Preparation method of oral bacteriostatic gel containing recombinant human collagen

Abstract

The application discloses a preparation method of an oral bacteriostatic gel containing recombinant human collagen. The method comprises the following steps: S1, taking materials; S2, taking deionized water A in S1, adding spilanthes acmella extract, panax notoginseng extract and radix scrophulariae extract into the deionized water and stirring until completely dissolved; S3, taking deionized water B in S1, adding cetylpyridinium chloride, tetrahydro methyl pyrimidine carboxylic acid, strontium chloride, sorbitol, glycerol and hydroxyethyl cellulose into the deionized water and completely dissolving; S4, pouring the solution in S2 into the solution in S3 in three batches and stirring; S5, adding hyaluronic acid and a modified additive into the solution in S4 and stirring; and S6, adding recombinant human collagen into the solution in S5 and stirring, and then performing radiation sterilization after defoaming to obtain a finished product. The product prepared by the method has the effects of inhibiting the growth of bacteria in the oral cavity and promoting oral cavity repair, and has a better bacteriostatic effect.

Description

Technical Field

[0001] This invention relates to the field of oral antibacterial gel technology, specifically to a method for preparing an oral antibacterial gel containing recombinant human collagen. Background Technology

[0002] The oral cavity is the beginning of the digestive tract. It connects to the outside through the oral fissure anteriorly and continues to the pharynx posteriorly via the pharyngeal isthmus. The oral cavity contains organs such as teeth and tongue. The anterior wall of the oral cavity is the lip, the lateral walls are the cheeks, the roof is the palate, and the floor consists of mucous membranes and muscles. The oral cavity is divided into the oral vestibule (anterolateral part) and the oral cavity proper (posteromedial part) by the upper and lower dental arches. When the upper and lower teeth are occlusal, the oral vestibule and the oral cavity proper communicate via the space behind the third molar. The oral cavity is one of the main routes for bacteria, viruses, fungi, and other foreign antigens to enter the human body. In addition to this exogenous burden, there are also a large number of endogenous microbial flora residing on the oral cavity, teeth, and mucosal surfaces. Microbial colony clusters can be divided into two main categories: the first category consists of free-form bacteria, mainly found in saliva; the second category consists of colony-form bacteria, mainly found in dental plaque or mucosal surfaces. Bacteria in the oral cavity include non-pathogenic and pathogenic bacteria, among which Staphylococcus aureus, Escherichia coli, and Candida albicans are three typical pathogenic bacteria. Existing oral antibacterial agents are not easy to adhere to the oral cavity wall in terms of morphology, have short contact time with the oral cavity, and have poor antibacterial effects. Moreover, the active ingredients of existing oral antibacterial agents are mostly antibiotics, which can lead to oral flora imbalance with long-term use. If they are traditional Chinese medicine types, the purity of the effective antibacterial substances is low, resulting in poor antibacterial effects. At the same time, due to lifestyle habits and traditional concepts, people do not pay much attention to oral hygiene care, leading to oral inflammation and ulceration. Therefore, a method for preparing an oral antibacterial gel containing recombinant human collagen is proposed. Summary of the Invention

[0003] The purpose of this invention is to provide a method for preparing an antibacterial oral gel containing recombinant human collagen, so as to solve the problems mentioned in the background art.

[0004] To achieve the above objectives, the present invention provides the following technical solution:

[0005] A method for preparing an oral antibacterial gel containing recombinant human collagen, wherein the method for preparing the oral antibacterial gel containing recombinant human collagen is as follows:

[0006] S1: Contains cetylpyridinium chloride, tetrahydromethylpyrimidine carboxylic acid, strontium chloride, sorbitol, purslane extract, Panax notoginseng extract, Scutellaria baicalensis extract, glycerin, hydroxyethyl cellulose, hyaluronic acid, recombinant human collagen, deionized water, and modifying additives.

[0007] S2: Take deionized water A from S1, add purslane extract, Panax notoginseng extract, and Scutellaria baicalensis extract to the deionized water and stir until completely dissolved;

[0008] S3: Take deionized water B from S1, and dissolve cetylpyridinium chloride, tetrahydromethylpyrimidine carboxylic acid, strontium chloride, sorbitol, glycerol, and hydroxyethyl cellulose completely in the deionized water;

[0009] S4: Pour the solution in S2 into the solution in S3 in three batches and stir.

[0010] S5: Add hyaluronic acid and modified additives to the S4 solution and stir.

[0011] S6: Add recombinant human collagen to the solution in S5 and stir. After degassing, sterilize by irradiation to obtain the finished product.

[0012] As a preferred option, the specific preparation steps are as follows:

[0013] S1: Contains cetylpyridinium chloride, tetrahydromethylpyrimidine carboxylic acid, strontium chloride, sorbitol, purslane extract, Panax notoginseng extract, Scutellaria baicalensis extract, glycerin, hydroxyethyl cellulose, hyaluronic acid, recombinant human collagen, deionized water, and modifying additives.

[0014] S2: Take deionized water A from S1, add purslane extract, Panax notoginseng extract, and Scutellaria baicalensis extract to the deionized water and stir until completely dissolved;

[0015] S3: Take deionized water B from S1, and dissolve cetylpyridinium chloride, tetrahydromethylpyrimidine carboxylic acid, strontium chloride, sorbitol, glycerol, and hydroxyethyl cellulose completely in the deionized water;

[0016] S4: Pour the solution in S2 into the solution in S3 in three batches and stir. First, add 1 / 2 at 300-400 rpm; second, add 1 / 4 at 100-200 rpm; third, add 1 / 4 at 100-200 rpm.

[0017] S5: Add hyaluronic acid and modified additives to the S4 solution and stir for 150-220 minutes at a speed of 600-900 rpm.

[0018] S6: Add recombinant human collagen to the solution in S5 and stir for 30-45 minutes at a speed of 400-600 rpm. After degassing, sterilize by irradiation at a dose of 14.1-20 KGy to obtain the finished product.

[0019] Preferably, the weight proportions of each component raw material in step S1 are as follows: 1-5 parts cetylpyridinium chloride, 2-3 parts tetrahydromethylpyrimidine carboxylic acid, 0.1-0.5 parts strontium chloride, 1-2 parts sorbitol, 1-3 parts purslane extract, 1-5 parts Panax notoginseng extract, 2-5 parts Scutellaria baicalensis extract, 5-10 parts glycerin, 1-2.5 parts hydroxyethyl cellulose, 3-5 parts hyaluronic acid, 0.5-2.5 parts recombinant human collagen, 50 parts deionized water A, 50 parts deionized water B, and 5-10 parts modifying additives.

[0020] Preferably, the preparation process of the modified additive in step S5 is as follows:

[0021] (1) Mix chitosan with deionized water at a ratio of 1:80-1:100 and stir for 1-2 hours at a temperature of 80-100℃ and a speed of 800-1000r / min to obtain a chitosan solution.

[0022] (2) Mix glacial acetic acid and chitosan solution in a homogenizer at a ratio of 1:80-1:100. Then add 0.2-0.3 times the mass of chitosan solution of eugenol and homogenize for 20-30 min at a speed of 18000-20000 r / min. Then add 0.2-0.3 times the mass of chitosan solution of sodium tripolyphosphate and stir for 3-6 min at a temperature of 30-50℃ and a speed of 1000-2000 r / min. After stirring, centrifuge, wash, and freeze dry to obtain pretreated powder.

[0023] (3) Mix the pretreated powder with deionized water at a mass ratio of 1:50-1:60, then add 0.01-0.02 times the mass of the pretreated powder dopamine solution, and then stir for 12-24 hours at a speed of 2000-3000 r / min. After stirring, centrifuge, wash and freeze dry to obtain the self-made additive.

[0024] Preferably, the chitosan in step (1) can also be homemade chitosan.

[0025] Preferably, the preparation steps of the self-made chitosan are as follows:

[0026] (1). Weigh 1g of CS into a 250ml beaker, add 100ml of deionized water and stir to disperse evenly;

[0027] (2). Weigh 800 mg HOBt and add it to the above solution, stirring until the solution is clear and transparent;

[0028] (3). Weigh 5.6g of NAC and add it to the above suspension. Add a certain concentration of EDC·HCl solution dropwise. Then use 1M NaOH solution to adjust the pH of the system to 5.0 and keep it unchanged. Stir continuously at room temperature in the dark for 3 hours.

[0029] (4) The reaction products were dialyzed using a dialysis bag (after treatment) with a molecular weight cutoff of 14K. The dialyzing sequence was as follows:

[0030] 1. Dialysis for 12 h under conditions of 5 mmol / L HCl and 1 μM EDTA;

[0031] 2. Dialysis for 12 hours under conditions of 5 mmol / L HCl and 1% NaCl;

[0032] 3. Dialysis for 12 hours under conditions of 5 mmol / L HCl and 1% NaCl;

[0033] 4. Dialysis for 12 hours under 1 mmol / L HCl conditions;

[0034] 5. Dialysis for 12 hours under 1 mmol / L HCl conditions;

[0035] (5) The solution after leaching was transferred to a petri dish with a diameter of 15 cm and frozen at -80°C. Then it was freeze-dried in a freeze dryer for 24 h. The final product was stored at 4°C in a dark and vacuum environment to avoid oxidation.

[0036] In summary, due to the adoption of the above-mentioned technologies, the beneficial effects of this invention are:

[0037] In this invention, firstly, eugenol is used as the core material, chitosan as the shell material, and sodium tripolyphosphate as the cross-linking agent to obtain a pretreated powder. The macromolecular chitosan can adsorb onto the surface of bacterial cell membranes, hindering the entry of nutrients. Simultaneously, the positive charge of chitosan interacts with the negative charge of bacteria, altering the permeability of the bacterial cell membrane. Secondly, eugenol acts on the lipids and proteins of the bacterial cell membrane, disrupting the cell membrane and causing its contents to leak out, thus inhibiting bacterial growth and achieving the effect of inhibiting and killing bacteria. Furthermore, the encapsulated eugenol can be slowly and continuously released, resulting in a better antibacterial effect. Finally, the addition of dopamine solution can neutralize the numerous hydroxyl groups present in the external structure of the pretreated powder, increasing the material's hydrophilicity. This study addresses the shortcomings of eugenol, such as its volatility and low water solubility, by improving its water solubility. The hydroxyl and amino groups of the self-made additive interact with the hydroxyl, fatty, and carboxyl groups of the oral cavity skin through hydrogen bonding, electrostatic attraction, and covalent Schiff base formation, exhibiting good adhesion. This allows it to adhere well to infected areas in the oral cavity, providing a sustained and highly effective antibacterial effect. Finally, due to the slightly acidic pH in the oral cavity, the acidic medium promotes better swelling and partial dissolution of the hydrophilic nanoparticles. This is caused by the ionic repulsion between the protonated free amino groups on a chitosan chain and adjacent chains, leading to greater diffusion of eugenol. The results indicate that acidic conditions are conducive to the sustained release of the antibacterial agent eugenol, further enhancing the product's antibacterial effect. Detailed Implementation

[0038] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention. Therefore, the following detailed description of the embodiments of the present invention is not intended to limit the scope of the claimed invention, but merely to represent selected embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0039] Example 1

[0040] S1: By weight, take 5 parts cetylpyridinium chloride, 2 parts tetrahydromethylpyrimidine carboxylic acid, 0.5 parts strontium chloride, 2 parts sorbitol, 3 parts purslane extract, 5 parts Panax notoginseng extract, 5 parts Scutellaria baicalensis extract, 5 parts glycerin, 2.5 parts hydroxyethyl cellulose, 5 parts hyaluronic acid, 0.5 parts recombinant human collagen, 50 parts deionized water A, 50 parts deionized water B, and 10 parts modifying additives;

[0041] S2: Take deionized water A from S1, add purslane extract, Panax notoginseng extract, and Scutellaria baicalensis extract to the deionized water and stir until completely dissolved;

[0042] S3: Take deionized water B from S1, and dissolve cetylpyridinium chloride, tetrahydromethylpyrimidine carboxylic acid, strontium chloride, sorbitol, glycerol, and hydroxyethyl cellulose completely in the deionized water;

[0043] S4: Pour the solution in S2 into the solution in S3 in three batches and stir. First, add 1 / 2 at 400 rpm; second, add 1 / 4 at 200 rpm; third, add 1 / 4 at 200 rpm.

[0044] S5: Add hyaluronic acid and modified additives to the S4 solution and stir for 220 minutes at 900 rpm.

[0045] S6: Add recombinant human collagen to the solution in S5 and stir for 45 minutes at 600 rpm. After degassing, sterilize by irradiation at a rate of 20 KGy to obtain the finished product.

[0046] The preparation process of the modified additive mentioned in step S5 is as follows:

[0047] (1) Mix the homemade chitosan with deionized water at a ratio of 1:80, and stir for 2 hours at a temperature of 100℃ and a speed of 1000r / min to obtain the homemade chitosan solution.

[0048] (2) Glacial acetic acid and homemade chitosan solution were mixed in a homogenizer at a ratio of 1:100. Then, eugenol was added at a ratio of 0.3 times the mass of the chitosan solution and homogenized at a speed of 18000 r / min for 30 min. Then, sodium tripolyphosphate was added at a ratio of 0.3 times the mass of the chitosan solution and stirred at a speed of 1000 r / min for 6 min. After centrifugation, washing, and freeze drying, pretreated powder was obtained.

[0049] (3) The pretreated powder and deionized water were mixed at a mass ratio of 1:60. Then, 0.02 times the mass of the pretreated powder was added to the dopamine solution. The mixture was stirred for 24 hours at a speed of 3000 r / min. After stirring, the mixture was centrifuged, washed, and freeze-dried to obtain the self-made additive.

[0050] The preparation steps of the self-made chitosan are as follows:

[0051] (1). Weigh 1g of CS into a 250ml beaker, add 100ml of deionized water and stir to disperse evenly;

[0052] (2). Weigh 800 mg HOBt and add it to the above solution, stirring until the solution is clear and transparent;

[0053] (3). Weigh 5.6g of NAC and add it to the above suspension. Add a certain concentration of EDC·HCl solution dropwise. Then use 1M NaOH solution to adjust the pH of the system to 5.0 and keep it unchanged. Stir continuously at room temperature in the dark for 3 hours.

[0054] (4) The reaction products were dialyzed using a dialysis bag (after treatment) with a molecular weight cutoff of 14K. The dialyzing sequence was as follows:

[0055] 1. Dialysis for 12 h under conditions of 5 mmol / L HCl and 1 μM EDTA;

[0056] 2. Dialysis for 12 hours under conditions of 5 mmol / L HCl and 1% NaCl;

[0057] 3. Dialysis for 12 hours under conditions of 5 mmol / L HCl and 1% NaCl;

[0058] 4. Dialysis for 12 hours under 1 mmol / L HCl conditions;

[0059] 5. Dialysis for 12 hours under 1 mmol / L HCl conditions;

[0060] (5) The solution after leaching was transferred to a petri dish with a diameter of 15 cm and frozen at -80°C. Then it was freeze-dried in a freeze dryer for 24 h. The final product was stored at 4°C in a dark and vacuum environment to avoid oxidation.

[0061] Example 2

[0062] S1: By weight, take 5 parts cetylpyridinium chloride, 2 parts tetrahydromethylpyrimidine carboxylic acid, 0.5 parts strontium chloride, 2 parts sorbitol, 5 parts glycerol, 2.5 parts hydroxyethyl cellulose, 5 parts hyaluronic acid, 0.5 parts recombinant human collagen, 50 parts deionized water B, and 10 parts modifying additives.

[0063] S2: Take deionized water B from S1, and dissolve cetylpyridinium chloride, tetrahydromethylpyrimidine carboxylic acid, strontium chloride, sorbitol, glycerol, and hydroxyethyl cellulose completely in the deionized water;

[0064] S3: Add hyaluronic acid and modified additives to the S2 solution and stir for 220 minutes at 900 rpm.

[0065] S4: Add recombinant human collagen to the solution in S3 and stir for 45 minutes at 600 rpm. After degassing, sterilize by irradiation at a rate of 20 KGy to obtain the finished product.

[0066] The preparation process of the modified additive mentioned in step S3 is as follows:

[0067] (1) Mix the homemade chitosan with deionized water at a ratio of 1:80, and stir for 2 hours at a temperature of 100℃ and a speed of 1000r / min to obtain the homemade chitosan solution.

[0068] (2) Glacial acetic acid and homemade chitosan solution were mixed in a homogenizer at a ratio of 1:100. Then, eugenol was added at a ratio of 0.3 times the mass of the chitosan solution and homogenized at a speed of 18000 r / min for 30 min. Then, sodium tripolyphosphate was added at a ratio of 0.3 times the mass of the chitosan solution and stirred at a speed of 1000 r / min for 6 min. After centrifugation, washing, and freeze drying, pretreated powder was obtained.

[0069] (3) The pretreated powder and deionized water were mixed at a mass ratio of 1:60. Then, 0.02 times the mass of the pretreated powder was added to the dopamine solution. The mixture was stirred for 24 hours at a speed of 3000 r / min. After stirring, the mixture was centrifuged, washed, and freeze-dried to obtain the self-made additive.

[0070] The preparation steps of the self-made chitosan are as follows:

[0071] (1). Weigh 1g of CS into a 250ml beaker, add 100ml of deionized water and stir to disperse evenly;

[0072] (2). Weigh 800 mg HOBt and add it to the above solution, stirring until the solution is clear and transparent;

[0073] (3). Weigh 5.6g of NAC and add it to the above suspension. Add a certain concentration of EDC·HCl solution dropwise. Then use 1M NaOH solution to adjust the pH of the system to 5.0 and keep it unchanged. Stir continuously at room temperature in the dark for 3 hours.

[0074] (4) The reaction products were dialyzed using a dialysis bag (after treatment) with a molecular weight cutoff of 14K. The dialyzing sequence was as follows:

[0075] 1. Dialysis for 12 h under conditions of 5 mmol / L HCl and 1 μM EDTA;

[0076] 2. Dialysis for 12 hours under conditions of 5 mmol / L HCl and 1% NaCl;

[0077] 3. Dialysis for 12 hours under conditions of 5 mmol / L HCl and 1% NaCl;

[0078] 4. Dialysis for 12 hours under 1 mmol / L HCl conditions;

[0079] 5. Dialysis for 12 hours under 1 mmol / L HCl conditions;

[0080] (5) The solution after leaching was transferred to a petri dish with a diameter of 15 cm and frozen at -80°C. Then it was freeze-dried in a freeze dryer for 24 h. The final product was stored at 4°C in a dark and vacuum environment to avoid oxidation.

[0081] Example 3

[0082] S1: By weight, take 3 parts of purslane extract, 5 parts of Panax notoginseng extract, 5 parts of Scutellaria baicalensis extract, 5 parts of hyaluronic acid, 0.5 parts of recombinant human collagen, 50 parts of deionized water A, and 10 parts of modified additives.

[0083] S2: Take deionized water A from S1, add purslane extract, Panax notoginseng extract, and Scutellaria baicalensis extract to the deionized water and stir until completely dissolved;

[0084] S3: Add hyaluronic acid and modified additives to the S2 solution and stir for 220 minutes at 900 rpm.

[0085] S4: Add recombinant human collagen to the solution in S3 and stir for 45 minutes at 600 rpm. After degassing, sterilize by irradiation at a rate of 20 KGy to obtain the finished product.

[0086] The preparation process of the modified additive mentioned in step S3 is as follows:

[0087] (1) Mix the homemade chitosan with deionized water at a ratio of 1:80, and stir for 2 hours at a temperature of 100℃ and a speed of 1000r / min to obtain the homemade chitosan solution.

[0088] (2) Glacial acetic acid and homemade chitosan solution were mixed in a homogenizer at a ratio of 1:100. Then, eugenol was added at a ratio of 0.3 times the mass of the chitosan solution and homogenized at a speed of 18000 r / min for 30 min. Then, sodium tripolyphosphate was added at a ratio of 0.3 times the mass of the chitosan solution and stirred at a speed of 1000 r / min for 6 min. After centrifugation, washing, and freeze drying, pretreated powder was obtained.

[0089] (3) The pretreated powder and deionized water were mixed at a mass ratio of 1:60. Then, 0.02 times the mass of the pretreated powder was added to the dopamine solution. The mixture was stirred for 24 hours at a speed of 3000 r / min. After stirring, the mixture was centrifuged, washed, and freeze-dried to obtain the self-made additive.

[0090] The preparation steps of the self-made chitosan are as follows:

[0091] (1). Weigh 1g of CS into a 250ml beaker, add 100ml of deionized water and stir to disperse evenly;

[0092] (2). Weigh 800 mg HOBt and add it to the above solution, stirring until the solution is clear and transparent;

[0093] (3). Weigh 5.6g of NAC and add it to the above suspension. Add a certain concentration of EDC·HCl solution dropwise. Then use 1M NaOH solution to adjust the pH of the system to 5.0 and keep it unchanged. Stir continuously at room temperature in the dark for 3 hours.

[0094] (4) The reaction products were dialyzed using a dialysis bag (after treatment) with a molecular weight cutoff of 14K. The dialyzing sequence was as follows:

[0095] 1. Dialysis for 12 h under conditions of 5 mmol / L HCl and 1 μM EDTA;

[0096] 2. Dialysis for 12 hours under conditions of 5 mmol / L HCl and 1% NaCl;

[0097] 3. Dialysis for 12 hours under conditions of 5 mmol / L HCl and 1% NaCl;

[0098] 4. Dialysis for 12 hours under 1 mmol / L HCl conditions;

[0099] 5. Dialysis for 12 hours under 1 mmol / L HCl conditions;

[0100] (5) The solution after leaching was transferred to a petri dish with a diameter of 15 cm and frozen at -80°C. Then it was freeze-dried in a freeze dryer for 24 h. The final product was stored at 4°C in a dark and vacuum environment to avoid oxidation.

[0101] Example 4

[0102] S1: By weight, take 5 parts cetylpyridinium chloride, 2 parts tetrahydromethylpyrimidine carboxylic acid, 0.5 parts strontium chloride, 2 parts sorbitol, 3 parts purslane extract, 5 parts Panax notoginseng extract, 5 parts Scutellaria baicalensis extract, 5 parts glycerin, 2.5 parts hydroxyethyl cellulose, 0.5 parts recombinant human collagen, 50 parts deionized water A, 50 parts deionized water B, and 10 parts modifying additives;

[0103] S2: Take deionized water A from S1, add purslane extract, Panax notoginseng extract, and Scutellaria baicalensis extract to the deionized water and stir until completely dissolved;

[0104] S3: Take deionized water B from S1, and dissolve cetylpyridinium chloride, tetrahydromethylpyrimidine carboxylic acid, strontium chloride, sorbitol, glycerol, and hydroxyethyl cellulose completely in the deionized water;

[0105] S4: Pour the solution in S2 into the solution in S3 in three batches and stir. First, add 1 / 2 at 400 rpm; second, add 1 / 4 at 200 rpm; third, add 1 / 4 at 200 rpm.

[0106] S5: Add the modified additive to the S4 solution and stir for 220 min at 900 rpm.

[0107] S6: Add recombinant human collagen to the solution in S5 and stir for 45 minutes at 600 rpm. After degassing, sterilize by irradiation at a rate of 20 KGy to obtain the finished product.

[0108] The preparation process of the modified additive mentioned in step S5 is as follows:

[0109] (1) Mix the homemade chitosan with deionized water at a ratio of 1:80, and stir for 2 hours at a temperature of 100℃ and a speed of 1000r / min to obtain the homemade chitosan solution.

[0110] (2) Glacial acetic acid and homemade chitosan solution were mixed in a homogenizer at a ratio of 1:100. Then, eugenol was added at a ratio of 0.3 times the mass of the chitosan solution and homogenized at a speed of 18000 r / min for 30 min. Then, sodium tripolyphosphate was added at a ratio of 0.3 times the mass of the chitosan solution and stirred at a speed of 1000 r / min for 6 min. After centrifugation, washing, and freeze drying, pretreated powder was obtained.

[0111] (3) The pretreated powder and deionized water were mixed at a mass ratio of 1:60. Then, 0.02 times the mass of the pretreated powder was added to the dopamine solution. The mixture was stirred for 24 hours at a speed of 3000 r / min. After stirring, the mixture was centrifuged, washed, and freeze-dried to obtain the self-made additive.

[0112] The preparation steps of the self-made chitosan are as follows:

[0113] (1). Weigh 1g of CS into a 250ml beaker, add 100ml of deionized water and stir to disperse evenly;

[0114] (2). Weigh 800 mg HOBt and add it to the above solution, stirring until the solution is clear and transparent;

[0115] (3). Weigh 5.6g of NAC and add it to the above suspension. Add a certain concentration of EDC·HCl solution dropwise. Then use 1M NaOH solution to adjust the pH of the system to 5.0 and keep it unchanged. Stir continuously at room temperature in the dark for 3 hours.

[0116] (4) The reaction products were dialyzed using a dialysis bag (after treatment) with a molecular weight cutoff of 14K. The dialyzing sequence was as follows:

[0117] 1. Dialysis for 12 h under conditions of 5 mmol / L HCl and 1 μM EDTA;

[0118] 2. Dialysis for 12 hours under conditions of 5 mmol / L HCl and 1% NaCl;

[0119] 3. Dialysis for 12 hours under conditions of 5 mmol / L HCl and 1% NaCl;

[0120] 4. Dialysis for 12 hours under 1 mmol / L HCl conditions;

[0121] 5. Dialysis for 12 hours under 1 mmol / L HCl conditions;

[0122] (5) The solution after leaching was transferred to a petri dish with a diameter of 15 cm and frozen at -80°C. Then it was freeze-dried in a freeze dryer for 24 h. The final product was stored at 4°C in a dark and vacuum environment to avoid oxidation.

[0123] Example 5

[0124] S1: By weight, take 5 parts cetylpyridinium chloride, 2 parts tetrahydromethylpyrimidine carboxylic acid, 0.5 parts strontium chloride, 2 parts sorbitol, 3 parts purslane extract, 5 parts Panax notoginseng extract, 5 parts Scutellaria baicalensis extract, 5 parts glycerin, 2.5 parts hydroxyethyl cellulose, 5 parts hyaluronic acid, 50 parts deionized water A, 50 parts deionized water B, and 10 parts modifying additives;

[0125] S2: Take deionized water A from S1, add purslane extract, Panax notoginseng extract, and Scutellaria baicalensis extract to the deionized water and stir until completely dissolved;

[0126] S3: Take deionized water B from S1, and dissolve cetylpyridinium chloride, tetrahydromethylpyrimidine carboxylic acid, strontium chloride, sorbitol, glycerol, and hydroxyethyl cellulose completely in the deionized water;

[0127] S4: Pour the solution in S2 into the solution in S3 in three batches and stir. First, add 1 / 2 at 400 rpm; second, add 1 / 4 at 200 rpm; third, add 1 / 4 at 200 rpm.

[0128] S5: Add hyaluronic acid and modified additives to the S4 solution and stir for 220 minutes at 900 rpm.

[0129] S6: Stir the solution in S5 for 45 minutes at 600 rpm. After degassing, sterilize by irradiation at a rate of 20 KGy to obtain the finished product.

[0130] The preparation process of the modified additive mentioned in step S5 is as follows:

[0131] (1) Mix the homemade chitosan with deionized water at a ratio of 1:80, and stir for 2 hours at a temperature of 100℃ and a speed of 1000r / min to obtain the homemade chitosan solution.

[0132] (2) Glacial acetic acid and self-made chitosan solution were mixed in a homogenizer at a ratio of 1:100. Then, eugenol with a mass of 0.3 times that of the chitosan solution was added and homogenized for 30 min at a speed of 18000 r / min. Then, sodium tripolyphosphate with a mass of 0.3 times that of the chitosan solution was added and stirred for 6 min at a temperature of 30℃ and a speed of 1000 r / min. After centrifugation, washing, and freeze drying, pretreated powder was obtained.

[0133] (3) The pretreated powder and deionized water were mixed at a mass ratio of 1:60. Then, 0.02 times the mass of the pretreated powder was added to a dopamine solution. The mixture was stirred for 24 hours at a speed of 3000 r / min. After stirring, the mixture was centrifuged, washed, and freeze-dried to obtain the self-made additive.

[0134] The preparation steps of the self-made chitosan are as follows:

[0135] (1). Weigh 1g of CS into a 250ml beaker, add 100ml of deionized water and stir to disperse evenly;

[0136] (2). Weigh 800 mg HOBt and add it to the above solution, stirring until the solution is clear and transparent;

[0137] (3). Weigh 5.6g of NAC and add it to the above suspension. Add a certain concentration of EDC·HCl solution dropwise. Then use 1M NaOH solution to adjust the pH of the system to 5.0 and keep it unchanged. Stir continuously at room temperature in the dark for 3 hours.

[0138] (4) The reaction products were dialyzed using a dialysis bag (after treatment) with a molecular weight cutoff of 14K. The dialyzing sequence was as follows:

[0139] 1. Dialysis for 12 h under conditions of 5 mmol / L HCl and 1 μM EDTA;

[0140] 2. Dialysis for 12 hours under conditions of 5 mmol / L HCl and 1% NaCl;

[0141] 3. Dialysis for 12 hours under conditions of 5 mmol / L HCl and 1% NaCl;

[0142] 4. Dialysis for 12 hours under 1 mmol / L HCl conditions;

[0143] 5. Dialysis for 12 hours under 1 mmol / L HCl conditions;

[0144] (5) The solution after leaching was transferred to a petri dish with a diameter of 15 cm and frozen at -80°C. Then it was freeze-dried in a freeze dryer for 24 h. The final product was stored at 4°C in a dark and vacuum environment to avoid oxidation.

[0145] Example 6

[0146] S1: By weight, take 5 parts cetylpyridinium chloride, 2 parts tetrahydromethylpyrimidine carboxylic acid, 0.5 parts strontium chloride, 2 parts sorbitol, 3 parts purslane extract, 5 parts Panax notoginseng extract, 5 parts Scutellaria baicalensis extract, 5 parts glycerin, 2.5 parts hydroxyethyl cellulose, 5 parts hyaluronic acid, 0.5 parts recombinant human collagen, 50 parts deionized water A, and 50 parts deionized water B;

[0147] S2: Take deionized water A from S1, add purslane extract, Panax notoginseng extract, and Scutellaria baicalensis extract to the deionized water and stir until completely dissolved;

[0148] S3: Take deionized water B from S1, and dissolve cetylpyridinium chloride, tetrahydromethylpyrimidine carboxylic acid, strontium chloride, sorbitol, glycerol, and hydroxyethyl cellulose completely in the deionized water;

[0149] S4: Pour the solution in S2 into the solution in S3 in three batches and stir. First, add 1 / 2 at 400 rpm; second, add 1 / 4 at 200 rpm; third, add 1 / 4 at 200 rpm.

[0150] S5: Add hyaluronic acid to the S4 solution and stir for 220 minutes at 900 rpm.

[0151] S6: Add recombinant human collagen to the solution in S5 and stir for 45 minutes at 600 rpm. After degassing, sterilize by irradiation at a rate of 20 KGy to obtain the finished product.

[0152] Comparative Example

[0153] S1: By weight, take 5 parts cetylpyridinium chloride, 2 parts tetrahydromethylpyrimidine carboxylic acid, 0.5 parts strontium chloride, 2 parts sorbitol, 3 parts purslane extract, 5 parts Panax notoginseng extract, 5 parts Scutellaria baicalensis extract, 5 parts glycerin, 2.5 parts hydroxyethyl cellulose, 50 parts deionized water A, and 50 parts deionized water B.

[0154] S2: Take deionized water A from S1, add purslane extract, Panax notoginseng extract, and Scutellaria baicalensis extract to the deionized water and stir until completely dissolved;

[0155] S3: Take deionized water B from S1, and dissolve cetylpyridinium chloride, tetrahydromethylpyrimidine carboxylic acid, strontium chloride, sorbitol, glycerol, and hydroxyethyl cellulose completely in the deionized water;

[0156] S4: Pour the solution in S2 into the solution in S3 in three batches and stir. Add 1 / 2 first, stirring at 400 rpm. Add 1 / 4 second, stirring at 200 rpm. Add 1 / 4 third, stirring at 200 rpm. After degassing, sterilize by irradiation at a dose of 20 KGy to obtain the finished product.

[0157] Detection method:

[0158] Common oral pathogens: Staphylococcus aureus, Streptococcus mutans, Porphyromonas gingivalis

[0159] Table 1

[0160]

[0161] Gingival Index (GI) Scoring Standards:

[0162] 0 = Healthy gums

[0163] 1 = Mild gingivitis: The gums show slight discoloration and mild edema, and do not bleed on probing.

[0164] 2 = Moderate gingivitis: Red, swollen, shiny gums, bleeding on probing.

[0165] 3 = Severe gingivitis: The gums are noticeably red and swollen or have ulcers, and there is a tendency for them to bleed spontaneously.

[0166] Take the average value of each group of experiments:

[0167] Table 2

[0168]

[0169]

[0170] Plaque Index (PLI) scoring criteria:

[0171] 0 = Plaque-free area at the gingival margin

[0172] 1. Thin plaque is present on the tooth surface at the gingival margin, but is not visible upon visual examination. Plaque can be seen by scraping the tooth surface with a probe tip. 2. Moderate amount of plaque is visible at the gingival margin or proximal surfaces.

[0173] 3. Extensive plaque buildup in the gingival sulcus, gingival margin, and adjacent surfaces.

[0174] Table 3

[0175]

[0176] As can be seen from Tables 1, 2 and 3, the effects are ranked (from best to worst): Example 1, Example 4, Example 5, Comparative Example, Example 2, Example 6, Example 3. The product prepared by this invention can inhibit the proliferation of bacteria in the oral cavity and promote oral repair; the antibacterial effect is better.

[0177] The above description is only a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any equivalent substitutions or modifications made by those skilled in the art within the scope of the technology disclosed in the present invention, based on the technical solution and inventive concept of the present invention, should be covered within the scope of protection of the present invention.

[0178] It should be noted that, in this document, relational terms such as "first" and "second" are used only to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such process, method, article, or apparatus.

Claims

1. A method for preparing an oral antibacterial gel containing recombinant human collagen, characterized in that, The preparation method of the oral antibacterial gel containing recombinant human collagen: S1: Contains cetylpyridinium chloride, tetrahydromethylpyrimidine carboxylic acid, strontium chloride, sorbitol, purslane extract, Panax notoginseng extract, Scutellaria baicalensis extract, glycerin, hydroxyethyl cellulose, hyaluronic acid, recombinant human collagen, deionized water, and modifying additives. S2: Take deionized water A from S1, add purslane extract, Panax notoginseng extract, and Scutellaria baicalensis extract to the deionized water and stir until completely dissolved; S3: Take deionized water B from S1, and dissolve cetylpyridinium chloride, tetrahydromethylpyrimidine carboxylic acid, strontium chloride, sorbitol, glycerol, and hydroxyethyl cellulose completely in the deionized water; S4: Pour the solution in S2 into the solution in S3 in three batches and stir. S5: Add hyaluronic acid and modified additives to the S4 solution and stir. S6: Add recombinant human collagen to the solution in S5 and stir. After degassing, sterilize by irradiation to obtain the finished product. The preparation process of the modified additive mentioned in step S5 is as follows: (1) Mix chitosan with deionized water at a ratio of 1:80 to 1:100 and stir for 1-2 hours at a temperature of 80-100℃ and a speed of 800-1000r / min to obtain a chitosan solution. (2) Mix glacial acetic acid and chitosan solution in a homogenizer at a ratio of 1:80-1:

100. Then add 0.2-0.3 times the mass of chitosan solution of eugenol and homogenize for 20-30 min at a speed of 18000-20000 r / min. Then add 0.2-0.3 times the mass of chitosan solution of sodium tripolyphosphate and stir for 3-6 min at a temperature of 30-50℃ and a speed of 1000-2000 r / min. After stirring, centrifuge, wash, and freeze dry to obtain pretreated powder. (3) Mix the pretreated powder with deionized water at a mass ratio of 1:50-1:60, then add 0.01-0.02 times the mass of the pretreated powder dopamine solution, and then stir for 12-24 hours at a speed of 2000-3000 r / min. After stirring, centrifuge, wash and freeze dry to obtain the self-made additive. In step (1), the chitosan is homemade chitosan; The preparation steps of the self-made chitosan are as follows: 1) Weigh 1g of CS into a 250ml beaker, add 100ml of deionized water and stir to disperse evenly; 2) Weigh 800 mg HOBt and add it to the above solution, stirring until the solution is clear and transparent; 3) Weigh 5.6g of NAC and add it to the above solution. Add a certain concentration of EDC·HCl solution dropwise. Then, use 1M NaOH solution to adjust the pH of the system to 5.0 and keep it unchanged. Stir continuously at room temperature in the dark for 3 hours. 4) Dialyze the reaction products using a dialysis bag with a molecular weight cutoff of 14K. The dialysis sequence is as follows: A. Dialysis for 12 h under conditions of 5 mmol / L HCl and 1 μM EDTA; B. Dialysis for 12 hours under conditions of 5 mmol / L HCl and 1% NaCl; C. Dialysis for 12 h under conditions of 5 mmol / L HCl and 1% NaCl; D. Dialysis for 12 hours under 1 mmol / L HCl conditions; Dialysis for 12 hours under E, 1 mmol / L HCl conditions; 5) Transfer the dialysis solution to a 15cm diameter petri dish and freeze at -80℃. Then freeze-dry in a freeze dryer for 24 hours. Store the final product at 4℃ in a dark, vacuum environment to prevent oxidation.

2. The method for preparing an oral antibacterial gel containing recombinant human collagen according to claim 1, characterized in that: The specific preparation steps are as follows: S1: Contains cetylpyridinium chloride, tetrahydromethylpyrimidine carboxylic acid, strontium chloride, sorbitol, purslane extract, Panax notoginseng extract, Scutellaria baicalensis extract, glycerin, hydroxyethyl cellulose, hyaluronic acid, recombinant human collagen, deionized water, and modifying additives. S2: Take deionized water A from S1, add purslane extract, Panax notoginseng extract, and Scutellaria baicalensis extract to the deionized water and stir until completely dissolved; S3: Take deionized water B from S1, and dissolve cetylpyridinium chloride, tetrahydromethylpyrimidine carboxylic acid, strontium chloride, sorbitol, glycerol, and hydroxyethyl cellulose completely in the deionized water; S4: Pour the solution in S2 into the solution in S3 in three batches and stir. First, add 1 / 2 at 300-400 rpm; second, add 1 / 4 at 100-200 rpm; third, add 1 / 4 at 100-200 rpm. S5: Add hyaluronic acid and modified additives to the S4 solution and stir for 150-220 minutes at a speed of 600-900 rpm. S6: Add recombinant human collagen to the solution in S5 and stir for 30-45 minutes at a speed of 400-600 rpm. After degassing, sterilize by irradiation at a dose of 14.1-20 KGy to obtain the finished product.

3. The method for preparing an oral antibacterial gel containing recombinant human collagen according to claim 2, characterized in that, The weight proportions of each component raw material in step S1 are as follows: 1-5 parts cetylpyridinium chloride, 2-3 parts tetrahydromethylpyrimidine carboxylic acid, 0.1-0.5 parts strontium chloride, 1-2 parts sorbitol, 1-3 parts purslane extract, 1-5 parts Panax notoginseng extract, 2-5 parts Scutellaria baicalensis extract, 5-10 parts glycerin, 1-2.5 parts hydroxyethyl cellulose, 3-5 parts hyaluronic acid, 0.5-2.5 parts recombinant human collagen, 50 parts deionized water A, 50 parts deionized water B, and 5-10 parts modifying additives.

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