Medical Recombinant Collagen Nursing Dressing for Sensitive Skin Wound Repair and Its Preparation Method

Abstract

The present invention relates to a medical recombinant collagen dressing for sensitive skin wound repair and its preparation method, belonging to the technical field of medical dressings. A medical recombinant collagen dressing for sensitive skin wound repair comprises a non-woven fabric and a recombinant collagen composition, and 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 according to a proportion, heating and stirring; then stirring and cooling, adding recombinant collagen, and stirring and mixing to obtain the product. By immobilizing the recombinant protein, the present invention enhances its stability, prolongs the service life of the dressing, reduces the replacement frequency, and lowers the medical cost. The stable recombinant collagen improves the biological activity and functionality, promotes wound healing, and reduces scarring.

Description

Technical Field

[0001] The present invention belongs to the technical field of medical dressings, and relates to a medical recombinant collagen care dressing for sensitive muscle wound repair 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 the rejection reaction with human tissues, lower the risks of allergy and infection, thus ensuring the safety of patients. Secondly, the structure and properties of recombinant collagen can be precisely regulated by genetic engineering techniques. This controllability enables us to design recombinant collagen dressings with specific functions according to specific medical needs, such as promoting wound healing, improving skin texture, etc.

[0003] However, the stability of recombinant collagen is poor because there are many non-covalent bonds in its molecular structure, such as hydrogen bonds, hydrophobic interactions and ionic bonds, etc. The breakage 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 protein immobilization materials, which 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, due to the low activity of epoxy groups under neutral conditions, the amount of protein that can be immobilized is relatively small. Summary of the Invention

[0005] The purpose of the present invention is to provide a medical recombinant collagen care dressing for sensitive muscle wound repair and a preparation method thereof. By immobilizing recombinant proteins, the present invention enhances their stability, extends the lifespan of the dressing, reduces the replacement frequency, and lowers the medical cost. Stabilizing recombinant collagen improves its biological activity and functionality, promotes wound healing, and reduces scarring. At the same time, it enhances the resistance to the environment, maintains stable performance in a complex medical environment, ensures the safety of patient recovery, 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 care dressing for sensitive muscle wound repair, comprising a non-woven fabric and a recombinant collagen composition. 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 preparation method of the recombinant collagen comprises the following steps:

[0009] 1) Place the polysaccharide carrier, dimethyl sulfoxide, epichlorohydrin, sodium hydroxide and sodium borohydride in a reaction kettle, stir and process, filter to obtain the solid matter, and after washing, obtain the prefabricated polysaccharide carrier;

[0010] 2) Place the prefabricated polysaccharide carrier in an amino acid solution, mix well, adjust the pH value, and then carry out low-temperature stirring to obtain the modified polysaccharide carrier;

[0011] 3) Immerse the modified polysaccharide carrier in the recombinant protein solution, carry out low-temperature treatment, add sodium chloride solution, stir and mix, filter by suction to obtain the solid matter, add ethanolamine solution, carry out low-temperature mixing, centrifuge, and after washing, obtain the recombinant collagen.

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

[0013] As a preferred technical solution of the present invention, in step 1), the dosage 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 - 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.

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

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

[0017] As a preferred technical solution of the present invention, in step 3), the dosage 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 from a recombinant protein and a 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: mixing agarose powder and cellulose powder in an acetic acid solution with a mass fraction of 4 - 5% at 60 °C and stirring for 1.2 - 1.5 h, then ultrasonicating to remove air bubbles to obtain a mixed solution. The mixed solution is dropped into a mixed solution composed of 2 mol / L NaOH and anhydrous ethanol in a ratio of 3 : 1 by extrusion method and soaked for 3 - 4 h to obtain the 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 preparation method of a medical recombinant collagen dressing for sensitive muscle wound repair, the preparation method comprising the following steps: sequentially adding purified water, carbomer, glycerol, and methylparaben into an emulsifier at a stirring speed of 25 - 30 r / min, heating to 80 - 90 °C, and holding for 20 - 30 min; then stirring and cooling to 35 - 45 °C, adding recombinant collagen, and stirring at a stirring speed of 25 - 30 r / min for 10 - 15 min to obtain a recombinant collagen composition, and wetting the recombinant collagen composition on the non-woven fabric to obtain the product.

[0021] The beneficial effects of the present invention:

[0022] (1) In the present invention, the polysaccharide carrier is prepared and subjected to epoxy and amino modification, so that the positively charged amino groups on the epoxy amino polysaccharide carrier adsorb the negatively charged regions on the recombinant protein through electrostatic force, enabling the recombinant protein to be physically adsorbed on the epoxy amino polysaccharide carrier. Then, the epoxy groups on the epoxy amino polysaccharide carrier react covalently with the amino groups on the recombinant protein close to the epoxy groups to fix the recombinant protein on the epoxy amino polysaccharide carrier. The covalent reaction between the epoxy group 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) By immobilizing the recombinant protein, the present invention enhances its stability, extends the lifespan of the dressing, reduces the replacement frequency, and lowers the medical cost. The stabilized recombinant collagen enhances biological activity and functionality, promotes wound healing, and reduces scarring. At the same time, it enhances the resistance to the environment, maintains stable performance in a complex medical environment, ensures the safety of patient recovery, and reduces the risk of complications. Detailed implementation manners

[0024] To further elaborate on the technical means and effects adopted by the present invention to achieve the predetermined invention purpose, the following, in combination with embodiments, details the specific implementation manners, structures, features, and their effects according to the present invention as follows.

[0025] Example 1

[0026] A medical recombinant collagen care dressing for sensitive skin wound repair comprises a non-woven fabric and a recombinant collagen composition. The recombinant collagen composition comprises the following components in parts by weight: 91 parts of purified water, 0.20 part of recombinant collagen, 0.20 part of carbomer, 8.0 parts of glycerol, and 0.05 part of methylparaben;

[0027] A preparation method of a medical recombinant collagen care dressing for sensitive skin wound repair comprises the following steps: sequentially feeding purified water, carbomer, glycerol, and methylparaben into an emulsifying machine according to a ratio, stirring at a speed of 25 r / min, heating to 80 °C, and keeping warm for 20 min; then stirring and cooling to 35 °C, adding recombinant collagen, and stirring at a stirring speed of 25 r / min for 10 min to obtain a recombinant collagen composition, and wetting the recombinant collagen composition on the non-woven fabric to obtain the product.

[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 for stirring treatment, filtering to obtain a solid, and washing to obtain a prefabricated polysaccharide carrier;

[0030] 2) Placing the prefabricated polysaccharide carrier in an amino acid solution for mixing, adjusting the pH value, and performing low-temperature stirring to obtain a modified polysaccharide carrier;

[0031] 3) Soaking the modified polysaccharide carrier in a recombinant protein solution for low-temperature treatment, adding a sodium chloride solution for stirring and mixing, filtering to obtain a solid, adding an ethanolamine solution for low-temperature mixing, centrifuging, and washing to obtain recombinant collagen.

[0032] In step 1), the stirring treatment is to stir at a temperature of 40°C and a rotation speed of 150 r / min for 2 h; the washing is to wash three times with distilled water; the dosage 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 dosage ratio of the prefabricated polysaccharide carrier and 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 adjustment is to adjust the pH value to 8.0 with hydrochloric acid; the low-temperature stirring is to stir at a temperature of 20°C for 10 h;

[0034] In step 3), the low-temperature treatment is to stir at a temperature of 25°C for 20 h; the stirring and mixing time is 30 min; the low-temperature mixing is to mix at a temperature of 20°C for 12 h; the cleaning is to clean 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 2.8 g:6 mL:1 mL:10 mL; the concentration of the sodium chloride solution is 1.5 mol / L; the concentration of the ethanolamine solution is 1 mol / L;

[0035] In step 3), the recombinant protein solution is prepared from recombinant protein and boric acid buffer in a mass ratio of 1.0:6; the molecular weight of the recombinant protein is 50 kDa; 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 a temperature of 60°C for 1.2 h and ultrasonicated to remove air bubbles to obtain a mixed solution, and the mixed solution is dropped into a mixed solution composed of 2 mol / L NaOH and anhydrous ethanol in a ratio of 3:1 by the extrusion method and soaked for 3 h to obtain the 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 care dressing for sensitive muscle wound repair includes a non-woven fabric and a recombinant collagen composition. The recombinant collagen composition includes the following components in parts by weight: 93 parts of purified water, 0.23 part of recombinant collagen, 0.22 part of carbomer, 8.1 parts of glycerol and 0.053 part of methylparaben;

[0039] A preparation method of a medical recombinant collagen dressing for sensitive skin wound repair comprises the following steps: sequentially adding purified water, carbomer, glycerol, and methylparaben into an emulsifying machine according to a ratio, stirring at a speed of 25 r / min, heating to 82 °C, and keeping warm for 22 min; then stirring and cooling to 38 °C, adding recombinant collagen, and stirring at a speed of 25 r / min for 11 min to obtain a recombinant collagen composition, and wetting the recombinant collagen composition on the non-woven fabric to obtain the product.

[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 for stirring treatment, filtering to obtain a solid, and washing to obtain a prefabricated polysaccharide carrier;

[0042] 2) Mixing the prefabricated polysaccharide carrier in an amino acid solution, adjusting the pH value, and performing low-temperature stirring to obtain a modified polysaccharide carrier;

[0043] 3) Immersing the modified polysaccharide carrier in a recombinant protein solution for low-temperature treatment, adding a sodium chloride solution for stirring and mixing, filtering to obtain a solid, adding an ethanolamine solution for low-temperature mixing, centrifuging, and washing to obtain recombinant collagen.

[0044] In step 1), the stirring treatment is stirring at a temperature of 40 °C and a speed of 160 r / min for 2.5 h; 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 12.5:112:12.4:0.26:0.004;

[0045] In step 2), the dosage ratio of the prefabricated polysaccharide carrier and 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 adjusting of the pH value is adjusting the pH value to 8.1 with hydrochloric acid; the low-temperature stirring is stirring at a temperature of 21 °C for 11 h;

[0046] In step 3), the low-temperature treatment is stirring at a temperature of 25 °C for 21 h; the time of stirring and mixing is 35 min; the low-temperature mixing is mixing at a temperature of 20 °C for 13 h; 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 2.85 g:6.2 mL:1.2 mL:10.5 mL; the concentration of the sodium chloride solution is 1.5 mol / L; the concentration of the ethanolamine solution is 1 mol / L;

[0047] In step 3), the recombinant protein solution is prepared from recombinant protein and 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 h and ultrasonicated to remove air bubbles to obtain a mixed solution. The mixed solution is dropped into a mixed solution composed of 2 mol / L NaOH and absolute ethanol in a ratio of 3:1 by extrusion method and soaked for 3.2 h to obtain the 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 dressing for sensitive skin wound repair comprises a non-woven fabric and a recombinant collagen composition. The recombinant collagen composition comprises the following components in parts by weight: 96 parts of purified water, 0.26 part of recombinant collagen, 0.23 part of carbomer, 8.2 parts of glycerol and 0.055 part of methylparaben.

[0051] A preparation method of a medical recombinant collagen dressing for sensitive skin wound repair comprises the following steps: sequentially adding purified water, carbomer, glycerol and methylparaben into an emulsifier according to a ratio, stirring at a speed of 28 r / min, heating to 85 °C and keeping warm for 25 min; then stirring and cooling to 40 °C, adding recombinant collagen, and stirring at a stirring speed of 28 r / min for 12 min to obtain the recombinant collagen composition. The recombinant collagen composition is moistened on the non-woven fabric to obtain the product.

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

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

[0054] 2) Placing the prefabricated polysaccharide carrier in an amino acid solution, mixing evenly, adjusting the pH value and then performing low-temperature stirring to obtain a modified polysaccharide carrier;

[0055] 3) Soaking the modified polysaccharide carrier in the recombinant protein solution for low-temperature treatment, adding a sodium chloride solution, stirring and mixing, filtering to obtain a solid, adding an ethanolamine solution for low-temperature mixing, centrifuging, and washing to obtain recombinant collagen.

[0056] In step 1), the stirring treatment is to stir at a temperature of 40°C and a rotation speed of 175 r / min for 2.5 h; the washing is to wash three times with distilled water; the dosage 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 dosage ratio of the prefabricated polysaccharide carrier and 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 adjustment is to adjust the pH value to 8.2 with hydrochloric acid; the low-temperature stirring is to stir at a temperature of 22°C for 12 h;

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

[0059] In step 3), the recombinant protein solution is prepared from recombinant protein and 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 a temperature of 60°C for 1.5 h and ultrasonicated to remove air bubbles to obtain a mixed solution, and the mixed solution is dropped into a mixed solution composed of 2 mol / L NaOH and anhydrous ethanol in a ratio of 3:1 by the extrusion method and soaked for 3.5 h to obtain the 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 dressing for sensitive skin wound repair includes a non-woven fabric and a recombinant collagen composition. The recombinant collagen composition includes the following components in parts by weight: 98 parts of purified water, 0.29 part of recombinant collagen, 0.24 part of carbomer, 8.3 parts of glycerol, and 0.057 part of methylparaben;

[0063] A preparation method of a medical recombinant collagen dressing for sensitive skin wound repair comprises the following steps: sequentially feeding purified water, carbomer, glycerol, and methylparaben into an emulsifying machine at a stirring speed of 30 r / min, heating to 88 °C, and holding for 28 min; then stirring and cooling to 42 °C, adding recombinant collagen, and stirring at a stirring speed of 30 r / min for 14 min to obtain a recombinant collagen composition, and wetting the recombinant collagen composition on the non-woven fabric to obtain the product.

[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 for stirring treatment, filtering to obtain a solid, and washing to obtain a prefabricated polysaccharide carrier;

[0066] 2) Placing the prefabricated polysaccharide carrier in an amino acid solution for mixing, adjusting the pH value, and performing low-temperature stirring to obtain a modified polysaccharide carrier;

[0067] 3) Soaking the modified polysaccharide carrier in a recombinant protein solution for low-temperature treatment, adding a sodium chloride solution for stirring and mixing, filtering to obtain a solid, adding an ethanolamine solution for low-temperature mixing, centrifuging, and washing to obtain recombinant collagen.

[0068] In step 1), the stirring treatment is stirring at a temperature of 40 °C and a speed of 190 r / min for 2.8 h; 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 13.5:118:12.8:0.29:0.004;

[0069] In step 2), the dosage ratio of the prefabricated polysaccharide carrier and 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 adjustment is adjusting the pH value to 8.4 with hydrochloric acid; the low-temperature stirring is stirring at a temperature of 24 °C for 13 h;

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

[0071] In step 3), the recombinant protein solution is prepared from a recombinant protein and a 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 comprises 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 h and ultrasonicated to remove air bubbles to obtain a mixed solution, and the mixed solution is dropped into a mixed solution composed of 2 mol / L NaOH and anhydrous ethanol in a ratio of 3:1 by the extrusion method, and soaked for 3.8 h to obtain the 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 dressing for sensitive skin wound repair comprises a non-woven fabric and a recombinant collagen composition. The recombinant collagen composition comprises the following components in parts by weight: 100 parts of purified water, 0.32 part of recombinant collagen, 0.26 part of carbomer, 8.4 parts of glycerol and 0.06 part of methylparaben;

[0075] A preparation method of a medical recombinant collagen dressing for sensitive skin wound repair comprises the following steps: sequentially adding purified water, carbomer, glycerol and methylparaben into an emulsifying machine at a stirring speed of 30 r / min, heating to 90 °C and keeping warm for 30 min; then stirring and cooling to 45 °C, adding recombinant collagen, and stirring at a stirring speed of 30 r / min for 15 min to obtain the recombinant collagen composition, and wetting the recombinant collagen composition on the non-woven fabric to obtain the product.

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

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

[0078] 2) Placing the prefabricated polysaccharide carrier in an amino acid solution, mixing evenly, adjusting the pH value, and performing low-temperature stirring to obtain a modified polysaccharide carrier;

[0079] 3) Soaking the modified polysaccharide carrier in the recombinant protein solution for low-temperature treatment, adding a sodium chloride solution, stirring and mixing, filtering to obtain a solid, adding an ethanolamine solution for low-temperature mixing, centrifuging, and washing to obtain recombinant collagen.

[0080] In step 1), the stirring treatment is to stir at a temperature of 40°C and a rotation speed of 200 r / min for 3 h; the washing is to wash 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 dosage ratio of the prefabricated polysaccharide carrier and 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 with hydrochloric acid; the low-temperature stirring is to stir at a temperature of 25°C for 14 h;

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

[0083] In step 3), the recombinant protein solution is prepared by mixing recombinant protein and boric acid buffer in a mass ratio of 1.2:7; the molecular weight of the recombinant protein is 100 kDa; 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 a temperature of 60°C for 1.5 h and ultrasonic degassing to obtain a mixed solution, and the mixed solution is dropped into a mixed solution composed of 2 mol / L NaOH and anhydrous ethanol in a ratio of 3:1 by the extrusion method and soaked for 4 h to obtain the 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, the difference in Comparative Example 3 is that no amino acid is used, and the other 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 respectively subjected to the following tests, and the test results are shown in Table 1.

[0094] Stability test

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

[0096] The calculation formula for the retention rate of recombinant collagen is as follows:

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

[0098] Wherein, C0 is the content of the active ingredient in the starting sample; C1 is the content of the 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] As can be seen from the test results in Table 1, compared with Comparative Examples 1-4, the recombinant collagen prepared by the present invention has excellent stability.

[0102] Wound surface repair test

[0103] Ninety healthy white rabbits (average body weight 4 kg) were randomly divided into 9 groups. After injection anesthesia, the hair on the back was shaved, and the skin was disinfected with 75% alcohol. An artificial wound surface was made on the back using a skin biopsy instrument. The shape of the wound surface was a 2 cm × 2 cm circle, and 0.2 cm deep tissue including skin, subcutaneous, and muscle tissues was excised within the circle. The wound surface was cleaned, and the medical recombinant collagen dressing prepared in Examples and Comparative Examples was respectively bandaged on the surface of the wound surface. The dressing was changed and re-bandaged every 2 days, and the wound was disinfected with iodophor. The area of the wound surface was measured using Imagel software, and it was observed for 5 days. The wound surface reduction rate (%) = (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 area 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] As can be seen from the test results in Table 2, compared with Comparative Examples 1-4, the medical recombinant collagen nursing dressings for sensitive muscle wound repair in Examples 1-5 of the present invention have significantly better effects in repairing damaged wound surfaces and promoting wound healing than Comparative Examples 1-4.

[0107] The above are only the preferred embodiments of the present invention, and do not impose any form of limitation on the present invention. Although the present invention has been disclosed as above with the preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art can make some changes or modifications to equivalent embodiments by using the disclosed technical content within the scope of the technical solution of the present invention. However, as long as it does not depart from the content of the technical solution of the present invention, any brief modifications, equivalent changes and modifications made to the above embodiments according to the technical essence of the present invention still fall 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 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; 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, filtering out solids, and washing 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; the low temperature stirring is stirring at a temperature of 20-25° C. for 10-14 hours; 3) After soaking the modified polysaccharide carrier in a recombinant protein solution and treating it at low temperature, adding a sodium chloride solution and stirring and mixing, filtering and taking out the solid matter, adding an ethanolamine solution and mixing it at low temperature, centrifuging, and washing, a recombinant collagen is obtained; the low temperature treatment is stirring at 25°C for 20-24h; the low temperature mixing is mixing at 20°C for 12-15h; the recombinant protein solution is prepared by mixing the recombinant protein and the boric acid buffer at a mass ratio of 1.0-1.2:6-7; the molecular weight of the recombinant protein is 50-100kDa; 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; The preparation method of the polysaccharide carrier comprises the following steps: agarose powder and cellulose powder are stirred in a 4-5% by mass 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 2mol / 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.

2. 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.

3. 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.

4. 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; and the solute in the amino acid solution is one or more of arginine, histidine and lysine.

5. 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.

6. The medical recombinant collagen nursing dressing for repairing sensitive skin wounds according to claim 1, characterized in that: In step 3), the stirring and mixing time is 30-40 minutes; and the washing is performed by washing with distilled water four times.

7. 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.

8. A method for preparing a medical recombinant collagen nursing dressing for repairing sensitive skin wounds as claimed in any one of claims 1 to 7, 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.