The method according to claim 64, wherein the composition comprises an rhPDGF-BB solution containing less than 500 μg / mL of rhPDGF-BB.

125. The method according to claim 64, wherein the composition comprises an rhPDGF-BB solution containing less than 400 μg / mL of rhPDGF-BB.

126. The method according to claim 64, wherein the composition comprises an rhPDGF-BB solution containing less than 300 μg / mL of rhPDGF-BB.

127. The method according to any one of claims 64 to 118, wherein the composition further comprises a carbokaine, lidocaine, epinephrine, or a combination thereof.

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