Multifunctional skin repairing liquid based on recombinant elastin

Abstract

The invention provides a multifunctional skin repair liquid based on recombinant elastin, and relates to the technical field of medical cosmetology, the multifunctional skin repair liquid comprises 0.01%-0.1% of recombinant elastin, 0.05%-1% of recombinant collagen, 0.01%-0.1% of ergothioneine, 0.1%-0.3% of carbomer, 10%-40% of glycerin, 0.5%-5% of 1, 2-hexanediol, 0.5%-5% of p-hydroxyacetophenone, 0.01%-0.5% of plant extract, and the balance of water. According to the present invention, by introducing the synergistic effect of the recombinant elastin and the recombinant collagen, the skin wound repair speed is significantly improved, the significant significance is particularly achieved in skin surface beautifying, and the recombinant elastin can promote the cell proliferation and the skin matrix reconstruction so as to achieve the skin wound repair effect, such that the skin wound repair effect is significantly improved, and the skin wound repair effect is significantly improved; and the recombinant collagen provides structural support for the skin and assists the recombinant elastin in improving the matrix elasticity, and the optimal proportion of the recombinant collagen and the recombinant elastin ensures the high efficiency of tissue regeneration and healing.

Description

Technical Field

[0001] The invention relates to the technical field of medical cosmetology, in particular to a multifunctional skin repairing liquid based on recombinant elastin. Background Art

[0002] According to the Chinese patent publication number "CN115894709A" disclosed a recombinant human elastin with skin damage repair effect and its application, it relates to a recombinant human elastin with skin damage repair effect and its application. The present invention provides a recombinant human elastin, the amino acid sequence of the recombinant human elastin is completely derived from human elastin, and has a secondary structure and reversible phase transition characteristics similar to natural elastin; the recombinant human elastin is efficiently expressed in Escherichia coli engineered bacteria.

[0003] According to a recombinant elastin protein, a preparation method and a use thereof disclosed in Chinese Patent Publication No. "CN117551184A", the amino acid sequence of the recombinant elastin protein comprises at least one of the following: (1) an amino acid sequence as shown in SEQ ID N.1 comprising N tandemly connected amino acids; N represents an integer ≥ 1; (2) an amino acid sequence having a homology of ≥ 45% with the amino acid sequence as shown in SEQ ID N.1; compared with elastin from animal sources, the recombinant elastin protein of the present invention has the characteristics of better water solubility and low immunogenicity, exhibits good biocompatibility and biological activity, can significantly promote the proliferation, adhesion and migration of human skin keratinocytes and fibroblasts, and has a repair function for scalded skin.

[0004] Elastin is an important extracellular matrix protein in animals. Its content in the body is second only to collagen. It plays an irreplaceable role in tissue expansion and contraction, resistance to external deformation, and its greatest feature is that it gives tissue elasticity. Elastin is widely used, but the production of natural elastin is extremely limited and the cost is high, which limits its application. The production process of natural elastin is time-consuming, the extraction process is complicated, and the final yield is extremely low. There are also safety issues.

[0005] At present, the research on recombinant proteins is increasing day by day, and the research on recombinant elastin and recombinant collagen is also increasing, and basic and clinical applications are being carried out. Recombinant proteins are obtained by splicing elastin or collagen gene sequences into plasmid vectors based on genetic engineering technology, transferring them to microbial, animal or plant recombinant expression systems, and then fermenting and separating and purifying them. This solves the problem that natural elastin and natural collagen, as heterologous proteins, have the risk of disease transmission and immunogenicity. Summary of the invention

[0006] Technical issues solved

[0007] In view of the deficiencies of the prior art, the present invention provides a multifunctional skin repair solution based on recombinant elastin, which solves the problem that natural elastin, as a heterologous protein, has the risk of disease transmission and immunogenicity.

[0008] Technical Solution

[0009] To achieve the above objectives, the present invention is implemented by the following technical scheme: a multifunctional skin repair solution based on recombinant elastin, wherein the multifunctional skin repair solution comprises the following components by mass percentage:

[0010] 0.01% to 0.1% of recombinant elastin, 0.05% to 1% of recombinant collagen, 0.01% to 0.1% of ergothioneine, 0.1% to 0.3% of carbomer, 10% to 40% of glycerol, 0.5% to 5% of 1,2-hexanediol, 0.5% to 5% of p-hydroxyacetophenone, 0.01% to 0.5% of plant extract, the plant extract comprising one or more of aloe extract, Centella asiatica extract or green tea polyphenols, 0.01% to 0.5% of edible-grade phospholipids, 0.01% to 0.1% of nano-hyaluronic acid, and the balance being water.

[0011] Preferably, the plant extract includes any combination of one or more of aloe vera extract, Centella asiatica extract or green tea polyphenols, with a content of 0.05% to 0.1%.

[0012] Preferably, 0.001% to 0.01% of microencapsulated liposomes are further added to the multifunctional skin repairing liquid, and the microencapsulated liposomes encapsulate the plant extract and nano hyaluronic acid.

[0013] Preferably, the multifunctional skin repairing liquid is further added with 0.001% to 0.01% of bio-electrode particles, and the bio-electrode particles generate bio-electric signals by contacting with the skin surface.

[0014] Preferably, the multifunctional skin repairing liquid further comprises 0.005% to 0.02% of a natural polysaccharide-based protective film component, wherein the natural polysaccharide comprises chitosan or xanthan gum, which is used to form a protective biofilm on the skin surface.

[0015] Preferably, the recombinant elastin and recombinant collagen in the multifunctional skin repair solution are pretreated by a biological enzyme cross-linking technology to form a three-dimensional network support structure after contacting the skin.

[0016] Preferably, the preparation process of the multifunctional skin repairing liquid is a triple emulsification process, and the triple emulsification process comprises the following steps:

[0017] Sp1, the first emulsification forms a stable water-in-oil system through high-speed shear emulsification technology;

[0018] Sp2, the second emulsification forms a water-in-oil-in-water system by adding edible grade phospholipids;

[0019] Sp3, the third emulsification achieves homogenized multiphase emulsion through nano homogenizer.

[0020] Preferably, the preparation process of the multifunctional skin repairing liquid is a two-phase emulsification process, which specifically comprises the following steps:

[0021] Sp1, add recombinant elastin, recombinant collagen, ergothioneine, p-hydroxyacetophenone, plant extract, glycerin and 1,2-hexanediol to part of the water and stir evenly to form the first phase;

[0022] Sp2, add edible lecithin to the remaining water, perform ultrasonic oscillation emulsification to form the second phase, and then add carbomer to form a gel;

[0023] Sp3, slowly add the first phase into the second phase, stirring at a high speed of 8000-12000 rpm, and after uniform mixing, process with a high-pressure homogenizer to obtain the repair liquid.

[0024] Preferably, the recombinant elastin and recombinant collagen are both derived from microbial fermentation and purification, and have 100% homology with the amino acid sequences of human elastin and type III collagen.

[0025] Beneficial Effects

[0026] The present invention provides a multifunctional skin repair solution based on recombinant elastin, which has the following beneficial effects:

[0027] 1. The present invention significantly improves the speed of skin wound repair by introducing the synergistic effect of recombinant elastin and recombinant collagen. Recombinant elastin accelerates collagen synthesis and tissue repair by promoting cell proliferation and skin matrix reconstruction, while recombinant collagen provides structural support for the skin and improves matrix elasticity. The optimal ratio of the two ensures the high efficiency of tissue regeneration and healing. In addition, a stable carrier system is formed in combination with carbomer, so that the active ingredients can act on the wound for a long time, further shortening the healing cycle of acute injuries. It is of great significance in medical cosmetology of the skin surface, and the skin surface can be repaired quickly and the skin recovery can be accelerated. At the same time, the rapid healing performance is also of great significance for the clinical treatment of acute injuries such as burns and postoperative skin repair. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 This is a comparison chart of the experimental results of the skin wound repairing liquid of the present invention. DETAILED DESCRIPTION

[0029] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention. Specific embodiment one:

[0031] As shown in the figure, the multifunctional skin repair solution based on recombinant elastin comprises the following components by mass percentage:

[0032] Recombinant elastin 0.01% to 0.1%, recombinant collagen 0.05% to 1%, ergothioneine 0.01% to 0.1%, carbomer 0.1% to 0.3%, glycerol 10% to 40%, 1,2-hexanediol 0.5% to 5%, p-hydroxyacetophenone 0.5% to 5%, plant extract 0.01% to 0.5%, plant extract includes one or more of aloe extract, Centella asiatica extract or green tea polyphenols, edible grade phospholipids 0.01% to 0.5%, nano hyaluronic acid 0.01% to 0.1%, and the balance is water. Recombinant elastin and recombinant collagen are both derived from microbial fermentation and purification, and have 100% homology with the amino acid sequence of human elastin and type III collagen. Plant extracts include any combination of one or more of aloe extract, Centella asiatica extract or green tea polyphenols, with a content of 0.05% to 0.1%. 0.001% to 0.01% of microencapsulated liposomes are further added to the multifunctional skin repair liquid, and the microencapsulated liposomes encapsulate plant extracts and nano hyaluronic acid. 0.001% to 0.01% of bioelectrode particles are further added to the multifunctional skin repair liquid, and the bioelectrode particles generate bioelectric signals by contacting the skin surface. The multifunctional skin repair liquid further includes 0.005% to 0.02% of natural polysaccharide-based protective film components, and natural polysaccharides include chitosan or xanthan gum, which are used to form a protective biofilm on the skin surface. The recombinant elastin and recombinant collagen in the multifunctional skin repair liquid are pretreated by bio-enzyme cross-linking technology to form a three-dimensional mesh support structure after contact with the skin.

[0033] Recombinant elastin (0.01% to 0.1%): As the main repair factor, it simulates natural elastin and has excellent skin affinity and tissue compatibility. It improves skin elasticity, promotes cell regeneration, and accelerates wound healing.

[0034] Recombinant collagen (0.05% to 1%): As a skin conditioning factor, it cooperates with recombinant elastin to form a skin-like matrix structure, significantly improving the stretchability and durability of the repair fluid. It also enhances the moisturizing effect and skin barrier function.

[0035] Ergothioneine (0.01% to 0.1%): A powerful antioxidant that can remove free radicals, reduce UV damage to the skin, inhibit inflammatory reactions, and improve the skin's ability to resist stress.

[0036] Carbomer (0.1% to 0.3%): As a thickener, it gives the repair solution a moderate viscosity and improves the application experience. It increases the residence time of other active ingredients on the skin surface and enhances absorption efficiency.

[0037] Glycerin (10% to 40%): A powerful moisturizer that locks in moisture. At high proportions, it can also relieve skin irritation and improve skin comfort.

[0038] 1,2-Hexanediol (0.5%~5%): Auxiliary moisturizer, enhances water retention. It also has antibacterial effect to prevent the repair fluid from being contaminated.

[0039] Parahydroxyacetophenone (0.5% to 5%): A naturally derived antibacterial and preservative ingredient that can work synergistically with 1,2-hexanediol to further enhance the preservative effect, and can work synergistically with glycerin and 1,2-hexanediol to further enhance the moisturizing effect.

[0040] Plant extracts (0.01% to 0.5%): including one or more of aloe vera extract, centella asiatica extract and green tea polyphenols. Aloe vera extract has a calming and anti-inflammatory effect and is suitable for sensitive skin; centella asiatica extract promotes wound healing; green tea polyphenols provide antioxidant and anti-aging protection.

[0041] Nano hyaluronic acid (0.01% to 0.1%): small molecular weight design, particle size of 50 to 100 nm, can penetrate the skin surface to the dermis, provide deep moisturizing effect, and promote the penetration and absorption of other active ingredients.

[0042] Edible lecithin (0.01% to 0.5%): As a natural emulsifier, it combines with carbomer to form a stable emulsification system, improving the skin's absorption rate and the stability of the repair solution.

[0043] Microencapsulated liposomes (0.001% to 0.01%): encapsulate plant extracts and nano-hyaluronic acid, and use controlled release technology to achieve long-term release. Form a slowly penetrating reservoir on the skin surface to continuously provide repair and moisturizing ingredients.

[0044] Bioelectrode particles (0.001% to 0.01%): generate weak bioelectric signals by contacting the skin, stimulating cell repair and regeneration, and significantly improving skin cell activity.

[0045] Natural polysaccharide-based protective film (0.005% to 0.02%): Chitosan or xanthan gum forms a protective biofilm on the skin surface to prevent external pollution and bacterial invasion, while reducing water evaporation.

[0046] Pretreatment with bio-enzyme cross-linking technology: Recombinant elastin and recombinant collagen are treated with enzymes to form a three-dimensional network support structure, ensuring their stable stay on the skin surface and improving the repair effect. Specific embodiment 2:

[0048] The preparation process of the multifunctional skin repairing liquid according to the specific embodiment 1 is a triple emulsification process, and the triple emulsification process includes the following steps:

[0049] Sp1, the first emulsification forms a stable water-in-oil system through high-speed shear emulsification technology;

[0050] Sp2, the second emulsification forms a water-in-oil-in-water system by adding edible grade phospholipids;

[0051] Sp3, the third emulsification achieves homogenized multiphase emulsion through nano homogenizer.

[0052] The triple emulsification process highly disperses multiple active ingredients through layer-by-layer emulsification to form a stable emulsified system, which significantly improves the absorption efficiency and stability of the skin repair fluid. Specifically, it includes the following steps: the first emulsification (Sp1) uses high-speed shear emulsification technology to form a water-in-oil (O / W) system to encapsulate the water-soluble active ingredients in the oil phase; the second emulsification (Sp2) forms a water-in-oil-in-water (W / O / W) system by adding edible-grade phospholipids, which protects the active ingredients while enhancing stability; the third emulsification (Sp3) uses a nano-homogenizer to homogenize the emulsion, further reduce the particle size and optimize the dispersibility of the emulsion.

[0053] First emulsification: Add recombinant elastin, recombinant collagen, nano hyaluronic acid and other active ingredients to the water phase, stir until completely dissolved, then slowly add them to the oil phase (such as vegetable oil), and use a high-speed shear emulsifier (shear speed 8000-12000rpm) to stir for 5-10 minutes to make the water-soluble ingredients form a stable oil-in-water (O / W) emulsion. The key to this step is the uniformity of the shear speed and the addition speed to prevent stratification or emulsion rupture.

[0054] Second emulsification: Add 0.01% to 0.5% of edible phospholipids to the O / W emulsion formed by the first emulsification, and stir at a low speed (300 to 500 rpm) for 5 minutes to evenly distribute the phospholipids, then slowly add them to the external aqueous phase (containing carbomer solution and other auxiliary ingredients), and stir at a medium speed (500 to 1000 rpm) for 10 minutes to form a stable water-in-oil-in-water (W / O / W) emulsion. This step stabilizes the water phase, oil phase and multifunctional active ingredients through the amphiphilic properties of phospholipids to avoid component loss or degradation.

[0055] The third emulsification: Place the W / O / W emulsion formed by the second emulsification in a nano homogenizer and process it 3 to 5 times at a working pressure of 500 to 1000 bar to control the particle size distribution of the emulsion between 100 and 200 nm. The homogenization process reduces large particles in the emulsion, improves dispersibility and stability, and enhances skin penetration and absorption efficiency. The temperature must be strictly controlled at 25°C to 50°C throughout the process to avoid inactivation of ingredients due to high temperature. At the same time, the ratio of water phase to oil phase should be maintained between 60:40 and 80:20 to ensure the success rate of emulsification.

[0056] The core of the triple emulsification process lies in the formation of a multi-layer emulsification structure and the homogenization of particle size, which significantly improves the physical and chemical stability of the repair fluid, allowing it to maintain its performance in both high and low temperature environments; enhances skin absorption efficiency, ensuring efficient penetration of reconstructed elastin, collagen and plant extracts; protects active ingredients from environmental influences, while extending the shelf life of the product. Specific embodiment three:

[0058] The preparation process of the multifunctional skin repairing liquid based on the specific embodiment 1 is prepared by a two-phase emulsification process, which specifically includes the following steps:

[0059] Sp1, add recombinant elastin, recombinant collagen, ergothioneine, p-hydroxyacetophenone, plant extract, glycerin and 1,2-hexanediol to part of the water and stir evenly to form the first phase;

[0060] Sp2, add edible lecithin to the remaining water, perform ultrasonic oscillation emulsification to form the second phase, and then add carbomer to form a gel;

[0061] Sp3, slowly add the first phase into the second phase, stirring at a high speed of 8000-12000 rpm, and after uniform mixing, process it with a high-pressure homogenizer to obtain a repair liquid.

[0062] The two-phase emulsification process forms a stable emulsified system by combining the dispersion and homogenization of water-soluble and oil-soluble components, thereby significantly improving the absorption efficiency and stability of the repair fluid. The specific steps are as follows: Step 1 (Sp1), prepare the first phase, add recombinant elastin, recombinant collagen, ergothioneine, parahydroxyacetophenone, plant extracts, glycerol and 1,2-hexanediol to part of the water, and use a medium-speed mixer to stir at a speed of 500-800rpm for 10-15 minutes to ensure that all ingredients are dissolved and evenly distributed to form a water-soluble transparent solution (first phase). The water temperature needs to be controlled at 25℃~30℃ at this stage to prevent the inactivation of the components. Step 2 (Sp2), prepare the second phase, add edible grade phospholipids to the remaining part of the water, and use ultrasonic emulsification equipment (20-40kHz) for 10 minutes to ensure the formation of a stable and uniform emulsified dispersion system (second phase). During the oscillation process, the temperature must be strictly controlled not to exceed 40℃ to avoid the reduction of emulsifier performance. The third step (Sp3) is two-phase mixing and high-pressure homogenization. The first phase is slowly added to the second phase, and mixed at a speed of 8000-12000rpm with a high-speed stirrer for 15-20 minutes to ensure that the two phases are fully integrated. The mixed emulsion is then treated with a high-pressure homogenizer, and the working pressure is set to 500-1000bar. The treatment is performed 3-5 times so that the particle size of the final emulsion is evenly distributed in the range of 100-200nm. The core of the entire process lies in the synergistic effect of the two-phase emulsification step. The first phase provides the basic dispersion of water-soluble active ingredients, and the second phase provides a protective barrier through a stable system formed by phospholipids and carbomer. High-pressure homogenization further improves the uniformity of particle size and skin absorption efficiency.

[0063] The solubility and activity of recombinant elastin and collagen are maintained as the core, and the multi-layer structure stability and dispersion uniformity of the emulsion are ensured through two emulsification and homogenization operations. The introduction of ultrasonic emulsification enables the second phase to achieve high uniformity in a short time, reducing the risk of emulsion stratification; high-pressure homogenization further enhances permeability and ingredient absorption by reducing particle size. The entire preparation process is simple and efficient, suitable for large-scale industrial production, especially for the development of complex formulas and high-end repair fluids. At the same time, by controlling the temperature and stirring speed of the emulsification system, the maximum utilization rate of the active ingredients is guaranteed, and the repair fluid finally prepared has excellent performance in terms of absorbency, moisturizing and stability. Specific embodiment four:

[0065] Based on the above specific embodiments, experiments were conducted to obtain the following experimental contents:

[0066] Test Example 1: A liquid dressing for skin wound repair, wherein the liquid dressing is made of the following raw materials in terms of weight percentage:

[0067] Recombinant elastin 0.01%, recombinant collagen 0.1%, ergothioneine 0.1%, carbomer 0.1%, glycerin 30%, 1,2-hexanediol 1%, p-hydroxyacetophenone 1%, and the balance is water.

[0068] The medical dressing prepared with the above-mentioned liquid dressing for skin wound repair is a facial mask, an eye mask, a dressing, a spray dressing or a liquid wound dressing, and the preparation method comprises the following steps:

[0069] (1) taking 30% of the prescription amount of purified water, adding and dissolving the recombinant elastin, recombinant collagen, ergothioneine and p-hydroxyacetophenone in the prescription amount in sequence, and then adding the prescription amount of glycerol and 1,2-hexanediol to make a mixed solution for standby use;

[0070] (2) Weigh carbomer and the remaining amount of purified water, add carbomer to the purified water, homogenize to obtain a mixture, and let it stand for later use;

[0071] (3) Add the mixed solution into the mixture, stir and homogenize to obtain a recombinant elastin repair solution.

[0072] The non-woven fabric is soaked in the recombinant elastin repair liquid obtained in the above (3) to obtain the recombinant elastin repair dressing.

[0073] Test Example 2: A liquid dressing for skin wound repair, wherein the liquid dressing is made of the following raw materials by weight percentage:

[0074] Recombinant elastin 0.005%, recombinant collagen 0.1%, ergothioneine 0.1%, carbomer 0.1%, glycerin 30%, 1,2-hexanediol 1%, p-hydroxyacetophenone 1%, and the balance is water.

[0075] The medical dressing prepared with the above-mentioned liquid dressing for skin wound repair is a facial mask, an eye mask, a dressing, a spray dressing or a liquid wound dressing, and the preparation method comprises the following steps:

[0076] (1) taking 30% of the prescription amount of purified water, adding and dissolving the recombinant elastin, recombinant collagen, ergothioneine and p-hydroxyacetophenone in the prescription amount in sequence, and then adding the prescription amount of glycerol and 1,2-hexanediol to make a mixed solution for standby use;

[0077] (2) Weigh carbomer and the remaining amount of purified water, add carbomer to the purified water, homogenize to obtain a mixture, and let it stand for later use;

[0078] (3) Add the mixed solution into the mixture, stir and homogenize to obtain a recombinant elastin repair solution.

[0079] The non-woven fabric is soaked in the recombinant elastin repair liquid obtained in the above (3) to obtain the recombinant elastin repair dressing.

[0080] Test Example 3: A liquid dressing for skin wound repair, wherein the liquid dressing is made of the following raw materials by weight percentage:

[0081] Recombinant elastin 0.05%, recombinant collagen 0.1%, ergothioneine 0.1%, carbomer 0.1%, glycerin 30%, 1,2-hexanediol 1%, p-hydroxyacetophenone 1%, and the balance is water.

[0082] The medical dressing prepared with the above-mentioned liquid dressing for skin wound repair is a facial mask, an eye mask, a dressing, a spray dressing or a liquid wound dressing, and the preparation method comprises the following steps:

[0083] (1) taking 30% of the prescription amount of purified water, adding and dissolving the recombinant elastin, recombinant collagen, ergothioneine and p-hydroxyacetophenone in the prescription amount in sequence, and then adding the prescription amount of glycerol and 1,2-hexanediol to make a mixed solution for standby use;

[0084] (2) Weigh carbomer and the remaining amount of purified water, add carbomer to the purified water, homogenize to obtain a mixture, and let it stand for later use;

[0085] (3) Add the mixed solution into the mixture, stir and homogenize to obtain a recombinant elastin repair solution.

[0086] The non-woven fabric is soaked in the recombinant elastin repair liquid obtained in the above (3) to obtain the recombinant elastin repair dressing.

[0087] Compared with the test example, the difference is that it does not contain recombinant elastin:

[0088] Recombinant collagen 0.1%, ergothioneine 0.1%, carbomer 0.1%, glycerin 30%, 1,2-hexanediol 1%, p-hydroxyacetophenone 1%, and the balance is water.

[0089] The preparation method comprises the following steps:

[0090] Take 30% of the prescribed amount of purified water, add and dissolve the recombinant collagen, ergothioneine and p-hydroxyacetophenone in the prescribed amount in sequence, then add the prescribed amount of glycerol and 1,2-hexanediol to prepare a mixed solution for standby use;

[0091] Weigh carbomer and the remaining amount of purified water, add carbomer to the purified water, homogenize to obtain a mixture, and let it stand for later use;

[0092] The mixed solution was added to the mixture and stirred to be homogenous to obtain a control solution without recombinant elastin.

[0093] The non-woven fabric is soaked with the comparative solution obtained in the above (3) to obtain the repair dressing without recombinant elastin.

[0094] Comparative Example 2:

[0095] Compared with the test example, the difference is that it does not contain recombinant elastin and recombinant collagen:

[0096] Ergothioneine 0.1%, carbomer 0.1%, glycerin 30%, 1,2-hexanediol 1%, p-hydroxyacetophenone 1%, and the balance is water.

[0097] The preparation method comprises the following steps:

[0098] Take 30% of the prescribed amount of purified water, add and dissolve the recombinant collagen, ergothioneine and p-hydroxyacetophenone in the prescribed amount in sequence, then add the prescribed amount of glycerol and 1,2-hexanediol to prepare a mixed solution for standby use;

[0099] Weigh carbomer and the remaining amount of purified water, add carbomer to the purified water, homogenize to obtain a mixture, and let it stand for later use;

[0100] The mixed solution was added to the mixture and stirred to be homogenous to obtain a control solution without recombinant elastin.

[0101] The non-woven fabric is soaked with the comparative solution obtained in the above (3) to obtain the repair dressing without recombinant elastin.

[0102] Effect test:

[0103] Mouse wound repair test:

[0104] 1.1 Model making (18 pieces):

[0105] 18 BALB / C mice, SPF, male, weighing 20±2g. 12 mice were shaved on the back, and then sodium sulfide was applied for hair removal, so that the back epidermis was completely exposed. The hair removal area was about 3×3cm. After the hair removal was completed, it was stabilized for 24 hours to eliminate the effect of sodium sulfide on epidermal damage. The damaged skin method was to gently rub with fine sandpaper until bleeding occurred, and obvious local redness and swelling occurred on the back of the mouse, indicating that acute skin inflammation was successfully induced in the mouse.

[0106] 1.2 Animal grouping (30 animals):

[0107] Six mice without modeling were selected as the blank group and 12 mice with successful modeling were randomly divided into the following groups:

[0108] Blank group: 6;

[0109] Control group: 6;

[0110] Model group: 6;

[0111] 1.3 Dosage and course of treatment:

[0112] Blank group: no treatment.

[0113] Control group: normal saline was applied to the wound, 6 times / 48h.

[0114] Model group: Apply the same medicine as in Test Example 1 to the wound, 6 times / 48 hours.

[0115] During the experiment, mice in each group had free access to food and water.

[0116] The skin of the mice in the blank group was in good shape within 1-7 days, with a smooth and flat surface and no red and swollen areas. The skin of the mice in the control group was rubbed with fine sandpaper until bleeding, and a large amount of local redness and swelling appeared on the epidermis on the first day; the red and swollen area further expanded on the third day, and irregular cracks appeared on the outer layer of the epidermis; the local redness and swelling improved on the fifth day; the local redness and swelling basically subsided on the seventh day, and the epidermal erythema began to scab, but it was still accompanied by a small amount of epidermal shedding. After the mice in the experimental group were rubbed with fine sandpaper until bleeding, the recombinant elastin repair fluid was applied. On the first day, the epidermis of the mice showed partial redness and swelling; on the third day, the area of ​​the red and swollen area decreased, and wrinkles appeared on the skin surface; on the fifth day, the redness and swelling basically subsided, and the skin wrinkles were significantly improved, accompanied by local epidermal shedding; on the seventh day, the skin of the mice recovered well, with a smooth and flat surface, and there was no significant difference from the skin of the mice in the blank group. These results show that the recombinant elastin repair fluid designed by this patent has a significant repair effect on acute skin damage in mice, can effectively reduce the degree of skin damage, and shorten the repair cycle of acutely damaged skin.

[0117] Human wound repair test:

[0118] (1) Experimental subjects:

[0119] 120 patients who had just completed microneedle surgery on their faces were randomly selected, including 18 males and 102 females, aged 18-54 years old, and were randomly divided into 6 groups, 20 cases in each group. There was no significant difference in the basic information of each patient, p>0.05, and comparison was possible;

[0120] (2) Treatment methods:

[0121] After cleaning the skin lesions every day, use about 5 mL of the skin wound dressing of each experimental group (Test Examples 1, 2 and 3, Comparative Examples 1 and 2, and the positive control commercial product "recombinant collagen dressing"), apply once a day at night during the 1st to 10th day, and apply once every other night during the 10th to 20th day, and each application time is 20 minutes;

[0122] (3) Treatment standards:

[0123] Ⅰ. Skin symptom scoring: erythema, edema, exudation, tingling, and pigmentation are scored as 0-3 points, with none, mild, moderate, and severe, respectively. The specific standards are as follows:

[0124] Erythema: bright red or dark red erythema is scored 3 points; obvious erythema is scored 2 points; light erythema is scored 1 point; no color change is scored 0 points.

[0125] Edema: Extensive dermal edema infiltration was scored as 3 points; multiple clear dermal edema was scored as 2 points; local dermal edema that was not easily palpable was scored as 1 point; no edema was scored as 0 points.

[0126] Exudation: If the exudation is very obvious and covers a large area, it will be scored 3 points; if the exudation is relatively obvious and covers a small area, it will be scored 2 points; if punctate exudation is seen after careful observation, it will be scored 1 point; if there is no exudation, it will be scored 0 points.

[0127] Tingling: Continuous tingling, unbearable, scored 3 points; continuous or intermittent tingling, tolerable, scored 2 points; occasional, mild tingling, scored 1 point; no tingling, scored 0 points.

[0128] Pigmentation: 3 points if the pigmentation is very obvious and covers a large area; 2 points if the pigmentation is relatively obvious and covers a small area; 1 point if slight pigmentation is visible after careful observation; 0 point if no pigmentation is present.

[0129] Efficacy evaluation standards: divided into four standards: cure, remarkable effect, effectiveness, and ineffectiveness.

[0130] Therapeutic index = [(pre-treatment score - post-treatment score) ÷ pre-treatment score] × 100%.

[0131] Cured: efficacy index ≥ 95%; markedly effective: efficacy index 70%-94%; effective: efficacy index 30%-69%; ineffective: efficacy index < 30%. Total effective rate = (number of cured cases + number of markedly effective cases + number of effective cases) ÷ total number of cases × 100%.

[0132] Ⅱ. Wound healing time: The average wound healing time (days) of patients in each experimental group and control group was calculated respectively;

[0133] Statistical test method: The results were analyzed using SPSS19.0 statistical software, and the data were expressed as mean ± standard deviation. One-way analysis of variance was used for comparison between groups, and p < 0.05 indicated statistically significant differences.

[0134] (4) Treatment results:

[0135] 1) The results of skin symptoms and average wound healing time are shown in the following table:

[0136] Table: Skin symptoms and average wound healing time

[0137]

[0138] The results in the table show that compared with Comparative Examples 1 and 2, the subjects using Test Examples 1 and 3 showed significant improvements in skin symptom scores and average wound healing time, which suggests that combining recombinant elastin with recombinant collagen in the skin wound repair liquid dressing of the present invention can increase the effect of the liquid dressing in improving skin symptom scores and average wound healing time.

[0139] When the ratio of recombinant elastin to recombinant collagen is 1:10 (Test Example 1), the effect is better than that of other ratios (1:20 and 1:2, Test Examples 2 and 3). The liquid dressing can significantly increase the repair effect on the skin wound, and its total effective rate can reach 100%, and the average wound healing time is only 4 days.

[0140] The skin symptoms and average wound healing time of the skin wound repair liquid dressing of the present invention (Test Examples 1, 2 and 3) were better than those of the positive control. Specific embodiment five:

[0142] Further based on the technical solutions of the above-mentioned specific embodiments 1 to 4, the following experiments are further conducted:

[0143] Experiment 1: Skin repair effect verification experiment

[0144] Purpose:

[0145] To evaluate the effect of a multifunctional skin repair solution based on recombinant elastin on skin repair.

[0146] Formulas for the experimental group and the control group:

[0147] Experimental group: Multifunctional skin repair solution based on recombinant elastin, prepared according to the following formula:

[0148] Recombinant elastin: 0.05%, recombinant collagen: 0.5%, ergothioneine: 0.05%, carbomer: 0.2%, glycerin: 20%, 1,2-hexanediol: 2%, p-hydroxyacetophenone: 2%, plant extract (aloe extract, Centella asiatica extract): 0.1%, edible grade phospholipids: 0.1%, nano hyaluronic acid: 0.05%, water: balance;

[0149] Control group: common repair solution on the market (without recombinant elastin, and mainly composed of glycerin, hyaluronic acid and carbomer).

[0150] Experimental design:

[0151] Experimental subjects: 30 healthy adult volunteers, aged 20-40 years old, with minor skin injuries (including small abrasions or scrapes).

[0152] Experimental period: 14 days, use skin repair solution twice a day.

[0153] Experimental indicators:

[0154] Skin injury recovery time: measures the number of days it takes for a wound to fully heal.

[0155] Skin elasticity recovery: The recovery rate was measured using a skin elasticity tester (including Cutometer).

[0156] Redness, swelling and pain reactions: Visual and tactile scoring is used to observe and record skin reactions.

[0157]

[0158] Experimental data:

[0159] Skin injury recovery time: The wound healing time in the experimental group was significantly shorter than that in the control group (6 days vs. 9 days).

[0160] Skin elasticity recovery rate: The skin elasticity recovery rate of the experimental group was higher (95% vs 85%).

[0161] Redness, swelling and pain response: The redness, swelling and pain scores of the experimental group were significantly lower than those of the control group.

[0162] in conclusion:

[0163] The multifunctional skin repair fluid based on recombinant elastin is superior to the control group in terms of skin repair effect, elasticity recovery and side effects, with better repair effect and less irritation reaction.

[0164] Experiment 2: Ingredient stability experiment

[0165] Purpose:

[0166] Verify the stability of a multifunctional skin repair solution based on recombinant elastin.

[0167] Formulas for the experimental group and the control group:

[0168] Experimental group: Multifunctional skin repair solution based on recombinant elastin, prepared according to the formula of Experiment 1.

[0169] Control group: common repair fluid available on the market (without recombinant elastin).

[0170] Experimental design:

[0171] Storage conditions: Store at room temperature (25°C), high temperature (40°C), and low temperature (5°C) for 3 months.

[0172] Observation indicators:

[0173] Appearance changes: Observe for precipitation, stratification or discoloration.

[0174] pH Change: Use a pH meter to measure the pH of the repair solution.

[0175] Changes in component content: Changes in the content of recombinant elastin and collagen were tested by HPLC or ELISA.

[0176] Experimental data:

[0177]

[0178] in conclusion:

[0179] The repair fluid showed good stability under different storage conditions. Although there was a slight loss of ingredients under high temperature conditions, it was still within the effective range. The appearance did not change much, indicating that the formula was stable.

[0180] Experiment 3: Skin permeability test

[0181] Purpose:

[0182] To evaluate the penetration of active ingredients in a multifunctional skin repair solution based on recombinant elastin at the skin surface and deep layers.

[0183] Formulas for the experimental group and the control group:

[0184] Experimental group: Multifunctional skin repair solution based on recombinant elastin, prepared according to the formula of Experiment 1.

[0185] Control group: common repair fluid available on the market (without recombinant elastin).

[0186] Experimental design:

[0187] Experimental subjects: 4 healthy adult volunteers were tested in the experimental group and the control group respectively.

[0188] Experimental method: Use a transdermal instrument to measure the penetration concentration of the repair solution on the skin surface and deep layers at different time points (0h, 2h, 6h, 12h).

[0189] Experimental data:

[0190]

[0191] Skin surface concentration: The concentration of the experimental group on the skin surface was significantly higher than that of the control group.

[0192] Deep skin concentration: The penetration concentration of the repair fluid in the experimental group in the deep skin layer was also significantly higher than that in the control group, indicating better permeability.

[0193] in conclusion:

[0194] The skin repair solution based on recombinant elastin has better penetration into the deep layers of the skin than ordinary commercially available repair solutions, indicating that its ingredients have better penetration properties.

[0195] Experiment 4: Skin irritation test

[0196] Purpose:

[0197] To evaluate the irritation of skin repair solutions to the skin.

[0198] Formulas for the experimental group and the control group:

[0199] Experimental group: Multifunctional skin repair solution based on recombinant elastin, prepared according to the formula of Experiment 1.

[0200] Control group: common repair fluid available on the market (without recombinant elastin).

[0201] Experimental design:

[0202] Experimental subjects: 20 healthy adult volunteers, skin patch test was performed.

[0203] Observation period: 48 hours, record redness, swelling, itching, burning and other reactions.

[0204] Experimental data:

[0205] Group Redness and swelling score (0-4) Itch score (0-4) Experimental Group 0.1±0.2 0.2±0.3 Control group 1.2±0.4 1.3±0.5

[0206] Redness, swelling, itching, and irritation: The scores of the experimental group in terms of redness, swelling, itching, and irritation were significantly lower than those of the control group, indicating better skin adaptability.

[0207] in conclusion:

[0208] The skin repair solution based on recombinant elastin is less irritating to the skin.

[0209] Virtually no allergic reaction or irritation.

[0210] Experiment 5: Polysaccharide-based protective film effect experiment

[0211] Purpose:

[0212] Verify the auxiliary effect of natural polysaccharide-based protective film on skin repair.

[0213] Formulas for the experimental group and the control group:

[0214] Experimental group: prepared according to the formula of Experiment 1, with the addition of a natural polysaccharide (chitosan or xanthan gum) based protective film.

[0215] Natural polysaccharides: 0.01% (chitosan or xanthan gum).

[0216] Control group: repair solution without natural polysaccharides.

[0217] Experimental design:

[0218] Experimental subjects: 20 healthy adult volunteers, who were treated with a repair solution containing natural polysaccharides or a repair solution without natural polysaccharides after skin damage.

[0219] Experimental indicators:

[0220] Skin Repair Speed: Regularly assess the progress of skin healing.

[0221] Skin Hydration: Skin moisture levels are measured by skin hydration testers, including the Corneometer.

[0222] Experimental data:

[0223] Group Repair time (days) Skin hydration (%) Experimental Group 5±1 98±3 Control group 7±2 88±5

[0224] Repair time: The repair time of the experimental group was significantly shorter than that of the control group.

[0225] Hydration: The skin hydration of the experimental group was significantly higher than that of the control group, indicating that the natural polysaccharide-based protective film helps to retain skin moisture.

[0226] in conclusion:

[0227] The natural polysaccharide-based protective film can effectively accelerate skin repair and enhance skin hydration, playing a positive role in the repair process.

[0228] These detailed experimental data demonstrate the superior performance of the multifunctional skin repair fluid based on recombinant elastin.

[0229] It should be noted that, in this article, relational terms such as first and second, etc. are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the terms "include", "comprise" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device. In the absence of further restrictions, the elements defined by the statement "comprising a reference structure" do not exclude the existence of other identical elements in the process, method, article or device including the elements.

[0230] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A multifunctional skin repair solution based on recombinant elastin, characterized in that: The multifunctional skin repairing liquid comprises the following components by mass percentage: Recombinant elastin 0.01%-0.1%, recombinant collagen 0.05%-1%, ergothioneine 0.01%-0.1%, carbomer 0.1%-0.3%, glycerol 10%-40%, 1,2-hexanediol 0.5%-5%, p-hydroxyacetophenone 0.5%-5%, plant extract 0.01%-0.5%, edible grade phospholipids 0.01%-0.5%, nano hyaluronic acid 0.01%-0.1%, and the balance is water.

2. The multifunctional skin repair solution based on recombinant elastin according to claim 1, characterized in that: The plant extract comprises any combination of one or more of aloe extract, Centella asiatica extract or green tea polyphenol, with a content of 0.05% to 0.1%.

3. The multifunctional skin repair solution based on recombinant elastin according to claim 1, characterized in that: 0.001% to 0.01% of microencapsulated liposomes are further added to the multifunctional skin repairing liquid, and the microencapsulated liposomes encapsulate the plant extract and the nano hyaluronic acid.

4. The multifunctional skin repair solution based on recombinant elastin according to claim 1, characterized in that: The multifunctional skin repairing liquid is further added with 0.001% to 0.01% of bio-electrode particles, and the bio-electrode particles generate bio-electric signals by contacting with the skin surface.

5. The multifunctional skin repair solution based on recombinant elastin according to claim 1, characterized in that: The multifunctional skin repairing liquid further comprises 0.005% to 0.02% of a natural polysaccharide-based protective film component.

6. The multifunctional skin repair solution based on recombinant elastin according to claim 1, characterized in that: The recombinant elastin and recombinant collagen in the multifunctional skin repair liquid are pretreated by a biological enzyme cross-linking technology to form a three-dimensional mesh support structure after contacting the skin.

7. The multifunctional skin repair solution based on recombinant elastin according to any one of claims 1 to 6, characterized in that: The preparation process of the multifunctional skin repairing liquid is a triple emulsification process, and the triple emulsification process includes the following steps: Sp1, the first emulsification forms a stable water-in-oil system through high-speed shear emulsification technology; Sp2, the second emulsification forms a water-in-oil-in-water system by adding edible grade phospholipids; Sp3, the third emulsification achieves homogenized multiphase emulsion through nano homogenizer.

8. The multifunctional skin repair solution based on recombinant elastin according to any one of claims 1 to 6, characterized in that: The preparation process of the multifunctional skin repairing liquid is a two-phase emulsification process, which specifically comprises the following steps: Sp1, add recombinant elastin, recombinant collagen, ergothioneine, p-hydroxyacetophenone, plant extract, glycerin and 1,2-hexanediol to part of the water and stir evenly to form the first phase; Sp2, add edible lecithin to the remaining water, perform ultrasonic oscillation emulsification to form the second phase, and then add carbomer to form a gel; Sp3, slowly add the first phase into the second phase, stirring at a high speed of 8000-12000 rpm, and after uniform mixing, process with a high-pressure homogenizer to obtain the repair liquid.

9. The multifunctional skin repair solution based on recombinant elastin according to claim 1, characterized in that: The recombinant elastin and recombinant collagen are both derived from microbial fermentation and purification, and have 100% homology with the amino acid sequences of human elastin and type III collagen.

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