Antibacterial composite periosteum for oral cavity and processing technology of antibacterial composite periosteum

Abstract

The invention discloses an antibacterial composite periosteum for an oral cavity and a processing technology of the antibacterial composite periosteum, and relates to the technical field of periosteum. The method specifically comprises the following steps: step 1, treating defatted pig peritoneum with a decellularized solution, cleaning and sterilizing to obtain a pig peritoneum decellularized matrix; 2, treating the pig peritoneal decellularized matrix with a glyoxal solution, cleaning, and sterilizing to obtain a pretreated pig peritoneal decellularized matrix; step 3, uniformly coating the surface of the pretreated porcine peritoneal acellular matrix with an antibacterial solution, and performing sterilization, standing and freeze-drying to obtain the antibacterial composite periosteum. Wherein the antibacterial solution is prepared from the following raw material components in concentration: 1 to 2 weight percent of modified chitosan, 0.2 to 0.4 weight percent of amino-terminated polyethylene glycol and 0.01 to 0.05 weight percent of p-toluenesulfonic acid, and a solvent is a PBS (Phosphate Buffer Solution). The antibacterial composite periosteum is good in biocompatibility and has excellent clinical application value in oral surgery.

Description

Technical Field

[0001] This invention relates to the field of periosteum technology, specifically to an antibacterial composite periosteum for oral cavity and its processing technology. Background Technology

[0002] The advantage of oral periosteum in dental implantology lies in its ability to be placed between the oral soft tissue and bone defects, thereby establishing a biological barrier. This helps maintain the position and morphology of bone graft material, preventing its loss, and inhibits soft tissue growth into the graft area while promoting the integration of bone graft material with surrounding bone tissue, thus improving implant stability. Therefore, in cases of insufficient alveolar bone volume or abnormal morphology, periosteum implantation is a necessary step to ensure the long-term stability and functional restoration of dental implants.

[0003] Existing periosteal materials have significant shortcomings: although simple natural decellularized matrix has excellent biocompatibility, it lacks targeted antibacterial ability and is difficult to resist postoperative infection caused by complex oral flora; synthetic polymer materials (such as unmodified chitosan) have acceptable antibacterial properties, but they have problems such as insufficient biocompatibility, high mechanical brittleness, and easy to cause local tissue irritation, which restricts the clinical application effect.

[0004] Therefore, developing a composite periosteum that combines mechanical stability adapted to the oral environment with good antibacterial and biocompatibility properties is of great value in clinical applications. Summary of the Invention

[0005] The purpose of this invention is to provide an antibacterial composite periosteum for oral cavity and its processing technology, so as to solve the problems mentioned in the background art.

[0006] To solve the above-mentioned technical problems, the present invention provides the following technical solution: A processing method for an antibacterial composite periosteum for oral cavity includes the following steps: Step 1: After being treated with decellularized solution, defatted porcine peritoneum is washed and sterilized to obtain decellularized porcine peritoneal matrix; Step 2: After treatment with glyoxal solution, the decellularized porcine peritoneum matrix is ​​washed and sterilized to obtain pretreated decellularized porcine peritoneum matrix; Step 3: The antibacterial solution is evenly coated on the surface of the pretreated porcine peritoneum decellularized matrix, sterilized, left to stand, and freeze-dried to obtain an antibacterial composite peritoneum.

[0007] Further, the method for preparing the defatted porcine peritoneum is as follows: (1) Porcine peritoneum with most of the fat removed and no obvious blood stains or other attachments is immersed in a 0.01M~0.1M sodium hydroxide aqueous solution and treated at 28~32℃ with a shaking speed of 130~140rpm for 3~5h, repeated 2~3 times, and then taken out; (2) Porcine peritoneum after defatting in (1) is immersed in pure water and treated at 28~32℃ with a shaking speed of 130~140rpm for 15~20min, repeated 4~6 times, and then taken out; then immersed in isopropanol and treated at 38~42℃ with a shaking speed of 130~140rpm for 7~9h, repeated 2~3 times, and then taken out to obtain defatted porcine peritoneum.

[0008] Further, the method for preparing the porcine peritoneal decellularized matrix is ​​as follows: immerse defatted porcine peritoneum in a decellularization solution and treat it at 28-32°C with a shaking speed of 130-140 rpm for 1-2 hours, then remove it; then immerse it in PBS buffer and treat it at 28-32°C with a shaking speed of 130-140 rpm for 20-40 minutes, then remove it and sterilize it to obtain the porcine peritoneal decellularized matrix.

[0009] Furthermore, the decellularization solution comprises the following raw material components at the following concentrations: Triton X-100 0.1~0.2 v / v%, trypsin 0.1~0.5 wt%, and PBS buffer as the solvent.

[0010] Further, the preparation method of the pretreated porcine peritoneal decellularized matrix is ​​as follows: the porcine peritoneal decellularized matrix is ​​immersed in glyoxal solution and treated at 28-32℃ with a shaking speed of 130-140 rpm for 20-60 min, then removed; it is then immersed in PBS buffer and treated at 28-32℃ with a shaking speed of 130-140 rpm for 10-20 min, then removed and sterilized to obtain the pretreated porcine peritoneal decellularized matrix.

[0011] Furthermore, the glyoxal solution is a PBS buffer solution of glyoxal with a concentration of 0.05~0.15wt%.

[0012] Furthermore, the antibacterial solution comprises the following raw material components at the following concentrations: 1-2 wt% modified chitosan, 0.2-0.4 wt% amino-terminated polyethylene glycol, 0.01-0.05 wt% p-toluenesulfonic acid, and PBS buffer as the solvent.

[0013] Further, the preparation method of the modified chitosan is as follows: (1) Chitosan is added to isopropanol and stirred and dispersed at 65~75℃ for 30~60min. Then, 2,3-epoxypropyltrimethylammonium chloride aqueous solution is added dropwise and stirred for 1~4h. After the reaction is stopped, the mixture is allowed to cool naturally to room temperature. Then, ethanol is added to the reaction system, the mixture is allowed to stand, the lower layer is collected, washed, and vacuum dried to obtain quaternized chitosan; (2) Serine is added to ethanol and stirred and mixed for 30~60min. Then, 98wt% concentrated sulfuric acid is added and stirred at 60~70℃ for 2~4h. After the reaction is stopped, the mixture is allowed to cool naturally to room temperature. Then, diethyl ether is added to the reaction system, the mixture is allowed to stand, the lower layer is collected, washed, and vacuum dried to obtain esterified serine. (3) Add quaternized chitosan, esterified serine and p-toluenesulfonic acid to purified water, stir and mix for 30-60 min, and stir and react in a high-pressure reactor at 135-145℃ for 10-30 min. After the reaction is stopped, let it cool naturally to room temperature, add acetone to the reaction system, let it stand, collect the lower layer, wash and vacuum dry to obtain modified chitosan esterified product; (4) Add the modified chitosan esterified product to an ethanol aqueous solution containing sodium hydroxide, stir and mix for 30-60 min, stir and react at 60-70℃ for 1-2 h, stop the reaction, let it cool naturally to room temperature, adjust the pH of the reaction system to 4.5-5, add acetone, let it stand, collect the lower layer, wash and vacuum dry to obtain modified chitosan.

[0014] Furthermore, the raw materials required for the preparation of the quaternized chitosan include the following components in parts by weight: 1-2 parts chitosan, 2-4 parts 2,3-epoxypropyltrimethylammonium chloride, 2-4 parts purified water, 20-30 parts isopropanol, and 40-60 parts ethanol.

[0015] Furthermore, the raw materials required for the preparation of esterified serine include the following components in parts by weight: 1.5 to 2.5 parts serine, 0.1 to 0.3 parts concentrated sulfuric acid, 15 to 20 parts ethanol, and 30 to 50 parts diethyl ether.

[0016] Furthermore, the raw materials required for the preparation of the modified chitosan esterification include the following components in parts by weight: 1-2 parts of quaternized chitosan, 1.5-2.5 parts of esterified serine, 0.03-0.05 parts of p-toluenesulfonic acid, 15-20 parts of purified water, and 35-50 parts of acetone.

[0017] Furthermore, the raw materials required for the preparation of the modified chitosan include the following components in parts by weight: 1-2 parts of modified chitosan ester, 0.4-0.6 parts of sodium hydroxide, 5-10 parts of ethanol, 5-10 parts of purified water, and 25-50 parts of acetone.

[0018] Furthermore, the amino-terminated polyethylene glycol has a molecular weight of 1000-3000.

[0019] Furthermore, the coating is applied by spraying, with a spraying amount of 0.2~0.5 g / cm³. 2 .

[0020] Furthermore, the sterilization is performed using Co60 irradiation sterilization, with an irradiation dose of 10~20kGy.

[0021] Furthermore, the settling time is 0.5 to 2 hours.

[0022] Compared with the prior art, the beneficial effects achieved by the present invention are: (1) In this invention, glyoxal is used to treat the decellularized porcine peritoneum matrix. The aldehyde groups on glyoxal can undergo a Schiff base reaction with the amino groups on the decellularized porcine peritoneum matrix, introducing aldehyde groups onto the decellularized porcine peritoneum matrix through covalent cross-linking. The introduced aldehyde groups can further undergo a Schiff base reaction with the amino groups on the subsequently sprayed antibacterial components, introducing antibacterial components onto the surface of the decellularized porcine peritoneum matrix through covalent cross-linking, thereby endowing it with excellent antibacterial properties, thus preparing an antibacterial composite peritoneum. That is, glyoxal acts as an intermediate bridge, connecting the decellularized porcine peritoneum matrix and the antibacterial components. Given the biotoxicity of glyoxal, its concentration needs to be controlled. Excessive concentration leads to over-crosslinking of the decellularized porcine peritoneum matrix. This results in residual aldehyde groups after subsequent crosslinking with antibacterial components, which are biotoxic. Furthermore, overly tight crosslinking affects the flexibility of the antibacterial composite periosteum, making it unsuitable for practical applications. Additionally, excessively high concentrations also increase the amount of free glyoxal remaining on the surface of the decellularized porcine peritoneum matrix, further increasing the risk of biotoxicity. Conversely, excessively low concentrations also result in a lower degree of crosslinking, affecting subsequent crosslinking with antibacterial components and preventing the antibacterial performance of the antibacterial composite periosteum from reaching the desired level. Therefore, in this invention, the concentration of glyoxal in the PBS buffer is controlled at 0.05–0.15 wt%.

[0023] (2) In this invention, chitosan is first quaternized and grafted with 2,3-epoxypropyltrimethylammonium chloride to obtain quaternized chitosan. Then, serine is grafted onto the quaternized chitosan via etherification. Considering that the carboxyl group on serine will compete with the hydroxyl group for esterification during the etherification process, serine is first esterified to obtain esterified serine. Subsequently, modified chitosan is prepared by grafting via etherification and deprotection via esterification. The modified chitosan contains a quaternary ammonium salt structure, which can further enhance the antibacterial properties of chitosan. The amino groups it contains can react with the aldehyde groups on the pretreated porcine peritoneum decellularized matrix via a Schiff base reaction, thereby endowing the porcine peritoneum decellularized matrix with good antibacterial properties.

[0024] (3) To enhance the flexibility of the antibacterial composite periosteum and improve its clinical application value, amino-terminated polyethylene glycol is further added to the invention. The amino groups in the amino-terminated polyethylene glycol, under the catalysis of p-toluenesulfonic acid, can undergo an amidation reaction with the carboxyl groups on the modified chitosan. The flexible segments of the amino-terminated polyethylene glycol can alleviate the brittleness of the composite periosteum caused by the modified chitosan spraying. The molecular weight of the amino-terminated polyethylene glycol should not be too large; otherwise, the antibacterial solution will be unevenly dispersed, leading to a decrease in the overall performance of the antibacterial composite periosteum.

[0025] (4) The antibacterial composite periosteum prepared by the present invention has good biocompatibility. When applied in oral surgery, it has good affinity with oral tissues and can reduce the occurrence of local inflammation.

[0026] In summary, this invention utilizes glyoxal to treat porcine peritoneal cell matrix and modifies chitosan, and combines this with amino-terminated polyethylene glycol to prepare an antibacterial solution, ultimately resulting in the comprehensive preparation of an antibacterial composite peritoneum with excellent antibacterial, biocompatibility, and flexibility properties. Detailed Implementation

[0027] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. 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.

[0028] It should be noted that the following quantities are by weight, and there are no special restrictions on the suppliers of all raw materials involved in this invention. Exemplary examples include: In the following examples, chitosan with a degree of deacetylation of 80%, amino-terminated polyethylene glycol with a molecular weight of 2000 and 4000, 2,3-epoxypropyltrimethylammonium chloride with a purity of 99%, and serine with a purity of 99% were all purchased from Merck Reagents Ltd.; other raw materials were commercially available; each part by weight in the following examples is 10g.

[0029] Preparatory steps: Prepare PBS buffer: Add purified water to 8g sodium chloride, 0.2g potassium chloride, 2.9g sodium phosphate, and 0.24g potassium phosphate, and bring the volume to 1L. Adjust the pH to 7.3±0.1 to obtain PBS buffer.

[0030] Example 1: A processing technology for an antibacterial composite periosteum for oral cavity: Step 1: Preparation of decellularized porcine peritoneum matrix: S11: (1) Immerse the pig peritoneum, which has been mostly degreased and free of obvious blood stains and other attachments, in a 0.5M sodium hydroxide aqueous solution and treat it at 30°C with a shaking speed of 135 rpm for 4 hours. Repeat this process 3 times and remove the peritoneum. (2) Immerse the defatted pig peritoneum from (1) in pure water and treat it at 30°C with a shaking speed of 135 rpm for 15 minutes. Repeat this process 5 times and remove the peritoneum. Then immerse it in isopropanol and treat it at 40°C with a shaking speed of 135 rpm for 8 hours. Repeat this process 3 times and remove the peritoneum to obtain defatted pig peritoneum. S12: Immerse defatted porcine peritoneum in decellularization solution and treat at 30°C with a shaking speed of 135 rpm for 1.5 h. Remove and then immerse in PBS buffer and treat at 30°C with a shaking speed of 135 rpm for 30 min. Remove and sterilize by Co60 irradiation with an irradiation dose of 15 kGy to obtain decellularized porcine peritoneum matrix. The decellularization solution includes the following raw material components at the following concentrations: Triton X-100 0.15 v / v%, trypsin 0.3 wt%, and PBS buffer as the solvent; Step 2: Preparation of pretreated porcine peritoneal decellularized matrix: The porcine peritoneal decellularized matrix was immersed in 0.1 wt% glyoxal in PBS buffer and treated at 30°C with a shaking speed of 135 rpm for 40 min. After removal, it was immersed in PBS buffer again and treated at 30°C with a shaking speed of 135 rpm for 15 min. After removal, it was sterilized by Co60 irradiation with an irradiation dose of 15 kGy to obtain the pretreated porcine peritoneal decellularized matrix. Step 3: Preparation of antibacterial composite periosteum: S31: Preparation of modified chitosan: (1) Add 1.5 parts of chitosan to 30 parts of isopropanol, stir and disperse at 70°C for 45 min, then add dropwise 2,3-epoxypropyltrimethylammonium chloride aqueous solution (prepared by 3 parts of 2,3-epoxypropyltrimethylammonium chloride and 3 parts of purified water), stir and react for 3 h, stop the reaction, let it cool naturally to room temperature, add 60 parts of ethanol to the reaction system, let it stand, collect the lower layer, wash and vacuum dry to obtain quaternized chitosan; (2) Add 2 parts of serine to 20 parts of ethanol, stir and mix for 45 min, then add 0.2 parts of 98wt% concentrated sulfuric acid, stir and react at 65°C for 3 h, stop the reaction, let it cool naturally to room temperature, add 50 parts of diethyl ether to the reaction system, let it stand, collect the lower layer, wash and vacuum dry to obtain esterified serine; 3) Add 1.5 parts of quaternized chitosan, 2 parts of esterified serine and 0.04 parts of p-toluenesulfonic acid to 20 parts of purified water, stir and mix for 45 min, and stir and react at 140°C for 20 min in a high-pressure reactor. After the reaction is stopped, let it cool naturally to room temperature, add 50 parts of acetone to the reaction system, let it stand, collect the lower layer, wash and vacuum dry to obtain modified chitosan ester; (4) Add 1.5 parts of modified chitosan ester to an ethanol aqueous solution containing sodium hydroxide (0.5 parts of sodium hydroxide are added to 10 parts of ethanol and 10 parts of purified water to prepare), stir and mix for 45 min, stir and react at 65°C for 1.5 h, stop the reaction, let it cool naturally to room temperature, adjust the pH of the reaction system to 5, add 50 parts of acetone, let it stand, collect the lower layer, wash and vacuum dry to obtain modified chitosan; S32: The antibacterial solution was evenly sprayed onto the surface of the pretreated porcine peritoneal decellularized matrix at a spraying rate of 0.4 g / cm³. 2 The tissue was sterilized by Co60 irradiation with a dose of 15 kGy, then left to stand for 1.5 h, and finally freeze-dried at -20℃ for 24 h to obtain an antibacterial composite periosteum. The antibacterial solution comprises the following raw material components at the following concentrations: 1.5 wt% modified chitosan, 0.3 wt% amino-terminated polyethylene glycol (molecular weight 2000), and 0.03 wt% p-toluenesulfonic acid, with PBS buffer as the solvent.

[0031] Example 2: A processing technology for an antibacterial composite periosteum for oral cavity: Example 2 is based on Example 1, except that the concentration of glyoxal in the PBS buffer is adjusted to 0.05 wt%, while other processes remain unchanged. Specifically: Step 2: Preparation of pretreated porcine peritoneal decellularized matrix: The porcine peritoneal decellularized matrix was immersed in 0.05wt% glyoxal in PBS buffer and treated at 30℃ with a shaking speed of 135 rpm for 40 min. After removal, it was immersed in PBS buffer again and treated at 30℃ with a shaking speed of 135 rpm for 15 min. After removal, it was sterilized by Co60 irradiation with an irradiation dose of 15 kGy to obtain the pretreated porcine peritoneal decellularized matrix.

[0032] Example 3: A processing technology for an antibacterial composite periosteum for oral cavity: Example 3 is based on Example 1, except that the concentration of glyoxal in the PBS buffer is adjusted to 0.15 wt%, while other processes remain unchanged. Specifically: Step 2: Preparation of pretreated porcine peritoneum decellularized matrix: The porcine peritoneum decellularized matrix was immersed in 0.15wt% glyoxal in PBS buffer and treated at 30℃ with a shaking speed of 135 rpm for 40 min. After removal, it was immersed in PBS buffer again and treated at 30℃ with a shaking speed of 135 rpm for 15 min. After removal, it was sterilized by Co60 irradiation with an irradiation dose of 15 kGy to obtain the pretreated porcine peritoneum decellularized matrix. Example 4: A processing technology for an antibacterial composite periosteum for oral cavity: Example 4 is based on Example 1, but with adjustments made to the raw material ratio of the antibacterial solution, while keeping other processes unchanged. Specifically: S32: The antibacterial solution was evenly sprayed onto the surface of the pretreated porcine peritoneal decellularized matrix at a spraying rate of 0.4 g / cm³. 2 The tissue was sterilized by Co60 irradiation with a dose of 15 kGy, then left to stand for 1.5 h, and finally freeze-dried at -20℃ for 24 h to obtain an antibacterial composite periosteum. The antibacterial solution comprises the following raw material components at the following concentrations: 1 wt% modified chitosan, 0.2 wt% amino-terminated polyethylene glycol (molecular weight 2000), 0.01 wt% p-toluenesulfonic acid, and PBS buffer as the solvent.

[0033] Example 5: A processing technology for an antibacterial composite periosteum for oral cavity: Example 5 is based on Example 1, but with adjustments made to the raw material ratio of the antibacterial solution, while keeping other processes unchanged. Specifically: S32: The antibacterial solution was evenly sprayed onto the surface of the pretreated porcine peritoneal decellularized matrix at a spraying rate of 0.4 g / cm³. 2 The tissue was sterilized by Co60 irradiation with a dose of 15 kGy, then left to stand for 1.5 h, and finally freeze-dried at -20℃ for 24 h to obtain an antibacterial composite periosteum. The antibacterial solution comprises the following raw material components at the following concentrations: 2 wt% modified chitosan, 0.4 wt% amino-terminated polyethylene glycol (molecular weight 2000), and 0.05 wt% p-toluenesulfonic acid, with PBS buffer as the solvent.

[0034] The following is a control experiment based on Example 1, with comparative examples 1 to 6, as detailed below: Comparative Example 1: Comparative Example 1 is based on Example 1, with the following adjustment: glyoxal was not used to treat the decellularized porcine peritoneum matrix, while other processes remained unchanged. Specifically: A processing technology for an antibacterial composite periosteum for oral cavity use: Step 1: Preparation of decellularized porcine peritoneum matrix: S11: (1) Immerse the pig peritoneum, which has been mostly degreased and free of obvious blood stains and other attachments, in a 0.5M sodium hydroxide aqueous solution and treat it at 30°C with a shaking speed of 135 rpm for 4 hours. Repeat this process 3 times and remove the peritoneum. (2) Immerse the defatted pig peritoneum from (1) in pure water and treat it at 30°C with a shaking speed of 135 rpm for 15 minutes. Repeat this process 5 times and remove the peritoneum. Then immerse it in isopropanol and treat it at 40°C with a shaking speed of 135 rpm for 8 hours. Repeat this process 3 times and remove the peritoneum to obtain defatted pig peritoneum. S12: Immerse defatted porcine peritoneum in decellularization solution and treat at 30°C with a shaking speed of 135 rpm for 1.5 h. Remove and then immerse in PBS buffer and treat at 30°C with a shaking speed of 135 rpm for 30 min. Remove and sterilize by Co60 irradiation with an irradiation dose of 15 kGy to obtain decellularized porcine peritoneum matrix. The decellularization solution includes the following raw material components at the following concentrations: Triton X-100 0.15 v / v%, trypsin 0.3 wt%, and PBS buffer as the solvent; Step 2: Preparation of antibacterial composite periosteum: S21: Preparation of modified chitosan: (1) Add 1.5 parts of chitosan to 30 parts of isopropanol, stir and disperse at 70°C for 45 min, then add dropwise 2,3-epoxypropyltrimethylammonium chloride aqueous solution (prepared by 3 parts of 2,3-epoxypropyltrimethylammonium chloride and 3 parts of purified water), stir and react for 3 h, stop the reaction, let it cool naturally to room temperature, add 60 parts of ethanol to the reaction system, let it stand, collect the lower layer material, wash and vacuum dry to obtain quaternized chitosan; (2) Add 2 parts of serine to 20 parts of ethanol, stir and mix for 45 min, then add 0.2 parts of 98wt% concentrated sulfuric acid, stir and react at 65°C for 3 h, stop the reaction, let it cool naturally to room temperature, add 50 parts of diethyl ether to the reaction system, let it stand, collect the lower layer material, wash and vacuum dry to obtain esterified serine; 3) Add 1.5 parts of quaternized chitosan, 2 parts of esterified serine and 0.04 parts of p-toluenesulfonic acid to 20 parts of purified water, stir and mix for 45 min, and stir and react at 140°C for 20 min in a high-pressure reactor. After the reaction is stopped, let it cool naturally to room temperature, add 50 parts of acetone to the reaction system, let it stand, collect the lower layer, wash and vacuum dry to obtain modified chitosan ester; (4) Add 1.5 parts of modified chitosan ester to an ethanol aqueous solution containing sodium hydroxide (0.5 parts of sodium hydroxide are added to 10 parts of ethanol and 10 parts of purified water to prepare), stir and mix for 45 min, stir and react at 65°C for 1.5 h, stop the reaction, let it cool naturally to room temperature, adjust the pH of the reaction system to 5, add 50 parts of acetone, let it stand, collect the lower layer, wash and vacuum dry to obtain modified chitosan; S22: Spray the antibacterial solution evenly onto the surface of the decellularized matrix of the porcine peritoneum at a spraying rate of 0.4 g / cm³. 2 The tissue was sterilized by Co60 irradiation with a dose of 15 kGy, then left to stand for 1.5 h, and finally freeze-dried at -20℃ for 24 h to obtain an antibacterial composite periosteum. The antibacterial solution comprises the following raw material components at the following concentrations: 1.5 wt% modified chitosan, 0.3 wt% amino-terminated polyethylene glycol (molecular weight 2000), and 0.03 wt% p-toluenesulfonic acid, with PBS buffer as the solvent.

[0035] Comparative Example 2: Comparative Example 2 is based on Example 1, except that the concentration of glyoxal in the PBS buffer was adjusted to 0.2 wt%, while other processes remained unchanged. Specifically: A processing technology for an antibacterial composite periosteum for oral cavity use: Step 1: Preparation of decellularized porcine peritoneum matrix: S11: (1) Immerse the pig peritoneum, which has been mostly degreased and free of obvious blood stains and other attachments, in a 0.5M sodium hydroxide aqueous solution and treat it at 30°C with a shaking speed of 135 rpm for 4 hours. Repeat this process 3 times and remove the peritoneum. (2) Immerse the defatted pig peritoneum from (1) in pure water and treat it at 30°C with a shaking speed of 135 rpm for 15 minutes. Repeat this process 5 times and remove the peritoneum. Then immerse it in isopropanol and treat it at 40°C with a shaking speed of 135 rpm for 8 hours. Repeat this process 3 times and remove the peritoneum to obtain defatted pig peritoneum. S12: Immerse defatted porcine peritoneum in decellularization solution and treat at 30°C with a shaking speed of 135 rpm for 1.5 h. Remove and then immerse in PBS buffer and treat at 30°C with a shaking speed of 135 rpm for 30 min. Remove and sterilize by Co60 irradiation with an irradiation dose of 15 kGy to obtain decellularized porcine peritoneum matrix. The decellularization solution includes the following raw material components at the following concentrations: Triton X-100 0.15 v / v%, trypsin 0.3 wt%, and PBS buffer as the solvent; Step 2: Preparation of pretreated porcine peritoneum decellularized matrix: The porcine peritoneum decellularized matrix was immersed in 0.2 wt% glyoxal in PBS buffer and treated at 30°C with a shaking speed of 135 rpm for 40 min. After removal, it was immersed in PBS buffer again and treated at 30°C with a shaking speed of 135 rpm for 15 min. After removal, it was sterilized by Co60 irradiation with an irradiation dose of 15 kGy to obtain the pretreated porcine peritoneum decellularized matrix. Step 3: Preparation of antibacterial composite periosteum: S31: Preparation of modified chitosan: (1) Add 1.5 parts of chitosan to 30 parts of isopropanol, stir and disperse at 70°C for 45 min, then add dropwise 2,3-epoxypropyltrimethylammonium chloride aqueous solution (prepared by 3 parts of 2,3-epoxypropyltrimethylammonium chloride and 3 parts of purified water), stir and react for 3 h, stop the reaction, let it cool naturally to room temperature, add 60 parts of ethanol to the reaction system, let it stand, collect the lower layer, wash and vacuum dry to obtain quaternized chitosan; (2) Add 2 parts of serine to 20 parts of ethanol, stir and mix for 45 min, then add 0.2 parts of 98wt% concentrated sulfuric acid, stir and react at 65°C for 3 h, stop the reaction, let it cool naturally to room temperature, add 50 parts of diethyl ether to the reaction system, let it stand, collect the lower layer, wash and vacuum dry to obtain esterified serine; 3) Add 1.5 parts of quaternized chitosan, 2 parts of esterified serine and 0.04 parts of p-toluenesulfonic acid to 20 parts of purified water, stir and mix for 45 min, and stir and react at 140°C for 20 min in a high-pressure reactor. After the reaction is stopped, let it cool naturally to room temperature, add 50 parts of acetone to the reaction system, let it stand, collect the lower layer, wash and vacuum dry to obtain modified chitosan ester; (4) Add 1.5 parts of modified chitosan ester to an ethanol aqueous solution containing sodium hydroxide (0.5 parts of sodium hydroxide are added to 10 parts of ethanol and 10 parts of purified water to prepare), stir and mix for 45 min, stir and react at 65°C for 1.5 h, stop the reaction, let it cool naturally to room temperature, adjust the pH of the reaction system to 5, add 50 parts of acetone, let it stand, collect the lower layer, wash and vacuum dry to obtain modified chitosan; S32: The antibacterial solution was evenly sprayed onto the surface of the pretreated porcine peritoneal decellularized matrix at a spraying rate of 0.4 g / cm³. 2 The tissue was sterilized by Co60 irradiation with a dose of 15 kGy, then left to stand for 1.5 h, and finally freeze-dried at -20℃ for 24 h to obtain an antibacterial composite periosteum. The antibacterial solution comprises the following raw material components at the following concentrations: 1.5 wt% modified chitosan, 0.3 wt% amino-terminated polyethylene glycol (molecular weight 2000), and 0.03 wt% p-toluenesulfonic acid, with PBS buffer as the solvent.

[0036] Comparative Example 3: Comparative Example 3 is based on Example 1, with the following adjustment: no modification treatment is performed on chitosan, while other processes remain unchanged. Specifically: A processing technology for an antibacterial composite periosteum for oral cavity use: Step 1: Preparation of decellularized porcine peritoneum matrix: S11: (1) Immerse the pig peritoneum, which has been mostly degreased and free of obvious blood stains and other attachments, in a 0.5M sodium hydroxide aqueous solution and treat it at 30°C with a shaking speed of 135 rpm for 4 hours. Repeat this process 3 times and remove the peritoneum. (2) Immerse the defatted pig peritoneum from (1) in pure water and treat it at 30°C with a shaking speed of 135 rpm for 15 minutes. Repeat this process 5 times and remove the peritoneum. Then immerse it in isopropanol and treat it at 40°C with a shaking speed of 135 rpm for 8 hours. Repeat this process 3 times and remove the peritoneum to obtain defatted pig peritoneum. S12: Immerse defatted porcine peritoneum in decellularization solution and treat at 30°C with a shaking speed of 135 rpm for 1.5 h. Remove and then immerse in PBS buffer and treat at 30°C with a shaking speed of 135 rpm for 30 min. Remove and sterilize by Co60 irradiation with an irradiation dose of 15 kGy to obtain decellularized porcine peritoneum matrix. The decellularization solution includes the following raw material components at the following concentrations: Triton X-100 0.15 v / v%, trypsin 0.3 wt%, and PBS buffer as the solvent; Step 2: Preparation of pretreated porcine peritoneal decellularized matrix: The porcine peritoneal decellularized matrix was immersed in 0.1 wt% glyoxal in PBS buffer and treated at 30°C with a shaking speed of 135 rpm for 40 min. After removal, it was immersed in PBS buffer again and treated at 30°C with a shaking speed of 135 rpm for 15 min. After removal, it was sterilized by Co60 irradiation with an irradiation dose of 15 kGy to obtain the pretreated porcine peritoneal decellularized matrix. Step 3: Preparation of antibacterial composite peritoneum: The antibacterial solution was uniformly sprayed onto the surface of the pretreated porcine peritoneal decellularized matrix at a spraying rate of 0.4 g / cm³. 2 The tissue was sterilized by Co60 irradiation with a dose of 15 kGy, then left to stand for 1.5 h, and finally freeze-dried at -20℃ for 24 h to obtain an antibacterial composite periosteum. The antibacterial solution comprises the following raw material components at the following concentrations: chitosan 1.5wt%, amino-terminated polyethylene glycol (molecular weight 2000) 0.3wt%, p-toluenesulfonic acid 0.03wt%, and PBS buffer as the solvent.

[0037] Comparative Example 4: Comparative Example 4 is based on Example 1, with the following adjustment: carboxyl protection of serine is not performed, while other processes remain unchanged. Specifically: A processing technology for an antibacterial composite periosteum for oral cavity use: Step 1: Preparation of decellularized porcine peritoneum matrix: S11: (1) Immerse the pig peritoneum, which has been mostly degreased and free of obvious blood stains and other attachments, in a 0.5M sodium hydroxide aqueous solution and treat it at 30°C with a shaking speed of 135 rpm for 4 hours. Repeat this process 3 times and remove the peritoneum. (2) Immerse the defatted pig peritoneum from (1) in pure water and treat it at 30°C with a shaking speed of 135 rpm for 15 minutes. Repeat this process 5 times and remove the peritoneum. Then immerse it in isopropanol and treat it at 40°C with a shaking speed of 135 rpm for 8 hours. Repeat this process 3 times and remove the peritoneum to obtain defatted pig peritoneum. S12: Immerse defatted porcine peritoneum in decellularization solution and treat at 30°C with a shaking speed of 135 rpm for 1.5 h. Remove and then immerse in PBS buffer and treat at 30°C with a shaking speed of 135 rpm for 30 min. Remove and sterilize by Co60 irradiation with an irradiation dose of 15 kGy to obtain decellularized porcine peritoneum matrix. The decellularization solution includes the following raw material components at the following concentrations: Triton X-100 0.15 v / v%, trypsin 0.3 wt%, and PBS buffer as the solvent; Step 2: Preparation of pretreated porcine peritoneal decellularized matrix: The porcine peritoneal decellularized matrix was immersed in 0.1 wt% glyoxal in PBS buffer and treated at 30°C with a shaking speed of 135 rpm for 40 min. After removal, it was immersed in PBS buffer again and treated at 30°C with a shaking speed of 135 rpm for 15 min. After removal, it was sterilized by Co60 irradiation with an irradiation dose of 15 kGy to obtain the pretreated porcine peritoneal decellularized matrix. Step 3: Preparation of antibacterial composite periosteum: S31: Preparation of modified chitosan: (1) Add 1.5 parts of chitosan to 30 parts of isopropanol, stir and disperse at 70°C for 45 min, then add 2,3-epoxypropyltrimethylammonium chloride aqueous solution (prepared by 3 parts of 2,3-epoxypropyltrimethylammonium chloride and 3 parts of purified water), stir and react for 3 h, stop the reaction, let it cool naturally to room temperature, add 60 parts of ethanol to the reaction system, let it stand, collect the lower layer material, wash and vacuum dry to obtain quaternized chitosan; (2) Add 1.5 parts of quaternized chitosan, 2 parts of serine and 0.04 parts of p-toluenesulfonic acid to 20 parts of purified water, stir and mix for 45 min, stir and react at 140°C for 20 min in a high-pressure reactor, stop the reaction, let it cool naturally to room temperature, add 50 parts of acetone to the reaction system, let it stand, collect the lower layer material, wash and vacuum dry to obtain modified chitosan; S32: The antibacterial solution was evenly sprayed onto the surface of the pretreated porcine peritoneal decellularized matrix at a spraying rate of 0.4 g / cm³. 2The tissue was sterilized by Co60 irradiation with a dose of 15 kGy, then left to stand for 1.5 h, and finally freeze-dried at -20℃ for 24 h to obtain an antibacterial composite periosteum. The antibacterial solution comprises the following raw material components at the following concentrations: 1.5 wt% modified chitosan, 0.3 wt% amino-terminated polyethylene glycol (molecular weight 2000), and 0.03 wt% p-toluenesulfonic acid, with PBS buffer as the solvent.

[0038] Comparative Example 5: Comparative Example 5 is based on Example 1, with the following adjustment: amino-terminated polyethylene glycol is not added to the antibacterial solution, while other processes remain unchanged. Specifically: A processing technology for an antibacterial composite periosteum for oral cavity use: Step 1: Preparation of decellularized porcine peritoneum matrix: S11: (1) Immerse the pig peritoneum, which has been mostly degreased and free of obvious blood stains and other attachments, in a 0.5M sodium hydroxide aqueous solution and treat it at 30°C with a shaking speed of 135 rpm for 4 hours. Repeat this process 3 times and remove the peritoneum. (2) Immerse the defatted pig peritoneum from (1) in pure water and treat it at 30°C with a shaking speed of 135 rpm for 15 minutes. Repeat this process 5 times and remove the peritoneum. Then immerse it in isopropanol and treat it at 40°C with a shaking speed of 135 rpm for 8 hours. Repeat this process 3 times and remove the peritoneum to obtain defatted pig peritoneum. S12: Immerse defatted porcine peritoneum in decellularization solution and treat at 30°C with a shaking speed of 135 rpm for 1.5 h. Remove and then immerse in PBS buffer and treat at 30°C with a shaking speed of 135 rpm for 30 min. Remove and sterilize by Co60 irradiation with an irradiation dose of 15 kGy to obtain decellularized porcine peritoneum matrix. The decellularization solution includes the following raw material components at the following concentrations: Triton X-100 0.15 v / v%, trypsin 0.3 wt%, and PBS buffer as the solvent; Step 2: Preparation of pretreated porcine peritoneal decellularized matrix: The porcine peritoneal decellularized matrix was immersed in 0.1 wt% glyoxal in PBS buffer and treated at 30°C with a shaking speed of 135 rpm for 40 min. After removal, it was immersed in PBS buffer again and treated at 30°C with a shaking speed of 135 rpm for 15 min. After removal, it was sterilized by Co60 irradiation with an irradiation dose of 15 kGy to obtain the pretreated porcine peritoneal decellularized matrix. Step 3: Preparation of antibacterial composite periosteum: S31: Preparation of modified chitosan: (1) Add 1.5 parts of chitosan to 30 parts of isopropanol, stir and disperse at 70°C for 45 min, then add dropwise 2,3-epoxypropyltrimethylammonium chloride aqueous solution (prepared by 3 parts of 2,3-epoxypropyltrimethylammonium chloride and 3 parts of purified water), stir and react for 3 h, stop the reaction, let it cool naturally to room temperature, add 60 parts of ethanol to the reaction system, let it stand, collect the lower layer, wash and vacuum dry to obtain quaternized chitosan; (2) Add 2 parts of serine to 20 parts of ethanol, stir and mix for 45 min, then add 0.2 parts of 98wt% concentrated sulfuric acid, stir and react at 65°C for 3 h, stop the reaction, let it cool naturally to room temperature, add 50 parts of diethyl ether to the reaction system, let it stand, collect the lower layer, wash and vacuum dry to obtain esterified serine; 3) Add 1.5 parts of quaternized chitosan, 2 parts of esterified serine and 0.04 parts of p-toluenesulfonic acid to 20 parts of purified water, stir and mix for 45 min, and stir and react at 140°C for 20 min in a high-pressure reactor. After the reaction is stopped, let it cool naturally to room temperature, add 50 parts of acetone to the reaction system, let it stand, collect the lower layer, wash and vacuum dry to obtain modified chitosan ester; (4) Add 1.5 parts of modified chitosan ester to an ethanol aqueous solution containing sodium hydroxide (0.5 parts of sodium hydroxide are added to 10 parts of ethanol and 10 parts of purified water to prepare), stir and mix for 45 min, stir and react at 65°C for 1.5 h, stop the reaction, let it cool naturally to room temperature, adjust the pH of the reaction system to 5, add 50 parts of acetone, let it stand, collect the lower layer, wash and vacuum dry to obtain modified chitosan; S32: The antibacterial solution was evenly sprayed onto the surface of the pretreated porcine peritoneal decellularized matrix at a spraying rate of 0.4 g / cm³. 2 The tissue was sterilized by Co60 irradiation with a dose of 15 kGy, then left to stand for 1.5 h, and finally freeze-dried at -20℃ for 24 h to obtain an antibacterial composite periosteum. The antibacterial solution comprises the following raw material components at the following concentrations: 1.5 wt% modified chitosan, 0.03 wt% p-toluenesulfonic acid, and PBS buffer as the solvent.

[0039] Comparative Example 6: Comparative Example 6 is based on Example 1, except that the molecular weight of the amino-terminated polyethylene glycol is adjusted to 4000, while other processes remain unchanged. Specifically: A processing technology for an antibacterial composite periosteum for oral cavity use: Step 1: Preparation of decellularized porcine peritoneum matrix: S11: (1) Immerse the pig peritoneum, which has been mostly degreased and free of obvious blood stains and other attachments, in a 0.5M sodium hydroxide aqueous solution and treat it at 30°C with a shaking speed of 135 rpm for 4 hours. Repeat this process 3 times and remove the peritoneum. (2) Immerse the defatted pig peritoneum from (1) in pure water and treat it at 30°C with a shaking speed of 135 rpm for 15 minutes. Repeat this process 5 times and remove the peritoneum. Then immerse it in isopropanol and treat it at 40°C with a shaking speed of 135 rpm for 8 hours. Repeat this process 3 times and remove the peritoneum to obtain defatted pig peritoneum. S12: Immerse defatted porcine peritoneum in decellularization solution and treat at 30°C with a shaking speed of 135 rpm for 1.5 h. Remove and then immerse in PBS buffer and treat at 30°C with a shaking speed of 135 rpm for 30 min. Remove and sterilize by Co60 irradiation with an irradiation dose of 15 kGy to obtain decellularized porcine peritoneum matrix. The decellularization solution includes the following raw material components at the following concentrations: Triton X-100 0.15 v / v%, trypsin 0.3 wt%, and PBS buffer as the solvent; Step 2: Preparation of pretreated porcine peritoneal decellularized matrix: The porcine peritoneal decellularized matrix was immersed in 0.1 wt% glyoxal in PBS buffer and treated at 30°C with a shaking speed of 135 rpm for 40 min. After removal, it was immersed in PBS buffer again and treated at 30°C with a shaking speed of 135 rpm for 15 min. After removal, it was sterilized by Co60 irradiation with an irradiation dose of 15 kGy to obtain the pretreated porcine peritoneal decellularized matrix. Step 3: Preparation of antibacterial composite periosteum: S31: Preparation of modified chitosan: (1) Add 1.5 parts of chitosan to 30 parts of isopropanol, stir and disperse at 70°C for 45 min, then add dropwise 2,3-epoxypropyltrimethylammonium chloride aqueous solution (prepared by 3 parts of 2,3-epoxypropyltrimethylammonium chloride and 3 parts of purified water), stir and react for 3 h, stop the reaction, let it cool naturally to room temperature, add 60 parts of ethanol to the reaction system, let it stand, collect the lower layer, wash and vacuum dry to obtain quaternized chitosan; (2) Add 2 parts of serine to 20 parts of ethanol, stir and mix for 45 min, then add 0.2 parts of 98wt% concentrated sulfuric acid, stir and react at 65°C for 3 h, stop the reaction, let it cool naturally to room temperature, add 50 parts of diethyl ether to the reaction system, let it stand, collect the lower layer, wash and vacuum dry to obtain esterified serine; 3) Add 1.5 parts of quaternized chitosan, 2 parts of esterified serine and 0.04 parts of p-toluenesulfonic acid to 20 parts of purified water, stir and mix for 45 min, and stir and react at 140°C for 20 min in a high-pressure reactor. After the reaction is stopped, let it cool naturally to room temperature, add 50 parts of acetone to the reaction system, let it stand, collect the lower layer, wash and vacuum dry to obtain modified chitosan ester; (4) Add 1.5 parts of modified chitosan ester to an ethanol aqueous solution containing sodium hydroxide (0.5 parts of sodium hydroxide are added to 10 parts of ethanol and 10 parts of purified water to prepare), stir and mix for 45 min, stir and react at 65°C for 1.5 h, stop the reaction, let it cool naturally to room temperature, adjust the pH of the reaction system to 5, add 50 parts of acetone, let it stand, collect the lower layer, wash and vacuum dry to obtain modified chitosan; S32: The antibacterial solution was evenly sprayed onto the surface of the pretreated porcine peritoneal decellularized matrix at a spraying rate of 0.4 g / cm³. 2 The tissue was sterilized by Co60 irradiation with a dose of 15 kGy, then left to stand for 1.5 h, and finally freeze-dried at -20℃ for 24 h to obtain an antibacterial composite periosteum. The antibacterial solution comprises the following raw material components at the following concentrations: 1.5 wt% modified chitosan, 0.3 wt% amino-terminated polyethylene glycol (molecular weight 4000), and 0.03 wt% p-toluenesulfonic acid, with PBS buffer as the solvent.

[0040] Performance testing: The antibacterial composite periosteum prepared in Examples 1-5 and Comparative Examples 1-6 were subjected to antibacterial tests, biocompatibility tests, and toughness tests. The specific test methods are as follows: 1. Antibacterial performance test: The antibacterial composite periosteum was cut into φ12mm circular pieces and placed in a solution of 15μL Staphylococcus aureus (1×10⁻⁶). 6The center of the culture medium (CFU / mL) was located, 2 mL of PBS buffer and 1 mL of LB culture medium were added to the culture medium, and then the medium was incubated in an incubator at 37°C for 24 h. After that, the diameter of the inhibition zone was measured. 2. Biocompatibility test: Mouse bone marrow mesenchymal stem cells were placed in DMEM medium containing 10 wt% fetal bovine serum. Antibacterial composite periosteum was cut into small pieces, and the surface area of ​​the antibacterial composite periosteum was divided into 3 cm³ pieces according to the volume ratio of the culture medium liquid to the surface area of ​​the antibacterial composite periosteum. 2 Add 1 mL of antibacterial composite periosteum to the aforementioned culture medium and soak at 37°C for 24 hours to obtain a cell suspension; then dilute the cell suspension at a rate of 1 × 10⁻⁶ cells per well. 4 Cells / well were seeded into 96-well plates and incubated at 37°C with 5% CO2 for 24 h to obtain periosteal extract. 5 mg / mL MTT reagent was added to each well at a rate of 40 μL / well, and the plates were incubated at 37°C for another 4 h. The MTT reagent was then removed, and 100 μL of dimethyl sulfoxide was added to each well. The plates were shaken slowly for 10 min, and the absorbance of each well was measured using a multi-mode microplate reader (570 nm). (3) Toughness test: The antibacterial composite periosteum was cut into 100mm (length) × 10mm (width) sizes, and then the toughness of the antibacterial composite periosteum was tested at 37℃ at a rate of 10mm / min.

[0041] The specific test results are shown in Table 1: Table 1

[0042] Conclusion: Comparing the data of Examples 1-5 and Comparative Examples 1-6 in Table 1 above, it can be seen that: In this invention, by treating porcine peritoneal cell matrix with glyoxal and modifying chitosan, and by preparing an antibacterial solution in conjunction with amino-terminated polyethylene glycol, an antibacterial composite peritoneum with excellent antibacterial, biocompatibility, and flexibility properties was finally prepared.

[0043] Finally, it should be noted that the above descriptions are merely preferred embodiments of the present invention and are not intended to limit the present invention. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or make equivalent substitutions for some of the technical features. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A processing technology for an antibacterial composite periosteum for oral cavity, characterized in that: Includes the following steps: Step 1: After being treated with decellularized solution, defatted porcine peritoneum is washed and sterilized to obtain decellularized porcine peritoneal matrix; Step 2: After treatment with glyoxal solution, the decellularized porcine peritoneum matrix is ​​washed and sterilized to obtain pretreated decellularized porcine peritoneum matrix; Step 3: The antibacterial solution is evenly coated on the surface of the pretreated porcine peritoneum decellularized matrix, sterilized, allowed to stand, and freeze-dried to obtain an antibacterial composite peritoneum; The antibacterial solution comprises the following raw material components at the following concentrations: 1-2 wt% modified chitosan, 0.2-0.4 wt% amino-terminated polyethylene glycol, and 0.01-0.05 wt% p-toluenesulfonic acid, with PBS buffer as the solvent.

2. The processing technology of an antibacterial composite periosteum for oral cavity according to claim 1, characterized in that: The modified chitosan is prepared by: (1) Chitosan was added to isopropanol and stirred and dispersed evenly at 65~75℃. Then, 2,3-epoxypropyltrimethylammonium chloride aqueous solution was added dropwise and stirred for 1~4h. The reaction was stopped, ethanol was added to the reaction system, and the mixture was allowed to stand. The lower layer was collected, washed and dried to obtain quaternized chitosan. (2) Add serine to ethanol, stir and mix evenly, then add 98wt% concentrated sulfuric acid, stir and react at 60~70℃ for 2~4h, stop the reaction, add diethyl ether to the reaction system, let stand, collect the lower layer, wash and dry to obtain esterified serine; (3) Add quaternized chitosan, esterified serine and p-toluenesulfonic acid to purified water, stir and mix evenly, and stir and react in a high-pressure reactor at 135~145℃ for 10~30min. After the reaction is stopped, add acetone to the reaction system, let it stand, collect the lower layer material, wash and dry it to obtain modified chitosan esterified product. (4) Add the modified chitosan ester to an ethanol aqueous solution containing sodium hydroxide, stir and mix evenly, stir and react at 60~70℃ for 1~2h, stop the reaction, adjust the pH of the reaction system to 4.5~5, add acetone, let stand, collect the lower layer material, wash and vacuum dry to obtain modified chitosan.

3. The processing technology of an antibacterial composite periosteum for oral cavity according to claim 2, characterized in that: The raw materials required for the preparation of the quaternized chitosan include the following components in parts by weight: 1-2 parts chitosan, 2-4 parts 2,3-epoxypropyltrimethylammonium chloride, 2-4 parts purified water, 20-30 parts isopropanol, and 40-60 parts ethanol. The raw materials required for the preparation of esterified serine include the following components in parts by weight: 1.5-2.5 parts serine, 0.1-0.3 parts concentrated sulfuric acid, 15-20 parts ethanol, and 30-50 parts diethyl ether. The raw materials required for the preparation of the modified chitosan esterification include the following components in parts by weight: 1-2 parts of quaternized chitosan, 1.5-2.5 parts of esterified serine, 0.03-0.05 parts of p-toluenesulfonic acid, 15-20 parts of purified water, and 35-50 parts of acetone. The raw materials required for the preparation of the modified chitosan include the following components in parts by weight: 1-2 parts of modified chitosan ester, 0.4-0.6 parts of sodium hydroxide, 5-10 parts of ethanol, 5-10 parts of purified water, and 25-50 parts of acetone.

4. The processing technology of an antibacterial composite periosteum for oral cavity according to claim 1, characterized in that: The amino-terminated polyethylene glycol has a molecular weight of 1000-3000.

5. The processing technology of an antibacterial composite periosteum for oral cavity according to claim 1, characterized in that: The method for preparing the defatted porcine peritoneum is as follows: (1) Immerse the pig peritoneum, which has been mostly cleaned of fat and has no obvious blood or other attachments, in a 0.01-0.1M sodium hydroxide aqueous solution and treat it at 28-32℃ with a shaking speed of 130-140rpm for 3-5 hours. Repeat this process 2-3 times and then remove the peritoneum. (2) Immerse the defatted porcine peritoneum from (1) in pure water and treat it at 28-32℃ with a shaking speed of 130-140 rpm for 15-20 minutes. Repeat this process 4-6 times. Then remove the peritoneum and immerse it in isopropanol. Treat it at 38-42℃ with a shaking speed of 130-140 rpm for 7-9 hours. Repeat this process 2-3 times. Remove the peritoneum to obtain defatted porcine peritoneum.

6. The processing technology of an antibacterial composite periosteum for oral cavity according to claim 1, characterized in that: The method for preparing the decellularized porcine peritoneum matrix is ​​as follows: immerse defatted porcine peritoneum in a decellularization solution and treat it at 28-32°C with a shaking speed of 130-140 rpm for 1-2 hours, then remove it; then immerse it in PBS buffer and treat it at 28-32°C with a shaking speed of 130-140 rpm for 20-40 minutes, then remove it and sterilize it to obtain the decellularized porcine peritoneum matrix; The decellularization solution includes the following raw material components at the following concentrations: Triton X-100 0.1~0.2 v / v%, trypsin 0.1~0.5 wt%, and PBS buffer as the solvent.

7. The processing technology of an antibacterial composite periosteum for oral cavity according to claim 1, characterized in that: The method for preparing the pretreated porcine peritoneal decellularized matrix is ​​as follows: immerse the porcine peritoneal decellularized matrix in glyoxal solution and treat it at 28-32℃ with a shaking speed of 130-140 rpm for 20-60 min, then remove it; then immerse it in PBS buffer and treat it at 28-32℃ with a shaking speed of 130-140 rpm for 10-20 min, then remove it and sterilize it to obtain the pretreated porcine peritoneal decellularized matrix; The glyoxal solution is a PBS buffer solution of glyoxal with a concentration of 0.05~0.15wt%.

8. The processing technology of an antibacterial composite periosteum for oral cavity according to claim 1, characterized in that: The coating is applied by spraying, with a coating amount of 0.2~0.5 g / cm³. 2 .

9. The processing technology of an antibacterial composite periosteum for oral cavity according to claim 1, characterized in that: The settling time is 0.5 to 2 hours.

10. An antibacterial composite periosteum for oral cavity obtained by a processing method according to any one of claims 1 to 9.

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