A kind of medical dressing with repairing effect and its preparation method and application

Abstract

The present invention provides a kind of medical dressing with repair effect and its preparation method and application, and relates to the field of medical repair. Taking the total weight of the medical dressing as 100%, the mass percentage of each component of the medical dressing is: glycerol 1-5%, carbomer homopolymer 0.1-0.2%, disodium edetate 0.05-0.2%, methylparaben 0.05-0.15%, triethanolamine 0.1-0.4%, recombinant humanized collagen 0.01-0.2%, and / or sodium hyaluronate 0.01-0.3%, and the remainder is purified water. By optimizing the auxiliary materials in the medical dressing, carbomer homopolymer, glycerol, disodium edetate, methylparaben and triethanolamine make the active component recombinant humanized collagen and / or sodium hyaluronate repair function better within a certain ratio range.

Description

Technical Field

[0001] The invention belongs to the technical field of medical repair, and in particular relates to a medical dressing with repairing effect and a preparation method and application thereof. Background Art

[0002] In daily life, we often see or encounter various kinds of trauma, which cause skin damage, cracks, tissue swelling, ruptured blood vessels and bleeding. For small crack wounds, physiological saline, hydrogen peroxide or potassium permanganate solution are mainly used for washing, and iodine or alcohol is used to disinfect the wound surface, which is relatively simple to operate. If the wound bleeds, Yunnan Baiyao and other external applications are mainly used, and then sterile dressings or relatively clean dressings are used for bandaging, which is relatively complicated to operate and has poor waterproofness. In modern clinical medicine, dressings are required for the sutures of surgical incisions, abrasions, scratches, burns or scalds. At present, gauze is mainly used to cover the wound, which is not only complicated to operate, but also has the effect of preventing infection, and does not have the function of promoting wound healing. In addition, due to blood coagulation, gauze is often tightly attached to the wound, and the patient is painful when changing the dressing, and it is easy to damage the new granulation tissue, which prolongs the wound healing time. In order to simplify the above operation, some products with comprehensive functions have emerged. Such as band-aids and adhesive plasters. However, Band-Aids are only used for minor wounds, the backing material has poor water absorption, which is not conducive to the drainage of wound secretions and pus, the use time should not be too long, and it has no antibacterial function. The processing temperature during the production of adhesive plasters is around 100°C, which causes the loss of drugs. There are still substances such as gasoline remaining in the plaster, which can easily cause skin irritation, it is easy to age, and it cannot be repeatedly used.

[0003] Recombinant humanized collagen and sodium hyaluronate are commonly used active ingredients for wound repair. However, collagen contains pathogens and has the risk of spreading viruses. Sodium hyaluronate is unstable and has a short residence time in skin tissue. Therefore, Chinese invention patent application CN113332175A discloses a medical moist repair patch, including a nutrient solution and a mask paper soaked in the nutrient solution. The nutrient solution is composed of a mixture of component A, component B and component C. Component A is calculated as the percentage of the total mass of the nutrient solution: Butanediol 2-6%, glycerol 2-6%, carbomer 0.5-2%, hydroxyethyl cellulose 1-4%, disodium edetate 0.01-1%, sodium propylparaben 0.1-1%, flavoring agent 0.1-1% and deionized water; component B is: triethanolamine 0.02-0.2%; component C is: recombinant human type III collagen 0.5-2%, sodium hyaluronate 0.05-2% and beta glucan 1-6%; the medical moist repair patch has the functions of moisturizing and repairing. However, there are many excipients in the application, and it does not pay attention to the proportion of each excipient that has a better transdermal effect.

[0004] Therefore, the present application optimizes the auxiliary materials in the medical dressing with repair function, and provides a medical dressing with repair function with better biological performance and transdermal properties, and a preparation method and application thereof. Summary of the invention

[0005] The present invention aims to solve the problems existing in the prior art and provides a medical dressing with better biological performance and transdermal properties and a preparation method and application thereof. By optimizing the auxiliary materials in the medical dressing, the active components recombinant humanized collagen and / or sodium hyaluronate have better repair function and transdermal effect by making the carbomer homopolymer, glycerol, disodium edetate, methylparaben and triethanolamine within a certain proportion range.

[0006] The first aspect of the present invention is to provide a medical dressing with repairing effect, the medical dressing comprising an active component and an auxiliary component, wherein the active component is recombinant humanized collagen and / or sodium hyaluronate, and the auxiliary component is glycerol, carbomer homopolymer, disodium edetate, methylparaben and triethanolamine. Based on the total weight of the medical dressing as 100%, the mass percentage of the auxiliary component is: 1-5% of glycerol, 0.1-0.2% of carbomer homopolymer, 0.05-0.2% of disodium edetate, 0.05-0.15% of methylparaben, and 0.1-0.4% of triethanolamine.

[0007] Preferably, the mass percentages of the auxiliary material components are: 2-4% glycerol, 0.12-0.18% carbomer homopolymer, 0.08-0.15% edetate disodium, 0.1-0.15% methylparaben, and 0.15-0.3% triethanolamine.

[0008] Preferably, the mass percentages of the auxiliary material components are: 3.384% glycerol, 0.1692% carbomer homopolymer, 0.1128% edetate disodium, 0.1128% methylparaben, and 0.1974% triethanolamine.

[0009] Preferably, the mass ratio of the carbomer homopolymer to glycerol is 1:10-25.

[0010] Preferably, the mass ratio of disodium edetate to methylparaben is 1-2:1-2.

[0011] Preferably, the mass ratio of disodium edetate to triethanolamine is 1:1-3.

[0012] Preferably, based on 100% of the total weight of the medical dressing, the weight percentage of the active components is: 0.01-0.2% recombinant humanized collagen, and / or 0.01-0.3% sodium hyaluronate.

[0013] In some embodiments, the medical dressing includes an active component and an auxiliary component, wherein the active component is recombinant humanized collagen and / or sodium hyaluronate, and the auxiliary components are glycerol, carbomer homopolymer, disodium edetate, methylparaben and triethanolamine. Based on the total weight of the medical dressing as 100%, the mass percentage of each component is: 1-5% glycerol, 0.1-0.2% carbomer homopolymer, 0.05-0.2% disodium edetate, 0.05-0.15% methylparaben, 0.1-0.4% triethanolamine, 0.01-0.2% recombinant humanized collagen, and / or 0.01-0.3% sodium hyaluronate, and the balance is purified water.

[0014] In some embodiments, based on the total weight of the medical dressing as 100%, the mass percentage of each component is: glycerol 2-4%, carbomer homopolymer 0.12-0.18%, edetate disodium 0.08-0.15%, methylparaben 0.1-0.15%, triethanolamine 0.15-0.3%, recombinant humanized collagen 0.05-0.15%, and the balance is purified water.

[0015] In some embodiments, based on the total weight of the medical dressing as 100%, the mass percentage of each component is: 3.384% glycerol, 0.1692% carbomer homopolymer, 0.1128% edetate disodium, 0.1128% methylparaben, 0.1974% triethanolamine, 0.1% recombinant humanized collagen, and the remainder is purified water.

[0016] In some embodiments, based on the total weight of the medical dressing as 100%, the mass percentage of each component is: glycerol 2.83%, carbomer homopolymer 0.19%, edetate disodium 0.12%, methylparaben 0.11%, triethanolamine 0.28%, sodium hyaluronate 0.05%, and the balance is purified water.

[0017] In some embodiments, based on the total weight of the medical dressing as 100%, the mass percentage of each component is: glycerol 2.8265%, carbomer homopolymer 0.1866%, edetate disodium 0.1244%, methylparaben 0.1131%, triethanolamine 0.2829%, recombinant humanized collagen 0.065%, sodium hyaluronate 0.3%, and the remainder is purified water.

[0018] In some embodiments, the sodium hyaluronate is compounded by high molecular weight sodium hyaluronate and low molecular weight sodium hyaluronate, and the mass ratio of the two is 1:8-12.

[0019] More preferably, the carbomer homopolymer is carbomer 980.

[0020] The second aspect of the present invention is to provide a method for preparing the above-mentioned medical dressing, the preparation method comprising the following steps:

[0021] (1) Add an appropriate amount of purified water to weigh glycerol, carbomer homopolymer, and edetate disodium, heat to 70-85° C., and stir to dissolve to obtain a mixture A;

[0022] (2) adding methylparaben to purified water and stirring to obtain a methylparaben solution;

[0023] (3) mixing the methylparaben solution with the mixture A, cooling the mixture to below 40° C. for standby use, and obtaining a mixture B;

[0024] (4) adding recombinant humanized collagen and / or sodium hyaluronate to purified water, stirring to dissolve, and then adding mixture B to obtain mixture C;

[0025] (5) At the same time, add triethanolamine to purified water, stir to dissolve, and obtain a triethanolamine solution. Add the triethanolamine solution to mixture C to obtain.

[0026] Preferably, after step (5), the final mixture is canned and sterilized to obtain a solution-type medical dressing.

[0027] Preferably, after step (5), the final mixture is poured into a sterile aluminum foil bag containing a non-woven fabric or silk fabric matrix, sealed and sterilized to obtain a dressing-type medical dressing.

[0028] The third aspect of the present invention relates to the use of the above-mentioned medical dressing or the medical dressing prepared by the above-mentioned preparation method in the preparation of products with wound repairing effect.

[0029] Compared with the prior art, the present invention has the following beneficial effects: by optimizing the auxiliary materials in the medical dressing, the carbomer homopolymer, glycerin, disodium edetate, methylparaben and triethanolamine are within a certain proportion range, so that the active components recombinant humanized collagen and / or sodium hyaluronate have better repair function and transdermal effect. DETAILED DESCRIPTION

[0030] It is worth noting that the raw materials used in the present invention are all common commercially available products, and their sources are not specifically limited.

[0031] Recombinant humanized collagen: Suzhou Jiangshan Juyuan Biotechnology Co., Ltd.;

[0032] Sodium hyaluronate: Bloomage Biotech Co., Ltd.

[0033] Carbomer 980: Shanghai Changwei Pharmaceutical Excipients Technology Co., Ltd.;

[0034] Edetate disodium: Chengdu Huayi Pharmaceutical Excipients Manufacturing Co., Ltd.;

[0035] Glycerin: Shenyang Baohua Trading Co., Ltd.;

[0036] Methylparaben: Hubei Gedian Renfu Pharmaceutical Excipients Co., Ltd.;

[0037] Triethanolamine: Chengdu Huayi Pharmaceutical Excipients Manufacturing Co., Ltd.

[0038] Example 1: The composition of the medical dressing is shown in Table 1.

[0039] Table 1: Composition of the medical dressing of Example 1

[0040]

[0041] Preparation method 1:

[0042] Add weighed glycerin, carbomer 980, and disodium edetate to an appropriate amount of purified water, heat to 80°C, and stir to dissolve until no particles are visible to the naked eye to obtain preparation a; add methylparaben to purified water, stir to dissolve until no particles are visible to the naked eye to obtain preparation b; add preparation b to preparation a, mix, and cool to below 40°C for standby use; add recombinant humanized collagen to purified water, stir to dissolve, add the cooled solutions of preparation a and preparation b, and continue to stir to obtain preparation c; add triethanolamine to purified water, stir to dissolve to obtain preparation d; slowly add preparation d to preparation c, bottle, and sterilize to obtain a solution-type medical dressing.

[0043] Preparation method 2:

[0044] Add weighed glycerin, carbomer 980, and disodium edetate to an appropriate amount of purified water, heat to 80°C, and stir to dissolve until no particles are visible to the naked eye to obtain preparation a; add methylparaben to purified water, stir to dissolve until no particles are visible to the naked eye to obtain preparation b; add preparation b to preparation a, mix, and cool to below 40°C for standby use; add recombinant humanized collagen to purified water, stir to dissolve, add the cooled solutions of preparation a and preparation b, and continue to stir to obtain preparation c; add triethanolamine to purified water, stir to dissolve to obtain preparation d; slowly add preparation d to preparation c, pour into a sterile aluminum foil bag filled with non-woven fabric, seal, and sterilize to obtain a dressing-type medical dressing.

[0045] Example 2: The composition of the medical dressing is shown in Table 2.

[0046] Table 2: Composition of the medical dressing of Example 2

[0047]

[0048] Preparation method 1:

[0049] Add weighed glycerin, carbomer 980, and disodium edetate to an appropriate amount of purified water, heat to 80°C, and stir to dissolve until no particles are visible to the naked eye to obtain preparation a; add methylparaben to purified water, stir to dissolve until no particles are visible to the naked eye to obtain preparation b; add preparation b to preparation a, mix, and cool to below 40°C for standby use; add sodium hyaluronate to purified water, stir to dissolve, add the cooled solutions of preparation a and preparation b, and continue stirring to obtain preparation c; add triethanolamine to purified water, stir to dissolve to obtain preparation d; slowly add preparation d to preparation c, bottle, and sterilize to obtain a solution-type medical dressing.

[0050] Preparation method 2:

[0051] Add weighed glycerin, carbomer 980, and disodium edetate to an appropriate amount of purified water, heat to 80°C, and stir to dissolve until no particles are visible to the naked eye to obtain preparation a; add methylparaben to purified water, stir to dissolve until no particles are visible to the naked eye to obtain preparation b; add preparation b to preparation a, mix, and cool to below 40°C for standby use; add sodium hyaluronate to purified water, stir to dissolve, add the cooled solutions of preparation a and preparation b, and continue to stir to obtain preparation c; add triethanolamine to purified water, stir to dissolve to obtain preparation d; slowly add preparation d to preparation c, sterilize, pour into a sterile aluminum foil bag filled with non-woven fabric, seal, and sterilize to obtain a dressing-type medical dressing.

[0052] Example 3: The composition of the medical dressing is shown in Table 3.

[0053] Table 3: Composition of the medical dressing of Example 3

[0054]

[0055] Preparation method 1:

[0056] Add weighed glycerin, carbomer homopolymer, and disodium edetate to an appropriate amount of purified water, heat to 80°C, and stir to dissolve until no particles are visible to the naked eye to obtain preparation a; add methylparaben to purified water, stir to dissolve until no particles are visible to the naked eye to obtain preparation b; add preparation b to preparation a, mix, and cool to below 40°C for standby use; add recombinant humanized collagen to purified water, stir to dissolve, add the cooled solutions of preparations a and b, and continue to stir to obtain preparation c; add sodium hyaluronate to purified water, stir to dissolve, add preparation c, and continue to stir to obtain preparation d; at the same time, add triethanolamine to purified water, stir to dissolve, and obtain preparation e; slowly add preparation e to preparation d, bottle, and sterilize to obtain a solution-type medical dressing.

[0057] Preparation method 2:

[0058] Add weighed glycerin, carbomer homopolymer, and disodium edetate to an appropriate amount of purified water, heat to 80°C, and stir to dissolve until no particles are visible to the naked eye to obtain preparation a; add methylparaben to purified water, stir to dissolve until no particles are visible to the naked eye to obtain preparation b; add preparation b to preparation a, mix, and cool to below 40°C for standby use; add recombinant humanized collagen to purified water, stir to dissolve, add the cooled solutions of preparations a and b, and continue to stir to obtain preparation c; add sodium hyaluronate to purified water, stir to dissolve, add preparation c, and continue to stir to obtain preparation d; at the same time, add triethanolamine to purified water, stir to dissolve, and obtain preparation e; slowly add preparation e to preparation d, sterilize, and pour into a sterile aluminum foil bag filled with non-woven fabric, seal, and sterilize to obtain a dressing-type medical dressing.

[0059] Comparative Example 1: The composition of the medical dressing is shown in Table 4.

[0060] Table 4: Composition of the medical dressing of Comparative Example 1

[0061]

[0062] The preparation method is the same as Preparation Method 1 and Preparation Method 2 in Example 1 to obtain a solution-type medical dressing and a dressing-type medical dressing.

[0063] Comparative Example 2: The composition of the medical dressing is shown in Table 5.

[0064] Table 5: Composition of the medical dressing of Comparative Example 2

[0065]

[0066] The preparation method is the same as Preparation Method 1 and Preparation Method 2 in Example 2 to obtain a solution-type medical dressing and a dressing-type medical dressing.

[0067] Comparative Example 3: The composition of the medical dressing is shown in Table 6.

[0068] Table 6: Composition of the medical dressing of Comparative Example 3

[0069]

[0070] The preparation method is the same as Preparation Method 1 and Preparation Method 2 in Example 3 to obtain a solution-type medical dressing and a dressing-type medical dressing.

[0071] Test Example 1: Cytotoxicity Test

[0072] 1. Materials

[0073] 1.1 Reagents

[0074] MEM culture medium, Gibco, lot number: 8123188

[0075] Fetal bovine serum, Zhejiang Tianhang Biotechnology Co., Ltd., batch number: 21070703

[0076] Digestive enzymes, Gibco, lot number: 2458323

[0077] PBS, Gibco, Lot No.: 8122732

[0078] MTT: sigma, lot number: MKBX8173V

[0079] DMSO: Sinopharm Chemical Reagent Co., Ltd., batch number: 20200923

[0080] Isopropyl alcohol, Sinopharm Chemical Reagent Co., Ltd., batch number: 20170926

[0081] 1.2 Solution preparation

[0082] Take 100 mg of MTT, add 100 mL of MEM culture medium, mix well to make a 1 mg / mL MTT solution.

[0083] 1.3 Sample preparation

[0084] Test sample group: Take 1 g of the test sample (the solution-type medical dressing prepared by Preparation Method 1 in Example 3), add 10 mL of MEM culture medium (containing serum) at a ratio of 0.1 g / mL, and extract at 37° C. with shaking for 24 h for use.

[0085] Blank control group: Take 10 mL of MEM culture medium (containing serum) and keep it at 37°C for 24 hours for later use.

[0086] Negative control group: weigh 2 g of high-density polyethylene, add 10 mL of MEM culture medium (containing serum) at a ratio of 0.2 g / mL, shake and extract at 37°C for 24 h, and set aside.

[0087] Positive control group: Take 1 mL of dimethyl sulfoxide (DMSO) and add 9 mL of MEM culture medium (containing serum) for later use.

[0088] 2. Test methods

[0089] 2.1 Preparation of cell suspension

[0090] The L929 cells (monolayer fibroblast L929, from the Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences) that have been cultured and grown vigorously were digested with digestive enzymes and then added with fresh culture medium and blown evenly. The cells were counted under a microscope using a hemocytometer, and the cell concentration was calculated and adjusted to make the final concentration of the cell suspension 1×105 Pieces / mL.

[0091] 2.2 Cell culture

[0092] Add 100 μL of culture medium to the outer wells of the 96-well culture plate, and add 100 μL of the prepared 1×10 5 / mL cell suspension, set up test group, blank control group, negative control group and positive control group, 2 columns in each group (12 wells in total). After incubation in a CO2 incubator (37℃, 5% CO2) for 24h, discard the original culture medium, add 100μL test solution, blank control solution, negative control solution or positive control solution to each corresponding well, and incubate in a CO2 incubator for 24h.

[0093] 2.3 Cytotoxicity evaluation

[0094] After adding the test solution and incubating for 24 hours, observe the cell morphology under a microscope. Discard the solution in the wells, add 50μL of MTT solution to each well, continue incubating in a 37℃ incubator for 2 hours, then discard the MTT solution, add 100μL of isopropanol solution to each well, shake the plate for 10 minutes, and measure the absorbance at 570nm (reference wavelength 650nm) on a microplate reader.

[0095] 2.4 Data Analysis

[0096] The decrease in survival rate was calculated by comparing with the blank equation as follows: Survival rate (%) = OD 570e / OD 570b ×100%

[0097] in,

[0098] OD 570e : Average optical density of the test sample extract;

[0099] OD 570b : Average value of blank optical density.

[0100] If the cell survival rate (%) of the test article group is less than 70% of the blank control group, it has potential cytotoxicity.

[0101] 3. Results

[0102] 3.1 Microscope observation

[0103] After adding the test solution and incubating for 24 hours, the cells were examined under a microscope. Discrete particles were found in the cytoplasm of the test group, blank control group and negative control group, without cell lysis and decreased cell proliferation. More than 70% of the cells in the positive control group were shrunken and had no cytoplasmic particles, and cell lysis and intercellular empty areas were clearly visible.

[0104] 3.2 Absorbance measurement

[0105] The results are shown in Table 7.

[0106] Table 7: Absorbance values ​​and viability of cells in the test group

[0107]

[0108] According to the results, under the experimental conditions, the cell survival rate of the test product was 85%, and there was no potential cytotoxicity.

[0109] Experimental Example 2: Skin Sensitization Test

[0110] 1. Test system

[0111] Experimental animals: Albino Dunkin Hartley guinea pigs, both male and female, 15, weight: 300-500 g.

[0112] Source of animals: Junqi Breeding Farm, Daoli District, Harbin City.

[0113] Laboratory Animal Production License Number: SCXK(Black)2021-004.

[0114] Feed source: Junqi Breeding Farm, Daoli District, Harbin City.

[0115] Animal feed production license number: SCXK (Black) 2021-004.

[0116] 2. Animal Management

[0117] Experimental Animal Use License Number: SYXK (Black) 2019007

[0118] Water: Drinking water is freely available

[0119] Housing conditions: Animals were housed in groups in plastic cages and identified by cage cards on which the experiment name, sample number, animal number, experiment leader, and experiment date were recorded.

[0120] Rearing environment: normal environment, temperature range 20-26℃, humidity range 40%-70%. Alternating light and dark during the day and night (12 / 12).

[0121] 3. Materials

[0122] 3.1 Reagents

[0123] Sodium chloride injection, Harbin Sanlian Pharmaceutical Co., Ltd., batch number: 221003F07

[0124] 3.2 Sample preparation

[0125] Test group: Take the dressing-type medical dressing obtained by Preparation Method 2 of Example 3, cut it into 2.5 cm×2.5 cm pieces, and use it as the test patch.

[0126] Negative control group: Cut the filter paper into 2.5cm×2.5cm, soak it with sodium chloride injection, and use it as a negative patch.

[0127] 4. Test methods

[0128] 4.1 Preparation before the test

[0129] Before the test, the hair of all the test parts of the guinea pigs was completely removed by shaving. The 15 guinea pigs were divided into 2 groups, 10 in the test group and 5 in the negative control group.

[0130] 4.2 Test methods

[0131] Induction phase: The test product was applied to the shaved left upper back of 10 guinea pigs. The bandage and patch were removed after 6 hours. This step was repeated for 3 consecutive days in 1 week. The same method was used for 3 weeks. The guinea pigs in the negative control group were operated in the same way using the corresponding patch.

[0132] Stimulation phase: 14 days after the last induction patch, all guinea pigs were stimulated. The test article was applied to the hairless area of ​​the abdomen of the guinea pigs in the test article group, and the bandage and patch were removed after 6 hours. The guinea pigs in the negative control group were operated in the same way using the corresponding patch.

[0133] Animal observation: 22 hours after the removal of the provocative patch, completely remove the hair of all the test areas of the guinea pigs by shaving. Wash and dry the hair-removed areas with warm water 2 hours later and 48 hours after the removal of the provocative patch. Observe and record the skin conditions of the provocative areas of the animals in the test group and the control group in the following table.

[0134] Table 8: Magnusson and Kligman classification

[0135]

[0136] 4.3 Results evaluation

[0137] According to the Magnusson and Kligman grading standard, when the grade of the negative control group animals is less than 1, and the grade in the test group is greater than or equal to 1, it generally indicates sensitization. If the grade of the negative control group animals is greater than or equal to 1, and the reaction of the test group animals exceeds the most severe reaction in the control group, it is considered sensitization. If it is a suspected reaction, re-challenge is recommended to confirm the results of the first challenge.

[0138] 5. Results

[0139] See the table below for specific results.

[0140] Table 9: Observation of sensitization test results

[0141]

[0142] According to the results, the reaction level of the test group was 0, and the sensitization rate was 0%; the reaction level of the negative control group was 0, and the sensitization rate was 0%. Under the conditions of this test, the test product had no skin sensitization reaction.

[0143] Test Example 3: Animal intradermal reaction test

[0144] 1. Test system

[0145] Experimental animals: Japanese white rabbits, both male and female, weight: 2.7-3.1 kg

[0146] Animal source: Junqi Breeding Farm, Daoli District, Harbin

[0147] Laboratory Animal Production License Number: SCXK (Black) 2021-004

[0148] Feed source: Junqi Breeding Farm, Daoli District, Harbin

[0149] Animal feed production license number: SCXK (black) 2021-004

[0150] 2. Animal Management

[0151] Experimental Animal Use License Number: SYXK (Black) 2019007

[0152] Water: Drinking water is freely available

[0153] Housing conditions: Animals were housed in groups in plastic cages and identified by cage cards on which the experiment name, sample number, animal number, experiment leader, and experiment date were recorded.

[0154] Rearing environment: normal environment, temperature range 20-26℃, humidity range 40%-70%. Alternating light and dark during the day and night (12 / 12).

[0155] 3. Materials

[0156] 3.1 Reagents

[0157] Sodium chloride injection, Harbin Sanlian Pharmaceutical Co., Ltd., batch number: 221003F07

[0158] Cottonseed oil: Source Leaf Biotechnology, batch number: J09D10S105262

[0159] 3.2 Sample preparation

[0160] Sodium chloride injection extract: weigh 1 g of the test sample (the solution-type medical dressing prepared by Preparation Method 1 in Example 3), add 10 mL of sodium chloride injection at a ratio of 0.1 g / mL, and shake and extract at 37°C for 72 h.

[0161] Cottonseed oil extract: weigh 1 g of the test sample (the solution-type medical dressing prepared by Preparation Method 1 in Example 3), add 10 mL of cottonseed oil at a ratio of 0.1 g / mL, and shake and extract at 37°C for 72 h.

[0162] Preparation of control solution: Prepare in the same manner using the same batch of sodium chloride injection / cottonseed oil.

[0163] 4. Test methods

[0164] 4.1 Preparation before the test

[0165] Three animals were selected. Four hours before the experiment, the hair on both sides of the spine of the rabbits' shoulders and backs was shaved off by clipping, with an area of ​​5×15 cm. 2 , for use when injecting.

[0166] 4.2 Test methods

[0167] 0.2 mL of extract prepared with sodium chloride injection was intradermally injected into 5 points on the left side of the spine of each rabbit, and 0.2 mL of sodium chloride control solution was intradermally injected into the last 5 points on the same side of the spine of each rabbit; 0.2 mL of extract prepared with cottonseed oil was intradermally injected into 5 points on the other side of the spine of each rabbit, and 0.2 mL of cottonseed oil control solution was intradermally injected into the last 5 points on the same side of the spine of each rabbit. The conditions of each injection site were observed and recorded immediately after injection and at 24h, 48h, and 72h. The tissue reactions of erythema and edema at each injection site during each observation period were scored according to the scoring system given in the table below, and the test results were recorded.

[0168] Table 10: Intradermal reaction scoring system

[0169]

[0170] 4.3 Results Evaluation

[0171] After scoring at (72±2)h, the total erythema and edema scores of the test article or blank control at (24±2)h, (48±2)h and (72±2)h for each animal are added together and then divided by 15[3 (scoring time point) × 5 (test article or blank control injection point)] to calculate the score of each animal test article or blank control. The total average score of each test article and the corresponding blank control is obtained by adding the scores of the three animals and dividing by 3. The final score of the test article is obtained by subtracting the blank control score from the test article score. If the final score of the test article is not greater than 1.0, it meets the test requirements. During any observation period, if the average reaction of the test article is suspected to be greater than the blank control reaction, 3 more rabbits should be retested. If the final score of the test article is not greater than 1.0, it meets the requirements.

[0172] 5. Results

[0173] The specific experimental results are shown in the table below.

[0174] Table 11: Intradermal reaction score table of sodium chloride control group

[0175]

[0176] Table 12: Intradermal reaction score table for the test sample sodium chloride extract group

[0177]

[0178] Table 13: Scoring table of intradermal reactions in cottonseed oil control group

[0179]

[0180] Table 14: Scoring table for intradermal reaction of cottonseed oil extract group

[0181]

[0182] According to the data in Tables 11 to 14, no irritation reactions such as erythema and edema occurred during the 72-hour observation period after intradermal injection of the test sample extract and solvent control. The average scores of the test sample extract group and the solvent control group were both 0, and no other abnormalities were observed at the injection site. Under the conditions of this test, the final score of the test sample was 0, and there was no intradermal irritation reaction.

[0183] Test Example 4: Material-mediated pyrogenicity

[0184] 1. Test system

[0185] Experimental animals: Japanese white rabbits, both male and female, weight: 3.2kg, 3.0kg, 3.3kg

[0186] Animal source: Junqi Breeding Farm, Daoli District, Harbin

[0187] Laboratory Animal Production License Number: SCXK (Black) 2021-004

[0188] Feed source: Junqi Breeding Farm, Daoli District, Harbin

[0189] Animal feed production license number: SCXK (black) 2021-004

[0190] 2. Animal Management

[0191] Experimental Animal Use License Number: SYXK (Black) 2019007

[0192] Water: Drinking water was provided freely available.

[0193] Housing conditions: Animals were kept individually in stainless steel cages and identified by cage cards, on which the test name, sample,

[0194] Number, animal number, person in charge of the experiment and the date of the experiment, etc.

[0195] Rearing environment: normal environment, temperature range 20-26℃, humidity range 40%-70%. Alternating light and dark during the day and night (12 / 12).

[0196] 3. Materials

[0197] 3.1 Reagents

[0198] Sodium chloride injection, Harbin Sanlian Pharmaceutical Co., Ltd., batch number: 221003F07

[0199] 3.2 Sample preparation

[0200] Test sample extract: Take 20 g of the test sample (the solution-type medical dressing prepared by Preparation Method 1 in Example 3), add 200 mL of sodium chloride injection at a ratio of 0.1 g / mL, and extract at 37°C with shaking for 72 h.

[0201] 4. Test methods

[0202] Take 3 rabbits that have passed the screening, and within 15 minutes after measuring their normal body temperature, slowly inject the test sample extract from the ear vein, and then measure their body temperature once every 30 minutes, for a total of 6 times. The highest temperature among the 6 times minus the normal body temperature is the temperature increase of the rabbit (℃). If the body temperature of 1 of the 3 rabbits increases by 0.6℃ or higher, or the total temperature increase of the 3 rabbits reaches 1.3℃ or higher, another 5 rabbits should be retested, and the inspection method is the same as above.

[0203] 5. Results Evaluation

[0204] In the 3 rabbits in the initial test, the temperature increase was lower than 0.6℃, and the total temperature increase of the 3 rabbits was lower than 1.3℃; or in the 5 rabbits in the retest, no more than 1 rabbit had a temperature increase of 0.6℃ or higher, and the total temperature increase of the 8 rabbits in the initial and retests was 3.5℃ or lower than 3.5℃, the pyrogen test of the test article was judged to be in compliance with the regulations.

[0205] If, among the three rabbits in the initial test, more than one rabbit had a body temperature increased by 0.6℃ or higher; or among the five rabbits in the retest, more than one rabbit had a body temperature increased by 0.6℃ or higher; or if the total body temperature increase of the eight rabbits in the initial and retests combined exceeded 3.5℃, the pyrogen test of the test article shall be judged as not meeting the requirements.

[0206] 6. Results

[0207] The test results are shown in the table below.

[0208] Table 15: Pyrogen test results

[0209]

[0210] After the test sample extract was injected into the ear vein, the body temperature of the three rabbits increased by less than 0.6°C, and the total body temperature increase of the three rabbits was less than 1.3°C. Under the conditions of this test, the test sample had no material-mediated pyrogenicity.

[0211] Test Example 5: Acute systemic toxicity test

[0212] 1. Test system

[0213] Experimental animals: Kunming mice, half male and half female, weight: 17g~23g

[0214] Animal source: Department of Experimental Animals, Harbin Medical University

[0215] Laboratory Animal Production License Number: SCXK (Black) 2019-001

[0216] Feed source: Beijing Keao Xieli Feed Co., Ltd.

[0217] Animal feed production license number: SCXK (Beijing) 2019-0003

[0218] 2. Animal Management

[0219] Experimental Animal Use License Number: SYXK (Black) 2019007

[0220] Water: Provide sterile water freely.

[0221] Housing conditions: Animals were housed in groups in plastic cages and identified by cage cards on which the experiment name, sample number, animal number, experiment leader, and experiment date were recorded.

[0222] Rearing environment: Barrier environment, temperature range 20-26℃, humidity range 40%-70%. Lighting alternating between day and night (12 / 12).

[0223] 3. Materials

[0224] 3.1 Reagents

[0225] Sodium chloride injection, Harbin Sanlian Pharmaceutical Co., Ltd., batch number: 221003F07

[0226] Cottonseed oil: Source Leaf Biotechnology, batch number: J09D10S105262

[0227] 3.2 Sample preparation

[0228] Sodium chloride extract: Take 1 g of the product (the solution-type medical dressing prepared by Preparation Method 1 in Example 3), add 10 mL of sodium chloride injection at a ratio of 0.1 g / mL, and extract at 37°C for 72 h.

[0229] Cottonseed oil extract: Take 1 g of the product (the solution-type medical dressing prepared by Preparation Method 1 in Example 3), add 10 mL of cottonseed oil at a ratio of 0.1 g / mL, and extract at 37°C for 72 h.

[0230] Preparation of control solution: Prepare in the same manner using the same batch of sodium chloride injection / cottonseed oil.

[0231] 4. Test methods

[0232] 4.1 Sodium Chloride Injection Extract Group:

[0233] Ten mice weighing 17g-23g were randomly divided into two groups, 5 mice in each group, and injected intravenously (iv) with the test sample sodium chloride extract and sodium chloride injection, respectively, at a dose of 50mL / kg and an injection time of 0.1mL / s.

[0234] 4.2 Cottonseed oil extract group:

[0235] Ten mice weighing 17g-23g were randomly divided into two groups, 5 mice in each group, and injected intraperitoneally (ip) with the test cottonseed oil extract and cottonseed oil at a dose of 50mL / kg.

[0236] 4.3 Results evaluation

[0237] After the injection, observe the immediate reaction of the mice, and observe and record the general condition, toxicity manifestations and number of dead animals in the test group and the control group at 4h, 24h, 48h and 72h. Refer to the table below to observe the degree of toxic reaction of the animals.

[0238] Table 16: Toxic reaction observation criteria

[0239]

[0240] 5. Test results

[0241] See the table below for specific results.

[0242] Table 17: Results of acute systemic toxicity test (sodium chloride extraction)

[0243]

[0244] Table 18: Results of acute systemic toxicity test (cottonseed oil extraction)

[0245]

[0246] Immediately after intravenous and intraperitoneal injection, mice in each group showed normal behavior and no abnormal reactions were observed. No toxic reactions were visible to the naked eye at 4, 24, 48, and 72 hours after injection, and the weight of animals in each group increased after the test. Under the conditions of this test, the test product had no acute systemic toxic reactions.

[0247] Test Example 6: Property Detection

[0248] The properties of the medical dressing prepared in Example 3 were tested, and the results are shown in the following table. The standard clauses were tested with reference to the product technical requirements.

[0249] Table 19: Test results of medical dressing properties of Example 3

[0250]

[0251] At the same time, the above method was used to test the corresponding indicators of the medical dressings obtained in Examples 1 and 2, and the results all met the corresponding standards.

[0252] Test Example 7:

[0253] 60 experimental rats were taken and scratched on their backs with an area of ​​about 1×2 cm to prepare a wound model. The rats were randomly divided into 6 groups, each with 10 rats. The wounds of the rats in each group were treated with the dressing-type medical dressings of Examples 1-3 and Comparative Examples 1-3, respectively, once a day for 8-10 hours each time, and the wound healing conditions of the rats were recorded 2, 4, 6, 8, and 10 days after treatment, the wound size of each rat was measured, the total wound area of ​​each group of rats was recorded, and the healing rate of the wounds of each group of rats after 10 days of treatment was calculated. The specific results are shown in the table below.

[0254] Wound healing rate % = (wound area before treatment - wound area after treatment) / wound area before treatment × 100%

[0255] Table 20: Wound healing rates of Examples 1-3 and Comparative Examples 1-3

[0256]

[0257] Note: Data were compared using Duncan's method. P < 0.05 indicated significant differences among the groups. Superscripts with different letters indicated significant differences.

[0258] According to the results in the table, under the premise of the total amount being unchanged, the mass ratio of carbomer homopolymer to glycerol, the mass ratio of disodium edetate to methylparaben, and the mass ratio of disodium edetate to triethanolamine will have a significant impact on the effect of the active components in the auxiliary materials. Among them, in terms of the healing rate of 10d, the effect of the Example 1-3 group is significantly better than that of the Comparative Example 1-3.

[0259] The above results show that the present application optimizes the auxiliary materials in the medical dressing with repair effect, so that the active components recombinant humanized collagen and / or sodium hyaluronate have better repair function within a certain proportion range of carbomer homopolymer, glycerol, disodium edetate, methylparaben and triethanolamine.

Claims

1. A medical dressing with repairing effect, comprising an active component and an auxiliary component, wherein the active component is recombinant humanized collagen and / or sodium hyaluronate, and the auxiliary component is glycerol, carbomer homopolymer, disodium edetate, methylparaben and triethanolamine, characterized in that Taking the total weight of the medical dressing as 100%, the mass percentages of the auxiliary material components are: glycerol 2-4%, carbomer homopolymer 0.12-0.18%, edetate disodium 0.08-0.15%, methylparaben 0.1-0.15%, triethanolamine 0.15-0.3%; wherein The mass ratio of the carbomer homopolymer to glycerol is 1:10-20, the mass ratio of the edetate disodium to methylparaben is 1-2:1, and the mass ratio of the edetate disodium to triethanolamine is 1:(1-7 / 3).

2. The medical dressing according to claim 1, characterized in that: Based on the total weight of the medical dressing as 100%, the recombinant humanized collagen is 0.01-0.2%, and / or the sodium hyaluronate is 0.01-0.3%.

3. The medical dressing according to any one of claims 1-2, characterized in that: The carbomer homopolymer is carbomer 980.

4. A method for preparing the medical dressing according to any one of claims 1 to 3, characterized in that: The preparation method comprises the following steps: (1) Add an appropriate amount of purified water to weigh glycerol, carbomer homopolymer, and edetate disodium, heat to 70-85° C., and stir to dissolve to obtain a mixture A; (2) adding methylparaben to purified water and stirring to obtain a methylparaben solution; (3) mixing the methylparaben solution with the mixture A, cooling the mixture to below 40° C. for standby use, and obtaining a mixture B; (4) adding recombinant humanized collagen and / or sodium hyaluronate to purified water, stirring to dissolve, and then adding mixture B to obtain mixture C; (5) At the same time, add triethanolamine to purified water, stir to dissolve, and obtain a triethanolamine solution. Add the triethanolamine solution to mixture C to obtain.

5. Use of the medical dressing according to any one of claims 1 to 3 or the medical dressing prepared by the preparation method according to claim 4 in preparing a product with a wound repairing effect.

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