A collagen material for facial filling

Abstract

The present invention relates to the field of biomaterials, specifically a collagen material comprising recombinant humanized collagen type 21 (ColpepA1 21) and its application in facial augmentation. This invention utilizes collagen to not only provide an effective solution to skin aging but also promotes the natural synthesis of collagen within the skin, optimizing skin hydration and barrier function, and enhancing the skin's self-repair capacity. The resulting collagen material is a facial augmentation material. Through local injection or external application, it rapidly smoothes and firms the facial skin, while also improving overall skin health with long-term use.

Description

Technical Field

[0001] The present invention belongs to the field of biomaterials, and in particular relates to a collagen material and its application in facial filling. Background Art

[0002] Collagen, the most abundant protein in the human body, not only makes up over 30% of the body's total protein, but is also a key element supporting the structure and function of many tissues. It is widely distributed in connective tissues such as skin, bones, tendons, and blood vessels, forming a three-dimensional network that provides tissues with the necessary mechanical strength and stability. Collagen plays a vital role in skin health, maintaining the skin's firmness and elasticity, as well as maintaining the smoothness and radiance of the skin's surface. However, with age, the natural production of collagen gradually slows. Combined with the effects of ultraviolet radiation, environmental pollution, and unhealthy lifestyle habits, existing collagen gradually degrades and becomes damaged, leading to the appearance of sagging, fine lines, and wrinkles on the skin. This not only affects the skin's appearance but can also negatively impact bone density, tendon elasticity, and vascular stability, ultimately affecting overall health.

[0003] In facial filler products, the use of collagen is intended to directly replenish the collagen needed by the skin to fill the volume lost due to collagen loss, restore skin firmness and elasticity, and reduce wrinkles and fine lines, thereby achieving a rejuvenating effect. Leveraging its natural role in the skin's structure and biocompatibility, collagen is an ideal choice for facial fillers and anti-aging treatments. However, current collagen facial filler products face challenges such as improving collagen's stability, extending its persistence in the skin, and enhancing its permeability and bioavailability. Furthermore, the precise matching and replenishment of different types of collagen required during the aging process is also a key focus for further optimization and improvement of existing products. Current scientific research and product development are dedicated to addressing these issues through innovative technologies and formula improvements to achieve more effective and long-lasting collagen replenishment and anti-aging effects.

[0004] In this context, in-depth research on specific collagen types, such as type 21 collagen (COL21A1), is particularly important. Type 21 collagen is a non-fibrillar collagen that is relatively rare in the adult human body, comprising less than 1% of the total collagen composition. Compared to other, more well-known collagen types, type 21 collagen has been relatively understudied. Type 21 collagen belongs to the small collagen family. As part of the FACIT (fibril-associated collagen with interrupted triple helices) family, type 21 collagen plays a crucial role in connecting extracellular matrix components. Studies by Chou MY and Li HC suggest that it may play a key role in the assembly of the extracellular matrix during angiogenesis [Chou MY, Li HC (March 2002)]. Furthermore, the gene expression of type 21 collagen is developmentally regulated, with elevated expression levels in the fetus, suggesting that it may play a crucial role in the development of various tissues. Surendran et al.'s study revealed its role in cardiac and aortic collagen formation, as well as its association with vascular remodeling and hypertension, further pointing to its potential value in promoting skin health and anti-aging [Surendran et al. (2016)]. This provides a new perspective for the study of COL21A1 in promoting skin health and anti-aging.

[0005] Although collagen materials on the market primarily rely on traditional collagen or its derivatives to replenish collagen loss in the skin, the effectiveness of these traditional products is often limited by the size of the collagen molecules and the skin's absorption rate, and they are difficult to optimize for the unique biological functions of specific types of collagen. However, based on the unique biological properties and mechanism of action of type 21 collagen, facial filler products developed are expected to surpass traditional solutions and provide more targeted, highly bioavailable, and multi-functional anti-aging therapies. These products not only improve the appearance of the skin, but also promote overall health and slow the aging process through their effects on extracellular matrix composition and vascular health, representing an important direction for the future development of facial filler technology. Summary of the Invention

[0006] The present invention relates to a collagen material and application thereof in facial filling.

[0007] In one aspect, the present invention provides a type 21 recombinant humanized collagen, wherein the amino acid sequence of the collagen is selected from SEQ ID No. 1:

[0008] GKPGLQGPKGDPGLPGNPGYPGQPGQDGKPGYQGIAGTPGVPGSPGIQGARGLPGYKGEPGRDGDKGDRGLPGFPGLHGMPGSKGEMGAKGDKGSPGFYGKKGAKGEKGNAGFPGL PGPAGEPGRHGKDGLMGSPGFKGEAGSPGAPGQDGTRGEPGIPGFPGNRGLMGQKGEIGPPGQQGKKGAPGMPGLMGSNGSPGQPGTPGSKGSKGEPGIQGMPGASGLKGEPGATG

[0009] This specific amino acid sequence of collagen is optimized to increase its bioavailability and efficacy in the skin.

[0010] In one embodiment, the collagen of the present invention can enhance skin moisture regulation and optimize barrier function.

[0011] In one embodiment, the collagen of the present invention can promote the expression of the water channel protein AQP3, thereby improving skin dryness and tightness by improving the skin's water retention capacity and reducing the water evaporation rate, thereby promoting overall skin health and anti-aging effects.

[0012] In one embodiment, the collagen described in the present invention can reduce fine lines and wrinkles on the skin, increase skin firmness, and provide key support for anti-aging and skin rejuvenation.

[0013] In one embodiment, the collagen of the present invention can promote the production of type III collagen and significantly increase the content of type III collagen. The increased type III collagen content can enhance skin elasticity and repair ability, especially showing significant benefits in promoting wound healing and combating skin aging.

[0014] In one embodiment, the collagen of the present invention enhances the structural integrity of the skin and improves elasticity by promoting the production of type I collagen and type III collagen, thereby reducing signs of aging and promoting skin health.

[0015] In one embodiment, the collagen of the present invention demonstrates potential as an active ingredient in facial filler products in an in vitro skin model after UVA and UVB irradiation.

[0016] In one embodiment, the collagen of the present invention exhibits protective and repairing effects on skin damage.

[0017] In one embodiment, the collagen of the present invention helps increase the thickness of the epidermal living cell layer.

[0018] In one embodiment, the collagen of the present invention helps to increase the density of collagen fibers.

[0019] In one embodiment, the collagen of the present invention helps to increase the content of type I collagen.

[0020] In one embodiment, the collagen of the present invention helps to increase the content of type III collagen.

[0021] In one embodiment, the collagen of the present invention helps to increase the content of type IV collagen.

[0022] In one embodiment, the collagen of the present invention helps to increase the content of type XVII collagen.

[0023] Another aspect of the present invention provides a collagen material for facial filling, wherein the collagen material for facial filling comprises recombinant humanized collagen type 21 (ColpepA1 21), the amino acid sequence of the collagen being selected from SEQ ID No. 1, and further comprises one or more auxiliary ingredients selected from the group consisting of a filling aid, a solvent, an antioxidant, an emulsifier, a moisturizer, a stabilizer, a pH adjuster, a thickener, and the like;

[0024] Preferably, the filling aid comprises hyaluronic acid, calcium hydroxyapatite, polylactic acid, platelet-rich plasma, polymethyl methacrylate microspheres, liquid injectable silicone, and the like.

[0025] In one embodiment, the collagen material of the present invention has a concentration of collagen of 0.0001% to 50% based on the total weight of the collagen material.

[0026] In one embodiment, the collagen material of the present invention, wherein the weight ratio of the collagen to the auxiliary components is selected from one or more of the following combinations:

[0027] In one embodiment, the ratio of collagen to antimicrobial agent is selected from 10:1 to 30:1; an appropriate concentration of antimicrobial agent can effectively inhibit bacterial growth, but too high a concentration may cause skin irritation and allergic reactions.

[0028] In one embodiment, the ratio of collagen to anti-inflammatory agent is selected from 15:1 to 100:1; the facial filling process may cause mild inflammation, so the appropriate use of anti-inflammatory agents can help alleviate these symptoms, but the dosage needs to be controlled to avoid inhibiting normal skin function.

[0029] In one embodiment, the ratio of collagen to growth factors is selected from 100:1 to 300:1; growth factors are very beneficial for promoting collagen synthesis and the skin's natural recovery process, but should be used with caution in facial fillers to avoid excessive growth or uneven skin texture.

[0030] In one embodiment, the ratio of collagen to moisturizer is selected from 1:1 to 8:1. Moisturizers are particularly important in facial fillers because they help maintain the skin's smoothness and plumpness. Increasing the moisturizer ratio can effectively improve the durability and natural feel of the filler.

[0031] In one embodiment, the ratio of collagen to antioxidant is selected from 10:1 to 50:1; the antioxidant can prevent oxidation of the preparation, but its dosage should be controlled so as not to affect the long-term stability of the product.

[0032] In one embodiment, the ratio of collagen to emulsifier or thickener is selected from 5:1 to 20:1; it helps to improve the texture and stability of the product, but should be used in moderation to ensure the applicability and comfort of the final product.

[0033] Another aspect of the present invention provides a medical composition, wherein the composition comprises type 21 recombinant humanized collagen, and the amino acid sequence of the collagen is selected from SEQ ID No. 1 or any of the aforementioned collagen materials.

[0034] In one embodiment, the preparation forms of the medical composition of the present invention include but are not limited to the following forms:

[0035] Injectable preparations, preferably including but not limited to injectable hydrogels, composite injections, etc., are used for direct injection into the skin through surgery to provide a deep facial filling effect.

[0036] Topical preparations, preferably including but not limited to topical gels, creams or lotions, are suitable for daily skin care to assist in reducing fine lines and restoring skin elasticity.

[0037] Solution preparations, preferably, include but are not limited to collagen solutions for microneedling or microneedle rollers, which promote the distribution and absorption of collagen in the skin in a minimally invasive manner.

[0038] In one embodiment, the product of the present invention may further contain a pharmaceutically, food-, health-care-, or dietary-acceptable carrier. For example, various compatible solid or liquid fillers, gels, or solvents may be used. These carriers should be suitable for human use, possess sufficient purity, and be low in toxicity. Common carriers include cellulose and its derivatives, polyols such as glycerol, and other pharmaceutical excipients such as talc and magnesium stearate.

[0039] Another aspect of the present invention provides a use of type 21 recombinant humanized collagen in the preparation of a medical composition for facial filling, wherein the amino acid sequence of the collagen comprises the amino acid sequence shown in SEQ ID No. 1.

[0040] Another aspect of the present invention provides a use of any one of the aforementioned collagen materials in preparing a medical composition for facial filling.

[0041] Another aspect of the present invention provides a method for preparing the collagen material for facial filling.

[0042] In one embodiment, the collagen material of the present invention is a collagen composite injection, wherein the raw materials used are 2%-10% (w / v) type 21 collagen and 0.1%-1% (w / v) hyaluronic acid; 0.05% (w / v) vitamin C is used as an antioxidant, and physiological saline is used as a solvent; the preparation process comprises the following steps:

[0043] Dissolved antioxidants;

[0044] Prepare collagen solution;

[0045] Added hyaluronic acid;

[0046] Mixing antioxidant solution;

[0047] pH and viscosity adjustment;

[0048] Sterile filtration and filling.

[0049] In one embodiment, the collagen material of the present invention is a collagen / hyaluronic acid blended injectable hydrogel; wherein the hydrogel structure is strengthened by using the crosslinking agent EDC (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide), and the optimal formula is determined by adjusting the mixture of collagen and hyaluronic acid in different proportions and its reaction with different concentrations of EDC; the raw materials used are 1%-5% (w / v) type 21 collagen and 0.5%-2% (w / v) hyaluronic acid; the formula also includes phosphate buffer as a dissolution base, and 1%-5% (w / v) glycerol is added as a moisturizer.

[0050] In one embodiment, the collagen material described herein is a collagen solution for microneedle / microneedle rollers. The collagen solution is suitable for use in microneedle or microneedle rollers, promoting the distribution and absorption of collagen into the skin through a minimally invasive approach. The formula contains a high collagen concentration of 3%-10% (w / v) to enhance absorption through the microneedle channels. Excipients include 0.1%-1% (w / v) hyaluronic acid to enhance moisturizing properties, 0.5%-2% (w / v) urea as a penetration enhancer to help collagen penetrate the skin, and a buffer adjusted to a pH close to that of skin (pH 5.5-7.0).

[0051] Advantages of the present invention

[0052] 1. Collagen type 21 can stimulate the production of type I and type III collagen, two key components of skin elasticity and structure. This ability offers a novel mechanism for collagen-based anti-aging strategies. By promoting the production of type I and type III collagen, collagen type 21 not only accelerates wound healing but also improves the quality of the repaired skin. This is particularly important for facial fillers, as high-quality skin repair results can provide a more natural and long-lasting aesthetic, reduce scarring, and improve skin elasticity and structural stability. Furthermore, by increasing the production of type I and type III collagen, collagen type 21 reduces fine lines and wrinkles and improves skin firmness and elasticity. In the plastic surgery and aesthetics industries, this offers a non-invasive skin rejuvenation solution, such as microneedling rollers, microneedling treatments, or topical products, that delivers collagen to deeper layers of the skin and stimulates the production of type I and type III collagen, achieving a filling effect without the need for injections.

[0053] 2. Because type 21 collagen occurs naturally in the human body, its biocompatibility and low immunogenicity are particularly prominent when used as a facial filler. This reduces the risk of immune reactions following injection, providing a safer treatment option for patients.

[0054] 3. In addition to being an effective facial filler, type 21 collagen offers numerous potential added benefits for promoting skin health, such as anti-aging and improving skin hydration. This allows its use not only as a filler but also as an active ingredient in skin care and beauty products, providing a more comprehensive beauty solution. The recombinant humanized type 21 collagen (ColpepA1 21) described in this invention demonstrates significant advantages in anti-aging. While providing essential collagen supplementation, ColpepA1 21's unique efficacy and optimized molecular properties make it stand out among other collagen materials. BRIEF DESCRIPTION OF THE DRAWINGS

[0055] Figure 1 The relative fluorescence intensity of type III collagen in cells of each experimental group is shown.

[0056] Figure 2 The bar graph shows the results of epidermal viable cell layer thickness, where ## indicates a significant difference compared with the blank control group (P<0.01), and ** indicates a significant difference compared with the negative control group (P<0.01). Figures 3 to 7 Same here.

[0057] Figure 3 A bar graph showing the relative area of ​​collagen fibers.

[0058] Figure 4 A histogram showing the relative integrated optical density (IOD) values ​​of Collagen I.

[0059] Figure 5 A histogram of the relative integrated optical density (IOD) values ​​of Collagen III is shown.

[0060] Figure 6 A histogram showing the relative integrated optical density (IOD) / Area values ​​of Collagen IV is shown.

[0061] Figure 7 A histogram showing the relative integrated optical density (IOD) / Area values ​​of Collagen XVII is shown.

[0062] Figure 8 The figure shows the effect of ColpepA1 21 compound injection on the cellular HYP content under oxidative stress environment.

[0063] Related definitions

[0064] Unless otherwise specified, the following terms used in the specification and claims have the following meanings:

[0065] As used herein, the term "collagen (COL)" refers to a vital structural protein in the human body, found in a variety of tissues and organs, such as skin, bones, tendons, ligaments, and cornea. The primary function of these proteins is to provide strength, stability, and elasticity to body tissues, ensuring the normal functioning of various physiological functions. Currently, there are at least 28 known types of collagen, each with a unique role and function in the body, forming a complex and finely regulated biological system.

[0066] As used herein, the term "type 21 collagen (COL21A1)" refers to a type of collagen found in the human body, belonging to the small molecule collagen family. As part of the FACIT (fibril-associated collagen with interrupted triple helix) family, type 21 collagen plays a crucial role in connecting components of the extracellular matrix. Through its unique molecular structure, this collagen facilitates interactions and linkages between different collagen types, thereby supporting the integrity and order of the extracellular matrix and its function in wound healing and tissue repair. Although less well-known than types I and III collagen, it plays a role in specific tissues and physiological processes, implicated in extracellular matrix composition and skin repair mechanisms. Although type 21 collagen is relatively rare in the adult human body, comprising less than 1% of total collagen, its potential role in cellular processes such as migration, proliferation, and differentiation, as well as its potential role in maintaining extracellular matrix structure and promoting tissue regeneration, remains an important area of ​​future research.

[0067] As used herein, the term "recombinant humanized collagen type 21 (ColpepA1 21)" refers to a protein expressed in Pichia pastoris via genetic recombination technology that mimics the type 21 collagen naturally found in the human body. This protein possesses similar biological functions and structural properties to natural type 21 collagen, with a specific amino acid sequence encoding specific properties, designed to enhance skin health and anti-aging capabilities.

[0068] As used herein, the term "HaCaT (human keratinocytes)" refers to an immortalized human keratinocyte cell line that is widely used to study skin biology, skin pathology, and drug toxicity testing. It is considered a valuable research tool because it can mimic the behavior of normal epidermal cells.

[0069] As used herein, the term "human foreskin fibroblasts (HFF)" refers to fibroblasts isolated from human fetal foreskin. They are an important component of skin structure, responsible for synthesizing collagen and other extracellular matrix proteins, and are commonly used to study skin aging, wound healing, and collagen synthesis.

[0070] As used herein, the term "human dermal fibroblasts (HDFs)" refers to fibroblasts isolated from the human dermis. These cells play a central role in the formation of skin structure, primarily responsible for the synthesis of collagen and other extracellular matrix proteins. HDFs are commonly used in research on skin aging, wound healing, disease modeling, and drug screening.

[0071] As used herein, the term "composition" refers to a preparation containing recombinant humanized collagen type 21 and possibly other auxiliary ingredients such as moisturizers, antioxidants or whitening ingredients, which work together to provide a more comprehensive skin care effect.

[0072] As used herein, the term "TGFβ1" (transforming growth factor β1) refers to a multifunctional cytokine that regulates cell proliferation, differentiation, and migration. In skin physiology, TGFβ1 has important influences on collagen synthesis, wound healing processes, and anti-inflammatory effects.

[0073] As used herein, the term "HPR (phthalein oxalate)" is a synthetic retinol derivative commonly used in skin care products for its ability to promote cell renewal, enhance skin repair, and improve the condition of aging skin.

[0074] As used herein, the term "ex vivo skin model" refers to a skin tissue sample extracted from a living body and maintained under laboratory conditions to simulate and study the biological properties of the skin in vitro and its response to various treatments.

[0075] As used herein, the terms "UVA and UVB radiation" refer to the two main types of ultraviolet radiation. UVA radiation (ultraviolet long waves) penetrates deep into the dermis and can cause long-term damage, such as skin aging. UVB radiation (ultraviolet medium waves) primarily affects the epidermis and is the primary cause of sunburn.

[0076] As used herein, the term "histiomorphology" refers to the microscopic structure of the skin, as determined by staining and microscopic observation, including the shape, size, arrangement, and interactions of cells. Changes in histological morphology, such as changes in epidermal thickness and dermal density, can directly reflect the skin's response to external factors, including damage from ultraviolet radiation, as well as the effectiveness of treatments and repair measures.

[0077] As used herein, the term "collagen fibers" refers to the fibrous structure of the skin's dermis, primarily composed of type I and type III collagen. Collagen fibers provide the skin with essential elasticity and firmness, and contribute to its overall appearance and feel. Changes in collagen fiber content and quality are key indicators of a facial filler's ability to strengthen the skin's support structure, improve skin laxity, and reduce wrinkles.

[0078] As used herein, the term "type I collagen" refers to the most abundant type of collagen in the human body. It is a fundamental building block of various tissues, including skin, bones, and tendons. Its primary function is to provide structural support and physical strength to these tissues, maintaining the skin's elasticity and firmness. A decrease in the natural production of type I collagen with aging is closely associated with skin laxity and wrinkle formation. In facial filler products, an increase in type I collagen is considered a key indicator of product effectiveness, particularly in anti-aging and improving skin structure.

[0079] As used herein, the term "type III collagen" refers to the type of collagen found alongside type I collagen in the skin's structure, particularly in younger skin. It plays a crucial role in maintaining skin elasticity and assisting in the healing process of damaged skin. Following skin injury, a temporary increase in type III collagen helps repair and rebuild the skin's structure. A decrease in type III collagen with age may affect the skin's elasticity and repair capacity. The increased content of type III collagen in facial fillers demonstrates the product's potential to promote natural skin repair and enhance skin elasticity.

[0080] As used herein, the term "type IV collagen" (Collagen IV) refers to a major component of the basement membrane, a specialized layer located between the epidermis and dermis that plays a key role in the skin's structural stability and barrier function. Type IV collagen forms the basement membrane's network structure, providing a support layer for skin cells to maintain physical stability and promote repair processes. With aging or external environmental damage, such as UV radiation, the function of the basement membrane may be impaired, leading to skin aging. Therefore, increasing the type IV collagen content in facial fillers can help restore and protect the skin's basement membrane, contributing to the overall health and appearance of the skin.

[0081] As used herein, the term "collagen XVII" refers to a transmembrane collagen protein primarily located in the basal cell membrane of the epidermis. By interacting with other components of the basal membrane, it participates in maintaining cell adhesion and the skin's barrier function. It plays a crucial role in maintaining skin integrity and responding to external stress. Damaged or aging skin may exhibit decreased expression of type XVII collagen, leading to weakened skin barrier function and sagging. Therefore, increasing the content of type XVII collagen in facial filler products can help strengthen the skin's self-repair ability and barrier function, reducing the impact of external environmental factors on the skin.

[0082] As used herein, the term "composition" or "preparation" refers to a mixture containing one or more collagen proteins described herein and other components, such as physiologically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to an organism, facilitating the absorption of the active ingredient and thereby exerting its biological activity.

[0083] As used herein, the term "antioxidant" refers to a class of substances that can neutralize free radicals and slow down or prevent cell damage, which plays an important role in reducing oxidative stress caused by environmental factors and preventing skin aging.

[0084] As used herein, the term "emulsifier" refers to a substance that can help oil and water mix to form a stable emulsion, and is used in skin care products to improve the texture and stability of the product.

[0085] As used herein, the term "humectant" refers to a substance that attracts or locks in water, helping the skin retain moisture and plays a vital role in maintaining the skin's hydration state and preventing dryness.

[0086] As used herein, the term "stabilizer" refers to a pharmaceutically acceptable excipient that protects the active pharmaceutical ingredient and / or formulation from chemical and / or physical degradation during manufacture, storage, and use. Stabilizers include, but are not limited to, sugars, amino acids, salts, polyols, and their metabolites as defined below, such as sodium chloride, calcium chloride, magnesium chloride, mannitol, sorbitol, sucrose, trehalose, arginine or its salts (e.g., arginine hydrochloride), glycine, alanine (α-alanine, β-alanine), betaine, leucine, lysine, glutamic acid, aspartic acid, proline, 4-hydroxyproline, sarcosine, gamma-aminobutyric acid (GABA), opines, alanine, octopine, strombine, and trimethylamine N-oxide (TMAO), human serum albumin (HSA), bovine serum albumin (BSA), α-casein, globulin, α-lactalbumin, LDH, lysozyme, myoglobin, ovalbumin, and RNase A. Some stabilizers, such as sodium chloride, calcium chloride, magnesium chloride, mannitol, sorbitol, sucrose, etc., can also play a role in controlling osmotic pressure. The stabilizers specifically used in the present invention are selected from one or more of polyols, amino acids, salts, and sugars. Preferred salts are sodium chloride, preferred sugars are sucrose and trehalose, and preferred polyols are sorbitol and mannitol. Preferred amino acids are arginine or its salts (such as arginine hydrochloride), glycine, and proline. Preferred stabilizers are sodium chloride, mannitol, sorbitol, sucrose, trehalose, arginine hydrochloride, glycine, proline, sodium chloride-sorbitol, sodium chloride-mannitol, sodium chloride-sucrose, sodium chloride-trehalose, arginine hydrochloride-mannitol, and arginine hydrochloride-sucrose.

[0087] As used herein, the term "hydroxyproline (HYP)" refers to a non-essential amino acid found primarily in collagen, where it forms a crucial component of the collagen tripeptide sequence. HYP plays a key role in collagen's structural stability and tensile strength, and is commonly used as a biomarker for assessing collagen content and synthesis activity.

[0088] As used herein, the term "H2O2 oxidative stress treatment" refers to an experimental technique that uses hydrogen peroxide (H2O2) to treat cells, simulating the oxidative stress experienced by cells under physiological or pathological conditions. This treatment is often used to study the mechanisms of cell response to oxidative damage, protective strategies, and the ability to survive under oxidative stress, particularly in the context of anti-aging, antioxidant, and disease model studies. DETAILED DESCRIPTION

[0089] In a specific embodiment, the materials and methods used are as follows:

[0090] Cell lines: HaCaT (human keratinocytes) and HFF-1 (human fibroblasts) were purchased from Fenghui Biotechnology.

[0091] Reagents: CCK-8 kit (Japan Dojin CK04) was used for cell viability assessment, and AQP3 (Shanghai ELISA) and type I collagen kits (Shanghai ELISA CMM2023H1) were used for related protein expression analysis.

[0092] Example 1: Production of Type III Collagen

[0093] This study aimed to evaluate the effectiveness of recombinant humanized collagen type 21 (ColpepA1 21) in stimulating the production of type III collagen. Type III collagen is a key component in maintaining skin structural stability and elasticity, and is crucial for facial skin rejuvenation and damage recovery. Through this study, we aimed to explore the potential of ColpepA1 21 as an effective ingredient in facial fillers and anti-aging collagen materials.

[0094] Experimental groups:

[0095] Blank control group (Control): No treatment was performed to evaluate the expression level of type III collagen in cells in an undamaged state.

[0096] Negative control group: cells were damaged by 5 J / cm² UVA to simulate the skin damage effect of sunlight.

[0097] Positive control group (HPR): 0.001% HPR (phthalein oxalate) was added after UVA damage, a therapeutic agent known to help skin repair.

[0098] ColpepA1 21 experimental group: 10 ppm and 50 ppm of type 21 recombinant humanized collagen were added after UVA damage.

[0099] HFF-1 fibroblasts were cultured separately and plated for 24 h. When the cell confluence reached 70-80%, UVA induction and sample treatment were added. The cells were cultured for 24 h. Immunofluorescence (IF) detection was used to quantitatively analyze the relative fluorescence intensity of type III collagen in the cells of each experimental group to detect the expression of type III collagen.

[0100] Data Collection and Analysis:

[0101] Figure 1 The relative fluorescence intensity of type III collagen obtained by immunofluorescence analysis is shown. As shown, the blank control group exhibited normal levels of type III collagen fluorescence intensity, representing baseline expression. The type III collagen fluorescence intensity in the UVA-negative control group was significantly lower than that in the control group, indicating damage to the extracellular matrix caused by UVA irradiation. The 0.001% HPR positive control group showed an increase relative to the UVA-negative control group, but still lower than the blank control group, indicating that HPR has a certain protective and repairing effect on the extracellular matrix. In the ColpepA1 21 experimental group, the fluorescence intensity of type III collagen increased significantly after the addition of 50 ppm ColpepA1 21, reaching a promotion rate of 25.42% compared to the UVA-negative control group, demonstrating that ColpepA1 21 significantly promotes type III collagen production.

[0102] The effect of recombinant humanized collagen type 21 in promoting the production of type III collagen significantly improves the skin's ability to repair damage and increases skin elasticity, which is very important in the anti-aging field. This effect not only supports the use of ColpepA1 21 in facial filler collagen materials, but also emphasizes its significant potential in promoting skin repair after damage, improving skin elasticity and combating aging. This provides valuable scientific evidence for facial filler collagen materials, demonstrates the unique value of ColpepA1 21 in improving skin health and delaying the aging process, and lays a solid foundation for the further development and commercial application of anti-aging collagen materials.

[0103] Example 2: In vitro skin model test

[0104] This study aimed to evaluate the effectiveness of ColpepA1 21 in improving skin elasticity and reducing wrinkles, specifically its repair and protection effects after UVA and UVB exposure. By simulating everyday UV exposure, the potential application of ColpepA1 21 in facial filler products was evaluated. By analyzing changes in ex vivo skin tissue after UV irradiation, the study examined changes in tissue morphology, collagen fibers, and the content of types I, III, IV, and XVII collagens. The study evaluated its effects on skin structure and its anti-aging potential, and analyzed the effectiveness of type 21 collagen in facial fillers.

[0105] Experimental groups:

[0106] Blank control group: no treatment was performed.

[0107] Negative control group: The specific treatment is not specified, but it generally refers to a group that is not added with the test sample and is only subjected to the same external conditions.

[0108] Positive control group: treated with a mixture of VC (vitamin C) and VE (vitamin E) known to have firming and anti-wrinkle effects as an effect control.

[0109] Sample group (type 21 collagen group, ColpepA1 21): treated with type 21 recombinant humanized collagen (ColpepA1 21) at a concentration of 0.0125% (v / v) to test its firming and anti-wrinkle effects.

[0110] Main reagents: in vitro skin tissue culture medium (Guangdong Boxi Biological), PBS (Solaibao), vitamin C (VC, Sigma), vitamin E (VE, Sigma), anhydrous ethanol (Sinopharm), xylene (Sinopharm), hematoxylin (Biyuntian), eosin (Biyuntian), Collagen I antibody (Proteintech), Collagen III antibody (Abcam), Collagen IV antibody (Abcam), Collagen XVII antibody (Abcam), and paraformaldehyde (Biosharp).

[0111] Main equipment: CO2 incubator (Thermo, 1501), clean bench (Suzhou Antai, SW-CJ-1F), upright microscope (Olympus, BX53), UVA irradiator (Philips), UVB irradiator (Philips), fluorescence microscope (Leica, DM2500).

[0112] 1. Tissue Processing

[0113] Freshly harvested skin tissue was washed in 75% alcohol for 30 seconds to remove surface impurities and microorganisms. The tissue was further cleaned and prepared by washing three times with sterile PBS buffer. The skin was cut into 24 ± 2 mm² pieces, with the epidermis facing up and the dermis facing down, and placed in a culture mold. The mold was placed in a 6-well plate, and 3.7 mL of culture medium was added to each well. The plates were incubated at 37°C and 5% CO2, with the culture medium changed daily.

[0114] 2. Irradiation and drug administration

[0115] After two days of culture, ex vivo skin tissues were irradiated and dosed according to the test group and corresponding treatment conditions. Irradiation doses were set at 30 J / cm² UVA and 50 J / cm² UVB, and irradiation was continued for four consecutive days. Fresh culture medium was replaced after each irradiation period, and drug treatment was administered. The positive control group received submerged drug administration, while the sample group received topical drug administration. Following four days of irradiation and drug administration, the ex vivo skin tissues were cultured for an additional three days, during which time only sample drug administration was performed without irradiation.

[0116] 3. Cleaning

[0117] After the incubation period, sterile PBS solution was used to clean the residual reagent on the surface of the model, and a sterile cotton swab was used to gently wipe away the residual liquid inside and outside the model.

[0118] 4. Tissue Morphology and Collagen Fiber Detection

[0119] After washing, the skin tissue was fixed with 4% paraformaldehyde, embedded, and sectioned. H&E staining and Masson staining were performed, respectively. The images were photographed under a microscope and analyzed using Image-Pro® Plus image processing software.

[0120] 5. Immunofluorescence Detection

[0121] The model to be tested was fixed with 4% paraformaldehyde for 24 hours, and then immunofluorescence detection was performed. The model was photographed under a microscope, and the images were collected and analyzed.

[0122] 6. Calculation of promotion rate

[0123] According to the immunofluorescence results, the following formula was used to calculate the improvement rate of each indicator in each experimental group to further evaluate the effect of type 21 collagen.

[0124] Improvement rate (%) = (experimental group - blank control group) / blank control group × 100%

[0125] 7. Statistical Analysis of Results

[0126] GraphPad Prism software was used for plotting, and the results were expressed as mean ± standard deviation. The differences among the groups were compared by t-test statistical analysis.

[0127] Example 3-1: Tissue morphology test

[0128] Histological morphology reflects the microscopic structure of the skin and is a key indicator for assessing skin health and aging. When evaluating facial filler products, observing changes in epidermal thickness and dermal density after treatment provides a direct understanding of the product's effectiveness in resisting skin damage, promoting skin repair, and improving skin firmness. When exposed to external factors (such as UV radiation), the skin's histological structure changes, such as thinning of the epidermis and a decrease in collagen fibers in the dermis, which are all signs of skin aging. Effective facial filler products should promote skin cell proliferation, restore or increase epidermal thickness, and enhance the density and elasticity of the dermis, thereby improving the skin's appearance and texture.

[0129] Figure 2 A bar graph shows the results of epidermal viable cell layer thickness. ## indicates a significant difference compared to the blank control group (P<0.01), and ** indicates a significant difference compared to the negative control group (P<0.01). As shown in the figure, the blank control group (BC) exhibits baseline epidermal viable cell layer thickness without any treatment. After UV irradiation, the negative control group (NC) showed a significant decrease in epidermal viable cell layer thickness, indicating that UV irradiation significantly damages the skin structure, leading to epidermal thinning. In the positive control group (PC), treatment with known anti-aging ingredients (such as VC+VE) resulted in a recovery in epidermal thickness, demonstrating that the positive control substance has some protective effect against UV damage. The sample group (type 21 collagen ColpepA121, 0.0125%) showed a significant increase in epidermal viable cell layer thickness (30.52%) compared to the negative control group, demonstrating that ColpepA121 significantly promotes epidermal thickness and protects against UV damage.

[0130] The results showed that ColpepA1 21 provides an effective protection mechanism against damage from UV radiation and increases skin elasticity and firmness. For facial filler products, this means it can effectively improve skin structure and restore or maintain the skin's youthful state, with potential therapeutic and cosmetic benefits.

[0131] Example 3-2: Collagen fiber test

[0132] Collagen fibers are the primary component of the skin's dermis, primarily composed of type I and type III collagen. They have a decisive influence on the skin's elasticity, firmness, and overall appearance. In the evaluation of facial filler products, measuring the content and quality of collagen fibers can be used to assess the product's ability to strengthen the skin's support structure, improve skin laxity, and reduce wrinkles. With aging or exposure to external factors such as ultraviolet rays, the collagen fibers in the skin gradually decrease and become damaged, leading to a loss of elasticity and firmness, and the appearance of wrinkles and sagging. By stimulating collagen production, facial fillers can help restore skin elasticity and firmness, reducing wrinkles.

[0133] Figure 3 The bar graph shows the results of the relative collagen fiber area. As shown, the blank control group (BC) received no treatment, and the relative collagen fiber area can be considered a reference value for normal levels. In the negative control group (NC), the relative collagen fiber area decreased significantly under UV irradiation, indicating that UV irradiation causes collagen fiber damage and deterioration of skin structure. In the positive control group (PC), skin samples treated with a substance known to have anti-aging effects showed some recovery of the relative collagen fiber area, indicating that the positive control substance can counteract UV-induced collagen fiber damage. Compared to the negative control group, the sample group (type 21 collagen ColpepA1 21, 0.0125%) showed a significant increase in relative collagen fiber area, with an increase of 43.48%. This suggests that ColpepA1 21 can significantly promote the increase of collagen fibers, improve skin firmness and elasticity, and combat the signs of aging.

[0134] The results showed that ColpepA1 21 as an active ingredient has significant skin firming and anti-aging effects for facial filler products, as evidenced by an increase in collagen fiber content, making it a valuable ingredient.

[0135] Example 3-3: Type I collagen (Collagen I) test

[0136] Type I collagen is the most abundant type of collagen in human skin, comprising approximately 80% of the total protein in the dermis. Its content is directly related to the skin's structural integrity and function. With aging, type I collagen production decreases, leading to sagging and wrinkling. Therefore, promoting type I collagen production is a key goal in the development of anti-aging and skin repair products. In the evaluation of facial fillers, an increase in type I collagen is considered a key indicator of improved skin structure, firmness, and elasticity.

[0137] Figure 4A bar graph shows the relative integrated optical density (IOD) values ​​of Collagen I. As shown, the blank control (BC) remains untreated and displays the baseline IOD value of Collagen I, providing a benchmark for subsequent comparisons. After UV irradiation, the IOD value of Collagen I in the negative control (NC) significantly decreased, indicating that UV irradiation significantly damages Collagen I, leading to a decrease in its content in the dermis. The IOD value of Collagen I in the positive control (PC) significantly increased compared to the negative control, suggesting that the VC+VE treatment in the PC helped restore Collagen I content and counteract the damaging effects of UV. The sample group (Collagen Type 21, ColpepA1 21, 0.0125%) showed Collagen I IOD values ​​similar to those in the positive control, but also significantly higher than those in the negative control, with an increase of 159.38%. This indicates that type 21 collagen effectively promotes the increase of Collagen I, which helps strengthen the skin's infrastructure and improve skin firmness and elasticity.

[0138] The results showed that ColpepA1 21 is an effective ingredient that can significantly increase the content of Collagen I in the skin. This is very helpful for preventing and treating UV-induced skin damage, reducing wrinkle formation, and improving skin firmness. This also provides a scientific basis for the use of ColpepA1 21 in facial filler products.

[0139] Example 3-4: Type III collagen (Collagen III) test

[0140] Type III collagen is another important collagen in the skin, especially found in higher concentrations in the skin of children and adolescents. Together with type I collagen, it forms the skin's main support structure. It plays a vital role in the skin's elasticity and repair process, especially during the healing process after skin injury, when type III collagen production temporarily increases to help repair damaged skin structure. With age, the proportion of type III collagen relative to type I collagen gradually decreases, affecting the skin's elasticity and repair ability. An increase in type III collagen reflects the product's potential to promote skin repair and enhance skin elasticity. For damaged or aging skin, products that can increase the content of type III collagen can help accelerate the skin's natural repair process and improve the skin's overall texture and appearance. An increase in type III collagen is seen as a facial filler product's ability to promote skin regeneration and restore the skin to a healthy state.

[0141] Figure 5A bar graph shows the relative integrated optical density (IOD) values ​​of Collagen III. As shown, the blank control group (BC) exhibited baseline levels of Collagen III IOD. In the negative control group (NC), after UV irradiation, the IOD value of type III collagen decreased significantly, indicating that UV irradiation significantly damaged type III collagen in the skin. In the positive control group (PC), after treatment with a known anti-aging agent, the content of type III collagen increased significantly, indicating that the positive control treatment helped restore collagen III levels after UV irradiation damage. The sample group (Collagen Type 21, ColpepA1 21, 0.0125%) showed a significantly higher IOD value of type III collagen than the negative control group, with an increase of 173.91%. This indicates that type 21 collagen significantly promotes the production of type III collagen, improves skin elasticity, and reduces wrinkle formation.

[0142] The results suggest that ColpepA1 21 could be an effective ingredient in facial fillers, particularly in promoting skin elasticity and anti-aging. The results highlighted its significant effect on increasing type III collagen content, which in turn suggests it can improve skin texture and appearance, providing scientific support for the efficacy of facial fillers.

[0143] Example 3-5: Type IV collagen (Collagen IV) test

[0144] Type IV collagen forms the network structure of the skin's basement membrane, providing a support layer to which surface cells firmly attach. This structure not only maintains the skin's physical stability but also plays a key role in the skin's repair process, nutrient transport, and barrier function. With aging or external environmental damage (such as UV radiation), the breakdown of type IV collagen impairs basement membrane function, accelerating the aging process. Therefore, the effect of facial fillers in increasing the content of type IV collagen can help restore and protect the skin's basement membrane, improving its overall health and appearance.

[0145] Figure 6A bar graph shows the relative integrated optical density (IOD) / Area values ​​of Collagen IV. As shown, the blank control (BC) represents the baseline level of Collagen IV in untreated skin samples. In the negative control (NC) under UV irradiation, the Collagen IV content decreased significantly, indicating that UV irradiation successfully induced skin aging conditions and damaged the basement membrane. The positive control (PC) showed a significant increase in Collagen IV content (marked with **) compared to the negative control, indicating that the treatment promoted Collagen IV synthesis or slowed its degradation, effectively counteracting UV damage. The Collagen IV content in the sample group (Collagen Type 21, 0.0125%) was also significantly higher than that in the negative control, with an increase of 102.78%, indicating that ColpepA1 21 significantly promoted the synthesis or stability of Collagen IV, helping to restore the structure and function of the basement membrane.

[0146] The results showed that the application of ColpepA1 21 showed positive effects in promoting skin health and preventing premature aging, providing scientific support for its use in facial fillers and skin care products. By strengthening the structural integrity and function of the basement membrane, ColpepA1 21 helps maintain the youthful appearance of the skin and is a valuable component of anti-aging strategies.

[0147] Example 3-6: Type XVII collagen (Collagen XVII) test

[0148] Type XVII collagen is a transmembrane collagen located on the cell membrane of epidermal basal cells. It interacts with other components of the basement membrane to stabilize cell attachment and facilitate signaling between cells. This adhesion is crucial for the integrity and barrier function of the skin. When the skin is damaged or aged, the expression of type XVII collagen may be affected, resulting in decreased epidermal cell adhesion and weakened skin barrier function. Therefore, facial filler products that increase the content of type XVII collagen through external application can help strengthen the skin's barrier function and promote skin health. In the evaluation of facial filler products, the increase in type XVII collagen is believed to enhance the skin's self-repair ability, reduce the effects of damage caused by external environmental factors, and combat skin aging.

[0149] Figure 7A bar graph shows the relative integrated optical density (IOD) / Area values ​​of Collagen XVII. As shown, the blank control (BC) exhibits normal Collagen XVII content without any treatment, serving as the baseline for the experiment. The negative control (NC) exhibited a significant decrease in Collagen XVII content under UV irradiation, indicating that the UV irradiation conditions in the experiment led to a decrease in collagen content, simulating skin damage or aging. The positive control (PC) was treated with a known anti-aging agent, resulting in a significant increase in Collagen XVII content. The sample (Collagen Type 21, ColpepA1 21, 0.0125%) showed a significant increase in Collagen XVII content of 50.94% compared to the negative control, demonstrating that ColpepA1 21 effectively increased Collagen XVII content.

[0150] The results showed that type 21 collagen can effectively increase the content of Collagen XVII in the skin, enhance the skin's self-repair ability, and resist the effects of external environmental factors, thereby combating aging. This provides scientific support for type 21 collagen as an effective facial anti-aging filler.

[0151] In summary, the positive effects demonstrated by ColpepA1 21 not only highlight its potential value in promoting skin repair, enhancing skin moisturizing, improving skin structural integrity, and combating photoaging, but also reveal its enormous market potential as an active ingredient in facial fillers and anti-aging collagen materials. These experimental results provide a solid scientific basis for the application of ColpepA1 21 in facial filler collagen materials, demonstrating its important role in improving skin appearance, promoting skin health, and delaying the aging process.

[0152] Therefore, ColpepA1 21 can not only serve as a highly effective facial filler ingredient to promote the skin's natural repair and regeneration, but can also play a key role in the development of future anti-aging products, providing innovative and effective solutions for consumers pursuing healthy, youthful skin.

[0153] Example 4: Preparation of Type 21 Collagen ColpepA1 21 Composite Injection

[0154] Material:

[0155] Type 21 collagen: 2% (w / v)

[0156] Normal saline: sufficient amount as solvent

[0157] Hyaluronic acid: 1% (w / v)

[0158] Vitamin C: 0.05% (w / v) as an antioxidant

[0159] Preparation steps:

[0160] Step 1: Dissolve the Antioxidant

[0161] Dissolve vitamin C in a small amount of saline solution to ensure complete dissolution.

[0162] Step 2: Prepare collagen solution

[0163] In a sterile container, slowly add the recombinant collagen powder to the remaining saline solution at the desired final concentration. Stir slowly at room temperature using a magnetic stirrer until the collagen is completely dissolved.

[0164] Step 3: Add Hyaluronic Acid

[0165] Add the pre-weighed amount of hyaluronic acid to the collagen solution and continue stirring to ensure uniform mixing until the hyaluronic acid is completely dissolved and a uniform viscous solution is formed.

[0166] Step 4: Mix the Antioxidant Solution

[0167] Slowly add the dissolved vitamin C solution into the solution containing collagen and hyaluronic acid, and continue stirring until the entire solution is evenly mixed.

[0168] Step 5: pH and viscosity adjustment

[0169] Check the pH of the solution and adjust it to a pH range suitable for injection (approximately 7.0-7.4) if necessary. Adjust the viscosity of the solution to ensure that it is suitable for subcutaneous injection and has good fluidity but is not too thick.

[0170] Step 6: Sterile Filtration and Filling

[0171] The solution was sterile filtered through a 0.22 μm sterile filter and the filtered solution was aseptically filled into pre-sterilized syringes.

[0172] Comparative Example 1: Preparation of Type III Collagen Composite Injection

[0173] Material:

[0174] Type III collagen: 2% (w / v)

[0175] Normal saline: sufficient amount as solvent

[0176] Hyaluronic acid: 1% (w / v)

[0177] Vitamin C: 0.05% (w / v) as an antioxidant

[0178] The preparation method is the same as Example 4.

[0179] Example 5: Effect of Type 21 Collagen Composite Injection on Human Collagen Synthesis

[0180] This study aimed to evaluate the ability of a type 21 collagen compound injection to enhance hydroxyproline (HYP) expression in human dermal fibroblasts (HDFs). HYP is a key component of collagen and reflects the total collagen content. Therefore, measuring HYP content can be used to assess the potential of this injection for facial augmentation applications.

[0181] Experimental groups:

[0182] Normal group (Control): No treatment was performed, representing the collagen expression level of cells in normal state.

[0183] H2O2 model group (NC): cells were treated with H2O2 to simulate oxidative damage caused by oxidative stress.

[0184] Sample group A: Based on the modeling group, damaged cells were treated with 0.5% ColpepA1 21.

[0185] Sample group B (positive control group): Based on the modeling group, an equal amount of 0.5% type III collagen type III composite injection prepared using the method of Comparative Example 1 was used to treat damaged cells.

[0186] Sample group C: Based on the modeling group, an equal amount of 0.5% ColpepA1 21 type 21 collagen composite injection prepared using the method of Example 4 was used to treat damaged cells.

[0187] HDF cell anti-aging test:

[0188] Cell seeding: dilute to 1×10 5 Cells were seeded into 12-well plates at a seeding density of 100 cells / ml, and 1 ml of cell dilution was used per well. The plates were incubated in a cell culture incubator (37°C, 5% CO2, 95% RH) for 24 ± 2 h. Three replicate wells were set up for each group according to the experimental grouping.

[0189] Liquid preparation:

[0190] Sample: Dissolve the sample in cell culture medium (add DMSO to dissolve the sample if it cannot be dissolved, but the final DMSO concentration should not exceed 0.5%), and then dilute it with cell culture medium;

[0191] H2O2 working solution: Prepare H2O2 working solution by using serum-free culture medium to prepare H2O2 mother solution.

[0192] H2O2 induction: When the cell confluence in the 12-well plate reaches 50%-70%, remove the cell culture medium from each group, wash once with PBS, discard the solution, and add H2O2 working solution.

[0193] Sample addition: After induction, wash twice with PBS. Add 1 ml of culture medium to each well of the normal group; add 1 ml of culture medium containing the corresponding sample concentration to each well of the sample group. After sample addition, place the 12-well plate in an incubator (37°C, 5% CO2, 95% RH) and incubate for 24 ± 2 h.

[0194] HYP content detection: After culturing cells for 24 ± 2 h, the cells were collected and the OD value was read at 560 nm according to the instructions of the HYP detection kit.

[0195] Figure 8 The data for HYP content in the normal group (Control), H2O2 modeling group (NC), and three sample groups are shown. The results showed that the HYP level in the normal group (Control) was the highest (22.71 ug / mL), serving as a baseline reference under normal physiological conditions. The HYP level in the H2O2 modeling group was significantly reduced to 6.98 ug / mL, indicating that H2O2 successfully simulated the reduced collagen synthesis caused by oxidative stress. The HYP level in sample group B (positive control group), which used a type III collagen compound injection (10.22 ug / mL), although higher than that in the modeling group, indicating a certain collagen recovery ability, was still significantly lower than that in the control group. The HYP content of sample group A, which was supplemented with 0.5% ColpepA1 21, was further increased compared with that of sample group B, reaching 12.01 ug / mL. The HYP level of sample group C, which was supplemented with the type 21 collagen composite injection prepared using the method of Example 4, reached 18.57 ug / mL, which was significantly higher than that of the other treatment groups and close to the level of the control group, demonstrating the best collagen recovery ability.

[0196] The ColpepA1 21 composite injection (Sample Group C) demonstrated the best collagen-boosting effect, particularly in recovery after simulated oxidative damage. Its unique formula contains not only collagen but also other ingredients that promote collagen synthesis or stabilization, such as hyaluronic acid and vitamin C, to help repair and protect the skin. The use of type 21 collagen composite injections, particularly in facial fillers and anti-aging treatments, has been shown to enhance the skin's collagen synthesis capacity, demonstrating its significant value in improving the recovery of damaged skin and maintaining its elasticity and structural stability. These findings provide strong scientific support for its clinical application and provide an experimental foundation for further development and optimization of anti-aging treatment strategies.

[0197] While the specific embodiments of the present invention have been described in detail above, these are merely exemplary and the present invention is not limited thereto. It will be apparent to those skilled in the art that any equivalent modifications and substitutions to the present invention fall within the scope of the present invention. Therefore, any equivalent changes and modifications made without departing from the spirit and scope of the present invention are encompassed within the scope of the present invention.

Claims

1. Use of a type 21 recombinant humanized collagen material in the preparation of a medical composition for facial filler for treating skin damage caused by UVA and / or UVB radiation, wherein the collagen material has an amino acid sequence selected from SEQ ID No. 1 and further comprises one or more auxiliary ingredients selected from the following: a filling aid, a solvent, an antioxidant, an emulsifier, a humectant, a stabilizer, a pH adjuster, and a thickener, wherein the collagen material achieves one or more of the following effects: (1) The collagen helps to increase the thickness of the epidermal living cell layer; (2) The collagen helps to increase the density of collagen fibers; (3) The collagen helps to increase the content of type I collagen; (4) The collagen helps to increase the content of type III collagen; (5) The collagen helps to increase the content of type IV collagen; (6) The collagen helps to increase the content of type XVII collagen.

2. The use according to claim 1, wherein Based on the total weight of the collagen material contained in the medical composition, the concentration of the collagen is 0.0001% to 50%.

3. The use according to claim 1, wherein: The weight ratio of the collagen and auxiliary components in the collagen material contained in the medical composition is selected from: Collagen and antimicrobial agent: 10:1 to 30:1; or Collagen and anti-inflammatory agent: 15:1 to 100:1; or Collagen to growth factor: 100:1 to 300:1; or Collagen and moisturizer: 1:1 to 8:1; or Collagen to antioxidants: 10:1 to 50:1; or Collagen and emulsifier or thickener: one or more combinations in a ratio of 5:1 to 20:

1.

4. The use according to any one of claims 1 to 3, wherein The preparation form of the medical composition includes one or more of an injectable preparation, a topical application preparation, and a solution preparation.

5. The use according to claim 4, wherein: The injectable preparation is selected from one or more of injectable hydrogels and compound injectables.

6. The use according to claim 5, wherein: The injection preparation is a collagen compound injection.

7. The use according to claim 4, wherein: The topical smearable preparation is selected from one or more of a smearable gel, a cream or an emulsion.

8. The use according to claim 4, wherein: The solution preparation is selected from collagen solution for microneedles or microneedle rollers.

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