Medical recombinant collagen nursing dressing for repairing skin-sensitive wound and preparation method of medical recombinant collagen nursing dressing

Abstract

The invention relates to a medical recombinant collagen nursing dressing for skin-sensitive wound repair and a preparation method of the medical recombinant collagen nursing dressing, and belongs to the technical field of medical dressings. The medical recombinant collagen nursing dressing for repairing the skin-sensitive wound comprises a non-woven fabric and a recombinant collagen composition, wherein the recombinant collagen composition comprises the following components: 91-100 parts of purified water, recombinant collagen, carbomer, glycerol and methylparaben. The preparation method comprises the following steps: sequentially feeding purified water, carbomer, glycerol and methylparaben into an emulsifying machine in proportion, heating and stirring; and then stirring and cooling, adding the recombinant collagen, and stirring and mixing to obtain the traditional Chinese medicine composition. By immobilizing the recombinant protein, the stability is enhanced, the service life of the dressing is prolonged, the replacement frequency is reduced, and the medical cost is reduced. The stable recombinant collagen improves biological activity and functionality, promotes wound healing and reduces scars.

Description

Technical Field

[0001] The invention belongs to the technical field of medical dressings, and relates to a medical recombinant collagen nursing dressing for repairing sensitive skin wounds and a preparation method thereof. Background Art

[0002] Recombinant collagen has high biocompatibility and low immunogenicity. This means that when it is used as a medical dressing, it can reduce rejection reactions with human tissues, reduce the risk of allergies and infections, and thus ensure the safety of patients. Secondly, the structure and properties of recombinant collagen can be precisely regulated through genetic engineering technology. This controllability allows us to design recombinant collagen dressings with specific functions according to specific medical needs, such as promoting wound healing and improving skin texture.

[0003] However, the stability of recombinant collagen is poor. This is because there are many non-covalent bonds in its molecular structure, such as hydrogen bonds, hydrophobic forces and ionic bonds. The breaking and recombination of these bonds are easily affected by external conditions. For example, changes in factors such as temperature, pH value, and ionic strength may cause changes in the structure of recombinant collagen, thereby affecting its stability and function. Therefore, in practical applications, a series of measures need to be taken to improve the stability of recombinant collagen.

[0004] Epoxy carriers are ideal materials for fixing proteins. They are stable under neutral conditions and can be stored for a long time at room temperature. Epoxy groups can form stable covalent bonds with nucleophilic groups on proteins. However, since the activity of epoxy groups is not high under neutral conditions, the amount of protein that can be fixed is relatively small. Summary of the invention

[0005] The purpose of the present invention is to provide a medical recombinant collagen nursing dressing for repairing sensitive skin wounds and a preparation method thereof. The present invention enhances the stability of the dressing, prolongs the life of the dressing, reduces the replacement frequency, and reduces medical costs by immobilizing the recombinant protein. Stable recombinant collagen enhances biological activity and functionality, promotes wound healing, and reduces scars. At the same time, it enhances environmental resistance, maintains stable performance in complex medical environments, ensures patient recovery safety, and reduces the risk of complications.

[0006] The purpose of the present invention can be achieved through the following technical solutions:

[0007] A medical recombinant collagen nursing dressing for repairing sensitive skin wounds comprises a non-woven fabric and a recombinant collagen composition, wherein the recombinant collagen composition comprises the following components in parts by weight: 91-100 parts of purified water, 0.20-0.32 parts of recombinant collagen, 0.20-0.26 parts of carbomer, 8.0-8.4 parts of glycerol and 0.05-0.06 parts of methylparaben.

[0008] As a preferred technical solution of the present invention, the method for preparing recombinant collagen comprises the following steps:

[0009] 1) placing a polysaccharide carrier, dimethyl sulfoxide, epichlorohydrin, sodium hydroxide and sodium borohydride in a reaction kettle, stirring the mixture, filtering out the solid, and washing the solid to obtain a prefabricated polysaccharide carrier;

[0010] 2) placing the prefabricated polysaccharide carrier in the amino acid solution and mixing, adjusting the pH value and stirring at low temperature to obtain a modified polysaccharide carrier;

[0011] 3) After the modified polysaccharide carrier is immersed in the recombinant protein solution and treated at low temperature, a sodium chloride solution is added and stirred and mixed, solids are taken out after suction filtration, ethanolamine solution is added and mixed at low temperature, centrifuged, and washed to obtain recombinant collagen.

[0012] As a preferred technical solution of the present invention, in step 1), the stirring treatment is stirring at 40°C and 150-200 r / min for 2-3 hours; the washing is washing three times with distilled water.

[0013] As a preferred technical solution of the present invention, in step 1), the usage ratio of the polysaccharide carrier, dimethyl sulfoxide, epichlorohydrin, sodium hydroxide and sodium borohydride is 12-14:110-120:12.2-13.0:0.25-0.30:0.004.

[0014] As a preferred technical solution of the present invention, in step 2), the dosage ratio of the prefabricated polysaccharide carrier and the amino acid solution is 5-6g:50-58mL; the concentration of the amino acid solution is 0.20-0.23mol / L; the solute in the amino acid solution is one or more of arginine, histidine and lysine.

[0015] As a preferred technical solution of the present invention, in step 2), the pH value is adjusted to 8.0-8.5 using hydrochloric acid; and the low-temperature stirring is stirring at a temperature of 20-25°C for 10-14h.

[0016] As a preferred technical solution of the present invention, in step 3), the low-temperature treatment is stirring at 25°C for 20-24h; the stirring and mixing time is 30-40min; the low-temperature mixing is mixing at 20°C for 12-15h; and the cleaning is cleaning four times with distilled water.

[0017] As a preferred technical solution of the present invention, in step 3), the usage ratio of the modified polysaccharide carrier, the recombinant protein solution, the sodium chloride solution and the ethanolamine solution is 2.8-3.0 g: 6-7 mL: 1-2 mL: 10-12 mL; the concentration of the sodium chloride solution is 1.5 mol / L; the concentration of the ethanolamine solution is 1 mol / L.

[0018] As a preferred technical solution of the present invention, in step 3), the recombinant protein solution is prepared by combining recombinant protein and boric acid buffer in a mass ratio of 1.0-1.2:6-7; the molecular weight of the recombinant protein is 50-100 kDa; the recombinant protein is one or more of recombinant humanized type I collagen, recombinant humanized type III collagen, recombinant humanized type IV collagen and recombinant humanized type VII collagen.

[0019] As a preferred technical solution of the present invention, the preparation of the polysaccharide carrier includes: stirring the agarose powder and cellulose powder in a 4-5% mass fraction acetic acid solution at 60°C for 1.2-1.5h and ultrasonically removing bubbles to obtain a mixed solution, and dripping the mixed solution drop by drop into a mixed solution composed of 2mol / L NaOH and anhydrous ethanol in a ratio of 3:1 by extrusion, and soaking for 3-4h to obtain a polysaccharide carrier; the mass ratio of the agarose powder, cellulose powder and acetic acid solution is 4-5:0.8-1.0:50-60.

[0020] A method for preparing a medical recombinant collagen nursing dressing for repairing sensitive skin wounds, the method comprising the following steps: sequentially adding purified water, carbomer, glycerin and methylparaben into an emulsifier in proportion, stirring at a speed of 25-30 r / min, heating to 80-90°C, and keeping warm for 20-30 min; stirring and cooling to 35-45°C, adding recombinant collagen, stirring at a speed of 25-30 r / min for 10-15 min, and wetting the recombinant collagen composition on the non-woven fabric to obtain the recombinant collagen composition.

[0021] Beneficial effects of the present invention:

[0022] (1) The present invention prepares a polysaccharide carrier and performs epoxy and amino modification on it, so that the positively charged amino groups on the epoxy amino polysaccharide carrier adsorb the negatively charged areas on the recombinant protein through electrostatic force, so that the recombinant protein is physically adsorbed on the epoxy amino polysaccharide carrier, and then the epoxy groups on the epoxy amino polysaccharide carrier covalently react with the amino groups close to the epoxy groups on the recombinant protein to fix the recombinant protein on the epoxy amino polysaccharide carrier. The covalent coupling reaction between the epoxy groups and the recombinant protein requires a long reaction time under alkaline conditions to achieve a stable connection. The present invention optimizes the curing effect by adjusting the appropriate pH value.

[0023] (2) The present invention enhances the stability of the dressing by immobilizing the recombinant protein, prolongs the life of the dressing, reduces the frequency of replacement, and reduces medical costs. Stabilizing the recombinant collagen improves the biological activity and functionality, promotes wound healing, and reduces scars. At the same time, it enhances environmental resistance, maintains stable performance in complex medical environments, ensures the safety of patient recovery, and reduces the risk of complications. DETAILED DESCRIPTION

[0024] In order to further illustrate the technical means and effects adopted by the present invention to achieve the predetermined invention purpose, the specific implementation methods, structures, features and effects of the present invention are described in detail below in combination with the embodiments.

[0025] Example 1

[0026] A medical recombinant collagen nursing dressing for repairing sensitive skin wounds comprises a non-woven fabric and a recombinant collagen composition, wherein the recombinant collagen composition comprises the following components by weight: 91 parts of purified water, 0.20 parts of recombinant collagen, 0.20 parts of carbomer, 8.0 parts of glycerol and 0.05 parts of methylparaben;

[0027] A preparation method of a medical recombinant collagen nursing dressing for repairing sensitive skin wounds comprises the following steps: sequentially adding purified water, carbomer, glycerin and methylparaben into an emulsifier in proportion, stirring at a speed of 25 r / min, heating to 80°C and keeping warm for 20 minutes; stirring and cooling to 35°C, adding recombinant collagen, stirring at a speed of 25 r / min for 10 minutes to obtain a recombinant collagen composition, and wetting the recombinant collagen composition on the non-woven fabric to obtain the dressing.

[0028] The preparation method of the recombinant collagen comprises the following steps:

[0029] 1) placing a polysaccharide carrier, dimethyl sulfoxide, epichlorohydrin, sodium hydroxide and sodium borohydride in a reaction kettle, stirring the mixture, filtering out the solid, and washing the solid to obtain a prefabricated polysaccharide carrier;

[0030] 2) placing the prefabricated polysaccharide carrier in the amino acid solution and mixing, adjusting the pH value and stirring at low temperature to obtain a modified polysaccharide carrier;

[0031] 3) After the modified polysaccharide carrier is immersed in the recombinant protein solution and treated at low temperature, a sodium chloride solution is added and stirred and mixed, solids are taken out after suction filtration, ethanolamine solution is added and mixed at low temperature, centrifuged, and washed to obtain recombinant collagen.

[0032] In step 1), the stirring treatment is stirring at 40°C and 150 r / min for 2 hours; the washing is washing three times with distilled water; the amount ratio of the polysaccharide carrier, dimethyl sulfoxide, epichlorohydrin, sodium hydroxide and sodium borohydride is 12:110:12.2:0.25:0.004;

[0033] In step 2), the ratio of the prefabricated polysaccharide carrier to the amino acid solution is 5 g:50 mL; the concentration of the amino acid solution is 0.20 mol / L; the solute in the amino acid solution is arginine; the pH value is adjusted to 8.0 using hydrochloric acid; the low-temperature stirring is stirring at 20° C. for 10 h;

[0034] In step 3), the low temperature treatment is stirring at 25°C for 20 hours; the stirring and mixing time is 30 minutes; the low temperature mixing is mixing at 20°C for 12 hours; the washing is washing four times with distilled water; the dosage ratio of the modified polysaccharide carrier, the recombinant protein solution, the sodium chloride solution and the ethanolamine solution is 2.8g:6mL:1mL:10mL; the concentration of the sodium chloride solution is 1.5mol / L; the concentration of the ethanolamine solution is 1mol / L;

[0035] In step 3), the recombinant protein solution is prepared by mixing the recombinant protein and boric acid buffer at a mass ratio of 1.0:6; the molecular weight of the recombinant protein is 50 kDa; and the recombinant protein is recombinant humanized type I collagen;

[0036] In step 1), the preparation of the polysaccharide carrier includes the following steps: agarose powder and cellulose powder are stirred in a 4% mass fraction acetic acid solution at 60°C for 1.2 hours and ultrasonically degassing bubbles to obtain a mixed solution, and the mixed solution is dripped dropwise into a mixed solution consisting of 2 mol / L NaOH and anhydrous ethanol in a ratio of 3:1 by an extrusion method, and soaked for 3 hours to obtain a polysaccharide carrier; the mass ratio of the agarose powder, cellulose powder and acetic acid solution is 4:0.8:50.

[0037] Example 2

[0038] A medical recombinant collagen nursing dressing for repairing sensitive skin wounds comprises a non-woven fabric and a recombinant collagen composition, wherein the recombinant collagen composition comprises the following components by weight: 93 parts of purified water, 0.23 parts of recombinant collagen, 0.22 parts of carbomer, 8.1 parts of glycerol and 0.053 parts of methylparaben;

[0039] A preparation method of a medical recombinant collagen nursing dressing for repairing sensitive skin wounds comprises the following steps: sequentially adding purified water, carbomer, glycerin and methylparaben into an emulsifier in proportion, stirring at a speed of 25 r / min, heating to 82°C, and keeping warm for 22 minutes; stirring and cooling to 38°C, adding recombinant collagen, stirring at a speed of 25 r / min for 11 minutes, and wetting the recombinant collagen composition on the non-woven fabric to obtain the recombinant collagen composition.

[0040] The preparation method of the recombinant collagen comprises the following steps:

[0041] 1) placing a polysaccharide carrier, dimethyl sulfoxide, epichlorohydrin, sodium hydroxide and sodium borohydride in a reaction kettle, stirring the mixture, filtering out the solid, and washing the solid to obtain a prefabricated polysaccharide carrier;

[0042] 2) placing the prefabricated polysaccharide carrier in the amino acid solution and mixing, adjusting the pH value and stirring at low temperature to obtain a modified polysaccharide carrier;

[0043] 3) After the modified polysaccharide carrier is immersed in the recombinant protein solution and treated at low temperature, a sodium chloride solution is added and stirred and mixed, solids are taken out after suction filtration, ethanolamine solution is added and mixed at low temperature, centrifuged, and washed to obtain recombinant collagen.

[0044] In step 1), the stirring treatment is stirring at 40°C and 160 r / min for 2.5 hours; the washing is washing three times with distilled water; the amount ratio of the polysaccharide carrier, dimethyl sulfoxide, epichlorohydrin, sodium hydroxide and sodium borohydride is 12.5:112:12.4:0.26:0.004;

[0045] In step 2), the ratio of the prefabricated polysaccharide carrier to the amino acid solution is 5.2 g:52 mL; the concentration of the amino acid solution is 0.21 mol / L; the solute in the amino acid solution is arginine; the pH value is adjusted to 8.1 using hydrochloric acid; the low-temperature stirring is stirring at 21° C. for 11 hours;

[0046] In step 3), the low temperature treatment is stirring at 25°C for 21 hours; the stirring and mixing time is 35 minutes; the low temperature mixing is mixing at 20°C for 13 hours; the washing is washing four times with distilled water; the dosage ratio of the modified polysaccharide carrier, the recombinant protein solution, the sodium chloride solution and the ethanolamine solution is 2.85g:6.2mL:1.2mL:10.5mL; the concentration of the sodium chloride solution is 1.5mol / L; the concentration of the ethanolamine solution is 1mol / L;

[0047] In step 3), the recombinant protein solution is prepared by mixing the recombinant protein and the boric acid buffer in a mass ratio of 1.05:6.2; the molecular weight of the recombinant protein is 70 kDa; the recombinant protein is recombinant humanized type I collagen;

[0048] In step 1), the preparation of the polysaccharide carrier includes the following steps: agarose powder and cellulose powder are stirred in a 4.2% mass fraction acetic acid solution at 60°C for 1.3 hours and ultrasonically degassing bubbles to obtain a mixed solution, and the mixed solution is dripped dropwise into a mixed solution consisting of 2 mol / L NaOH and anhydrous ethanol in a ratio of 3:1 by an extrusion method, and soaked for 3.2 hours to obtain a polysaccharide carrier; the mass ratio of the agarose powder, cellulose powder and acetic acid solution is 4.2:0.85:52.

[0049] Example 3

[0050] A medical recombinant collagen nursing dressing for repairing sensitive skin wounds comprises a non-woven fabric and a recombinant collagen composition, wherein the recombinant collagen composition comprises the following components by weight: 96 parts of purified water, 0.26 parts of recombinant collagen, 0.23 parts of carbomer, 8.2 parts of glycerol and 0.055 parts of methylparaben;

[0051] A preparation method of a medical recombinant collagen nursing dressing for repairing sensitive skin wounds comprises the following steps: sequentially adding purified water, carbomer, glycerin and methylparaben into an emulsifier in proportion, stirring at a speed of 28 r / min, heating to 85°C and keeping warm for 25 minutes; stirring and cooling to 40°C, adding recombinant collagen, stirring at a speed of 28 r / min for 12 minutes to obtain a recombinant collagen composition, and wetting the recombinant collagen composition on the non-woven fabric to obtain the recombinant collagen composition.

[0052] The preparation method of the recombinant collagen comprises the following steps:

[0053] 1) placing a polysaccharide carrier, dimethyl sulfoxide, epichlorohydrin, sodium hydroxide and sodium borohydride in a reaction kettle, stirring the mixture, filtering out the solid, and washing the solid to obtain a prefabricated polysaccharide carrier;

[0054] 2) placing the prefabricated polysaccharide carrier in the amino acid solution and mixing, adjusting the pH value and stirring at low temperature to obtain a modified polysaccharide carrier;

[0055] 3) After the modified polysaccharide carrier is immersed in the recombinant protein solution and treated at low temperature, a sodium chloride solution is added and stirred and mixed, solids are taken out after suction filtration, ethanolamine solution is added and mixed at low temperature, centrifuged, and washed to obtain recombinant collagen.

[0056] In step 1), the stirring treatment is stirring at 40°C and 175 r / min for 2.5 hours; the washing is washing three times with distilled water; the amount ratio of the polysaccharide carrier, dimethyl sulfoxide, epichlorohydrin, sodium hydroxide and sodium borohydride is 13:115:12.6:0.28:0.004;

[0057] In step 2), the ratio of the prefabricated polysaccharide carrier to the amino acid solution is 5.5 g:54 mL; the concentration of the amino acid solution is 0.22 mol / L; the solute in the amino acid solution is arginine; the pH value is adjusted to 8.2 using hydrochloric acid; the low-temperature stirring is stirring at 22° C. for 12 h;

[0058] In step 3), the low temperature treatment is stirring at 25°C for 22 hours; the stirring and mixing time is 35 minutes; the low temperature mixing is mixing at 20°C for 13.5 hours; the washing is washing four times with distilled water; the dosage ratio of the modified polysaccharide carrier, the recombinant protein solution, the sodium chloride solution and the ethanolamine solution is 2.9g:6.5mL:1.5mL:11mL; the concentration of the sodium chloride solution is 1.5mol / L; the concentration of the ethanolamine solution is 1mol / L;

[0059] In step 3), the recombinant protein solution is prepared by mixing the recombinant protein and the boric acid buffer in a mass ratio of 1.1:6.5; the molecular weight of the recombinant protein is 80 kDa; the recombinant protein is recombinant humanized type I collagen;

[0060] In step 1), the preparation of the polysaccharide carrier includes the following steps: agarose powder and cellulose powder are stirred in a 4.5% mass fraction acetic acid solution at 60°C for 1.5 hours and ultrasonically degassing bubbles to obtain a mixed solution, and the mixed solution is dripped dropwise into a mixed solution consisting of 2 mol / L NaOH and anhydrous ethanol in a ratio of 3:1 by an extrusion method, and soaked for 3.5 hours to obtain a polysaccharide carrier; the mass ratio of the agarose powder, cellulose powder and acetic acid solution is 4.5:0.9:55.

[0061] Example 4

[0062] A medical recombinant collagen nursing dressing for repairing sensitive skin wounds comprises a non-woven fabric and a recombinant collagen composition, wherein the recombinant collagen composition comprises the following components by weight: 98 parts of purified water, 0.29 parts of recombinant collagen, 0.24 parts of carbomer, 8.3 parts of glycerol and 0.057 parts of methylparaben;

[0063] A preparation method of a medical recombinant collagen nursing dressing for repairing sensitive skin wounds comprises the following steps: sequentially adding purified water, carbomer, glycerin and methylparaben into an emulsifier in proportion, stirring at a speed of 30 r / min, heating to 88°C, and keeping warm for 28 minutes; stirring and cooling to 42°C, adding recombinant collagen, stirring at a speed of 30 r / min for 14 minutes, and wetting the recombinant collagen composition on the non-woven fabric to obtain the recombinant collagen composition.

[0064] The preparation method of the recombinant collagen comprises the following steps:

[0065] 1) placing a polysaccharide carrier, dimethyl sulfoxide, epichlorohydrin, sodium hydroxide and sodium borohydride in a reaction kettle, stirring the mixture, filtering out the solid, and washing the solid to obtain a prefabricated polysaccharide carrier;

[0066] 2) placing the prefabricated polysaccharide carrier in the amino acid solution and mixing, adjusting the pH value and stirring at low temperature to obtain a modified polysaccharide carrier;

[0067] 3) After the modified polysaccharide carrier is immersed in the recombinant protein solution and treated at low temperature, a sodium chloride solution is added and stirred and mixed, solids are taken out after suction filtration, ethanolamine solution is added and mixed at low temperature, centrifuged, and washed to obtain recombinant collagen.

[0068] In step 1), the stirring treatment is stirring at 40°C and 190 r / min for 2.8 hours; the washing is washing three times with distilled water; the amount ratio of the polysaccharide carrier, dimethyl sulfoxide, epichlorohydrin, sodium hydroxide and sodium borohydride is 13.5:118:12.8:0.29:0.004;

[0069] In step 2), the ratio of the prefabricated polysaccharide carrier to the amino acid solution is 5.8 g:56 mL; the concentration of the amino acid solution is 0.23 mol / L; the solute in the amino acid solution is arginine; the pH value is adjusted to 8.4 using hydrochloric acid; the low-temperature stirring is stirring at 24°C for 13 hours;

[0070] In step 3), the low temperature treatment is stirring at 25°C for 23h; the stirring and mixing time is 38min; the low temperature mixing is mixing at 20°C for 14h; the washing is washing four times with distilled water; the dosage ratio of the modified polysaccharide carrier, the recombinant protein solution, the sodium chloride solution and the ethanolamine solution is 2.95g:6.8mL:1.8mL:11.5mL; the concentration of the sodium chloride solution is 1.5mol / L; the concentration of the ethanolamine solution is 1mol / L;

[0071] In step 3), the recombinant protein solution is prepared by mixing the recombinant protein and the boric acid buffer in a mass ratio of 1.15:6.8; the molecular weight of the recombinant protein is 90 kDa; the recombinant protein is recombinant humanized type I collagen;

[0072] In step 1), the preparation of the polysaccharide carrier includes the following steps: agarose powder and cellulose powder are stirred in a 4.8% mass fraction acetic acid solution at 60°C for 1.4 hours and ultrasonically degassing bubbles to obtain a mixed solution, and the mixed solution is dripped dropwise into a mixed solution consisting of 2 mol / L NaOH and anhydrous ethanol in a ratio of 3:1 by an extrusion method, and soaked for 3.8 hours to obtain a polysaccharide carrier; the mass ratio of the agarose powder, cellulose powder and acetic acid solution is 4.8:0.95:58.

[0073] Example 5

[0074] A medical recombinant collagen nursing dressing for repairing sensitive skin wounds comprises a non-woven fabric and a recombinant collagen composition, wherein the recombinant collagen composition comprises the following components by weight: 100 parts of purified water, 0.32 parts of recombinant collagen, 0.26 parts of carbomer, 8.4 parts of glycerol and 0.06 parts of methylparaben;

[0075] A preparation method of a medical recombinant collagen nursing dressing for repairing sensitive skin wounds comprises the following steps: sequentially adding purified water, carbomer, glycerin and methylparaben into an emulsifier in proportion, stirring at a speed of 30 r / min, heating to 90°C and keeping warm for 30 minutes; stirring and cooling to 45°C, adding recombinant collagen, stirring at a speed of 30 r / min for 15 minutes, and wetting the recombinant collagen composition on the non-woven fabric to obtain the recombinant collagen composition.

[0076] The preparation method of the recombinant collagen comprises the following steps:

[0077] 1) placing a polysaccharide carrier, dimethyl sulfoxide, epichlorohydrin, sodium hydroxide and sodium borohydride in a reaction kettle, stirring the mixture, filtering out the solid, and washing the solid to obtain a prefabricated polysaccharide carrier;

[0078] 2) placing the prefabricated polysaccharide carrier in the amino acid solution and mixing, adjusting the pH value and stirring at low temperature to obtain a modified polysaccharide carrier;

[0079] 3) After the modified polysaccharide carrier is immersed in the recombinant protein solution and treated at low temperature, a sodium chloride solution is added and stirred and mixed, solids are taken out after suction filtration, ethanolamine solution is added and mixed at low temperature, centrifuged, and washed to obtain recombinant collagen.

[0080] In step 1), the stirring treatment is stirring at 40°C and 200 r / min for 3 hours; the washing is washing three times with distilled water; the dosage ratio of the polysaccharide carrier, dimethyl sulfoxide, epichlorohydrin, sodium hydroxide and sodium borohydride is 14:120:13.0:0.30:0.004;

[0081] In step 2), the ratio of the prefabricated polysaccharide carrier to the amino acid solution is 6 g:58 mL; the concentration of the amino acid solution is 0.23 mol / L; the solute in the amino acid solution is arginine; the pH value is adjusted to 8.5 using hydrochloric acid; the low-temperature stirring is stirring at 25°C for 14 hours;

[0082] In step 3), the low temperature treatment is stirring at 25°C for 24 hours; the stirring and mixing time is 40 minutes; the low temperature mixing is mixing at 20°C for 15 hours; the washing is washing four times with distilled water; the dosage ratio of the modified polysaccharide carrier, the recombinant protein solution, the sodium chloride solution and the ethanolamine solution is 3.0g:7mL:2mL:12mL; the concentration of the sodium chloride solution is 1.5mol / L; the concentration of the ethanolamine solution is 1mol / L;

[0083] In step 3), the recombinant protein solution is prepared by mixing the recombinant protein and the boric acid buffer in a mass ratio of 1.2:7; the molecular weight of the recombinant protein is 100 kDa; and the recombinant protein is recombinant humanized type I collagen;

[0084] In step 1), the preparation of the polysaccharide carrier includes the following steps: agarose powder and cellulose powder are stirred in a 5% mass fraction acetic acid solution at 60°C for 1.5 hours and ultrasonically degassing bubbles to obtain a mixed solution, and the mixed solution is dripped dropwise into a mixed solution consisting of 2 mol / L NaOH and anhydrous ethanol in a ratio of 3:1 by an extrusion method, and soaked for 4 hours to obtain a polysaccharide carrier; the mass ratio of the agarose powder, cellulose powder and acetic acid solution is 5:1.0:60.

[0085] Comparative Example 1

[0086] Compared with Example 2, the difference in Comparative Example 1 is that agarose is used instead of cellulose, and the other components, preparation steps and parameters are the same.

[0087] Comparative Example 2

[0088] Compared with Example 2, the difference in Comparative Example 2 is that cellulose is used instead of agarose, and the other components, preparation steps and parameters are the same.

[0089] Comparative Example 3

[0090] Compared with Example 2, Comparative Example 3 is different in that no amino acid is used, and the remaining components, preparation steps and parameters are the same.

[0091] Comparative Example 4

[0092] Compared with Example 2, the difference in Comparative Example 4 is that a polysaccharide carrier is used instead of an epoxy polysaccharide carrier, and the other components, preparation steps and parameters are the same.

[0093] Examples 1-5 and Comparative Examples 1-4 were tested as follows, and the test results are shown in Table 1.

[0094] Stability test

[0095] The different samples of Examples 1-5 and Comparative Examples 1-4 were stored at 55° C. for 20 days, and the retention rates of the recombinant collagen therein were measured to evaluate the stability of the recombinant collagen prepared by the present invention. The test results are shown in Table 1.

[0096] The retention rate of recombinant collagen is calculated as follows:

[0097] Retention rate of recombinant collagen (%) = C1 / C0 × 100%;

[0098] Wherein, C0 is the content of active ingredient in the initial sample; C1 is the content of active ingredient in the sample after storage for 15 days.

[0099] Table 1

[0100] sample Retention rate of recombinant collagen (%) Example 1 85.3 Example 2 86.8 Example 3 85.7 Example 4 85.8 Example 5 86.3 Comparative Example 1 73.8 Comparative Example 2 75.4 Comparative Example 3 81.6 Comparative Example 4 80.8

[0101] From the test results in Table 1, it can be seen that compared with Comparative Examples 1-4, the recombinant collagen prepared in Examples 1-5 of the present invention has excellent stability.

[0102] Wound repair test

[0103] 90 healthy rabbits (average weight 4kg) were selected and randomly divided into 9 groups. After injection anesthesia, hair was cut on the back, and the skin was wiped and disinfected with 75% alcohol, an artificial wound surface was made on the back using a skin biopsy device. The wound surface was in the shape of a 2cm×2cm circle, and 0.2cm deep tissue was removed within the circle, including skin, subcutaneous and muscle tissue. The wound surface was cleaned, and the medical recombinant collagen nursing dressings prepared in the embodiments and comparative examples were bandaged on the surface of the wound surface, and the dressing was changed and re-bandaged every 2 days, and the wound was disinfected with iodine tincture. The area of ​​the wound surface was measured using Imagel software, and observed for 5 days, and the wound surface reduction rate (%) was calculated as (S0-S1) / S0×100, where S0 is the area of ​​the wound surface before treatment, and S1 is the area of ​​the wound surface after treatment;

[0104] Table 2

[0105] sample Wound surface reduction rate (%) Example 1 88.3 Example 2 91.6 Example 3 87.2 Example 4 90.1 Example 5 90.4 Comparative Example 1 69.7 Comparative Example 2 70.5 Comparative Example 3 76.8 Comparative Example 4 73.3

[0106] From the test results in Table 2, it can be seen that compared with Comparative Examples 1-4, the medical recombinant collagen care dressings for repairing sensitive skin wounds of Examples 1-5 of the present invention are significantly better than Comparative Examples 1-4 in repairing damaged wound surfaces and promoting wound healing.

[0107] The above description is only a preferred embodiment of the present invention and does not limit the present invention in any form. Although the present invention has been disclosed as a preferred embodiment as above, it is not used to limit the present invention. Any technical personnel in this field can make some changes or modify the technical contents disclosed above into equivalent embodiments without departing from the scope of the technical solution of the present invention. However, any brief modifications, equivalent changes and modifications made to the above embodiments based on the technical essence of the present invention without departing from the content of the technical solution of the present invention are still within the scope of the technical solution of the present invention.

Claims

1. A medical recombinant collagen nursing dressing for repairing sensitive skin wounds, comprising a non-woven fabric and a recombinant collagen composition, characterized in that: The recombinant collagen composition comprises the following components in parts by weight: 91-100 parts of purified water, 0.20-0.32 parts of recombinant collagen, 0.20-0.26 parts of carbomer, 8.0-8.4 parts of glycerol and 0.05-0.06 parts of methylparaben.

2. The medical recombinant collagen nursing dressing for repairing sensitive skin wounds according to claim 1, characterized in that: The preparation method of the recombinant collagen comprises the following steps: 1) placing a polysaccharide carrier, dimethyl sulfoxide, epichlorohydrin, sodium hydroxide and sodium borohydride in a reaction kettle and stirring the mixture, filtering out the solid matter, and washing the solid matter to obtain a prefabricated polysaccharide carrier; 2) placing the prefabricated polysaccharide carrier in the amino acid solution and mixing, adjusting the pH value and stirring at low temperature to obtain a modified polysaccharide carrier; 3) After the modified polysaccharide carrier is immersed in the recombinant protein solution and treated at low temperature, a sodium chloride solution is added and stirred and mixed, solids are taken out after suction filtration, ethanolamine solution is added and mixed at low temperature, centrifuged, and washed to obtain recombinant collagen.

3. The medical recombinant collagen nursing dressing for repairing sensitive skin wounds according to claim 1, characterized in that: In step 1), the stirring treatment is stirring at 40° C. and 150-200 r / min for 2-3 hours; and the washing is washing three times with distilled water.

4. The medical recombinant collagen nursing dressing for repairing sensitive skin wounds according to claim 1, characterized in that: In step 1), the usage ratio of the polysaccharide carrier, dimethyl sulfoxide, epichlorohydrin, sodium hydroxide and sodium borohydride is 12-14:110-120:12.2-13.0:0.25-0.30:0.

004.

5. The medical recombinant collagen nursing dressing for repairing sensitive skin wounds according to claim 1, characterized in that: In step 2), the usage ratio of the prefabricated polysaccharide carrier and the amino acid solution is 5-6 g: 50-58 mL; the concentration of the amino acid solution is 0.20-0.23 mol / L; the solute in the amino acid solution is one or more of arginine, histidine and lysine.

6. The medical recombinant collagen nursing dressing for repairing sensitive skin wounds according to claim 1, characterized in that: In step 2), the pH value is adjusted to 8.0-8.5 using hydrochloric acid; and the low-temperature stirring is stirring at a temperature of 20-25° C. for 10-14 hours.

7. The medical recombinant collagen nursing dressing for repairing sensitive skin wounds according to claim 1, characterized in that: In step 3), the low temperature treatment is stirring at 25°C for 20-24 hours; the stirring and mixing time is 30-40 minutes; the low temperature mixing is mixing at 20°C for 12-15 hours; and the washing is washing four times with distilled water.

8. The medical recombinant collagen nursing dressing for repairing sensitive skin wounds according to claim 1, characterized in that: In step 3), the usage ratio of the modified polysaccharide carrier, the recombinant protein solution, the sodium chloride solution and the ethanolamine solution is 2.8-3.0 g: 6-7 mL: 1-2 mL: 10-12 mL. The concentration of the sodium chloride solution is 1.5 mol / L; the concentration of the ethanolamine solution is 1 mol / L; In step 3), the recombinant protein solution is prepared by mixing recombinant protein and boric acid buffer at a mass ratio of 1.0-1.2:6-7; the molecular weight of the recombinant protein is 50-100 kDa; the recombinant protein is one or more of recombinant humanized type I collagen, recombinant humanized type III collagen, recombinant humanized type IV collagen and recombinant humanized type VII collagen.

9. The medical recombinant collagen nursing dressing for repairing sensitive skin wounds according to claim 1, characterized in that: The preparation method of the polysaccharide carrier comprises the following steps: agarose powder and cellulose powder are stirred in a 4-5% mass fraction acetic acid solution at 60°C for 1.2-1.5h and bubbles are removed by ultrasound to obtain a mixed solution, and the mixed solution is dripped dropwise into a mixed solution consisting of 2 mol / L NaOH and anhydrous ethanol in a ratio of 3:1 by an extrusion method, and soaked for 3-4h to obtain a polysaccharide carrier; the mass ratio of the agarose powder, cellulose powder and acetic acid solution is 4-5:0.8-1.0:50-60.

10. A method for preparing a medical recombinant collagen nursing dressing for repairing sensitive skin wounds as claimed in any one of claims 1 to 9, characterized in that: The preparation method comprises the following steps: sequentially adding purified water, carbomer, glycerin and methylparaben into an emulsifier in proportion, stirring at a speed of 25-30 r / min, heating to 80-90° C., and keeping the temperature for 20-30 min; stirring and cooling to 35-45° C., adding recombinant collagen, stirring at a speed of 25-30 r / min for 10-15 min, and wetting the recombinant collagen composition on the non-woven fabric to obtain the product.