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 present invention relates to the technical field of collagen dressings, and specifically to a moisturizing and repairing collagen dressing and a preparation method thereof. Background Art

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

[0003] The patent with the patent number CN115779140B discloses a wound dressing containing collagen and a preparation method thereof, including the following components: benzimidazole quaternary ammonium salt modified silicone oil, type III recombinant human collagen, humectant, water, phosphotungstic acid, triethanolamine.

[0004] Traditional dressings basically do not have a moisturizing function, and currently, efforts are mainly focused on researching medical dressings based on polymer materials and composite materials, such as hydrocolloid dressings. However, the use of dressings is not limited to medical applications but has also extended to the skin care field and the adjuvant treatment of some skin diseases, such as sensitive skin, dermatitis, etc. Most of the existing technologies treat such skin diseases by topical application of hormones, but hormones have certain side effects, and long-term use will 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 purpose of the present invention is to provide a moisturizing and repairing collagen dressing and a preparation method thereof to solve the problems raised in the above background art.

[0006] To solve the above technical problems, the present invention provides the following technical solution: A moisturizing and repairing collagen dressing includes the following mass components: 10 - 30 parts of recombinant human fibronectin, 10 - 20 parts of recombinant collagen, 5 - 15 parts of β - glucan, 5 - 10 parts of compounded hexylene glycol, 40 - 50 parts of carbomer, 5 - 15 parts of flavoring agent, 80 - 120 parts of purified water, 10 - 30 parts of modified recombinant human type III collagen solution, 10 - 20 parts of modified chitosan solution.

[0007] A preparation method of a moisturizing and repairing collagen dressing includes the following steps:

[0008] Mix recombinant human fibronectin, recombinant collagen, β - glucan, compounded hexylene glycol, carbomer, flavoring agent, purified water, modified recombinant human type III collagen solution, and modified chitosan solution, pass nitrogen for 20 min, and then irradiate to obtain the collagen dressing.

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

[0010] In the above technical solution, the irradiated dressing has high moisture retention performance, air permeability and biocompatibility. At the same time, irradiation can eliminate pathogenic microorganisms in the hydrogel, and no additional cross-linking agent is required. The prepared dressing has stable mechanical properties and higher safety.

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

[0012] Furthermore, hexylene glycol is sourced from Beijing Huayudrun Technology Co., Ltd.

[0013] Ethylhexylglycerol is sourced from Jinan Jinbang Environmental Protection Technology Co., Ltd.

[0014] Furthermore, recombinant human fibronectin is sourced from Jilin Ruijin Haoye Biotechnology Co., Ltd.

[0015] Recombinant collagen is sourced from Bloomage Biotechnology Co., Ltd.

[0016] β-glucan, sourced from Ganoderma lucidum, is sourced from Jilin Ruijin Haoye Biotechnology Co., Ltd.

[0017] Carbomer is sourced from Guangzhou Tianci High-Tech Materials Co., Ltd.

[0018] The flavoring agent is compound cyclodextrin and is sourced from Jilin Ruijin Haoye Biotechnology Co., Ltd.

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

[0020] Place the recombinant human type III collagen in a low-temperature refrigerator for freezing, then transfer it to a freeze dryer and freeze-dry it to complete dehydration under vacuum. Place the dried collagen in a 1-ethyl-(3-dimethylaminopropyl)carbodiimide solution, and then add an N-hydroxysuccinimide solution. Adjust the pH value to 4 - 6 with a sodium fatty acid methyl ester sulfonate solution and place it at 2 - 6°C for 18 - 24 h to obtain the modified recombinant human type III collagen solution.

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

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

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

[0024] Furthermore, 1-ethyl-(3-dimethylaminopropyl)carbodiimide, CAS number: 1892-57-5,

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

[0026] N-hydroxysuccinimide, CAS number: 6066-82-6,

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

[0028] 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), all sourced from Merck reagents.

[0029] Furthermore, sodium methyl ester sulfonate, CAS number: 1266615-59-1,

[0030] The concentration of the sodium methyl ester sulfonate solution is 15 to 25 mmol / L, sourced from Merck reagents.

[0031] In the above technical solution, hexylene glycol has good moisturizing and antibacterial properties, and ethylhexylglycerin can be used as both a moisturizer and a skin feel regulator. The combination of the two can enhance the antibacterial effect and moisturizing performance of the dressing. In addition, ethylhexylglycerin also has a deodorizing effect, inhibiting the growth of odor-causing bacteria without affecting the skin flora beneficial to the human body; recombinant human fibronectin can deposit at the injury site 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 skin roughness, damage, etc.; β-glucan can produce cytokines, improve skin cell activity, promote metabolism, and can promote the healing of laser-dependent dermatitis or barrier-damaged skin, achieving anti-allergic and anti-inflammatory effects;

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

[0033] Furthermore, the modified chitosan solution is obtained by crosslinking a chitosan solution with a modified hyaluronic acid hydrogel. The specific process is as follows:

[0034] Mix the chitosan solution and the modified hyaluronic acid hydrogel evenly by stirring, then add the genipin solution and stir 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] Furthermore, genipin, CAS number: 6902-77-8,

[0037] The concentration of the genipin solution is 0.85 - 0.90 mol / L and it is sourced from Merck reagents.

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

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

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

[0041] Furthermore, chitosan is sourced from Jilin Ruijin Haoye Biotechnology Co., Ltd.;

[0042] The acetic acid solution, CAS number: 64-19-17, has a concentration of 0.15 - 0.20 mol / L and is sourced from Merck reagents.

[0043] Furthermore, the modified hyaluronic acid hydrogel is prepared by grafting graphene oxide onto 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 h, the resulting solution was placed in a dialysis bag with a molecular weight cut-off of 1400 and dialyzed for 5 h 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: with a particle size of 10 - 20 μm, sourced from Jiangsu Xianfeng Nano Materials Technology Co., Ltd;

[0047] Hyaluronic acid, sourced from Merck reagent.

[0048] Furthermore, 4-dimethylaminopyridine, CAS number: 1122 - 58 - 3;

[0049] N,N'-diisopropylcarbodiimide, CAS number: 693 - 13 - 0;

[0050] All are sourced from Merck reagent.

[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. While maintaining good biocompatibility, it 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 from pure chitosan has weak water retention ability. Combining chitosan with the modified hyaluronic acid hydrogel can provide a moist tissue environment for the skin and has an inhibitory effect on bacteria, thereby accelerating wound healing.

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

[0053] 1. By modifying the recombinant human type III collagen solution with 1-ethyl-(3-dimethylaminopropyl)carbodiimide and N-hydroxysuccinimide, the amino group on 1-ethyl-(3-dimethylaminopropyl)carbodiimide binds to the carboxyl group on the side chain of collagen, introducing an amino group, improving the activity of collagen, further promoting cell proliferation and differentiation, and promoting wound healing. N-hydroxysuccinimide can form a stable hydrazide structure with the amino group, further forming a strong coupling, improving the stability of collagen. Compared with unmodified collagen, it can repair the skin barrier more quickly. In addition, the modified collagen has a reduced repellency to mammalian cells, reduces the adhesion ability of bacteria, and improves 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 not only maintains good biocompatibility but also improves the ability of hyaluronic acid to load and release drugs. The hydrogel prepared from pure chitosan has weak water-holding capacity. Combining chitosan with modified hyaluronic acid can provide a moist tissue environment for the skin and has an inhibitory effect on bacteria, thus accelerating wound healing.

[0055] 3. The dressing prepared by the irradiation method does not require the addition of extra cross-linking agents during the process, avoiding the possible toxicity or irritation to the skin or wound caused by the residue of chemical cross-linking agents. In addition, the prepared dressing has good moisture retention performance, 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 implementation manners

[0056] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0057] In the following detailed implementation manners,

[0058] Hexylene glycol, sourced from Beijing Huayudrun Technology Co., Ltd.;

[0059] Ethylhexylglycerin, sourced from Jinan Jinbang Environmental Protection Technology Co., Ltd.;

[0060] Carbomer, sourced from Guangzhou Tianci High-Tech Materials Co., Ltd.;

[0061] The flavoring agent is compound cyclodextrin, sourced from Jilin Ruijin Haoye Biotechnology Co., Ltd.;

[0062] Recombinant collagen, sourced from Bloomage Biotechnology Co., Ltd.;

[0063] β-glucan, sourced from Ganoderma lucidum, sourced from Jilin Ruijin Haoye Biotechnology Co., Ltd.;

[0064] Recombinant human fibronectin, sourced from Jilin Ruijin Haoye Biotechnology Co., Ltd.;

[0065] The mass fraction of recombinant human type III collagen is 3-7%, sourced from Beijing Absin Biotechnology Co., Ltd.;

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

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

[0068] Sodium methyl ester sulfonate of fatty acid: CAS No.: 1266615-59-1;

[0069] Chitosan, sourced from Jilin Ruijin Haoye Biotechnology Co., Ltd.;

[0070] Acetic acid solution, CAS No.: 64-19-17, with a concentration of 0.15 - 0.20 mol / L;

[0071] Genipin solution with a concentration of 0.85 - 0.90 mol / L, CAS No.: 6902-77-8, all of the above are sourced from Merck reagents;

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

[0073] Hyaluronic acid, sourced from Merck reagents.

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

[0075] N,N-Diisopropylcarbodiimide: CAS No.: 693-13-0, all sourced from Merck reagents.

[0076] Example 1: A preparation method of a moisturizing and repairing collagen dressing, including the following processes:

[0077] Mix recombinant human fibronectin, recombinant collagen, β-glucan, compounded hexylene glycol, carbomer, flavoring agent, and purified water in a mass ratio of 10:10:5:5:40:5:80, pass nitrogen for 20 min, and then irradiate to obtain a collagen dressing; the irradiation dose rate is 40 Gy / min, and the total dose is 2400 Gy.

[0078] Example 2: A preparation method of a moisturizing and repairing collagen dressing, including the following processes:

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

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

[0082] Chitosan was dissolved in an acetic acid solution with a concentration of 0.15 mol / L and stirred evenly at 25°C to obtain a chitosan solution; the volume ratio of chitosan to acetic acid solution was 1:50;

[0083] Graphene oxide and hyaluronic acid were dissolved in deionized water, 4-dimethylaminopyridine and N,N'-diisopropylcarbodiimide were added, and it was stirred evenly at 20°C. After reacting for 24 h, the obtained solution was put into a dialysis bag with a cut-off relative molecular mass of 1400 for dialysis for 5 h to obtain a modified hyaluronic acid hydrogel. The temperature of the freeze dryer was -100°C and the freezing time was 12 h; 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 it was placed on a magnetic stirring rack for stirring 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 the collagen dressing:

[0086] Recombinant human fibronectin, recombinant collagen, β-glucan, compounded hexylene glycol, 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, and nitrogen was passed through for 20 min, and then irradiated to obtain a collagen dressing; the irradiation dose rate was 40 Gy / min and the total dose was 2400 Gy.

[0087] Example 3: A preparation method of a moisturizing and repairing collagen dressing, including the following processes:

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

[0089] Place the recombinant human type III collagen in a low-temperature refrigerator at -80°C and freeze it for 12 hours. Then transfer it to a freeze dryer at -105°C and freeze-dry it for 24 hours until completely dehydrated under vacuum. Place the dried collagen in a 1-ethyl-(3-dimethylaminopropyl)carbodiimide solution with a concentration of 0.08 mol / L, and then add an N-hydroxysuccinimide solution with a concentration of 0.06 mol / L. Use a sodium fatty acid methyl ester sulfonate solution with a concentration of 20 mmol / L as a buffer to adjust the pH value to 5, and place it at 4°C for 21 hours to obtain the 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 is 4:2:2.5;

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

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

[0092] Dissolve graphene oxide and hyaluronic acid in deionized water, add 4-dimethylaminopyridine and N,N'-diisopropylcarbodiimide, stir evenly at 25°C, and after reacting for 30 hours, place the obtained solution in a dialysis bag with a cut-off relative molecular mass of 1400 and dialyze for 5 hours to obtain the modified hyaluronic acid hydrogel. The temperature of the freeze dryer is -105°C and the freezing time is 12 hours. The mass ratio of graphene oxide, hyaluronic acid gel, 4-dimethylaminopyridine, and N,N'-diisopropylcarbodiimide is 20:65:1:1;

[0093] Mix the chitosan solution and the modified hyaluronic acid hydrogel evenly, and then add a genipin solution with a concentration of 0.87 mol / L, and place it on a magnetic stirrer to stir to obtain the modified chitosan solution. The volume ratio of the chitosan solution, the modified hyaluronic acid hydrogel, and the genipin solution is 5:1:0.1;

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

[0095] Mix recombinant human fibronectin, recombinant collagen, β-glucan, compounded hexylene glycol, carbomer, flavoring agent, purified water, modified recombinant human type III collagen solution, and modified chitosan solution according to the mass ratio of 20:15:10:7:45:10:100:20:15, pass nitrogen for 20 minutes, and then irradiate to obtain the collagen dressing. The irradiation dose rate is 50 Gy / min and the total dose is 3000 Gy.

[0096] Example 4: A preparation method of a moisturizing and repairing collagen dressing, comprising the following processes:

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

[0098] Place the recombinant human type III collagen in a low-temperature refrigerator at -85°C and freeze it for 12 h, then transfer it to a freeze dryer at -110°C and freeze-dry it for 24 h. Freeze-dry it to complete dehydration under a vacuum state. Place the dried collagen in a 1-ethyl-(3-dimethylaminopropyl)carbodiimide solution with a concentration of 0.13 mol / L, and then add an N-hydroxysuccinimide solution with a concentration of 0.08 mol / L. Use a sodium fatty acid methyl ester sulfonate solution with a concentration of 25 mmol / L as a buffer to adjust the pH value to 6, and place it at 6°C for 24 h to obtain a modified recombinant human type III collagen solution; the mass ratio of the 1-ethyl-(3-dimethylaminopropyl)carbodiimide solution, the N-hydroxysuccinimide solution, and the recombinant human type III collagen is 4:3:2;

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

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

[0101] Dissolve graphene oxide and hyaluronic acid in deionized water, add 4-dimethylaminopyridine and N,N'-diisopropylcarbodiimide, stir evenly at 30°C, and after reacting for 36 h, put the obtained solution into a dialysis bag with a cut-off relative molecular mass of 1400 and dialyze for 5 h to obtain a modified hyaluronic acid hydrogel. The temperature of the freeze dryer is -110°C and the freezing time is 12 h; the mass ratio of graphene oxide, hyaluronic acid gel, 4-dimethylaminopyridine, and N,N'-diisopropylcarbodiimide is 20:80:1:1;

[0102] Mix and stir evenly the chitosan solution and the modified hyaluronic acid hydrogel, and then add a genipin solution with a concentration of 0.90 mol / L, and place it on a magnetic stirring rack and stir to obtain a modified chitosan solution; the volume ratio of the chitosan solution, the modified hyaluronic acid hydrogel, and the genipin solution is 5:5:0.3.

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

[0104] Recombinant human fibronectin, recombinant collagen, β-glucan, compounded hexylene glycol, carbomer, flavoring agent, purified water, modified recombinant human type III collagen solution, and modified chitosan solution were mixed at a mass ratio of 30:20:15:10:50:15:120:30:20. Nitrogen was passed through for 20 min, and then irradiation was carried out to obtain a collagen dressing. The irradiation dose rate was 60 Gy / min, and the total dose was 3000 Gy.

[0105] Comparative Example 1: Taking Example 2 as a control, the recombinant human type III collagen solution was not modified. A preparation method of a moisturizing and repairing collagen dressing includes the following processes:

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

[0107] Chitosan was dissolved in an acetic acid solution with a concentration of 0.15 mol / L and stirred 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, and the mixture was stirred evenly at 20 °C. After reacting for 24 h, the obtained solution was put into a dialysis bag with a cut-off relative molecular mass of 1400 and dialyzed for 5 h to obtain a modified hyaluronic acid hydrogel. The temperature of the freeze dryer was -110 °C, and the freezing time was 12 h; 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 stirred on a magnetic stirrer 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, compounded hexylene glycol, carbomer, flavoring agent, purified water, recombinant human type III collagen solution, and modified chitosan solution were mixed at a mass ratio of 10:10:5:5:40:5:80:10:10. Nitrogen was passed through for 20 min, and then irradiation was carried out to obtain a collagen dressing. The irradiation dose rate was 40 Gy / min, and the total dose was 2400 Gy.

[0112] Comparative Example 2: Taking Example 2 as a control, the chitosan solution was not modified. A preparation method of 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 frozen in a low-temperature refrigerator at -75°C for 12 h, and then transferred to a freeze dryer at -100°C for freeze-drying for 24 h. It was freeze-dried to complete dehydration under vacuum. The dried collagen was placed in a 1-ethyl-(3-dimethylaminopropyl)carbodiimide solution with a concentration of 0.03 mol / L, and then a N-hydroxysuccinimide solution with a concentration of 0.05 mol / L was added. A sodium fatty acid methyl ester sulfonate solution with a concentration of 15 mmol / L was used as a buffer to adjust the pH value to 4, and it was placed at 2°C for 18 h to obtain a modified recombinant human type III collagen solution; the mass ratio of the 1-ethyl-(3-dimethylaminopropyl)carbodiimide solution, the N-hydroxysuccinimide solution, and the recombinant human type III collagen was 4:1:2;

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

[0116] The recombinant human fibronectin, recombinant collagen, β-glucan, compounded hexylene glycol, 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, nitrogen was passed through for 20 min, and then irradiation was carried out to obtain a collagen dressing. The irradiation dose rate was 40 Gy / min, and the total dose was 2400 Gy.

[0117] Comparative Example 3: Taking Example 2 as a control, the recombinant human type III collagen solution and the chitosan solution were not modified. A preparation method of a moisturizing and repairing collagen dressing includes the following process:

[0118] The recombinant human fibronectin, recombinant collagen, β-glucan, compounded hexylene glycol, 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, nitrogen was passed through for 20 min, and then irradiation was carried out to obtain a collagen dressing. The irradiation dose rate was 40 Gy / min, and the total dose was 2400 Gy.

[0119] Experiment:

[0120] The collagen dressings obtained in Examples 1 to 4 and Comparative Examples 1 to 3 were taken to prepare specimens, and their performances were respectively detected and the detection results were recorded:

[0121] Repair effect test: 120 subjects aged 20 - 50 with hormone dermatitis were selected and evenly divided into 6 groups, with the same number of people in each group of the same age range. The groups were numbered. Groups 1 - 3 used the collagen dressings prepared in Examples 1 - 4, and groups 4 - 6 used the collagen dressings prepared in Comparative Examples 1 - 3. After washing the face and wiping it dry, without using any cosmetics or drugs, the dressings were directly applied once every 3 days for 1 consecutive month, and the repair effect was evaluated.

[0122] The evaluation criteria are as follows:

[0123] Marked effect: After using the dressing, upon examination of the skin, the redness and itching symptoms completely disappear, and there is no skin inflammation.

[0124] Effective: The inflammation is reduced after use.

[0125] Ineffective: The symptoms show no change or worsen after use.

[0126] Moisturizing effect test: Using the skin moisture meter Corneometer CM 825, the water content of the skin stratum corneum before using the collagen dressing was detected. After removing the dressing 15 minutes after use, the water content of the stratum corneum was tested, and the water content of the stratum corneum was tested again 8 hours later.

[0127]

[0128] Based on the data in the above table, the following conclusions can be clearly obtained:

[0129] A comparison was made between the collagen dressings obtained in Examples 1 - 4 and those obtained in Comparative Examples 1 - 3. From the test results, it can be seen that

[0130] Compared with Example 1, in Comparative Example 1, the recombinant human type III collagen solution was not modified, and amino groups could not be introduced into the side chain of collagen, so the activity of collagen could not be increased, and the proliferation and differentiation rate of cells slowed down. Compared with the modified collagen, the repair effect on the skin decreased.

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

[0132] Compared with Example 1, in Comparative Example 3, neither the recombinant human type III collagen solution nor the chitosan solution was modified, and its repair and moisturizing effects decreased significantly.

[0133] Compared with Examples 2-4, the water content of the stratum corneum of the skin after 8 hours of using the dressing prepared in Example 1 decreased significantly. The reason is that the modified recombinant human type III collagen solution and the modified chitosan solution were not added. Although the immediate moisturizing ability is good, the long-term moisturizing ability is weakened;

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

[0135] Finally, it should be noted that the above are only the preferred embodiments of the present invention and are not intended to limit the present invention. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments, or perform equivalent replacements on some of the technical features. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. A collagen dressing for moisturizing and repair, characterized in that: It includes the following mass components: 10 - 30 parts of recombinant human fibronectin, 10 - 20 parts of recombinant collagen, 5 - 15 parts of β-glucan, 5 - 10 parts of compounded hexylene glycol, 40 - 50 parts of carbomer, 5 - 15 parts of flavoring agent, and 80 - 120 parts of purified water.

2. The collagen dressing for moisturizing and repair according to claim 1, wherein: The collagen dressing further includes 10 - 30 parts of modified recombinant human type III collagen solution and 10 - 20 parts of modified chitosan solution.

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

4. A preparation method of a moisturizing and repairing collagen dressing, characterized in that: It includes the following processes: Mix recombinant human fibronectin, recombinant collagen, β-glucan, compounded hexylene glycol, carbomer, flavoring agent, purified water, modified recombinant human type III collagen solution, and modified chitosan solution, and obtain the collagen dressing after irradiation.

5. The preparation method of 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: Place recombinant human type III collagen in a low-temperature refrigerator for freezing, then transfer it to a freeze dryer and freeze-dry it under vacuum until completely dehydrated. Place the dried collagen in a 1-ethyl-(3-dimethylaminopropyl)carbodiimide solution, and then add an N-hydroxysuccinimide solution. Adjust the pH value to 4 - 6 with a sodium fatty acid methyl ester sulfonate solution, and place it at 2 - 6°C for 18 - 24 h to obtain the modified recombinant human type III collagen solution.

6. The preparation method of a moisturizing and repairing collagen dressing according to claim 4, characterized in that: The modified chitosan solution is prepared by the following process: Mix the chitosan solution and the modified hyaluronic acid hydrogel evenly by stirring, then add a genipin solution, and place it on a magnetic stirring rack for stirring to obtain the modified chitosan solution.

7. The preparation method of a moisturizing and repairing collagen dressing according to claim 6, characterized in that: The modified hyaluronic acid hydrogel is prepared by the following process: Dissolve graphene oxide and hyaluronic acid in deionized water, add 4-dimethylaminopyridine and N,N-diisopropylcarbodiimide, stir evenly at 20 - 30°C, react for 24 - 36 h, and then put the obtained solution into a dialysis bag with a cut-off relative molecular mass of 1400 for dialysis for 5 h to obtain the modified hyaluronic acid hydrogel.

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

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