Moisturizing and repairing collagen dressing and preparation method thereof

Abstract

The invention relates to the technical field of collagen dressings, in particular to a moisturizing and repairing collagen dressing and a preparation method thereof. The moisturizing and repairing collagen dressing is prepared from the following components in parts by mass: 10 to 30 parts of recombinant human fibronectin, 10 to 20 parts of recombinant collagen, 5 to 15 parts of beta-glucan, 5 to 10 parts of compound hexanediol, 40 to 50 parts of carbomer, 5 to 15 parts of a flavoring agent, 80 to 120 parts of purified water, 10 to 30 parts of a modified recombinant human III type collagen solution and 10 to 20 parts of a modified chitosan solution. The recombinant human fibronectin disclosed by the invention can be deposited at an injured part to form a blood clot, so that the aim of stopping bleeding is fulfilled, and the subcutaneous tissue is protected; the recombinant collagen has high biological activity and can inhibit inflammation and improve the conditions of pachulosis, damage and the like of the skin; the beta-glucan can generate cytokines, promote healing of skin with laser-dependent dermatitis or barrier damage, and realize anti-allergic and anti-inflammatory effects.

Description

Technical Field

[0001] The invention relates to the technical field of collagen dressings, in particular to a moisturizing and repairing collagen dressing and a preparation method thereof. Background Art

[0002] Collagen is the main component of animal connective tissue and the most abundant and widely distributed functional protein in mammals, accounting for 25% to 30% of the total protein. Collagen has good biocompatibility, degradability and biological activity, so it has been widely used and paid attention to in the fields of food, medicine, cosmetics, tissue engineering, etc.

[0003] Patent No. CN115779140B discloses a wound dressing containing collagen and a preparation method, which includes the following components: benzimidazole quaternary ammonium salt modified silicone oil, type III recombinant human collagen, moisturizer, water, phosphotungstic acid, and triethanolamine.

[0004] Traditional dressings basically have no moisturizing function, and currently the main focus is on the research of medical dressings based on polymer materials and composite materials, such as hydrocolloid dressings. However, the use of dressings is no longer limited to medical use, but also extends to the field of skin care and the auxiliary treatment of some skin diseases, such as sensitive skin and dermatitis. Most of the existing technologies treat such skin diseases through topical hormones, but hormones have certain side effects. Long-term use can cause skin hormone dermatitis and thinning of the skin barrier. Therefore, we propose a moisturizing and repairing collagen dressing and a preparation method thereof. Summary of the invention

[0005] The object of the present invention is to provide a moisturizing and repairing collagen dressing and a preparation method thereof, so as to solve the problems raised in the above-mentioned background technology.

[0006] In order to solve the above technical problems, the present invention provides the following technical solutions: a moisturizing and repairing collagen dressing, comprising the following mass components: 10 to 30 parts of recombinant human fibronectin, 10 to 20 parts of recombinant collagen, 5 to 15 parts of β-glucan, 5 to 10 parts of compound hexylene glycol, 40 to 50 parts of carbomer, 5 to 15 parts of flavoring agents, 80 to 120 parts of purified water, 10 to 30 parts of modified recombinant human type III collagen solution, and 10 to 20 parts of modified chitosan solution.

[0007] A method for preparing a moisturizing and repairing collagen dressing comprises the following steps:

[0008] Recombinant human fibronectin, recombinant collagen, β-glucan, compound hexylene glycol, carbomer, flavoring agent, purified water, modified recombinant human type III collagen solution, and modified chitosan solution were mixed, nitrogen was passed through for 20 minutes, and then irradiated to prepare collagen dressing.

[0009] Furthermore, the irradiation dose rate is 40 to 60 Gy / min, and the total dose is 2400 to 3600 Gy.

[0010] In the above technical scheme, the irradiated dressing has high moisturizing properties, air permeability and biocompatibility. At the same time, irradiation can eliminate pathogenic microorganisms in the hydrogel, and no additional cross-linking agent needs to be added. The prepared dressing has stable mechanical properties and higher safety.

[0011] Furthermore, the compound hexylene glycol is prepared by mixing hexylene glycol and ethylhexylglycerin; the mass ratio of hexylene glycol to ethylhexylglycerin is 0.3:0.7.

[0012] Further, hexanediol: CAS number 920-22-5, purity 98%, sourced from Henan Weiyuan Biotechnology Co., Ltd.;

[0013] Ethylhexylglycerin, CAS number 70445-33-9, purity 99%, comes from Jinan Jinbang Environmental Protection Technology Co., Ltd.

[0014] Further, recombinant human fibronectin: CAS number 86088-83-7, purity 98%, from Hubei Ruiboxin Chemical Co., Ltd.;

[0015] Recombinant collagen: CAS number is 9007-34-5, purity is 99%, from Wuhan Huaxiang Kejie Biotechnology Co., Ltd.

[0016] β-Glucan: CAS number is 9012-72-0, purity is 99%, from Shenzhen Love Biotechnology Co., Ltd.

[0017] Carbomer: CAS number is 9007-20-9, purity is 99%, sourced from Guangdong Hongyou Biotechnology Co., Ltd.

[0018] The flavoring agent is vanillin, CAS number is 121-33-5, which comes from Xi'an Jinxiang Pharmaceutical Excipients Co., Ltd.

[0019] Furthermore, the modified recombinant human type III collagen solution is prepared by cross-linking a recombinant human type III collagen solution with 1-ethyl-(3-dimethylaminopropyl)carbodiimide and N-hydroxysuccinimide, and the specific process is as follows:

[0020] The recombinant human type III collagen is frozen in a low-temperature refrigerator, then transferred to a freeze dryer, and freeze-dried under vacuum until completely dehydrated. The dried collagen is placed in a 1-ethyl-(3-dimethylaminopropyl) carbodiimide solution, and then N-hydroxysuccinimide solution is added. The pH value is adjusted to 4-6 with sodium fatty acid methyl ester sulfonate solution as a buffer, and placed at 2-6°C for 18-24 hours to obtain a modified recombinant human type III collagen solution.

[0021] Furthermore, the quality fraction of recombinant human type III collagen is 3-7%, and is sourced from Beijing Aibixin Biotechnology Co., Ltd.

[0022] Furthermore, the temperature of the low-temperature refrigerator is -75~-85℃, model: GY-A250WN, sourced from Wuxi Guanya Constant Temperature Refrigeration Technology Co., Ltd., and the freezing time is 12h.

[0023] Furthermore, the freeze drying machine temperature is -100 to -110°C, model: FD-12A-110, sourced from Shanghai Shunzhi Instrument Manufacturing Co., Ltd., and the freeze drying time is 24h.

[0024] Further, 1-ethyl-(3-dimethylaminopropyl)carbodiimide, CAS No.: 1892-57-5,

[0025] The concentration of 1-ethyl-(3-dimethylaminopropyl)carbodiimide solution is 0.03-0.13 mol / L;

[0026] N-Hydroxysuccinimide, CAS No.: 6066-82-6,

[0027] The concentration of N-hydroxysuccinimide solution is 0.05-0.08 mol / L;

[0028] The mass ratio of 1-ethyl-(3-dimethylaminopropyl)carbodiimide solution, N-hydroxysuccinimide solution, and recombinant human type III collagen is 4:(1-3):(2-3), all of which are from Merck reagents.

[0029] Further, sodium fatty acid methyl ester sulfonate, CAS number: 1266615-59-1,

[0030] The concentration of sodium fatty acid methyl ester sulfonate solution is 15-25 mmol / L and is derived from Merck reagent.

[0031] In the above technical scheme, hexylene glycol has good moisturizing and antibacterial properties, ethylhexylglycerol can be used as a moisturizer and skin feel regulator at the same time, and the combination of the two can enhance the antibacterial effect and moisturizing properties of the dressing. In addition, ethylhexylglycerol also has a deodorizing effect, which inhibits the growth of odor-causing bacteria without affecting the skin flora that is beneficial to the human body; recombinant human fibronectin can be deposited at the site of injury to form a blood clot, thereby achieving the purpose of hemostasis, protecting the subcutaneous tissue, and further repairing the damaged tissue; recombinant collagen has high biological activity, can inhibit inflammation, and improve rough and damaged skin; β-glucan can produce cytokines, increase skin cell activity, promote metabolism, and promote the healing of laser-dependent dermatitis or skin with damaged barriers, achieving anti-allergic and anti-inflammatory effects;

[0032] The amino group on 1-ethyl-(3-dimethylaminopropyl) carbodiimide combines with the carboxyl group on the side chain of collagen to introduce an amino group, which improves the activity of collagen, further promotes cell proliferation and differentiation, promotes wound healing, and can repair the skin barrier more quickly than unmodified collagen. N-hydroxysuccinimide can form a stable hydrazide structure with the amino group, further forming a strong coupling, improving the stability of collagen. In addition, the modified collagen has a reduced repulsion to mammalian cells, reduces the adhesion of bacteria, and improves the biocompatibility and antibacterial properties of collagen.

[0033] Furthermore, the modified chitosan solution is obtained by cross-linking a chitosan solution with a modified hyaluronic acid hydrogel, and the specific process is as follows:

[0034] The chitosan solution and the modified hyaluronic acid hydrogel are mixed and stirred evenly, and then the genipin solution is added and stirred evenly to obtain the modified chitosan solution.

[0035] Furthermore, the volume ratio of the chitosan solution, the modified hyaluronic acid hydrogel, and the genipin solution is 5:(1-5):(0.1-0.3).

[0036] Further, genipin, CAS No.: 6902-77-8,

[0037] The concentration of genipin solution is 0.85-0.90 mol / L and is derived from Merck reagent.

[0038] Further, the chitosan solution is prepared by the following process:

[0039] Dissolve chitosan in acetic acid solution and stir evenly at 25°C to obtain a chitosan solution.

[0040] Furthermore, the volume ratio of chitosan to acetic acid solution is 1:50.

[0041] Further, chitosan, CAS number: 9012-76-4, was obtained from Merck reagents;

[0042] Acetic acid solution, CAS number: 64-19-17, concentration 0.15-0.20 mol / L, from Merck reagent.

[0043] Furthermore, the modified hyaluronic acid hydrogel is prepared by grafting graphene oxide with hyaluronic acid and then repeatedly freezing. The specific process is as follows:

[0044] Graphene oxide and hyaluronic acid were dissolved in deionized water, 4-dimethylaminopyridine and N,N-diisopropylcarbodiimide were added, and the mixture was stirred evenly at 20-30°C. After reacting for 24-36 hours, the obtained solution was placed in a dialysis bag with a relative molecular mass cutoff of 1400 and dialyzed for 5 hours to obtain a modified hyaluronic acid hydrogel.

[0045] Furthermore, the mass ratio of graphene oxide, hyaluronic acid gel, 4-dimethylaminopyridine, and N,N-diisopropylcarbodiimide is 20:(50-80):1:1;

[0046] Graphene oxide: particle size 10-20 μm, sourced from Jiangsu Xianfeng Nanomaterial Technology Co., Ltd.;

[0047] Hyaluronic acid, from Merck reagent.

[0048] Further, 4-dimethylaminopyridine, CAS No.: 1122-58-3;

[0049] N,N-Diisopropylcarbodiimide, CAS No.: 693-13-0;

[0050] All reagents were from Merck.

[0051] In the above technical solution, graphene oxide is grafted with hyaluronic acid, and the oxygen-containing functional groups on the surface of graphene oxide form a more stable structure with hyaluronic acid, which not only maintains good biocompatibility, but also improves the ability of hyaluronic acid to load and release drugs. Chitosan is a polymer material with good biocompatibility and antibacterial properties, but the hydrogel prepared by chitosan alone has weak water holding capacity. Combining chitosan with modified hyaluronic acid hydrogel can provide a moist tissue environment for the skin and inhibit bacteria, thereby accelerating wound healing.

[0052] Compared with the prior art, the beneficial effects achieved by the present invention are:

[0053] 1. The recombinant human type III collagen solution was modified by 1-ethyl-(3-dimethylaminopropyl) carbodiimide and N-hydroxysuccinimide, so that the amino group on 1-ethyl-(3-dimethylaminopropyl) carbodiimide was combined with the carboxyl group on the side chain of collagen, and the amino group was introduced, which improved the activity of collagen, further promoted cell proliferation and differentiation, and promoted wound healing. N-hydroxysuccinimide can form a stable hydrazide structure with the amino group, further forming a strong coupling, improving the stability of collagen, and can repair the skin barrier more quickly than unmodified collagen. In addition, the modified collagen has reduced repulsion to mammalian cells, reduced the adhesion ability of bacteria, and improved the biocompatibility and antibacterial properties of collagen.

[0054] 2. By grafting graphene oxide with hyaluronic acid, a more stable structure is formed on the surface of hyaluronic acid, which maintains good biocompatibility while improving the ability of hyaluronic acid to load and release drugs. The hydrogel prepared by chitosan alone has weak water holding capacity. Combining chitosan with modified hyaluronic acid can provide a moist tissue environment for the skin and inhibit bacteria, thereby accelerating wound healing.

[0055] 3. The dressing made by irradiation does not require the addition of additional cross-linking agents during the process, avoiding the possibility of chemical cross-linking agent residues being toxic or irritating to the skin or wounds. In addition, the prepared dressing has good moisturizing properties, air permeability and biocompatibility. At the same time, irradiation can eliminate pathogenic microorganisms in the hydrogel, making the dressing have better mechanical properties and higher safety. DETAILED DESCRIPTION

[0056] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. 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.

[0057] In the following specific implementations,

[0058] Hexanediol: CAS number is 920-22-5, purity is 98%, sourced from Henan Weiyuan Biotechnology Co., Ltd.

[0059] Ethylhexylglycerin, CAS No. 70445-33-9, purity 99%, sourced from Jinan Jinbang Environmental Protection Technology Co., Ltd.;

[0060] Carbomer: CAS number is 9007-20-9, purity is 99%, sourced from Guangdong Hongyou Biotechnology Co., Ltd.

[0061] The flavoring agent is vanillin, CAS No. 121-33-5, from Xi'an Jinxiang Pharmaceutical Excipients Co., Ltd.

[0062] Recombinant collagen: CAS number is 9007-34-5, purity is 99%, from Wuhan Huaxiang Kejie Biotechnology Co., Ltd.

[0063] β-Glucan: CAS number is 9012-72-0, purity is 99%, from Shenzhen Love Biotechnology Co., Ltd.

[0064] Recombinant human fibronectin: CAS number 86088-83-7, purity 98%, from Hubei Ruiboxin Chemical Co., Ltd.

[0065] The quality fraction of recombinant human type III collagen is 3-7%, which is from Beijing Aibixin Biotechnology Co., Ltd.

[0066] 1-Ethyl-(3-dimethylaminopropyl)carbodiimide: CAS No.: 1892-57-5;

[0067] N-Hydroxysuccinimide: CAS No.: 6066-82-6;

[0068] Sodium fatty acid methyl ester sulfonate: CAS number: 1266615-59-1;

[0069] Chitosan: CAS No.: 9012-76-4;

[0070] Acetic acid solution, CAS number: 64-19-17, concentration 0.15~0.20mol / L;

[0071] The concentration of genipin solution is 0.85-0.90 mol / L, CAS number: 6902-77-8, all of which are from Merck reagents;

[0072] Graphene oxide: particle size 10-20 μm, sourced from Jiangsu Xianfeng Nanomaterial Technology Co., Ltd.;

[0073] Hyaluronic acid, from Merck reagent.

[0074] 4-Dimethylaminopyridine: CAS No.: 1122-58-3;

[0075] N,N-Diisopropylcarbodiimide: CAS number: 693-13-0, all from Merck reagents.

[0076] Example 1: A method for preparing a moisturizing and repairing collagen dressing, comprising the following process:

[0077] Recombinant human fibronectin, recombinant collagen, β-glucan, compound hexylene glycol, carbomer, flavoring agent, and purified water were mixed in a mass ratio of 10:10:5:5:40:5:80, passed nitrogen for 20 minutes, and then irradiated to obtain a collagen dressing; the irradiation dose rate was 40 Gy / min, and the total dose was 2400 Gy.

[0078] Example 2: A method for preparing a moisturizing and repairing collagen dressing, comprising the following process:

[0079] Step (1) Preparation of modified recombinant human type III collagen solution:

[0080] The recombinant human type III collagen was placed in a -75°C low-temperature refrigerator for 12 hours, and then transferred to a -100°C freezer for freeze drying for 24 hours. It was freeze dried under vacuum until completely dehydrated. The dried collagen was placed in a 0.03 mol / L 1-ethyl-(3-dimethylaminopropyl) carbodiimide solution, and then a 0.05 mol / L N-hydroxysuccinimide solution was added. The pH value was adjusted to 4 using a 15 mmol / L sodium fatty acid methyl ester sulfonate solution as a buffer, and placed at 2°C for 18 hours to obtain a modified recombinant human type III collagen solution. The mass ratio of 1-ethyl-(3-dimethylaminopropyl) carbodiimide solution, N-hydroxysuccinimide solution, and recombinant human type III collagen was 4:1:2.

[0081] Step (2) Preparation of modified chitosan solution:

[0082] Dissolve chitosan in 0.15 mol / L acetic acid solution and stir evenly at 25°C to obtain chitosan solution; the volume ratio of chitosan to acetic acid solution is 1:50;

[0083] Graphene oxide and hyaluronic acid were dissolved in deionized water, 4-dimethylaminopyridine and N,N-diisopropylcarbodiimide were added, and the mixture was stirred evenly at 20°C. After reacting for 24 hours, the obtained solution was placed in a dialysis bag with a relative molecular mass of 1400 and dialyzed for 5 hours to obtain a modified hyaluronic acid hydrogel. The freeze dryer temperature was -100°C and the freezing time was 12 hours. The mass ratio of graphene oxide, hyaluronic acid gel, 4-dimethylaminopyridine and N,N-diisopropylcarbodiimide was 20:50:1:1.

[0084] The chitosan solution and the modified hyaluronic acid hydrogel were mixed and stirred evenly, and then a genipin solution with a concentration of 0.85 mol / L was added, and the mixture was placed on a magnetic stirring rack and stirred to obtain a modified chitosan solution; the volume ratio of the chitosan solution, the modified hyaluronic acid hydrogel, and the genipin solution was 5:1:0.1;

[0085] Step (3) Preparation of collagen dressing:

[0086] Recombinant human fibronectin, recombinant collagen, β-glucan, compound hexanediol, carbomer, flavoring agent, purified water, modified recombinant human type III collagen solution, and modified chitosan solution were mixed in a mass ratio of 10:10:5:5:40:5:80:10:10, passed nitrogen for 20 minutes, and then irradiated to obtain collagen dressing; the irradiation dose rate was 40 Gy / min, and the total dose was 2400 Gy.

[0087] Example 3: A method for preparing a moisturizing and repairing collagen dressing, comprising the following process:

[0088] Step (1) Preparation of modified recombinant human type III collagen solution:

[0089] The recombinant human type III collagen was placed in a -80°C low-temperature refrigerator for 12 hours, and then transferred to a -105°C freeze dryer for freeze drying for 24 hours. The dried collagen was placed in a 0.08 mol / L 1-ethyl-(3-dimethylaminopropyl) carbodiimide solution, and then a 0.06 mol / L N-hydroxysuccinimide solution was added. The pH value was adjusted to 5 using a 20 mmol / L sodium fatty acid methyl ester sulfonate solution as a buffer, and the solution was placed at 4°C for 21 hours to obtain a modified recombinant human type III collagen solution. The mass ratio of 1-ethyl-(3-dimethylaminopropyl) carbodiimide solution, N-hydroxysuccinimide solution, and recombinant human type III collagen was 4:2:2.5.

[0090] Step (2) Preparation of modified chitosan solution:

[0091] Dissolve chitosan in acetic acid solution with a concentration of 0.17 mol / L, and stir evenly at 25°C to obtain a chitosan solution; the volume ratio of chitosan to acetic acid solution is 1:50;

[0092] Graphene oxide and hyaluronic acid were dissolved in deionized water, 4-dimethylaminopyridine and N,N-diisopropylcarbodiimide were added, and the mixture was stirred evenly at 25°C. After reacting for 30 hours, the obtained solution was placed in a dialysis bag with a relative molecular mass of 1400 and dialyzed for 5 hours to obtain a modified hyaluronic acid hydrogel. The freeze dryer temperature was -105°C and the freezing time was 12 hours. The mass ratio of graphene oxide, hyaluronic acid gel, 4-dimethylaminopyridine and N,N-diisopropylcarbodiimide was 20:65:1:1.

[0093] The chitosan solution and the modified hyaluronic acid hydrogel were mixed and stirred evenly, and then a genipin solution with a concentration of 0.87 mol / L was added, and the mixture was placed on a magnetic stirring rack and stirred to obtain a modified chitosan solution; the volume ratio of the chitosan solution, the modified hyaluronic acid hydrogel, and the genipin solution was 5:1:0.1;

[0094] Step (3) Preparation of collagen dressing:

[0095] Recombinant human fibronectin, recombinant collagen, β-glucan, compound hexanediol, carbomer, flavoring agent, purified water, modified recombinant human type III collagen solution, and modified chitosan solution were mixed in a mass ratio of 20:15:10:7:45:10:100:20:15, passed nitrogen for 20 minutes, and then irradiated to obtain collagen dressing with an irradiation dose rate of 50 Gy / min and a total dose of 3000 Gy.

[0096] Example 4: A method for preparing a moisturizing and repairing collagen dressing, comprising the following process:

[0097] Step (1) Preparation of modified recombinant human type III collagen solution:

[0098] The recombinant human type III collagen was placed in a -85°C low-temperature refrigerator for 12 hours, and then transferred to a -110°C freeze dryer for freeze drying for 24 hours. The dried collagen was placed in a 0.13 mol / L 1-ethyl-(3-dimethylaminopropyl) carbodiimide solution, and then a 0.08 mol / L N-hydroxysuccinimide solution was added. The pH value was adjusted to 6 using a 25 mmol / L sodium fatty acid methyl ester sulfonate solution as a buffer, and the solution was placed at 6°C for 24 hours to obtain a modified recombinant human type III collagen solution. The mass ratio of 1-ethyl-(3-dimethylaminopropyl) carbodiimide solution, N-hydroxysuccinimide solution, and recombinant human type III collagen was 4:3:2.

[0099] Step (2) Preparation of modified chitosan solution:

[0100] Dissolve chitosan in 0.20 mol / L acetic acid solution and stir evenly at 25°C to obtain a chitosan solution;

[0101] Graphene oxide and hyaluronic acid were dissolved in deionized water, 4-dimethylaminopyridine and N,N-diisopropylcarbodiimide were added, stirred evenly at 30°C, reacted for 36 hours, and the resulting solution was placed in a dialysis bag with a relative molecular mass of 1400 for 5 hours to obtain a modified hyaluronic acid hydrogel. The freeze dryer temperature was -110°C and the freezing time was 12 hours. The mass ratio of graphene oxide, hyaluronic acid gel, 4-dimethylaminopyridine, and N,N-diisopropylcarbodiimide was 20:80:1:1.

[0102] The chitosan solution and the modified hyaluronic acid hydrogel were mixed and stirred evenly, and then a 0.90 mol / L genipin solution was added, and the mixture was placed on a magnetic stirring rack and stirred to obtain a modified chitosan solution; the volume ratio of the chitosan solution, the modified hyaluronic acid hydrogel, and the genipin solution was 5:5:0.3.

[0103] Step (3) Preparation of collagen dressing:

[0104] Recombinant human fibronectin, recombinant collagen, β-glucan, compound hexanediol, carbomer, flavoring agent, purified water, modified recombinant human type III collagen solution, and modified chitosan solution were mixed in a mass ratio of 30:20:15:10:50:15:120:30:20, passed nitrogen for 20 minutes, and then irradiated to obtain collagen dressing with an irradiation dose rate of 60 Gy / min and a total dose of 3000 Gy.

[0105] Comparative Example 1: Taking Example 2 as a control, a method for preparing a moisturizing and repairing collagen dressing without modifying the recombinant human type III collagen solution includes the following processes:

[0106] Step (1) Preparation of modified chitosan solution:

[0107] Dissolve chitosan in 0.15 mol / L acetic acid solution and stir evenly at 25°C to obtain a chitosan solution;

[0108] Graphene oxide and hyaluronic acid were dissolved in deionized water, 4-dimethylaminopyridine and N,N-diisopropylcarbodiimide were added, stirred evenly at 20°C, reacted for 24 hours, and the resulting solution was placed in a dialysis bag with a relative molecular mass of 1400 for 5 hours to obtain a modified hyaluronic acid hydrogel. The freeze dryer temperature was -110°C and the freezing time was 12 hours. The mass ratio of graphene oxide, hyaluronic acid gel, 4-dimethylaminopyridine, and N,N-diisopropylcarbodiimide was 20:50:1:1.

[0109] The chitosan solution and the modified hyaluronic acid hydrogel were mixed and stirred evenly, and then a genipin solution with a concentration of 0.85 mol / L was added, and the mixture was placed on a magnetic stirring rack and stirred to obtain a modified chitosan solution; the volume ratio of the chitosan solution, the modified hyaluronic acid hydrogel, and the genipin solution was 5:1:0.1;

[0110] Step (2) Preparation of collagen dressing:

[0111] Recombinant human fibronectin, recombinant collagen, β-glucan, compound hexanediol, carbomer, flavoring agent, purified water, recombinant human type III collagen solution, and modified chitosan solution were mixed in a mass ratio of 10:10:5:5:40:5:80:10:10, passed nitrogen for 20 minutes, and then irradiated to obtain collagen dressing with an irradiation dose rate of 40 Gy / min and a total dose of 2400 Gy.

[0112] Comparative Example 2: Using Example 2 as a control, without modifying the chitosan solution, a method for preparing a moisturizing and repairing collagen dressing includes the following processes:

[0113] Step (1) Preparation of modified recombinant human type III collagen solution:

[0114] The recombinant human type III collagen was placed in a -75°C low-temperature refrigerator for 12 hours, and then transferred to a -100°C freezer for freeze drying for 24 hours. It was freeze dried under vacuum until completely dehydrated. The dried collagen was placed in a 0.03 mol / L 1-ethyl-(3-dimethylaminopropyl) carbodiimide solution, and then a 0.05 mol / L N-hydroxysuccinimide solution was added. The pH value was adjusted to 4 using a 15 mmol / L sodium fatty acid methyl ester sulfonate solution as a buffer, and placed at 2°C for 18 hours to obtain a modified recombinant human type III collagen solution. The mass ratio of 1-ethyl-(3-dimethylaminopropyl) carbodiimide solution, N-hydroxysuccinimide solution, and recombinant human type III collagen was 4:1:2.

[0115] Step (2) Preparation of collagen dressing:

[0116] Recombinant human fibronectin, recombinant collagen, β-glucan, compound hexanediol, carbomer, flavoring agent, purified water, modified recombinant human type III collagen solution, and chitosan solution were mixed in a mass ratio of 10:10:5:5:40:5:80:10:10, passed nitrogen for 20 minutes, and then irradiated to obtain collagen dressing with an irradiation dose rate of 40 Gy / min and a total dose of 2400 Gy.

[0117] Comparative Example 3: Taking Example 2 as a control, a method for preparing a moisturizing and repairing collagen dressing is described, wherein the recombinant human type III collagen solution and the chitosan solution are not modified, and the method comprises the following processes:

[0118] Recombinant human fibronectin, recombinant collagen, β-glucan, compound hexanediol, carbomer, flavoring agent, purified water, recombinant human type III collagen solution, and chitosan solution were mixed in a mass ratio of 10:10:5:5:40:5:80:10:10, passed nitrogen for 20 minutes, and then irradiated to obtain collagen dressing with an irradiation dose rate of 40 Gy / min and a total dose of 2400 Gy.

[0119] experiment:

[0120] The collagen dressings obtained in Examples 1 to 4 and Comparative Examples 1 to 3 were taken to prepare samples, and their properties were tested respectively and the test results were recorded:

[0121] Repair effect test: 120 subjects aged 20 to 50 years with hormonal dermatitis were selected and divided into 6 groups on average. The number of people in the same age group in each group was the same. The groups were numbered. No. 1 to 3 used the collagen dressings prepared in Examples 1 to 4, and No. 4 to 6 used the collagen dressings prepared in Comparative Examples 1 to 3. After washing and wiping the face clean, the dressing was directly applied without using any cosmetics or medicines. It was used once every 3 days for 1 month, and the repair effect was evaluated.

[0122] The evaluation criteria are as follows:

[0123] Significantly effective: After applying the dressing, the skin was examined and the redness and itching symptoms completely disappeared, and there was no skin inflammation;

[0124] Effective: Inflammation is reduced after use;

[0125] Ineffective: Symptoms remain unchanged or worsen after use;

[0126] Moisturizing effect test: Use the skin moisture meter Corneometer CM 825 to test the water content of the stratum corneum before using the collagen dressing. Remove the dressing 15 minutes after use and test the water content of the stratum corneum. Test the water content of the stratum corneum again after 8 hours.

[0127]

[0128] According to the data in the above table, we can clearly draw the following conclusions:

[0129] The collagen dressings obtained in Examples 1-4 were compared with the collagen dressings obtained in Comparative Examples 1-3. The test results show that:

[0130] Compared with Example 1, Comparative Example 1 does not modify the recombinant human type III collagen solution, fails to introduce amino groups on the side chains of collagen, fails to improve the activity of collagen, slows down the proliferation and differentiation of cells, and has a reduced skin repair effect compared with the modified collagen.

[0131] Compared with Example 1, in Comparative Example 2, the chitosan solution was not modified, and the hydrogel prepared by pure chitosan had weak water holding capacity, and the skin moisturizing effect was reduced. At the same time, graphene oxide failed to be grafted with hyaluronic acid, and its drug loading and release capacity were reduced, resulting in a decrease in the repair effect on the skin.

[0132] Compared with Example 1, Comparative Example 3 does not modify the recombinant human type III collagen solution and chitosan solution, and its repair and moisturizing effects are significantly reduced;

[0133] Compared with Examples 2-4, the water content of the stratum corneum of the skin decreased significantly after 8 hours of using the dressing prepared in Example 1. The reason is that no modified recombinant human type III collagen solution and modified chitosan solution were added. Although the immediate moisturizing ability was good, the long-term moisturizing ability was weakened;

[0134] It can be seen that the setting of the materials used in this application can promote the improvement of the moisturizing and repairing effects of the prepared collagen dressing.

[0135] Finally, it should be noted that the above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art can still modify the technical solutions described in the aforementioned embodiments or replace some of the technical features therein by equivalents. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A moisturizing and repairing collagen dressing, characterized in that: The invention comprises the following components by mass: 10 to 30 parts of recombinant human fibronectin, 10 to 20 parts of recombinant collagen, 5 to 15 parts of beta-glucan, 5 to 10 parts of compound hexylene glycol, 40 to 50 parts of carbomer, 5 to 15 parts of flavoring agent and 80 to 120 parts of purified water.

2. The moisturizing and repairing collagen dressing according to claim 1, characterized in that: The collagen dressing also includes 10 to 30 parts of modified recombinant human type III collagen solution and 10 to 20 parts of modified chitosan solution.

3. The moisturizing and repairing collagen dressing according to claim 2, characterized in that: The modified recombinant human type III collagen solution comprises 1-ethyl-(3-dimethylaminopropyl)carbodiimide, N-hydroxysuccinimide and recombinant human type III collagen.

4. A method for preparing a moisturizing and repairing collagen dressing, characterized in that: Including the following processes: Recombinant human fibronectin, recombinant collagen, β-glucan, compound hexylene glycol, carbomer, flavoring agent, purified water, modified recombinant human type III collagen solution and modified chitosan solution are mixed and irradiated to obtain collagen dressing.

5. The method for preparing a moisturizing and repairing collagen dressing according to claim 4, characterized in that: The modified recombinant human type III collagen solution is prepared by the following process: The recombinant human type III collagen is frozen in a low-temperature refrigerator, then transferred to a freeze dryer, and freeze-dried under vacuum until completely dehydrated. The dried collagen is placed in a 1-ethyl-(3-dimethylaminopropyl) carbodiimide solution, and then N-hydroxysuccinimide solution is added. The pH value is adjusted to 4-6 with sodium fatty acid methyl ester sulfonate solution as a buffer, and placed at 2-6°C for 18-24 hours to obtain a modified recombinant human type III collagen solution.

6. The method for preparing a moisturizing and repairing collagen dressing according to claim 4, characterized in that: The modified chitosan solution is prepared by the following process: The chitosan solution and the modified hyaluronic acid hydrogel are mixed and stirred evenly, and then the genipin solution is added, and the mixture is placed on a magnetic stirring rack and stirred to obtain a modified chitosan solution.

7. The method for preparing a moisturizing and repairing collagen dressing according to claim 6, characterized in that: The modified hyaluronic acid hydrogel is prepared by the following process: Graphene oxide and hyaluronic acid were dissolved in deionized water, 4-dimethylaminopyridine and N,N-diisopropylcarbodiimide were added, and the mixture was stirred evenly at 20-30°C. After reacting for 24-36 hours, the obtained solution was placed in a dialysis bag with a relative molecular mass cutoff of 1400 and dialyzed for 5 hours to obtain a modified hyaluronic acid hydrogel.

8. The method for preparing a moisturizing and repairing collagen dressing according to claim 4, characterized in that: The irradiation process conditions are: irradiation dose rate 40-60 Gy / min, total dose 2400-3600 Gy.

9. The method for preparing a moisturizing and repairing collagen dressing according to claim 5, characterized in that: The mass ratio of the 1-ethyl-(3-dimethylaminopropyl)carbodiimide solution, N-hydroxysuccinimide solution and recombinant human type III collagen is 4:(1-3):(2-3).

10. The method for preparing a moisturizing and repairing collagen dressing according to claim 6, characterized in that: The volume ratio of the chitosan solution, the modified hyaluronic acid hydrogel and the genipin solution is 5:(1-5):(0.1-0.3).

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