WO2025064646A1

WO2025064646A1 - Compositions and methods for wound healing and hair growth using polyelectrolyte complex (pec) films

Info

Publication number: WO2025064646A1
Application number: PCT/US2024/047445
Authority: WO
Inventors: Dhruv Patel; Suneel Kumar; Noshir A. LANGRANA; Francois Berthiaume; Rene S. Schloss; Priya MISTRY; Hwan June KANG
Original assignee: Rutgers State University of New Jersey
Current assignee: Rutgers State University of New Jersey
Priority date: 2023-09-19
Filing date: 2024-09-19
Publication date: 2025-03-27
Anticipated expiration: 2026-03-19

Abstract

Compositions and methods for the treatment of chronic wounds, including diabetic ulcers, and hair loss using PEC films are provided.

Description

COMPOSITIONS AND METHODS FOR WOUND HEALING AND HAIR GROWTH USING POLYELECTROLYTE COMPLEX (PEC) FILMS By Suneel Kumar Noshir A. Langrana Francois Berthiaume Rene S. Schloss Priya Mistry Dhruv Patel Hwan June Kang Cross-Reference to Related Applications This application claims priority to US Provisional Application Number 63/583,685 filed September 19, 2023, the entire contents being incorporated herein by reference as though set forth in full. Field of the Invention The invention relates to compositions and methods for wound healing and hair growth. More specifically, the invention provides methods and compositions effective for wound treatment and/or promotion of hair growth using polyelectrolyte complex (PEC) films. Background of the Invention Several publications and patent documents are cited throughout the specification in order to describe the state of the art to which this invention pertains. Each of these citations is incorporated herein by reference as though set forth in full. Chronic wound care is an important medical issue. Chronic wounds, such as diabetic, venous, or pressure ulcers place an immense financial cost on the healthcare system, while also causing pain and physical stress on the afflicted patients. Frequently, in order to heal chronic wounds a bandage or wound dressing, i.e., a bandage with healing factors applied in it, is applied to the wound in combination with antibiotic treatments to avoid, or combat infection. Such treatments are lengthy and are in many cases not particularly efficacious and thus may take months or even years to heal such chronic wounds, during which time the quality of life for the patient (and their family) is reduced. The burden on the healthcare system for these treatments is also quite high as many patients suffering from chronic wounds are elderly with compromised immune systems. In these patients, such wounds are associated with an increased risk of infection, thus leading to further complications such as sepsis and sometimes death. Other types of catastrophic wounds, e.g., from car accidents, gun shots and severe bed sores can also be associated with similar complications and in such cases, the importance of reducing infection in the wounds is equally important. Chronic, non-healing wounds of the skin are common complications in ~25% of diabetic patients. Current treatment focuses on preventing infection while simultaneously managing blood glucose. To date approved treatments for promoting healing of diabetic ulcerations are not yet available. Thus, there is an urgent need for the development of targeted treatments to treat chronic wounds. Additionally, chronic wounds can damage hair follicles which can lead to hair loss in the absence of hair follicles repair. Other causes of hair loss including overactive male hormones, excessive sebum secretion, poor blood circulation, peroxides, hypofunction of scalp due to bacteria, genetic factors, aging, and stress are also known. Drugs currently used as hair growth and hair tonic agents include vasodilators to promote increased blood flow to the scalp, female hormones to inhibit the action of male hormones, and anti-androgen to inhibit 5α-reductase which converts testosterone to 5-dihydrotestosterone (5-DHT). The vasodilators include carpronium chloride, minoxidil, and various plant extracts. The female hormones include estrogen, estradiol, and progesterone, while the anti-androgens includes finasteride and pentadecanoic acid. However, due to insufficient results or side effects of the hair loss treatment, development of more effective and safer treatments for all types of hair loss is required. Furthermore, in a modern society that values appearance, the need for prevention and treatment of both wound related hair loss and age-related hair loss is becoming more important. Summary of the Invention The present invention comprises methods of treating a chronic wound in a patient in need thereof comprising providing a complex of polyelectrolytes (PEC), which form a film, and introducing the film into a wound site for reducing wound healing time. In certain embodiments, said PEC complex forming a film comprises chitosan and polygalacturonic acid present in less than 50:50 ratio. In certain embodiments, said PEC complex forming a film comprises chitosan and polygalacturonic acids in a 40%-60% ratio. In certain embodiments, said film has a thickness between 75-250 microns, about 100 microns, or about 180 microns. In certain embodiments, said film exhibits anti-inflammatory properties. In certain embodiments, the composite is pre-formed prior to contacting the wound or forms in situ upon contact with the wound. In certain embodiments, said complex further comprises pectin or dextran sulphate. In certain embodiments, said complex comprises chitosan-polygalacturonic acid-pectin or chitosan-galacturonic acid- dextran sulphate. In certain embodiments, the complex comprises about 40% chitosan, about 55% polygalacturonic acid and about 5% pectin. In certain embodiments, said complex comprises about 40% chitosan, about 40% galacturonic acid and about 20% dextran sulphate. In certain embodiments, a second wound therapy is not introduced to said wound site. Another aspect of the invention comprises methods of treating hair loss in a patient in need thereof, comprising providing a complex of polyelectrolytes (PEC), which form a film, and introducing the film to a site exhibiting undesirable hair loss wherein the film promotes hair growth at the site. In certain embodiments, the methods further comprise introducing an agent that promotes hair growth to the site of hair loss. In certain embodiments, the agent that promotes hair growth is at least one that promotes blood circulation, antiandrogen, and corticosteroid. In certain embodiments, the agent that promotes hair growth is at least one of minoxidil, finasteride, cromakalin, pinacidil, naminidil, diphenylcyclopropenone, Tricomin therapeutic, cosmeceutical, cyproterone acetate, danazol, flutamide; a 5-alpha reductase inhibitor, turosteride, LY-191704, MK-306, dutasteride, s-triazines, benzopyrans, pyridinopyrans, thiane-1-oxides, Spironolactone (Aldactone), Cimetidine (Tagamet), Cyproterone Acetate, estrogen, progesterone, Ketoconazole (Nizoral), Finasteride (Propecia, Proscar), Dutasteride (Avodart), and Cyproterone Acetate with Ethinyloestradiol (Diane 35, Diane 50), clobetasol propionate, betamethasone valerate, triamcinolone acetonide, mometasone furoate, and hydrocortisone acetate. In certain embodiments, said PEC complex forming a film comprises chitosan and polygalacturonic acid present in less than 50:50 ratio. In certain embodiments, said PEC complex forming a film comprises chitosan and polygalacturonic acids in a 40%-60% ratio. In certain embodiments, said film has a thickness between 75-250 microns, about 100 microns, or about 180 microns. In certain embodiments, said film exhibits hair growth properties. In certain embodiments, the composite is performed prior to contacting the site of hair loss or in situ upon contacting the site. In certain embodiments, said complex further comprises pectin or dextran sulphate. In certain embodiments, said complex comprises chitosan-polygalacturonic acid-pectin or chitosan-galacturonic acid- dextran sulphate. In certain embodiments the complex comprises about 40% chitosan, about 55% polygalacturonic acid and about 5% pectin. In certain embodiments, said complex comprises about 40% chitosan, about 40% galacturonic acid and about 20% dextran sulphate. Another aspect of the invention includes hair growth compositions comprising a complex of polyelectrolytes, and at least one agent that promotes hair growth. In certain embodiments, the agent that promotes hair growth is at least one agent that promotes blood circulation, an antiandrogen, and a corticosteroid. In certain embodiments, the agent that promotes hair growth is at least one of minoxidil, finasteride, cromakalin, pinacidil, naminidil, diphenylcyclopropenone, Tricomin therapeutic, cosmeceutical, cyproterone acetate, danazol, flutamide; a 5-alpha reductase inhibitor, turosteride, LY-191704, MK-306, dutasteride, s- triazines, benzopyrans, pyridinopyrans, thiane-1-oxides, Spironolactone (Aldactone), Cimetidine (Tagamet), Cyproterone Acetate, estrogen, progesterone, Ketoconazole (Nizoral), Finasteride (Propecia, Proscar), Dutasteride (Avodart), and Cyproterone Acetate with Ethinyloestradiol (Diane 35, Diane 50), clobetasol propionate, betamethasone valerate, triamcinolone acetonide, mometasone furoate, and hydrocortisone acetate. In certain embodiments, the PEC comprises about 40% chitosan and about 60% galacturonic acid having a thickness between 150 and 200 microns. In certain embodiments, the PEC comprises about 40% chitosan, about 55% polygalacturonic acid and about 5% pectin and having a thickness of about 150-200 microns. In certain embodiments, the PEC comprises about 40% chitosan, about 40% galacturonic acid and about 20% dextran sulphate and having a thickness of about 150-200 microns. Another aspect of the invention includes wound healing compositions comprising a complex of polyelectrolytes. Another aspect of the invention consists of wound healing compositions consisting of a complex of polyelectrolytes. In certain embodiments, the composition does not include a second wound therapy. In certain embodiments, the composition includes a second wound therapy. said second wound therapy is at least one of cytokines, cells, antibiotic or antibacterial use, debridement, irrigation, negative pressure wound therapy, warming, oxygenation, moist wound healing, removing mechanical stress, adding cells, and an agent that enhances levels of healing factors. In certain embodiments, the agent that enhances levels of at least one of vascular endothelial growth factor (VEGF), monocyte chemoattractant protein-1 (MCP-1), alpha-melanocyte-stimulating hormone (α-MSH), fibroblast growth factor-2 (FGF-2), insulin-like growth factor (IGF), platelet-derived growth factor (PDGF), transforming growth factor-β (TGF-β), and epidermal growth factor (EGF). In certain embodiments, the composition further comprises at least one of a diluent, carrier, buffer, stabilizer, antioxidant, preservative. In certain embodiments, the PEC comprises about 40% chitosan and about 60% galacturonic acid having a thickness between 150 and 200 microns. In certain embodiments, the PEC comprises about 40% chitosan, about 55% polygalacturonic acid and about 5% pectin and having a thickness of about 150-200 microns. In certain embodiments, the PEC comprises about 40% chitosan, about 40% galacturonic acid and about 20% dextran sulphate and having a thickness of about 150-200 microns. In another aspect of the invention, methods of treating a pressure wound in a patient in need thereof are provided. In certain embodiments, the methods include providing a complex of polyelectrolytes (PEC), said complex forming a film; and introducing said film into a pressure wound site, said film being effective to reduce healing time of the pressure wound. In certain embodiments, said PEC complex forming a film comprises chitosan and polygalacturonic acid present in less than 50:50 ratio. In certain embodiments, said PEC complex forming a film comprises chitosan and polygalacturonic acids in a 40%-60% ratio. In certain embodiments, said film has a thickness between 75-250 microns, about 100 microns, or about 180 microns. In certain embodiments, said film exhibits anti-inflammatory properties. In certain embodiments, the composite is pre-formed prior to contacting the wound or forms in situ upon contact with the wound. In certain embodiments, said complex further comprises pectin or dextran sulphate. In certain embodiments, said complex comprises chitosan-polygalacturonic acid-pectin or chitosan-galacturonic acid- dextran sulphate. In certain embodiments, the complex comprises about 40% chitosan, about 55% polygalacturonic acid and about 5% pectin. In certain embodiments, said complex comprises about 40% chitosan, about 40% galacturonic acid and about 20% dextran sulphate. In certain embodiments, a second wound therapy is not introduced to said wound site. In certain embodiments, the pressure wound is a pressure ulcer or a skin lesion. In certain embodiments the wound is associated with or caused by a spinal cord injury (SCI). In certain embodiments, the patient has at least one pressure wound risk factor selected from paralysis, sensory impairment, changes in collagen metabolism, muscle atrophy, and altered circulation. In certain embodiments, the patient has at least one spinal cord comorbidity of infection, sepsis, autonomic dysreflexia, and spasticity. In certain embodiments, further comprising at least one secondary intervention. In certain embodiments, the secondary intervention is selected from psychosocial support, pressure relief, nutrition supplementation, infection eradication, and surgical intervention. Still other aspects and advantages of these compositions and methods for making the compositions and using the compositions are described further in the following detailed description of the preferred embodiments thereof. Brief Description of the Drawings FIGs.1A-C: Role of PEC films with or without nanoparticles in diabetic wound closure in mice. FIG.1A. Representative images are shown of skin wounds at different time points (left to right, days 0-42). Blue arrows indicate the wound contraction/closing and the appearance of hair around the wound in PEC and PEC+vRAGE-ELP groups only. The area of the wound was measured at studied time-points and presented as percent wound closure (FIGs.1B and 1C). Data are represented as mean ± SEM (n=6/each group). Data are analyzed statistically using repeated measures of two-way ANOVA followed by post hoc Tukey’s HSD test. G1-F-T gel; G2- F- T+vRAGE-ELP gel; G3-PEC film; G4- PEC+vRAGE-ELP film; NS-not significant. FIGs.2A-2D: The representative images (FIG.2A) show the progression of skin wound closure in diabetic full-thickness wounds at the study endpoint (day 42) in different treatment groups. Two red lines on the left and right indicate the wound width (first appearance of hair follicles). The top two groups have open wounds as there is no epidermis while the two red triangles indicate a better representation of hair follicles in the bottom two groups treated with PEC films. Scale bar = 2.6 mm. The graphs represent wound width (FIG. 2B), epidermis, and dermis thickness (FIGs.2C-2D) in PEC-treated groups only. Data are represented as mean ± SEM (n=6/each group). Statistical analysis was done using one-way ANOVA followed by Tukey’s HSD test and student t-test. ns-not significant. FIG.3: The effect of PEC on hair growth in diabetic wounds. Data show the total number of hair follicles 1 mm to the left and right side of wounds from the first appearance of hair follicle as shown in Figure 2 by red triangles. Data are represented as mean ± SEM (n=6/each group). Statistical analysis was done using one-way ANOVA followed by Tukey’s HSD test. *p<0.05. FIG.4: Representative from male mice images showing the progression of wound healing and hair growth around spinal cord injury in both PEC and PEC+ polyhethylene glycol (PEG) groups over 20 days. Both PEC and PEG groups showed healing faster than the control group. PEG showed the fastest healing with wound contraction/closing beginning within 3 days. FIG. 5: Graphical representation showing the healing of a spinal cord injury-induced wounds in male mice after treatment with a PEC film, PEG film, or no film (control). The results depict that PEC and PEG film show a statistically significant increase in wound healing over normal wound healing. The data was graphed as the mean ± SEM (n=4 for Control and PEC, n=5 for PEG). Statistical analysis was done using two-way ANOVA followed by Tukey’s multiple comparison test. *p<0.05, **p<0.01, ***p<0.001, and ****p<0.0001. The blue color asterisk represents the comparison between control and PEG while the black color asterisk represents the comparison between Control and PEC/PEG. FIG. 6A-6B: (FIG. 6A) Representative images of wound histology (trichrome staine) show wound closure in the male SCI mice. Edges of the wounds are bound by red lines. The dark blue lines in the PEC and PEG images indicate the regeneration of epidermis layer of skin. The double-headed white arrows in those images indicate the dermis. The image for the control group shows an open wound with a thin dermal layer and no epidermal layer formed yet. (FIG.6B) The trichrome stain images in FIG. 6A were analyzed using ImageJ to determine the wound width, epidermal thickness, and dermal thickness. The data was graphed as the mean ± SEM (n=4 for Control and PEC, n=5 for PEG). Statistical analysis was done using one-way ANOVA followed by Tukey’s multiple comparison test. * p<0.05, ** p<0.01; ns-not significant. FIG.7: Angiogenesis marker (CD31) representative immunostained images of wound center show the formation of new and more blood vessels in treated groups. Significant blood vessels are outlined in red. Scale bar = 200 µm. Quantitative data in graphs showing the count of CD31+ cells per square millimeter, percent area of CD31+ staining, and average length of the ten most outstanding blood vessels of each tissue sample in µm. Data shows the superior effect of PEC and PEG films compared to no film control in blood vessels formation while PEG film outperform the PEC films as well in certain parameters. * p<0.05, ** p<0.01. FIG.8: Progressive wound site images in the varying treatment groups in the SCI female mouse model showing the faster wound closure with PEC and PEG films over no film control. Scale bar = 1 cm. FIG.9: The wound site images were quantized using ImageJ to determine the wound area at each timepoint. Percent wound closure was calculated to normalize the wounds to the respective day 0 images. The data was graphed as the mean ± SEM (n=4 for Control, n=6 for PEC, and n=5 for PEG). Statistical analysis was done using two-way ANOVA followed by Tukey’s multiple comparison test. The blue color asterisk represents the comparison between control and PEG while the black color asterisk represents the comparison between Control and PEC/PEG. *p<0.05 and **p<0.01 Detailed Description of the Invention Disclosed herein are methods and compositions using polyelectrolyte complex (PEC) film to help promote wound healing and hair growth in patients, including patients suffering from diabetic ulcers. To achieve this, we prepared 25 genetically modified male diabetic mice with a 10 mm full-thickness excisional skin wound as a model of a diabetic ulcer. These wounds were immediately covered with one of four films and observed over six weeks with weekly data collection. The results show that the PEC film has a significant impact on wound healing for diabetic mice. The PEC film was also shown to initiate hair growth as early as 21 days after treatment. Thus, PEC agents enhance wound healing and initiate hair growth, thereby improving clinical and survivability outcomes for patients, particularly diabetic patients. The stimulation of hair growth was surprising and provides a new avenue for treating undesirable hair loss. Thus, PEC film comprising compositions and methods of use thereof for ameliorating hair loss are provided. Definitions: The following definitions are provided to facilitate an understanding of the present invention. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Generally, conventional methods of molecular biology, microbiology, recombinant DNA techniques, cell biology, and virology within the skill of the art are employed in the present invention. Such techniques are explained fully in the literature. For purposes of the present invention, "a" or "an" entity refers to one or more of that entity; for example, "an agent" refers to one or more agents or at least one agent. As such, the terms "a" or "an," "one or more" and "at least one" can be used interchangeably herein. It is also noted that the terms "comprising," "including," and "having" can be used interchangeably. Furthermore, a compound "selected from the group consisting of" refers to one or more of the compounds in the list that follows, including mixtures (i.e., combinations) of two or more of the compounds. The terms “subject,” “individual,” and “patient” are used interchangeably herein, and refer to an animal, for example a human, to whom treatment, including prophylactic treatment, with the PEC composition according to the present invention, is provided. The term “subject” as used herein refers to human and non-human animals. The terms “non-human animals” and “non- human mammals” are used interchangeably herein and include all vertebrates, e.g., mammals, such as non-human primates, (particularly higher primates), sheep, dog, rodent, (e.g., mouse or rat), guinea pig, goat, pig, cat, rabbits, cows, horses and non-mammals such as reptiles, amphibians, chickens, and turkeys. The term does not denote a particular age or sex. As used herein, the terms “component,” “composition,” “composition of compounds,” “compound,” “drug,” “pharmacologically active agent,” “active agent,” “therapeutic,” “therapy,” “treatment,” or “medicament” are used interchangeably herein to refer to a compound or compounds or composition of matter which, when administered to a subject (human or animal) induces a desired pharmacological and/or physiologic effect by local and/or systemic action. The terms “agent” and “test compound” denote a chemical compound, a mixture of chemical compounds, a biological macromolecule, or an extract made from biological materials such as bacteria, plants, fungi, or animal (particularly mammalian) cells or tissues. It is also contemplated that the term “compound” or “compounds” refers to the compounds discussed herein and includes precursors and derivatives of the compounds, and pharmaceutically acceptable salts of the compounds, precursors, and derivatives. The invention also includes prodrugs of the compounds, pharmaceutical compositions including the compounds and a pharmaceutically acceptable carrier, and pharmaceutical compositions including prodrugs of the compounds and a pharmaceutically acceptable carrier. The phrase "consisting essentially of" when referring to a particular nucleotide or amino acid means a sequence having the properties of a given SEQ ID NO. For example, when used in reference to an amino acid sequence, the phrase includes the sequence per se and molecular modifications that would not affect the functional and novel characteristics of the sequence. PEC Films PECs primarily consist of at least two oppositely charged polymers. Polyelectrolyte complex biomaterials, especially polysaccharide based, have a potential for wound healing, drug delivery, and tissue-engineering applications. PEC materials can be formed using different methods. Among the most common method is co-acervation of polymer solutions. In this method, one of the polymer solutions is added drop-wise to its complementary charged polymer solution under rapid agitation. This results in formation of a nano-colloidal PEC suspension in unreacted polymer mixer. The driving force for such complexation is entropy and strong electrostatic attraction between the complementary charged polymers. The parameters that affect the structure of the PEC particles are pH, ionic strength, and polymer mixing order. By comparison with current products, polyelectrolyte complex (PEC) based films provide the benefits of ease of handling when wet or dry and are strong. Furthermore, since the product is based on chitosan, it offers the additional physiological properties of homeostasis and accelerated healing of the chronic wounds. Polygalacturonic acid (PgA) is another biocompatible and biodegradable polymer, which provides negative surface charge to PEC. Another important aspect of these films is their anti-inflammatory property. The anti-inflammatory characteristics of chitosan/PgA films provide an additional mechanism to enhance chronic wound healing. Chitosan and PgA form polyelectrolyte complexes in solution. By varying their proportion, e.g., use of between 30%, 40%, 50%, 60% or 70% chitosan with proportional PgA, films with specific charge densities have been created. These PEC films have adequate mechanical strength and are useful to enhance wound healing and hair growth. PgA is a product of pectin degradation. Pectin is a plant polysaccharide primarily obtained from edible plants. Pectin contains poly(d-galacturonic acid) bonded via glycosidic linkage. Pectin also contains neutral sugars, which are either inserted in or attached to the main chains. Since PgA and Pectin share chemical similarity, in certain embodiments of this invention, PgA can be replaced by pectin. The molecular weights of chitosan and PgA polymers are design parameters. We can fabricate films with high to low molecular weights of polymers based on the film properties desired. Technically any water soluble polysaccharide or protein can be added to chitosan/polygalacturonic acid films. These polymers include for example alginate, dextran sulfate, collagen, gelatin, chitin, polylysine, heparin, hyaluronic acid, methoxy cellulose, and oxidized regenerated cellulose. The polymers can be added individually or in combination to alter the properties. Other useful polymers can be employed to create the films of the invention. These include, without limitation, 1. Pectin/chitosan/hydroxypropyl methylcellulose (Macleod G S, et al. Int J Pharm.1999; 187(2):251-7) 2. Pectin/Chitosan/Alginate (Yu C Y et al. Colloids and Surfaces B: Biointerfaces.2009; 68(2):245-9) 3. Pectin/Chitosan/Gelatin (Li J, et al. Journal of Biomedical Materials Research Part B: Applied Biomaterials.2010; 95(2):308-19) and 4. Pectin/Chitosan/Poly-l-lysine (Marudova M, et al. Carbohydr Res.2005; 340(13):2144-9). Polyelectrolyte complexes (PEC) primarily consist of at least two oppositely charged polymers. By keeping the concentration of polyanions (PgA) higher than polycations (chitosan), a film with an overall negative charge can be created. Chitosan/PgA films with lower than 50:50 ratios, are effective in achieving wound healing and hair growth. In certain embodiments, the ratio of Chitosan/PgA in the PEC films is between approximately 10:90-90:10, 20:80-80:20, 30:70-70:30, 40:60-70:30, or 40:60-60:40. In certain embodiments the ratio of Chitosan/PgA is 60:40. Additional polymers useful for the practice of the invention include three material composites, such as chitosan-PgA-pectin and chitosan-PgA dextran sulfate. These three material composites should possess improved properties such as mild stickiness. In certain embodiments, the chitosan will be kept between 10%-90% including at approximately 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%. In particularly preferred composites chitosan will be kept at approximately 40%. In certain embodiments, pectin and dextran sulphate will range from 0 to 20% and the remainder will be PgA. Accordingly, in certain embodiments, the PgA will be kept between 10%-90% including at approximately 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%. In particularly preferred composites PgA will be kept at approximately 60%. In another preferred embodiment, a chitosan (40%)-PgA (55%)-Pectin (5%) composite can be prepared. In yet another embodiment, a chitosan (40%)-PgA (40%)-Dextran sulphate (20%) can be prepared. The PEC film may include a pharmaceutically acceptable carrier. The term “pharmaceutically acceptable” means a material that is not biologically or otherwise undesirable, i.e., the material can be administered to a subject along with the selected compound without causing any undesirable biological effects or interacting in a deleterious manner with any of the other components of the pharmaceutical composition in which it is contained. Examples of the carrier may include alcohol, oil, surfactants, fatty acid, silicone oil, preservatives, humectants, moisturizers, viscosity modifiers, emulsions, stabilizers, sunscreens, color developers, fragrances, and diluents. Since specific compounds or compositions that may be used as the alcohol, oil, surfactants, fatty acid, silicone oil, preservatives, humectants, moisturizers, viscosity modifiers, emulsions, stabilizers, sunscreens, color developers, fragrances, and diluents are already known in the art, those skilled in the art may select and use the appropriate compounds or compositions. In certain embodiments, the PEC film is covered after it is administered. In certain embodiments, the PEC film is covered by a bandage. Compositions Comprising PEC Films for Hair Loss Also provided herein are compositions comprising PEC films for the treatment of hair loss or baldness. In certain embodiments the compositions comprising PEC films promotes the thickening of hair. Any of the PEC films discussed above can be used in these compositions. In certain embodiments, the composition contains PEC film without any other compound or agent. In certain embodiments, the composition further comprises at least one agent that promotes hair growth, such as minoxidil. The hair growth-promoting activity of the PEC may add and synergize with that of the agent that promotes hair growth, thereby resulting in a more pronounced effect, and/or the ability to use a lower dose of the agent thus decreasing side effects. Furthermore, the use of PEC provides for controlled release over a period of days, so that the user does not have to apply the agent on a daily basis, as is currently being done. Agents that promote hair growth, may also be delivered over time using the PEC film. The time scale of release can be varied by packaging the bioactive compounds into nanoparticles through self- assembly or incorporating the bioactive compound in manufactured nano-and micro-spheres. The phrase “agent that promotes hair growth” refers to any one of the numerous drugs may also be used for hair growth. Typical hair growth agents include agents that promote blood circulation, antiandrogens, corticosteroids and other agents. Agents that promote blood circulation include, minoxidil (also known as Rogaine) (6- amino-1,2-dihydro-1-hydroxy-2-imino-4-phenoxypyrimidine) disclosed in U.S. Pat. No. 5,215,894 and finasteride disclosed in U.S. Pat. No.5,215,894. Other agents include cromakalin, pinacidil, naminidil, diphenylcyclopropenone, Tricomin therapeutic, cosmeceutical, cyproterone acetate, danazol, flutamide; 5-alpha reductase inhibitors selected from the group consisting of turosteride, LY-191704, MK-306 and dutasteride; s-triazines, benzopyrans, pyridinopyrans and thiane-1-oxides. These agents help people with many different types of hair loss, including: male and female pattern hair loss, which is also known as androgenetic alopecia; alopecia areata, an autoimmune disease in which the body’s immune system attacks healthy tissues, including the hair follicles; telogen effluvium, in which hair falls out all over the scalp due to an interruption in the body’s cycle of hair production; anagen effluvium, or rapid hair loss resulting from medical treatment; hypotrichosis, which is a rare condition in which very little hair grows on the scalp and body; and some forms of cicatricial, or scarring, alopecia. Antiandrogens are medications that inhibit sex hormones that damage or destroy hair follicles and are particularly helpful in stopping female pattern baldness. Antiandrogens medications include, without limitation, Spironolactone (Aldactone), Cimetidine (Tagamet), Cyproterone Acetate, estrogen, progesterone, Ketoconazole (Nizoral), Finasteride (Propecia, Proscar), Dutasteride (Avodart), and Cyproterone Acetate with Ethinyloestradiol (Diane 35, Diane 50) . Corticosteroids are medications that are immune system suppressors that can counteract the effects of an autoimmune disease and allow hair to grow. Corticosteroids are particularly useful in the treatment of hair loss is associated with or caused by an autoimmune condition. Exemplary corticosteroids include, without limitation, clobetasol propionate, betamethasone valerate, triamcinolone acetonide, mometasone furoate, and hydrocortisone acetate. Other agents that promote hair growth are known in the art. For example, protein or peptide drugs are being used to treat hair loss. See, e.g., US Patent Application No. 2021/0244793 A1. In certain embodiments, the at least one agent is delivered over time using the PEC film. The time scale of release can be varied by packaging the bioactive compounds into nanoparticles through self-assembly or incorporating the bioactive compound in manufactured nano-and micro-spheres. The hair loss compositions of the present disclosure may be prepared in any formulation conventionally prepared in the art, and provided in the formulation selected from the group consisting of hair tonic, hair conditioner, hair essence, hair lotion, hair nutrition lotion, hair shampoo, hair conditioner, hair treatment, hair cream, hair nourishment cream, hair moisture cream, hair massage cream, hair wax, hair aerosol, hair pack, hair nourishment pack, hair soap, hair cleansing foam, hair oil, hair dryer, hair preservative, hair dye, hair wave agent, hair bleach, hair gel, hair glaze, hair dresser, hair lacquer, hair moisturizer, hair mousse, and hair spray, but is not limited thereto. In certain embodiments, the PEC film and the at least one agent that promotes hair growth act synergistically. The term “synergy” or “synergistic” refers to the interaction or cooperation of two or more substances, or other agents to produce a combined effect greater than the sum of their separate effects. Methods of Treating The term “preventing” as used herein refers to administering a compound prior to the onset of clinical symptoms of a disease or conditions so as to prevent a physical manifestation of aberrations associated with the disease or condition. The term “in need of treatment” as used herein refers to a judgment made by a caregiver (e.g., physician, nurse, nurse practitioner, or individual in the case of humans; veterinarian in the case of animals, including non-human mammals) that a subject requires or will benefit from treatment. This judgment is made based on a variety of factors that are in the realm of a care giver's expertise, but that includes the knowledge that the subject is ill, or will be ill, as the result of a condition that is treatable by the disclosed compounds. By “treatment” and “treating” is meant the medical management of a subject with the intent to cure, ameliorate, or stabilize, a pathological condition or disorder, particularly chronic wounds. This term includes active treatment, that is, treatment directed specifically toward the improvement of a disease, pathological condition, or disorder, and also includes causal treatment, that is, treatment directed toward removal of the cause of the associated disease, pathological condition, or disorder. In addition, this term includes palliative treatment, that is, treatment designed for the relief of symptoms from the chronic wound, rather than the curing of the wound, pathological condition, or disorder; and supportive treatment, that is, treatment employed to supplement another specific therapy directed toward the improvement of the associated disease, pathological condition, or disorder. It is understood that treatment, while intended to cure, ameliorate, or stabilize, a disease, pathological condition, or disorder, need not actually result in the cure, amelioration, or stabilization. The effects of treatment can be measured or assessed as described herein and as known in the art as is suitable for the disease, pathological condition, or disorder involved. Such measurements and assessments can be made in qualitative and/or quantitative terms. Thus, for example, characteristics or features of a disease, pathological condition, or disorder and/or symptoms of a disease, pathological condition, or disorder can be reduced to any effect or to any amount. In certain embodiments, administration of the compositions described herein modulate hair growth by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100%. As used herein, the terms "modulate", "modulating" or "modulation" refer to changing the rate at which a particular process occurs, inhibiting or promoting a particular process, reversing a particular process, and/or preventing the initiation of a particular process. Accordingly, if the particular process is hair loss, the term "modulation" includes, without limitation, decreasing the rate at which hair loss occurs; inhibiting hair loss; reversing hair loss (including thickening hair or increasing the number of hair follicles) and/or preventing further hair loss. Hair Loss Provided herein are methods of treatment of hair loss and/or regrowing lost hair. The methods provided herein are also methods of preventing further hair loss. The methods described herein comprise administering an effective amount at least one of the PEC films, or hair loss compositions comprising a PEC film, described above to the patient. The “effective amount” of the present invention, as related to methods of treating hair loss, refers to an amount that, when administered to an individual, represents an improvement, treatment, or prevention effect of promoting hair growth or reducing hair loss. The phrase “hair loss” refers to a condition that is marked by a slow loss of hair over time, which becomes more pronounced during the latter years of a person's life. This results in a thinning of the person's hair, which is undesirable because it is aesthetically less appealing. Signs of hair loss may include: gradual thinning of ones hair on top of one's head and circular or patchy bald spots. Hair loss may be a sign of age or a symptom of a more serious disorder. Cancer patients may experience a loss of hair due to chemotherapy treatment, or some patients may suffer from alopecia areata. Or a person may be genetically predisposed to hair loss with age, such as male pattern baldness. Some patients may suffer from trichotillomania, or an autoimmune disease, or the like, that causes hair loss. Other hair loss factors include, a family history, hormonal changes and medical conditions, medications and supplements, radiation therapy, stress, excessive hair treatments, significant weight loss and poor nutrition. But irrespective of the actual reasons causing the person's hair loss, it is usually desirable to regrow hair that has been lost. In certain embodiments, the method of treatment effectively suppresses symptoms associated with hair loss. In certain embodiments, symptoms of hair loss include, gradual thinning of hair on top of head, circular or patchy bald spots, sudden loosening of hair, and/or full-body hair loss. In certain embodiments, the PEC films presented above are administered topically to the patient at the site of hair loss. In certain embodiments, the PEC film is covered after administration to the patient. In certain embodiments, the PEC film administered in conjunction with micro-needling, thereby adding to the efficacy of the micro-needling treatment. Micro-needling is sometimes used as a method to stimulate hair growth and enhance the transport of drugs into the skin. In certain embodiments, micro-needling is not used in conjunction with the PEC film as skin injury is not necessary for the PEC film to promote hair growth. Chronic Wounds Provided herein are methods of treatment of chronic wounds. The methods described herein comprise administering an effective amount at least one of the PEC films described above to the patient. The “effective amount” of the present invention, as related to methods of treating chronic wounds, refers to an amount that, when administered to an individual, represents an improvement or treatment of the wound. Such amounts may be determined by with consideration of the age and/or weight of a patient, and further by the size, condition, location, and/or severity of the wound or wounds to be treated. The term “wound” is intended to refer to an injury to living tissue of a subject. Unless otherwise noted, embodiments referring to a wound include embodiments where the wound is one in which the skin is cut or broken. Further, embodiments referring to a wound include embodiments where the epidermis is broken. Still further, embodiments referring to a wound include embodiments where the dermis is cut or broken. In still other embodiments referring to a wound, the wound is a tissue other than of the skin. Unless otherwise noted, embodiments referring to a wound include embodiments where the wound is an ulcer. Still further, a wound may also be a burn. The burn may be of the third degree. The term “acute wound” refers to a wound which heals consistent with the timing or process conventional to the type and severity of the wound for the species of the subject. The term “chronic wound” refers to a wound which does not heal consistent with the timing or process conventional to the type and severity of the wound for the species of the subject. An exemplary chronic wound is a diabetic wound. The term “diabetic wound” refers to any wound in an individual having diabetes. The terms “hard-to-heal” or “refractory” refer to a wound which does not heal using conventional therapies available as of the filing date of this application. The terms “disease”, “disorder”, or “condition” are used herein to refer to any manifestations, symptoms, or combination of manifestations or symptoms, recognized or diagnosed as connected with a chronic wound, hard-to-heal wound, or diabetic wound. In certain embodiments, the method of treatment effectively reduces the healing time or increases the healing rate of the wound. In some embodiments, the method decreases the inflammation associated with wound or healing of the wound. In some embodiment, the method decreases the magnitude or extent of scarring. In certain embodiments, the healing time of the wound (e.g., the length of one or more of the inflammatory, proliferative, or remodeling phase of wound healing) is reduced by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 95%, compared to the healing time of the wound (e.g., the length of one or more of the inflammatory, proliferative, or remodeling phase of wound healing) that has not been treated. In an embodiment, the wound healing rate (e.g., the absolute area healed per day, the percentage of initial area healed per day, or the greatest average wound margin distance from the wound centre divided by the time to complete wound closure) is increased by at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 fold, compared to the healing rate of the wound that has not been treated. In certain embodiments, the PEC films presented above are administered topically to the patient at the site of injury. In certain embodiments, the PEC film is covered after administration to the patient. In certain embodiments, the PEC is administered alone. In certain embodiments, the PEC film is may be administered alone, as pharmaceutical compositions in combination with diluents and/or carriers and/or buffers and/or other components. In one embodiment, they may be administered in saline. In another embodiment, in a hydrogel. Compositions may include stabilizers, antioxidants, and/or preservatives. Compositions may include, e.g., neutral buffered saline or phosphate buffered saline. In certain embodiments, the PEC is administered without another wound therapy, such as polyethylene glycol, cytokines, cells, antibiotic or antibacterial use, debridement, irrigation, negative pressure wound therapy (vacuum-assisted closure), warming, oxygenation, moist wound healing, removing mechanical stress, and/or adding cells (e.g., keratinocytes) or other materials (e.g., artificial skin substitutes that have fibroblasts and/or keratinocytes in a matrix of collagen) to secrete or enhance levels of healing factors (e.g., vascular endothelial growth factor (VEGF), monocyte chemoattractant protein-1 (MCP-1), alpha-melanocyte-stimulating hormone (α-MSH), fibroblast growth factor-2 (FGF-2), insulin- like growth factor (IGF), platelet-derived growth factor (PDGF), transforming growth factor-β (TGF-β), and epidermal growth factor (EGF) or other agents. In certain embodiments a second wound therapy is used. In certain embodiments, the second wound therapy is polyethylene glycol (PEG). In certain embodiments, the PEG is present in an amount between 0.1M-0.2M. In certain embodiments, the PEG is present in in approximately 0.1M. Pressure wounds Provided herein are methods of treatment of a pressure wound in a patient. In certain embodiments, the pressure wound is a skin lesion or a pressure ulcer. In certain embodiments, the pressure wound is associated with or caused by a spinal cord injury (SCI). The methods described herein comprise administering an effective amount at least one of the PEC films described above to the patient. The “effective amount” of the present invention, as related to methods of treating a wound caused by a spinal cord injury in a patient, refers to an amount that, when administered to an individual, represents an improvement or treatment or in reduction in healing time of the wound. Such amounts may be determined by with consideration of the age and/or weight of a patient, and further by the size, condition, location, and/or severity of the wound or wounds to be treated. The term “wound” is intended to refer to an injury to living tissue of a subject. Unless otherwise noted, embodiments referring to a wound include embodiments where the wound is one in which the skin is cut or broken. Further, embodiments referring to a wound include embodiments where the epidermis is broken. Still further, embodiments referring to a wound include embodiments where the dermis is cut or broken. In still other embodiments referring to a wound, the wound is a tissue other than of the skin. Unless otherwise noted, embodiments referring to a wound include embodiments where the wound is an ulcer. The term “pressure ulcer” (also known as “bed sore,” “pressure sore,” or “decubitus ulcer”) is a tissue abnormality or lesion resulting from pressure imposed upon soft tissue underlying skin, fat, fascia, muscle, bone, or any combination thereof. Following prolonged periods of loading in compression, tension and/or shear, the soft tissue positioned between a bony prominence (e.g. the ischial tuberosities, trochanter, shoulder blades, sacrum) and an external surface (e.g. bed, wheelchair) begins to deform and break down. The sustained deformation of tissue and the occlusion of capillaries and ischemic reduction of blood flow to the loaded tissue region leads to a reduction of oxygen, nutrients, and removal of metabolic waste products, resulting in soft tissue breakdown. While the majority of pressure ulcers are located on the torso bony prominences, other locations such as the wrist, ankle malleolus, clavicles, ears, and nose can also be affected by pressure ulcers, particularly from medical devices attached to these locations (e.g., masks that include elastic and straps that apply pressure to ears and noses). Pressure ulcers are typically associated with individuals having compromised mobility or lack of sensation, such as the infirm, elderly and people suffering from spinal cord injury, stroke, bone and joint disease, vascular pathologies, tumors, and diabetes. People in intensive care units, hospital wards, or undergoing long surgical procedures are also at risk of developing pressure ulcers. Some medical devices on the skin can also apply sufficient sustained force to cause a pressure ulcer. For example, a patient lying on a return electrode pad for an extended surgery is at risk of developing a pressure ulcer. Similarly, a brace, splint, cast, collar, or strap can apply pressure to the body and cause a pressure ulcer. In certain embodiments, the method of treatment effectively reduces the healing time or increases the healing rate of the wound. In some embodiments, the method decreases the inflammation associated with wound or healing of the wound. In some embodiment, the method decreases the magnitude or extent of scarring. In certain embodiments, the healing time of the wound (e.g., the length of one or more of the inflammatory, proliferative, or remodeling phase of wound healing) is reduced by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 95%, compared to the healing time of the wound (e.g., the length of one or more of the inflammatory, proliferative, or remodeling phase of wound healing) that has not been treated. In an embodiment, the wound healing rate (e.g., the absolute area healed per day, the percentage of initial area healed per day, or the greatest average wound margin distance from the wound centre divided by the time to complete wound closure) is increased by at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 fold, compared to the healing rate of the wound that has not been treated. In certain embodiments, the PEC films presented above are administered topically to the patient at the site of injury. In certain embodiments, the PEC film is covered after administration to the patient. In certain embodiments, the PEC is administered alone. In certain embodiments, the PEC film is may be administered alone, as pharmaceutical compositions in combination with diluents and/or carriers and/or buffers and/or other components. In one embodiment, they may be administered in saline. In another embodiment, in a hydrogel. Compositions may include stabilizers, antioxidants, and/or preservatives. Compositions may include, e.g., neutral buffered saline or phosphate buffered saline. In certain embodiments, the PEC is administered without another wound therapy, such as polyethylene glycol, cytokines, cells, antibiotic or antibacterial use, debridement, irrigation, negative pressure wound therapy (vacuum-assisted closure), warming, oxygenation, moist wound healing, removing mechanical stress, and/or adding cells (e.g., keratinocytes) or other materials (e.g., artificial skin substitutes that have fibroblasts and/or keratinocytes in a matrix of collagen) to secrete or enhance levels of healing factors (e.g., vascular endothelial growth factor (VEGF), monocyte chemoattractant protein-1 (MCP-1), alpha-melanocyte-stimulating hormone (α-MSH), fibroblast growth factor-2 (FGF-2), insulin- like growth factor (IGF), platelet-derived growth factor (PDGF), transforming growth factor-β (TGF-β), and epidermal growth factor (EGF) or other agents. In certain embodiments a second wound therapy, including without limitation a therapy listed above, is used. In certain embodiments, the second wound therapy is polyethylene glycol (PEG). In certain embodiments, the PEG is present in an amount between 0.1M-0.2M. In certain embodiments, the PEG is present in in approximately 0.1M. Kits and Articles of Manufacture Any of the aforementioned products can be incorporated into a kit which may contain a PEC film, an agent that promotes hair growth, a pharmaceutically acceptable carrier, instructions for use, a container, a vessel for administration, or any combination thereof. Materials and Methods The following materials and methods are provided to facilitate the practice of the present invention. Manufacturing of Polyelectrolyte Complexes (PEC) Films: PEC films were made from a combination of medical-grade chitosan (98% deacetylated, ChitoLytic Inc.) and polygalacturonic acid (PgA) solutions (Sigma-Aldrich). To prepare the solutions, 300 mg of chitosan and Pga was added into 30 mL of double-deionized Picopure water in separate 50 ml centrifuge tubes.1.5 mL of HCl (1M) was added to the chitosan solution while 1 mL of NaOH (1M) was added to the PgA solution. Both the solutions were vortex quickly for 10 seconds each and then left shaking at 37°C on a rotating nutator for a minimum of 18 hours. After that, the pH of the chitosan and PgA solutions were recorded using a calibrated pH meter. The chitosan and PgA solutions were then combined in a 60:40 volumetric ratio and sonicated at 60% amplitude for three minutes. The pH of the combined solution was recorded using a calibrated pH meter again. The solution was then poured into a circular 37 mm radius PTFE (Teflon) evaporating dish and placed on a hot plate set at 41℃ for 32-36 hours. For incorporating nanoparticles, chitosan and PgA combined solution was prepared as mentioned above for PEC films. Following sonication, the targeted amount of protein nanoparticles (vRAGE-ELP, 2 µM/cm²) was introduced to the solution and sonicated at 60% amplitude similarly once (20 seconds) in ice-cold environment to prevent the protein nanoparticle degradation. The combined mixture was then poured similarly into the circular 37 mm radius evaporating dish and placed on a hot plate set at 41℃ for 32-36 hours. All the PEC films were then sterilized in an Ultraviolet (UV) Crosslinker for 15 minutes on each side and stored in a sterile pouch until animal experiments. In Vivo Animal Testing: All animal studies were done following a protocol approved by the Institutional Animal Care and Use Committee (IACUC) at Nelson Laboratories at Rutgers University. Genetically modified diabetic male mice (BKS. Cg-Dock7m+/+ Leprdb/J, Jackson Laboratory, Bar Harbor, ME, USA) were used at the age of 10 weeks. The day before surgery, mice were anesthetized by isoflurane inhalation. The dorsum was shaved using a clipper, and Nair™ cream was applied to remove residual hair. On the day of surgery, a full-thickness excisional skin wound was created on the back of mice. Briefly, the mice were anesthetized, and betadine scrub and 70% ethanol were applied alternatively on the back three times. Full-thickness skin was then excised by using 10 mm diameter biopsy punch and surgical scissors. A wound photograph was captured immediately after wounding (day 0) and mice were divided into the following treatment groups: F-T gel: After wounding, human fibrin (F, 80 µl of 6.25 mg/ml, Sigma Aldrich) and thrombin (T, 20 µl of 12.5 U/ml, Sigma Aldrich) solution were mixed and immediately applied onto the wound (total volume = 100 µl), then waited for 2 min for the gel to settle, and then the wound was covered with a transparent Tegaderm dressing. F-T+vRAGE-ELP gel: After wounding F-T gel was mixed along with vRAGE-ELP nanoparticles (2 µM of vRAGE-ELP included in the total volume of 100 µl) and immediately applied onto the wound, then waited for 2 min for the gel to settle, and then the wound was covered with transparent Tegaderm dressing. PEC film: Already prepared PEC film (1 cm X 1 cm) was quickly dipped in PBS for a fraction of a second, and immediately applied to the wound, and then the wound was covered with transparent Tegaderm dressing. PEC+vRAGE-ELP film: Already prepared PEC film (1 cm X 1 cm) laden with vRAGE-ELP (2 µM/cm²) was quickly dipped in PBS for a fraction of a second, and applied immediately to the wound, and then the wound was covered with transparent Tegaderm dressing. Animals were then returned to their respective cages and wound photographs were taken on post-wounding days 3, 7, 14, 21, 28, 35, and 42. In the end, animals were sacrificed, and wound tissues were harvested for histology. The photographs were used as qualitative data and ImageJ analysis of the wound area for each timepoint was used as quantitative data. Wound closure percentage was calculated using the formula 100 - (wound area at time point/ initial wound area *100) where day 0 data served as the initial wound area for each mouse. Histological Analysis: At the end of the study (day 42), the wound tissues were collected from each animal including the scar area, fixed in 10% formalin for ~72 hours. Samples were trimmed and cut at the center of the wound and processed at Rutgers Research Pathology Services center for histological analysis. Tissue samples were cut into 5 µm thin sections after being paraffin- embedded and stained with hematoxylin and eosin (H&E) along with trichrome stain. Tissue morphology was analyzed and photographed on glass slides under a light microscope. The wounds were studied for epidermal thickness, dermal thickness if wounds are closed otherwise not in open wounds where the epidermis is missing, and wound width (between the first appearance of hair follicles left and right) were measured using ImageJ. Hair follicles were counted on both sides of wounds from the 1 mm of the first hair follicle appearance. The following examples are provided to illustrate certain embodiments of the invention. They are not intended to limit the invention in any way. EXAMPLE I TREATMENT OF DIABETIC WOUNDS AND HAIR LOSS IN MICE As discussed in the methods above, 25 genetically modified male diabetic mice were shaved and given a 10 mm full-thickness excisional skin wound as a model of a diabetic ulcer. This study was conducted over six weeks with weekly data collection. These diabetic full-thickness wounds were treated once with different treatments immediately after wounding. Treatments include F-T gel, F-T gel + vRAGE-ELP, PEC film and PEC fil + vRAGE-ELP (FIG.1) The results show that the PEC films with or without nanoparticles have a significant impact on wound healing (p<0.001, days 21-42) as compared to either gel control (FIG.1). vRAGE-ELP in gel or PEC film shows an added trend for a slight improvement compared to their respective controls at a used concentration of 2 µM, however, the benefit was not statistically significant. On the other hand, vehicle groups (F-T and F- T+vRAGE-ELP gel) only healed up to 50% at day 42. Histological analysis showed the progression of skin wound closure in diabetic full- thickness wound after PEC film application (FIG.2). Wound closure was indicated by regeneration of the epidermis and fully formed dermis on day 42. Wound closure was only shown in wounds treated with PEC films with or without vRAGE-ELP nanoparticles. Comparatively, the wounds of the control group (F-T gel with or without vRAGE-ELP nanoparticles) are still open as indicated by the open epidermis wound. Therefore, the epidermis and dermis thickness was not calculated in these groups. The PEC films treated groups have a decreased wound width indicating better wound healing and less scar area. When the PEC film- treated groups were compared for the epidermis and dermis thickness, there is a better trend of thickness in the PEC+vRAGE-ELP film group as compared to PEC film. Along with the better wound healing phenomenon in the PEC film groups, we also observed the hair growth around the wounds from day 21 onward compared to the gel controls (FIG.1A). The hair growth is primarily localized to the skin area that was under the PEC films during treatment. Similarly, these changes were noticed in the histological analysis and prominent preservation of hair follicles in PEC-treated groups on both sides of wounds (FIG 2C). Therefore, we calculated hair follicles on both sides of wounds 1 mm from the first appearance of a hair follicle. We found that the total number of hair follicles is significantly increased in the PEC film-treated animals as compared to gel-treated controls (p<0.05). We found the effect of vRAGE-ELP nanoparticles added a slight effect on PEC film-treated animals (FIG.3). EXAMPLE II TREATMENT OF WOUNDS AND HAIR LOSS CAUSED BY SPINAL CORD INJURY (SCI) IN BOTH MALE AND FEMALE MICE The current example relates to the use of a PEC film to promote wound healing in patients with SCI. The PEC film is composed of 60% Chitosan, 40% polygalactouronic acid, optionally combined with 0.1M PEG. About one-third of people with new spinal cord injuries develop pressure ulcers during their initial hospitalization. A study that used the National Model Systems SCI database reported new pressure ulcers among 7.9% of persons in the first year after SCI and 8.9% in the second year. About 7–8% of deaths in the SCI population are related to a pressure sore related sepsis. There are several SCI related risk factors that put a patient at risk of developing pressure ulcers such as paralysis and sensory impairment, changes in collagen metabolism, muscle atrophy, and altered circulation. Once a pressure injury occurs, early wound care is essential to promote timely wound-healing. Comorbidities of infection, sepsis, autonomic dysreflexia and spasticity makes these wounds especially dangerous to persons with SCI. Current treatment is a multidisciplinary approach of fixing the cause of the pressure injury, psychosocial support, pressure relief, nutrition, eradicate infections, or surgical interventions. An exemplary PEC film used in this experiment comprises Chitosan, Chi (60%), Polygalacturonic acid, PgA (40%), 1M HCL (1.5 mL), 1M NaOH(1 mL), and, optionally, 0.1 M polyethylene glycol (PEG). These components were mixed using sonication and dried in a closed oven at 41ºC for 36-72 hours. Drying time was dependent on the amount of film being dried. In this experiment, a mouse model was created by introducing a spinal cord injury in the both male and female mice at the T9-T10 vertebrae. Next, an 8 mm wound was induced in the mouse model. Immediately after wound induction, the wound was then treated with, nothing (control), the PEC film dressing or a dressing comprising the combination of a PEC film with 0.1M PEG (PEG film). The dressing was kept over the wound throughout the study period. Wound closure and hair growth were then monitored for up to 20 days (FIG.4-5). The PEC film showed faster wound healing than the control group, with the combination of PEC film with PEG (PEG film) showing wound healing and hair growth immediately. Both the PEC and PEG showed almost complete healing by Day 14, and complete healing by Day 20. In contrast, the control group only showed ~50% wound closure by Day 20 (FIG.4-5). This study shows that a PEC film alone, or in combination with PEG significantly enhances the spinal cord injury-induced chronic wound healing and preserve the hair growth around the scar. Histological analysis of wound scar showed that the PEC and PEG films completely healed the SCI-induced pressure wound while in no film control wound remained open without epidermis (FIG.6A). In addition, PEG film significantly reduced the wound width and increased the thickness of epidermal and dermal layer of skin along with PEC film (FIG.6B). This data further emphasizes the improved wound healing in SCI mice using PEC and/or PEG films. Immunohistochemistry of these skin wound at 20 days suggests significant improvement in angiogenesis using the PEG film along with PEC film over the non-film control (FIG.7). PEG film improved the formation of more and larger blood vessels during the wound healing process. In addition, other markers such as for macrophages (F4/80), proliferation (Ki67), and remodeling connective tissue (Alpha-smooth muscle actin, α-SMA) show the improved wound healing processes using PEC and PEG film in SCI mice (data not shown). In the SCI female mice, wound healing was almost completed by day 10. Wound healing was complete by Day 14. In contrast, the control group only showed ~55% wound closure by Day 14 (FIG.8-9). This study shows that a PEC film alone, or in combination with PEG significantly enhances the spinal cord injury-induced chronic wound healing and preserve the hair growth around the scar in female SCI mice. However, PEG film showed superior response on wound healing starting from day 10. EXAMPLE III DIAGNOSIS/ TREATMENT OF PATIENT WITH CHRONIC WOUNDS OR PRESSURE WOUNDS The information herein above can be applied clinically to patients for therapeutic intervention as part of a treatment of for chronic wounds, such as diabetic ulcers, burns, and scar tissue; or pressure wounds, such as pressure ulcers. In cases of skin burns that extend deep into the dermis, the scar tissue does not reconstitute skin appendages, including hair follicles. The use of the film can be used to help restore these features, thus improving appearance as well as the thermoregulatory function of the skin. A preferred embodiment of the invention comprises clinical application of the PEC film recited herein to a patient. This can occur after a patient arrives in the clinic and presents with symptoms of a chronic wound. In certain embodiments, the chronic wound is a diabetic wound, burn, or an ulcer. In certain embodiments, the pressure wound is a pressure ulcer. The PEC film can be applied directly to the wound and covered with transparent dressing to observe wound healing. In certain embodiments, the PEC films were prepared from a 60:40 chitosan to polygalacturonic acid (PgA) solution ratio. In certain embodiments, administration of the PEC film does not include a wound healing drug. In certain embodiments, administration of the PEC film does not include administration of a drug-packed nanoparticle. In certain embodiments, administration of the PEC film does not include the delivery of elastin-like polypeptides (ELPs) or ELPs conjugated with stromal cell-derived growth factor-1. In certain embodiments, administration of the PEC film includes delivery of PEG. In certain embodiments, the treatment may include administering an additional wound healing factor, including without limitation, those listed hereinabove. In certain embodiments, administration of the PEC film enhances wound healing and results in a reduction in wound size. After administration, patients may be assessed daily, weekly, every 2 weeks, every 4 weeks, and/or every two months. This assessment may continue for 1 year or longer. Furthermore, the information herein above can be applied clinically to patients for diagnosing the presence of, or an increased susceptibility for developing a chronic wound, such as a diabetic ulcer. A preferred embodiment of the invention comprises clinical application of the information described herein to a patient. This can occur after a patient arrives in the clinic and is diagnosed with a chronic wound. Kits for performing the method of the invention are also provided herein. After diagnosis, the patient can be treated as discussed above. EXAMPLE IV DIAGNOSIS ANDTREATMENT OF PATIENT WITH HAIR LOSS The information herein above can be applied clinically to patients for therapeutic intervention as part of a treatment for hair loss. The use of the film can be used to help restore or thicken hair, thus improving appearance as well as the thermoregulatory function of the skin. A preferred embodiment of the invention comprises clinical application of the PEC film recited herein to a patient having hair loss. This can occur after a patient arrives in the clinic and presents with symptoms of hair loss. In certain embodiments, the hair loss is age-related hair loss. The PEC film can be applied directly to the wound and covered with transparent dressing to observe hair growth. In certain embodiments, the PEC films were prepared from a 60:40 chitosan to polygalacturonic acid (PgA) solution ratio. Due to the PEC film's ability to release bioactive compounds, the PEC could also be used to deliver agents that promote hair growth, such as minoxidil. The hair growth-promoting activity of the PEC may add and synergize with that of the minoxidil, thus resulting in a more pronounced effect, and/or the ability to use a lower dose of the minoxidil thus decreasing side effects. Furthermore, the use of PEC may provide for controlled release over a period of days, so that the user does not have to apply the minoxidil on a daily basis, as is currently being done. Other bioactive compounds, such as growth factors and chemokines, may also be delivered over time using the PEC film. The time scale of release can be varied by packaging the bioactive compounds into nanoparticles through self-assembly (as in the case of elastin peptide-based nanoparticles) or incorporating the bioactive compound in manufactured nano-and micro-spheres (for example in poly-lactic-glycolic acid particles). In certain embodiments, the PEC film is used in conjunction with micro-needling, which may add to the efficacy of the micro-needling treatment. Micro-needling is sometimes used as a method to stimulate hair growth and enhance the transport of drugs into the skin. In certain embodiments, micro-needling is not used in conjunction with the PEC film as skin injury is not necessary for the PEC film to promote hair growth. In certain embodiments, administration of the PEC film enhances hair growth and/or decreases the rate of balding. After administration, patients may be assessed daily, weekly, every 2 weeks, every 4 weeks, and/or every two months. This assessment may continue for 1 year or longer. Furthermore, the information herein above can be applied clinically to patients for diagnosing the presence of, or an increased susceptibility for developing hair loss. A preferred embodiment of the invention comprises clinical application of the information described herein to a patient. This can occur after a patient arrives in the clinic and is diagnosed with hair loss or a propensity for developing hair loss. Kits for performing the method of the invention are also provided herein. After diagnosis, the patient can be treated as discussed above. While certain features of the invention have been described herein, many modifications, substitutions, changes, and equivalents will now occur to those of ordinary skill in the art. It is, therefore, to be understood that the appended claims are intended to cover all such modifications and changes as fall within the true spirit of the invention.

Claims

What is claimed is: 1. A method of treating a chronic wound in a patient in need thereof comprising: a. providing a complex of polyelectrolytes (PEC), said complex forming a film; and b. introducing said film into a wound site, said film being effective to reduce wound healing time.

2. The method of claim 1, wherein said PEC complex forming a film comprises chitosan and polygalacturonic acid present in less than 50:50 ratio.

3. The method of claim 1, wherein said PEC complex forming a film comprises chitosan and polygalacturonic acids in a 40%-60% ratio.

4. The method of any one of claims 1-3 wherein said film has a thickness between 75-250 microns.

5. The method of claim 4, wherein said film has a thickness of about 100 microns.

6. The method of claim 4, wherein said film has a thickness of about 180 microns.

7. The method of anyone of claims 1-6, wherein said complex exhibits anti-inflammatory properties.

8. The method of anyone of claims 1-7, wherein the composite is preformed prior to contacting the wound.

9. The method of anyone of claims 1-7, wherein the composite is formed in situ upon contact with the wound.

10. The method of anyone of claims 1-9, wherein said complex further comprises pectin or dextran sulphate.

11. The method of claim 10, wherein said complex comprises chitosan-polygalacturonic acid-pectin.

12. The method of claim 11, wherein said complex comprises about 40% chitosan, about 55% polygalacturonic acid and about 5% pectin.

13. The method of claim 10, wherein said complex comprises chitosan-galacturonic acid- dextran sulphate.

14. The method of claim 13, wherein said complex comprises about 40% chitosan, about 40% galacturonic acid and about 20% dextran sulphate.

15. The method of any one of claims 1-14, wherein a second wound therapy is not introduced to said wound site.

16. A method of treating hair loss in a patient in need thereof comprising: a. providing a complex of polyelectrolytes (PEC), said complex forming a film; and b. introducing said film to a site of hair loss, said film being effective to promote hair growth.

17. The method of claim 16, further comprising introducing an agent that promotes hair growth at said site.

18. The method of claim 17, wherein the agent that promotes hair growth is at least one of a blood circulation promoting agent, an antiandrogens, and a corticosteroid.

19. The method of claim 16 or claim 17, wherein the agent that promotes hair growth is at least one of minoxidil, finasteride, cromakalin, pinacidil, naminidil, diphenylcyclopropenone, Tricomin therapeutic, cosmeceutical, cyproterone acetate, danazol, flutamide; a 5-alpha reductase inhibitor, turosteride, LY-191704, MK-306, dutasteride, s-triazines, benzopyrans, pyridinopyrans, thiane-1-oxides, Spironolactone (Aldactone), Cimetidine (Tagamet), Cyproterone Acetate, estrogen, progesterone, Ketoconazole (Nizoral), Finasteride (Propecia, Proscar), Dutasteride (Avodart), and Cyproterone Acetate with Ethinyloestradiol (Diane 35, Diane 50), clobetasol propionate, betamethasone valerate, triamcinolone acetonide, mometasone furoate, and hydrocortisone acetate.

20. The method of claim 16 or claim 17, wherein the agent that promotes hair growth is at least one of minoxidil and finasteride.

21. The method of anyone of claims 16-21, wherein said PEC complex forming a film comprises chitosan and polygalacturonic acid present in less than 50:50 ratio.

22. The method of anyone of claims 16-21, wherein said PEC complex forming a film comprises chitosan and polygalacturonic acids in a 40%-60% ratio.

23. The method of claim 22 wherein said film has a thickness between 75-250 microns.

24. The method of claim 23, wherein said film has a thickness of about 100 microns.

25. The method of claim 23, wherein said film has a thickness of about 180 microns.

26. The method of anyone of claims 16-25, wherein said complex exhibits hair growth properties.

27. The method of anyone of claims 16-26, wherein the composite is preformed prior to contacting the site of hair loss.

28. The method of anyone of claims 16-26, wherein the composite is formed in situ upon contact with the site of hair loss.

29. The method of anyone of claims 16-28, wherein said complex further comprises pectin or dextran sulphate.

30. The method of claim 29, wherein said complex comprises chitosan-polygalacturonic acid-pectin.

31. The method of claim 30, wherein said complex comprises about 40% chitosan, about 55% polygalacturonic acid and about 5% pectin.

32. The method of claim 29, wherein said complex comprises chitosan-galacturonic acid- dextran sulphate.

33. The method of claim 32, wherein said complex comprises about 40% chitosan, about 40% galacturonic acid and about 20% dextran sulphate.

34. A hair growth composition comprising a complex of polyelectrolytes, and at least one agent that promotes hair growth.

35. The hair growth composition of claim 34, wherein the agent that promotes hair growth is at least one selected from agents that promote blood circulation, antiandrogens, and corticosteroids.

36. The hair growth composition of claim 34, wherein the agent that promotes hair growth is at least one of minoxidil, finasteride, cromakalin, pinacidil, naminidil, diphenylcyclopropenone, Tricomin therapeutic, cosmeceutical, cyproterone acetate, danazol, flutamide; a 5-alpha reductase inhibitor, turosteride, LY-191704, MK-306, dutasteride, s-triazines, benzopyrans, pyridinopyrans, thiane-1-oxides, Spironolactone (Aldactone), Cimetidine (Tagamet), Cyproterone Acetate, estrogen, progesterone, Ketoconazole (Nizoral), Finasteride (Propecia, Proscar), Dutasteride (Avodart), and Cyproterone Acetate with Ethinyloestradiol (Diane 35, Diane 50), clobetasol propionate, betamethasone valerate, triamcinolone acetonide, mometasone furoate, and hydrocortisone acetate.

37. The hair growth composition of claim 34, wherein the agent that promotes hair growth is at least one of minoxidil and finasteride.

38. The hair growth composition of any one of claims 34-37, wherein the PEC comprises about 40% chitosan and about 60% galacturonic acid having a thickness between 150 and 200 microns.

39. The hair growth composition of anyone of claims 34-37, wherein the PEC comprises about 40% chitosan, about 55% polygalacturonic acid and about 5% pectin and having a thickness of about 150-200 microns.

40. The hair growth composition of any one of claims 34-37, wherein the PEC comprises about 40% chitosan, about 40% galacturonic acid and about 20% dextran sulphate and having a thickness of about 150-200 microns.

41. A wound healing composition comprising a complex of polyelectrolytes.

42. A wound healing composition consisting of a complex of polyelectrolytes.

43. The wound healing composition of claim 41, wherein said composition does not include a second wound therapy.

44. The wound healing composition of claim 41, wherein said composition includes a second wound therapy.

45. The wound healing composition of claim 44, wherein said second wound therapy is at least one of cytokines, cells, antibiotic or antibacterial use, debridement, irrigation, negative pressure wound therapy, warming, oxygenation, moist wound healing, removing mechanical stress, adding cells, and an agent that enhances levels of healing factors.

46. The wound healing composition of claim 45, wherein the agent enhances levels of at least one of vascular endothelial growth factor (VEGF), monocyte chemoattractant protein-1 (MCP-1), alpha-melanocyte-stimulating hormone (α-MSH), fibroblast growth factor-2 (FGF-2), insulin-like growth factor (IGF), platelet-derived growth factor (PDGF), transforming growth factor-β (TGF-β), and epidermal growth factor (EGF).

47. The wound healing composition of any one of claims 41-46, further comprising at least one of a diluent, carrier, buffer, stabilizer, antioxidant, preservative.

48. The wound healing composition of any one of claims 41-47, wherein the PEC comprises about 40% chitosan and about 60% galacturonic acid having a thickness between 150 and 200 microns.

49. The wound healing composition of any one of claims 41-48, wherein the PEC comprises about 40% chitosan, about 55% polygalacturonic acid and about 5% pectin and having a thickness of about 150-200 microns.

50. The wound healing composition of any one of claims 41-49, wherein the PEC comprises about 40% chitosan, about 40% galacturonic acid and about 20% dextran sulphate and having a thickness of about 150-200 microns.

51. A method of treating a pressure wound in a patient in need thereof comprising: a. providing a complex of polyelectrolytes (PEC), said complex forming a film; and b. introducing said film into a pressure wound site, said film being effective to reduce healing time of the pressure wound.

52. The method of claim 51, wherein said PEC complex forming a film comprises chitosan and polygalacturonic acid present in less than 50:50 ratio.

53. The method of claim 51, wherein said PEC complex forming a film comprises chitosan and polygalacturonic acids in a 40%-60% ratio.

54. The method of any one of claims 51-53 wherein said film has a thickness between 75-250 microns.

55. The method of claim 54, wherein said film has a thickness of about 100 microns.

56. The method of claim 54, wherein said film has a thickness of about 180 microns.

57. The method of any one of claims 51-56, wherein said complex exhibits anti- inflammatory properties.

58. The method of any one of claims 51-57, wherein the composite is preformed prior to contacting the pressure wound.

59. The method of any one of claims 51-58, wherein the composite is formed in situ upon contact with the wound.

60. The method of claim 51-59, wherein said complex further comprises pectin or dextran sulphate.

61. The method of claim 60, wherein said complex comprises chitosan-polygalacturonic acid-pectin.

62. The method of claim 61, wherein said complex comprises about 40% chitosan, about 55% polygalacturonic acid and about 5% pectin.

63. The method of any one of claims 60-62, wherein said complex comprises chitosan- galacturonic acid- dextran sulphate.

64. The method of claim 63, wherein said complex comprises about 40% chitosan, about 40% galacturonic acid and about 20% dextran sulphate.

65. The method of claim 61, wherein a second wound therapy is not introduced to said wound site.

66. The method of any one of preceding claims 51-65, wherein the pressure wound is a pressure ulcer or a skin lesion.

67. The method of any one of claims 51-66, wherein said wound is an injury is associated with or caused by a spinal cord injury.

68. The method of any one of claims 51-67, wherein the patient has at least one pressure wound risk factor selected from paralysis, sensory impairment, changes in collagen metabolism, muscle atrophy, and altered circulation.

69. The method of claim any one of claims 51-67, wherein the patient has at least one spinal cord comorbidity of infection, sepsis, autonomic dysreflexia, and spasticity.

70. The method of any one of claims 51-68, further comprising at least one secondary intervention.

71. The method of claim 70, wherein the at least one secondary intervention is selected from psychosocial support, pressure relief, nutrition supplementation, infection eradication, and surgical intervention.