Use of hair stem cells in wound healing

Abstract

A method comprising (a) harvesting a hair follicle by applying wax to a first area of ​​skin from which the hair follicle protrudes, and (b) removing the wax from the first area and applying the wax to a wound in a second area of ​​skin such that the hair follicle and its keratinocyte stem cells are in contact with the wound. Related compositions, products, and medical devices are also disclosed.

Description

Technical Field

[0001] Cross - Reference to Related Applications This PCT application was filed on July 9, 2023, and claims the benefit under 35 U.S.C. § 119(e) of U.S. Provisional Patent Application No. 63 / 512,628, which has the same invention title and inventors as this application. This provisional patent application is hereby incorporated by reference in its entirety into this specification.

[0002] Technical Field The present invention is in the field of stem - cell - based grafts or transplants.

Background Art

[0003] Wound healing is a major problem in modern medicine.

[0004] More than 13 million people worldwide suffer from chronic wounds annually. Many factors can lead to healing disorders. The main factors are hypoxia, bacterial colonization, ischemia, reperfusion injury, changes in cell responses, and collagen synthesis deficiency.

[0005] Abnormal wound healing is rarely seen in normal healthy subjects. Abnormal wound healing is usually associated with co - morbidities (such as diabetes, autoimmune diseases, peripheral arterial disease), an increase in body mass index, anatomical location, and drug therapy.

[0006] Chronic wounds can be defined as a common type of skin injury that is physiologically impaired due to a breakdown in the wound - healing cycle, more specifically, when this healing process stops at the inflammatory stage. A wound is considered chronic if it does not heal within 6 weeks and shows signs of long - term inflammation, defective wound matrix, and failure of re - epithelialization. In this case, the duration of wound healing varies depending on the type and severity of the wound, and some wounds may not heal completely. Local treatments are applied to reduce pain and itching from the wound, infection, and bleeding, and to treat excessive exudate that can cause an unpleasant odor, which is a very troublesome factor for patients.

[0007] Chronic non-healing wounds such as diabetic foot ulcers, venous leg ulcers, and pressure ulcers present unique challenges.

[0008] Acute skin wounds, such as burns, present a distinct but equally challenging set of problems.

[0009] Wound healing is a natural physiological response to tissue damage that typically lasts 4 to 6 weeks. The healing process can be divided into four stages: the hemostatic phase, the inflammatory phase, the proliferative phase, and the remodeling phase.

[0010] The healing process involves numerous cell types, cytokines, mediators, and the vascular system. An initial cascade of vasoconstriction and platelet aggregation is designed to stop bleeding. This cascade is followed by an influx of various inflammatory cells, starting with neutrophils. These inflammatory cells then release various mediators and cytokines to promote angiogenesis, thrombus formation, and re-epithelialization. Fibroblasts then place extracellular components that function as a scaffold.

[0011] For several days, the inflammatory phase is introduced to limit further damage, close the wound, remove cellular debris and bacteria, and promote cell migration. This stage is characterized by increased hemostasis, chemotaxis, and vascular permeability. Mediators, cytokines, and other factors secreted by neutrophils, monocytes, endothelial cells, and platelets play a role in promoting collagen degradation, fibroblast transformation, new blood vessel growth, cellular permeability, and re-epithelialization.

[0012] Simultaneously, the proliferative phase began with the formation of granulation tissue, re-epithelialization, and angiogenesis.

[0013] Collagen production by fibroblasts occurs along with the migration of cells from the periphery and adjacent edges of the wound, enabling re-epithelialization. Neoangiogenesis, neovascularization, and vascularization occur by forming new blood vessels from existing ones.

[0014] The remodeling phase is a stage in which excess collagen is broken down and wound contraction occurs, causing the wound to reach its maximum strength, which is approximately 80% of the tensile strength of normal skin.

[0015] U.S. Patent No. 7,419,661 describes the use of dermal sheath tissue and / or cells derived therefrom in wound healing systems. The relevant characteristics of this tissue type are utilized to provide novel wound healing materials, which may be applied in the provision of novel therapeutic compositions and novel surgical dressings when wound closure and minimal scarring are desired.

[0016] International Publication No. 2011 / 104030 describes a method for promoting the healing or functional restoration of superficial wounds of the skin and / or skin appendages of a second skin region, comprising the steps of: applying cells obtained from the hair follicle sheath of a first skin region of a donor to a second skin region of a recipient. This application further relates to cells, preparations, uses of cells, and methods for producing preparations.

[0017] International Publication No. 2011 / 050948 describes a tape, particularly an adhesive tape, for the treatment of skin disorders, comprising at least one base layer having a first surface facing away from the skin during use and a second surface facing towards the skin during use, and at least one first film, particularly a soluble film, located on the side of the second surface and covering at least a portion of the second surface of the base layer, wherein the first film, particularly the soluble film, has a first surface facing away from the skin during use and a second surface facing towards the skin during use, and comprises at least one enzyme, particularly a protease.

[0018] European Patent No. 2343079B1 describes a method and / or apparatus suitable for producing a transplantable cell suspension of living tissue suitable for transplantation into a patient. In the application of this method and / or use of this apparatus, donor tissue is collected, subjected to cell dissociation, cells suitable for transplantation back into the patient are collected and dispersed in a solution suitable for immediate dispersion on the recipient transplant site.

[0019] International Publication No. 2016 / 131096A1 describes methods and products for delivering cells to biological sites. The present application intends to provide a product comprising an alkylamine-functionalized substrate and cells bound to the functionalized substrate for delivery to a biological site, wherein the alkylamine-functionalized substrate comprises a surface density having a primary amine-to-carbon atomic ratio greater than 0.005, and a method for delivering cells to a biological site, comprising applying the product to a biological site to enable the movement of cells from the product to the biological site, thereby delivering the cells to the site.

[0020] European Patent No. 1198557B1 describes the treatment of skin defects with organ cultured autologous keratinocytes isolated from the outer root sheath of hair in the growth phase or growing stage. Disclosed herein are methods for primary organ-type cultures (i.e., epidermal equivalents) and subsequent organ-type cultures in a fully defined medium supplemented with substances isolated from autologous human serum and blood components, along with minimal allogeneic biological supplements. [Overview of the project] [Means for solving the problem]

[0021] A broad aspect of some embodiments of the present invention relates to the application of cells to wounds without prior culture or proliferation.

[0022] One aspect of several embodiments of the present invention relates to the use of wax as a transport medium for keratinocyte stem cells without cell culture or proliferation. In some embodiments, the elimination of cell culture / proliferation contributes to a shorter treatment period. Alternatively or additionally, the use of wax as a transport medium contributes to reduced invasiveness and / or eliminates the need for surgery. In some exemplary embodiments of the present invention, the use of wax as a transport medium contributes to reducing patient pain and / or promoting faster wound healing and / or enabling patients to resume normal daily activities more quickly.

[0023] Another aspect of some embodiments of the present invention relates to transplanting keratinocyte stem cells located at the tip of a donor's plucked hair into an open wound without culturing the keratinocyte stem cells. In some exemplary embodiments of the present invention, wax is used as a transfer medium. In some exemplary embodiments of the present invention, the transplant is autologous. In some exemplary embodiments of the present invention, necrotic fragments or non-growing tissue are removed before transplantation onto the wound. Alternatively or additionally, in some embodiments, the wound bed is prepared to receive a skin graft or flap, or for closure as necessary.

[0024] Another aspect of some embodiments of the present invention relates to the use of wax as a transfer medium for harvesting hair follicles at a first location and transferring them to a wound bed at a second location. In various exemplary embodiments of the present invention, the wax is applied to the first location as a liquid (melted), paste, or solid (sheet or strip) to remove hair from normal skin. The wax is then removed and applied to the wound so that keratinocyte stem cells located at the tip of the plucked hair are in contact with the wound bed. In some embodiments, the wax is applied using a dressing material that covers the wax while holding the wax in place on the wound bed. In various exemplary embodiments of the present invention, the first location includes the legs and / or neck and / or arms. Alternatively or additionally, in various exemplary embodiments of the present invention, the hair is applied to a second location of the subject from which the hair was provided (autologous graft).

[0025] In various exemplary embodiments of the present invention, the wax is impregnated with growth factors and / or antibiotics and / or cytokines and / or extracellular matrix molecules. In some exemplary embodiments of the present invention, hair follicles harvested on the wax surface are treated with one or more biologically active agents before being applied to a wound. In various exemplary embodiments of the present invention, this treatment includes, for example, immersion or spraying with a solution of a biologically active agent. In various exemplary embodiments of the present invention, biologically active agents include antibacterial compounds such as silver, betaine, chitin, polyhexamethylene biguanide, antibiotics, and / or antifungal substances.

[0026] Alternatively or additionally, in some embodiments, biologically active factors include epidermal growth factor (EGF) and / or fibroblast growth factor (FGF) and / or transforming growth factor β (TGF-β) and / or platelet-derived growth factor (PDGF), and / or vascular endothelial growth factor (VEGF) and / or interleukin (e.g., IL1 and / or IL6) and / or tumor necrosis factor α (TNF-α). In some exemplary embodiments of the present invention, one or more factors contribute to reducing perturbations of growth factors and cytokines that occur in chronic wounds.

[0027] In various exemplary embodiments of the present invention, the treatment enhances cell growth and / or differentiation, and / or removes debris, and / or inactivates unwanted microorganisms.

[0028] For the purposes of this specification and the appended claims, the term "wax" is to be construed broadly to include not only long-chain fatty acids bonded to long-chain alcohols via ester oxygen, but also other polymers such as thermosensitive polymers and / or UV-sensitive polymers and / or carbohydrate polymers, or other materials having sufficient adhesiveness to allow hair removal from normal skin areas and being biocompatible for application to chronic wounds.

[0029] Although embodiments are presented separately above, in various exemplary embodiments of the present invention, two or three of the embodiments and / or features thereof are combined to create additional embodiments of the present invention.

[0030] The various embodiments described above will be understood to address technical challenges related to reducing the invasiveness of stem cell harvesting.

[0031] Alternatively or additionally, it will be understood that the various embodiments described above relate to solving technical challenges associated with reducing the time required to prepare autografts.

[0032] Alternatively or additionally, it will be understood that the various embodiments described above relate to solving technical challenges associated with the treatment of dermatological injuries in humans and / or animals, including but not limited to chronic injuries, including, but not limited to, non-healing or infectious surgical or traumatic wounds, venous ulcers, arterial ulcers, pressure ulcers (e.g., bedsores, supine ulcers, or compression ulcers), diabetic foot ulcers, and ischemic ulcers, epidermolysis bullosa, pyoderma gangrenosum, wounds after excision of giant congenital nevi, and acute wounds.

[0033] Alternatively or additionally, it will be understood that the various embodiments described above relate to solving technical challenges related to the rejuvenation of photodamaged skin after the removal of tattoos, nevi, skin cancer, papillomas, amputations, or skin resurfacing.

[0034] Alternatively or additionally, the various embodiments described above will be understood to relate to solving technical challenges associated with improving scarring by providing keratinocyte stem cells to wounds.

[0035] In some exemplary embodiments of the present invention, a method is provided comprising the steps of (a) collecting hair follicles containing keratinocyte stem cells by applying wax to a first area of ​​skin from which the hair follicles protrude; and (b) removing the wax from the first area and applying it to a wound in a second area of ​​skin so that the hair follicles and their keratinocyte stem cells are in contact with the wound. In some embodiments, the method includes priming the hair follicles containing keratinocyte stem cells before application to the wound. Alternatively or additionally, in some embodiments, the method includes washing the wax and / or hair follicles before application to the wound. Alternatively or additionally, in some embodiments, the method includes treating the hair follicles containing wax and / or keratinocyte stem cells with one or more antibiotics before application to the wound.

[0036] In some exemplary embodiments of the present invention, a wax composition to which hair follicles containing keratinocyte stem cells are attached is provided for use in a wound treatment method, which comprises applying the composition to a wound so that hair follicles containing keratinocyte stem cells come into contact with the wound. In some embodiments, the composition for use comprises hair follicles containing keratinocyte stem cells that remain attached to the composition during application. Alternatively or additionally, in some embodiments, the method comprises priming the hair follicles containing keratinocyte stem cells before application to the wound. Alternatively or additionally, in some embodiments, the method comprises washing the wax and / or hair follicles containing keratinocyte stem cells before application to the wound. Alternatively or additionally, in some embodiments, the method comprises treating the wax and / or hair follicles containing keratinocyte stem cells with one or more antibiotics before application to the wound.

[0037] In some exemplary embodiments of the present invention, a composition is provided comprising a wax impregnated with at least one selected from the group consisting of antibiotics, growth factors, cytokines, and extracellular matrix molecules.

[0038] In some exemplary embodiments of the present invention, a product is provided comprising a hair removal wax strip or sheet having perforations distributed on a surface that comes into contact with the skin during use.

[0039] In some exemplary embodiments of the present invention, a medical device is provided comprising a wax composition to which hair follicles containing keratinocyte stem cells are attached for treating dermatological injuries. In some embodiments, the dermatological injuries are selected from the group consisting of non-healing or infectious surgical or traumatic wounds, venous ulcers, arterial ulcers, pressure ulcers such as bedsores; supine ulcers; or pressure ulcers, diabetic foot ulcers, ischemic ulcers, epidermolysis bullosa, pyoderma gangrenosum, and wounds following excision of giant congenital nevi in ​​humans and animals. Alternatively or additionally, in some embodiments, the dermatological injuries are selected from the group consisting of treatment of postoperative injuries after removal of tattoos, nevi, skin cancer, papillomas, rejuvenation of photo-damaged skin after amputation and skin resurfacing.

[0040] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art in which the present invention pertains. Suitable methods and materials are described below, but similar or equivalent methods and materials may be used in the practice of the present invention. In case of any conflict, the patent specification, including the definitions, shall prevail. All materials, methods, and examples are illustrative and not intended to limit the scope of the invention.

[0041] As used herein, the terms “comprising” and “including” or their grammatical variations should be interpreted as specifying the inclusion of a described feature, integer, action, or component without precluding the addition of one or more additional features, integers, actions, components, or groups thereof. This term is broader than and includes the terms “consisting of” and “consisting essentially of” as defined in the U.S. Patent and Trademark Office's Patent Examination Manual of Procedure. Accordingly, any statement that an embodiment “includes” or “comprises” a feature is a specific statement that a subordinate embodiment “consisting essentially of” and / or “consisting of” the described feature.

[0042] Where used herein, the phrase “essentially from” or its grammatical variation thereof should be interpreted as identifying a described feature, integer, step, or component, but not precluding the addition of one or more additional features, integers, steps, components, or groups thereof, except in cases where such additional features, integers, steps, components, or groups thereof do not substantially alter the basic and novel features of the claimed composition, apparatus, or method.

[0043] As used herein and in the appended claims, the phrase “adapted to” imposes additional structural limitations on the components listed above.

[0044] The term “method” refers to any configuration, means, techniques, and procedures for accomplishing a given task, and includes, but is not limited to, any configurations, means, techniques, and procedures known to or readily developed from configurations, means, techniques, and procedures known by practitioners of architecture and / or computer science.

[0045] Unless otherwise specified, the percentage (%) of chemical substances is expressed in wt / w (weight / weight). [Brief explanation of the drawing]

[0046] To understand the present invention and to see how it can be put into practice, embodiments are described here only as non-limiting examples, with reference to the accompanying drawings. In the drawings, identical and similar structures, elements or parts thereof appearing in multiple drawings are generally labeled with the same or similar reference numerals in the drawings in which they appear. The dimensions of the components and features shown in the drawings have been selected primarily for convenience and clarity of presentation and are not necessarily to scale. The accompanying drawings are as follows:

[0047] [Figure 1] Figure 1 is a simplified flowchart of the method according to several embodiments of the present invention; and

[0048] [Figure 2] Figure 2 is a schematic diagram of a product according to several embodiments of the present invention. [Modes for carrying out the invention]

[0049] Embodiments of the present invention relate to methods for harvesting hair follicles containing keratinocyte stem cells and applying those keratinocyte stem cells to wounds, as well as related compositions and products.

[0050] Specifically, some embodiments of the present invention can be used to promote wound healing by transplanting uncultured keratinocyte stem cells into a wound.

[0051] The principles and operation of methods, compositions, and products according to exemplary embodiments of the present invention can be better understood by referring to the drawings and accompanying descriptions.

[0052] Before describing in detail at least one embodiment of the present invention, it should be understood that the present invention is not limited in its application to the details described below or illustrated by the examples. Other embodiments of the present invention are possible, or can be carried out or implemented in various ways. It should also be understood that the expressions and terms used herein are for illustrative purposes only and should not be considered limiting.

[0053] Exemplary Method Figure 1 is a simplified flowchart of a method, generally shown as 100, for preparing a skin graft in situ according to some embodiments of the present invention.

[0054] An illustrated exemplary method includes the steps of: 110 harvesting keratinocyte stem cells by applying wax to a first area of ​​skin where hair follicles protrude; 120 removing the wax from the first area; and 130 applying the wax to a wound in a second area of ​​skin so that the hair follicles and their keratinocyte stem cells attached to the wax are in contact with the wound bed. In these embodiments, the wax is left on the wound for several days or weeks to allow the keratinocyte stem cells to grow and settle in the wound. The proliferating population of keratinocyte stem cells functions as a skin graft covering the wound.

[0055] In some embodiments, method 100 includes a step 140 of priming keratinocyte stem cells located on hair follicles prior to step 130 of application. Priming 140 prepares the cells to be more effective for healing. In some exemplary embodiments of the present invention, priming 140 is performed prior to 3, 4, 5, 7, 14, 21, 28, or more days, or an intermediate number of days. One method of priming includes inducing cell proliferation by rubbing the skin daily for a period of time. In various exemplary embodiments of the present invention, the period is 5, 10, 15, 20, 25, or 30 minutes, or an intermediate or longer period, at a frequency of once, twice, three or more times per day.

[0056] Alternatively or additionally, in some embodiments, method 100 includes a step of preparing the wound before step 130 to be applied. In various exemplary embodiments of the present invention, wound repair includes washing and / or granulation tissue formation and / or removal of fibrinous tissue.

[0057] Alternatively or additionally, in some embodiments, Method 100 includes a step 150 of washing the hair follicles containing wax and / or keratinocyte stem cells before step 130 to be applied.

[0058] Alternatively or additionally, in some embodiments, Method 100 includes a step 160 in which hair follicles containing wax and / or keratinocyte stem cells are treated with one or more antibiotics and / or growth factors and / or cytokines and / or extracellular matrix molecules prior to step 130 in which the method is applied.

[0059] In some exemplary embodiments of the present invention, hair follicles collected on a wax surface are treated with one or more biologically active agents before being applied to a wound. In various exemplary embodiments of the present invention, this treatment includes, for example, immersion in a solution of the biologically active agent or spraying with the same. In various exemplary embodiments of the present invention, the biologically active agent includes antimicrobial compounds such as silver, betaine, chitin, polyhexamethylene biguanide, antibiotics, and / or antifungal substances.

[0060] Alternatively or additionally, in some embodiments, the biologically active agents include epidermal growth factor (EGF) and / or fibroblast growth factor (FGF) and / or transforming growth factor β (TGF-β) and / or platelet-derived growth factor (PDGF), and / or vascular endothelial growth factor (VEGF) and / or interleukins (e.g., IL1 and / or IL6) and / or tumor necrosis factor α (TNF-α). In some exemplary embodiments of the present invention, one or more factors contribute to reducing the growth factor and cytokine perturbations that occur in chronic wounds.

[0061] In many exemplary embodiments of Method 100, the first area of ​​skin from which hair follicles containing keratinocyte stem cells are harvested and the second area of ​​skin where the wound is located are on the body of the same person. In these embodiments, Methods 100 and 200 are autologous skin grafting methods.

[0062] Exemplary compositions for use. In some exemplary embodiments of the present invention, a wax composition for use in a wound treatment method is provided, comprising: collecting hair follicles containing keratinocyte stem cells by adhering the hair follicles to a composition; and applying the collected hair follicles containing keratinocyte stem cells to a wound. This method itself is described above and illustrated in Figure 1.

[0063] In some embodiments of the composition for use, hair follicles containing keratinocyte stem cells remain attached to the composition during application to the wound.

[0064] Alternatively or additionally, in some embodiments of the composition for use, the method includes priming the hair follicles containing keratinocyte stem cells before application to the wound. The priming is as described above. Alternatively or additionally, in some embodiments of the composition for use, the method includes washing the hair follicles containing wax and / or keratinocyte stem cells before application to the wound. Alternatively or additionally, in some embodiments of the composition for use, the method includes treating the hair follicles containing wax and / or keratinocyte stem cells with one or more antibiotics before application to the wound.

[0065] In some exemplary embodiments of the present invention, a composition is provided comprising a wax impregnated with an antibiotic, a growth factor, an extracellular matrix component, an autologous growth factor / molecule such as PRP, fibrinogen, cytokines, extracellular matrix molecules, silver, betaine, chitin, polyhexamethylene biguanide, an antifungal substance, epidermal growth factor (EGF), fibroblast growth factor (FGF), transforming growth factor β (TGFβ), platelet-derived growth factor (PDGF), vascular endothelial growth factor (VEGF) and / or interleukins (e.g., IL1 and / or IL6), and tumor necrosis factor α (TNF-α).

[0066] Extracellular matrix components include, but are not limited to, collagen, fibrin, laminin, GAG, and HA. In some exemplary embodiments of the present invention, the composition is provided as a pre-formed strip or sheet. In some embodiments, these units are provided as a continuous roll with perforations between the units to facilitate the separation of one unit from another. In other exemplary embodiments of the present invention, the wax is provided as a block that is melted before application.

[0067] Figure 2 is a schematic diagram of a product, shown as 300 in whole, according to several embodiments of the present invention. In the illustrated embodiments, the product includes a hair removal wax strip or sheet having perforations 320 distributed on a surface 310 that comes into contact with the skin during use. Such a strip or sheet is useful for carrying out methods 100 and / or 200 described above. The perforations 320 contribute to the breathability and / or moisture permeability of the strip / sheet 300.

[0068] Exemplary advantages In some exemplary embodiments of the present invention, the need to culture keratinocyte stem cells is eliminated.

[0069] Alternatively or additionally, in some embodiments, the need to provide fibroblast stem cells or mesenchymal stem cells (MSCs) to prepare skin grafts is avoided.

[0070] Alternatively or additionally, harvesting keratinocyte stem cells by plucking hair is a relatively non-invasive method of stem cell harvesting. Alternatively or additionally, since hair regenerates quickly and automatically, repeated harvesting is possible over time as needed.

[0071] Exemplary wax properties In some exemplary embodiments of the present invention, the wax is biocompatible. For the purposes of this specification and the appended claims, the term “biocompatible” means not harmful to the human body and human cells.

[0072] Alternatively or additionally, in some embodiments, the wax is biodegradable or degrades at a predetermined rate. In these embodiments, the wax does not need to be removed from the wound. In some embodiments, it takes about a week for the cells / tissue to engraft, and progressive degradation of the wax occurs during and / or beyond this period.

[0073] Alternatively or additionally, in some embodiments, the wax is permeable to air and / or water. Permeability can be achieved, for example, by providing small perforations in the wax sheet or strip. According to these embodiments, increasing the size or density (number per unit area) of the perforations contributes to increased permeability. In some embodiments, permeability contributes to an increased wound healing rate by allowing CO2 and water vapor released from cellular respiration in the healing wound to escape into the environment.

[0074] Exemplary wax composition In various exemplary embodiments of the present invention, different wax compositions are used.

[0075] In some embodiments, the wax comprises rosin (containing resin acids) and / or modified rosin and / or beeswax and / or paraffin and / or elastomer and / or other thermoplastic materials.

[0076] In other embodiments, the wax is a low-temperature self-supporting wax, such as one known to those skilled in the art from European Patent No. 029816. The low-temperature wax is heated to change from a solid consistency to a paste or fluid consistency.

[0077] In some exemplary embodiments of the present invention, a sugar wax is used. The sugar wax comprises polysaccharides or monosaccharides (sucrose, glucose, fructose, dextrose, maltose, honey, maple syrup, invert sugar, or a combination thereof), water, and citric acid. In some embodiments, the sugar wax is caramelized.

[0078] One exemplary wax composition comprises 60–85% by weight of rosin-type wax, 25–40% by weight of an aqueous solution, and 0.2–3% by weight of a surfactant or a mixture of surfactants. In some exemplary embodiments of the present invention, the rosin-type wax comprises a mixture of water and starch or cellulose pulp. In other exemplary embodiments of the present invention, the rosin-type wax comprises a mixture of modified rosin, liquid paraffin, beeswax, and microcrystalline wax. Alternatively or additionally, in some embodiments, a wax based on natural pine resin is used.

[0079] Alternatively or additionally, in some embodiments, the wax composition further comprises 0 to 10% by weight of one or more additives selected from the group consisting of fragrances, essential oils, emulsions, preservatives, analgesics, antibiotics, dyes, plasticizers, salts, emollients, coconut oil, and honey.

[0080] Alternatively or additionally, in some embodiments, the wax contains one or more surfactants. Surfactants are amphiphilic substances having affinity for both the lipophilic and hydrophilic phases of the emulsion, and by being positioned around the dispersed droplets of the emulsion, they can make the two phases miscible. Therefore, the addition of one or more surfactants, particularly water-in-oil surfactants, in appropriate proportions is advantageous in enabling the creation of a stable mixture. In some exemplary embodiments of the present invention, plant-derived glycolipid emulsifiers are used.

[0081] Exemplary dressing In some exemplary embodiments of the present invention, the wax is protected by a dressing material. In some embodiments, the dressing material includes a low-adhesion dressing material (e.g., gauze) and / or a semi-permeable film (e.g., TEGADERM). TM Contains chlorhexidine gluconate.

[0082] Alternatively or additionally, in some embodiments, the dressing material includes a hydrophilic colloid and / or hydrogel. The hydrophilic colloid and / or hydrogel contributes to the dressing material's ability to absorb exudate and / or maintain a moist environment. Examples of hydrophilic colloids and / or hydrogels include, but are not limited to, alginates, nonwoven fibers derived from seaweed, and collagen. Nonwoven fibers derived from seaweed and collagen are suitable for use in highly exudative wounds because of their ability to absorb large amounts of fluid.

[0083] Exemplary Usage Scenario Various exemplary embodiments of the present invention are used to treat chronic wounds in humans and / or animals, including but not limited to non-healing or infectious surgical or traumatic wounds, venous ulcers, arterial ulcers, pressure ulcers (e.g., bedsores, supine ulcers, or compression ulcers), diabetic foot ulcers, ischemic ulcers, epidermolysis bullosa, pyoderma gangrenosum, wounds after excision of giant congenital nevi, and acute wounds.

[0084] Alternatively or additionally, various exemplary embodiments of the present invention are used for rejuvenation of photodamaged skin after tattoo, birthmark, skin cancer, papilloma removal, post-mutilation, or skin resurfacing.

[0085] Alternatively or additionally, various exemplary embodiments of the present invention contribute to scar improvement by providing keratinocyte stem cells to wounds.

[0086] In general, this technique is suitable for any case where epithelial coverage of a wound is required.

[0087] Alternatively or additionally, in some embodiments, the graft / transplant provides pigmentation. In some embodiments, melanocytes contained in the hair follicle migrate to the depigmented area and re-establish themselves. In some embodiments, the depigmented area is prepared by deepithelialization to allow melanocytes to cover the wound with a pigmented surface layer.

[0088] During the term of this patent, it is anticipated that many new wax formulations will be developed, and the scope of the present invention is intended to a priori include all such new technologies.

[0089] As used herein, the term “approximately” means ±10%.

[0090] While the present invention has been described in conjunction with its specific embodiments, it is evident that many alternatives, modifications, and variations will be apparent to those skilled in the art. Therefore, it is intended to encompass all such alternatives, modifications, and variations that fall within the spirit and broad scope of the appended claims.

[0091] Specifically, various numerical indicators have been used. It should be understood that these numerical indicators may further vary based on the various engineering principles, materials, intended applications, and designs incorporated into the various embodiments of the present invention. Furthermore, components and / or operations belonging to exemplary embodiments of the present invention and shown as single units may be divided into subunits. Conversely, components and / or operations belonging to exemplary embodiments of the present invention and shown as subunits / individual operations may be combined into a single unit / operation having the described / illustrated function.

[0092] Alternatively, or additionally, features used to describe a method may be used to characterize an apparatus, and features used to describe an apparatus may be used to characterize a method.

[0093] Furthermore, it should be understood that the individual features and / or embodiments described above can be combined in all possible and partial combinations to produce further embodiments of the invention. The examples above are illustrative in nature and do not limit the scope of the invention, which is defined solely by the following claims.

[0094] Each enumeration of embodiments of the present invention that includes a specific feature, part, component, module, or process is an explicit statement that additional embodiments of the present invention exist that do not include the enumerated feature, part, component, module, or process.

[0095] Alternatively or additionally, various exemplary embodiments of the present invention exclude any particular features, parts, components, modules, processes, or elements not specifically disclosed herein.

[0096] In particular, the present invention is described in the context of autografts.

[0097] All publications, references, patents, and patent applications referenced herein are incorporated herein in whole by reference to the same extent that each individual publication, patent, or patent application is specifically and individually indicated as being incorporated herein by reference. Furthermore, any citation or identification of any reference in this application should not be construed as an admission that such reference is available as prior art to the present invention.

[0098] As used herein, the terms "include" and "have," as well as their conjugations, mean "to include, but not necessarily limit."

[0099] Further objectives, advantages, and novel features of various embodiments of the present invention will become apparent to those skilled in the art by considering the following examples, which are not intended to be limiting. Furthermore, each of the various embodiments and aspects of the present invention described above and in the following claims will find experimental support in the following examples. [Examples]

[0100] Herein, along with the above description, the following examples illustrating the present invention in a non-limiting manner will be referenced.

[0101] Example 1: Xenotransplantation from mouse to mouse To provide proof of concept, hair follicles were harvested from the backs of normal (hairy) mice using a commercially available wax formulation for female hair removal. The wax formulation contained rosin wax, beeswax, paraffin and film-forming agents, coconut oil and fragrance. The wax containing the harvested hair follicles was transplanted into open wounds created on the backs of nude mice. For comparison, wax containing no hair at all was also transplanted into wounds on the backs of other mice. Three weeks after transplantation, the healed wounds were harvested, and new epidermis, created from the migration of stem cells from the transplanted epidermis, was identified by histological analysis of the tissue.

[0102] This example illustrates that newly harvested hair follicles can spontaneously proliferate and form grafts in laboratory culture without prior cell proliferation. Nude mice were used to eliminate the possibility of an immune response. Elimination of potential immune responses allowed for testing the ability of allogeneically transplanted cells to "take" on the wound and contribute to the healing process. Between days 3 and 7, a moderate amount of exudate was observed on the compresses of mice in both experimental groups. Subsequently, the presence of exudate gradually decreased in mice transplanted with wax containing hair follicles, while in the other groups, the presence of exudate gradually increased until day 14 and then gradually decreased until day 28. Throughout the experimental period, no signs of infection / maceration were observed in mice transplanted with wax containing hair follicles. In mice transplanted with wax alone, signs of infection / maceration were observed between days 7 and 21, peaking on day 14. Less exudate and infection were obtained when using wax containing hair follicles.

[0103] Example 2: Xenotransplantation from human to mouse To confirm the results of Example 1, hair follicles were collected from human subjects using a commercially available wax formulation, as in Example 1. Again, the wax containing the collected hair follicles was transplanted into an open wound created on the back of a nude mouse. Three weeks after transplantation, the healed wound was collected, and the new epidermis produced from the migration of stem cells from the transplanted epidermis was identified by histological analysis of the tissue.

[0104] This example confirms the results of Example 1 and suggests that interspecies transplantation using the described method is feasible.

Claims

1. The following steps: (a) A step of collecting hair follicles by applying wax to a first area of ​​skin in which hair follicles protrude, and (b) The step of removing the wax from the first area and applying the wax to the wound in the second area of ​​the skin so that the hair follicle and its keratinocyte stem cells come into contact with the wound, Methods that include...

2. The method according to claim 1, comprising priming the hair follicle containing keratinocyte stem cells before applying it to the wound.

3. The method according to claim 1 or 2, comprising washing the hair follicle containing the wax and / or keratinocyte stem cells before applying it to the wound.

4. The method according to any one of claims 1 to 3, comprising treating the hair follicle containing the wax and / or keratinocyte stem cells with one or more antibiotics before applying it to the wound.

5. A wax composition to which hair follicles containing keratinocyte stem cells are attached, for use in a wound treatment method comprising applying the composition to a wound so that hair follicles containing keratinocyte stem cells come into contact with the wound.

6. The composition for use according to claim 5, wherein the hair follicle containing the keratinocyte stem cell remains attached to the composition during the application.

7. The composition for use according to claim 5 or 6, wherein the method comprises priming the hair follicle containing keratinocyte stem cells before applying it to the wound.

8. The composition for use according to any one of claims 5 to 7, wherein the method comprises washing the hair follicle containing the wax and / or keratinocyte stem cells before applying it to the wound.

9. The composition for use according to any one of claims 5 to 8, wherein the method comprises treating the hair follicle containing the wax and / or keratinocyte stem cells with one or more antibiotics before applying it to the wound.

10. A composition comprising a wax impregnated with at least one substance selected from the group consisting of antibiotics, growth factors, cytokines, and extracellular matrix molecules.

11. A product comprising a hair removal wax strip or sheet having perforations distributed on a surface that comes into contact with the skin during use.

12. A medical device comprising a wax composition to which hair follicles containing keratinocyte stem cells are attached, for the treatment of dermatological damage.

13. The medical device according to claim 12, characterized in that the dermatological injury is selected from the group consisting of non-healing or infectious surgical or traumatic wounds, venous ulcers, arterial ulcers, bedsores; pressure ulcers; or pressure ulcers, diabetic foot ulcers, ischemic ulcers, epidermolysis bullosa, pyoderma gangrenosum, and wounds following excision of giant congenital nevi.

14. The medical device according to claim 12, characterized in that the dermatological injury is selected from the group consisting of treatment of postoperative injuries after tattoo removal, birthmarks, skin cancer, and papilloma removal, as well as rejuvenation of photodamaged skin after amputation and skin resurfacing.

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