Moisture-resistant antibacterial bone adhesive based on pig collagen and preparation method of moisture-resistant antibacterial bone adhesive

Abstract

The invention relates to the field of biomedical materials, and discloses a moisture-resistant antibacterial bone adhesive based on pig collagen, the moisture-resistant antibacterial bone adhesive based on pig collagen comprises the following components by weight: 0.01-100 g of pig collagen, 0.01-100 g of tannic acid, 0.01-100 g of nano-hydroxyapatite, 0.01-100 g of nano-hydroxyapatite, 0.01-100 g of calcium carbonate, 0.01-100 g of calcium carbonate, 0.01-100 g of calcium carbonate, 0.01-100 g of calcium carbonate and 0.01-100 g of calcium carbonate. The preparation method comprises the following steps: S1, preparation of a pig collagen aqueous solution: dissolving pig collagen in deionized water to obtain the pig collagen aqueous solution; s2, preparation of a tannic acid aqueous solution: dissolving tannic acid powder in deionized water to obtain the tannic acid aqueous solution; s3, preparation of a tannic acid-nano hydroxyapatite mixed solution; and S4, mixing, oscillating and forming. A bionic microenvironment can be provided for bone cells through the pig collagen aqueous solution, cell adhesion and proliferation and new bone tissue formation are promoted, the bone adhesive force is achieved under the humid condition, meanwhile, high commercial feasibility is achieved, and the cost should be economical.

Description

Technical Field

[0001] The invention relates to the field of biomedical materials, in particular to a moisture-resistant antibacterial bone adhesive based on porcine collagen. Background Art

[0002] Fractures are a common and critical traumatic diagnosis in clinical medicine. According to statistics, approximately 178 million new fractures occur worldwide each year. The femur, humerus, and tibia are the most common fracture sites, accounting for 3%, 14%, and 24% of adult fractures, respectively. The core of fracture treatment lies in repositioning and fixing the broken bone segments, restoring their original position and tightly uniting them, while relying on endogenous bone formation to promote healing. In ancient times, external plaster immobilization was a common conservative treatment. However, starting in the late 19th century, internal fixation techniques began to emerge and gradually developed. Today, the principles established by the Society for Bone and Joint Replacement are considered the gold standard by orthopedic surgeons worldwide, emphasizing the importance of stable fixation when treating comminuted and complex fractures. This makes internal fixation the preferred treatment for long bone fractures.

[0003] To overcome the inherent defects of traditional internal fixation technology, more and more medical workers and researchers have begun to explore alternative fracture treatment methods, among which bone adhesives are considered to be a promising option.

[0004] However, the production cost of traditional bone adhesives is too high, making them inconvenient for commercial production. Summary of the Invention

[0005] In view of the shortcomings of the existing technology, the present invention provides a moisture-resistant antibacterial bone adhesive based on porcine collagen, which solves the problem that the required manufacturing cost is too high and it is inconvenient for commercial manufacturing.

[0006] To achieve the above objectives, the present invention is implemented through the following technical scheme: a moisture-resistant antibacterial bone adhesive based on porcine collagen, comprising porcine collagen, tannic acid and nano-hydroxyapatite, 0.01 to 100 grams of porcine collagen, 0.01 to 100 grams of tannic acid, and 0.01 to 100 grams of nano-hydroxyapatite.

[0007] Preferably, a method for preparing a moisture-resistant antibacterial bone adhesive based on porcine collagen is used for the moisture-resistant antibacterial bone adhesive based on porcine collagen according to claim 1, the method comprising the following steps: S1. Preparation of porcine collagen aqueous solution: dissolving porcine collagen in deionized water to obtain a porcine collagen aqueous solution; S2. Preparation of tannic acid aqueous solution: dissolving tannic acid powder in deionized water to obtain a tannic acid aqueous solution; S3, preparation of tannic acid-nanohydroxyapatite mixed solution: dissolving hydroxyapatite powder in deionized water and adding the solution to the tannic acid aqueous solution obtained in S2 to obtain a tannic acid-nanohydroxyapatite mixed solution; S4, mixing and oscillating molding: the porcine collagen aqueous solution obtained in S1 and the tannic acid-nanohydroxyapatite mixed solution obtained in S3 are mixed to obtain a moisture-resistant antibacterial bone adhesive based on porcine collagen.

[0008] The S4 step also includes an ethylene oxide sterilization step, with an ethylene oxide concentration of 600-1000 mg / L, a sterilization temperature of 45-55°C, and a sterilization time of 2-5 hours.

[0009] Preferably, in step S1, the amount of porcine collagen is 0.01 to 100 grams, the amount of deionized water is 0.01 to 100 grams, and the stirring time during dissolution is 20 to 60 minutes.

[0010] Preferably, the amount of tannic acid in step S2 is 0.01 to 100 grams, the amount of deionized water is 0.01 to 100 grams, and the stirring time during dissolution is 10 to 30 minutes.

[0011] Preferably, in the step S3, the amount of hydroxyapatite powder is 0.01 to 100 g, the amount of deionized water is 0.01 to 100 g, the stirring time during dissolution is 10 to 30 min, and a nano-hydroxyapatite suspension is formed. The volume ratio of the tannic acid aqueous solution obtained in the step S2 to the nano-hydroxyapatite suspension is 1:1, and the stirring time during mixing is 15 to 30 min.

[0012] Preferably, in step S4, the volume ratio of the porcine collagen aqueous solution obtained in step S1 to the tannic acid-nanohydroxyapatite mixed solution obtained in step S3 is 1:1, the oscillation time during mixing is 5 to 15 minutes, and the standing time after oscillation is 10 to 30 minutes.

[0013] Preferably, the concentration of the porcine collagen aqueous solution obtained in step S1 is 5 to 50 wt %, and the concentration of the tannic acid aqueous solution obtained in step S2 is 5 to 50 wt %.

[0014] Preferably, the concentration of the nano-hydroxyapatite suspension obtained in step S3 is 5 to 50 wt %, and the particle size of the nano-hydroxyapatite powder is 20 to 100 nm.

[0015] Preferably, the bone adhesive forms adhesion on the wet bone tissue surface within 1 minute, has an adhesion strength of not less than 0.5 MPa, and has an inhibition rate against Staphylococcus aureus of ≥90%.

[0016] The present invention provides a moisture-resistant antibacterial bone adhesive based on porcine collagen. It has the following beneficial effects: 1. In the present invention, the porcine collagen aqueous solution can provide a bionic microenvironment for bone cells, promote cell adhesion, proliferation and new bone tissue formation, have bone adhesion under moist conditions and have high commercial feasibility at a relatively economical cost.

[0017] 2. In the present invention, porcine collagen, tannic acid and nano-hydroxyapatite are rapidly combined through hydrogen bonds and coordination bonds to form a bone adhesive with wet bone tissue adhesion, thereby improving the immediate adhesion of the adhesive on the wet bone tissue surface.

[0018] 3. In the present invention, by selecting nano-hydroxyapatite as the main inorganic component of bone tissue and coordinating with the ethylene oxide sterilization step, while promoting bone formation and bone ingrowth, it can also ensure excellent antibacterial properties to prevent infection in the fracture area.

[0019] 4. In the present invention, by controlling the concentration of the prepared solution, the adhesion strength is guaranteed to be ≥0.5MPa under a wet environment, and it can also be solidified within 1 minute to bond the bones. It is convenient to prepare and apply to the fracture site within a time range acceptable to doctors, and is suitable for a variety of common clinical fracture types. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 The infrared spectra of porcine collagen, tannic acid, nano-hydroxyapatite, and bone adhesive (porcine collagen-tannic acid-hydroxyapatite); Figure 2 This is a photo of the bone adhesive of the present invention bonding bovine bone slices; Figure 3 is an adhesion performance curve diagram of the lap-shear adhesion test of the bone adhesive of the present invention; Figure 4 is an adhesion performance curve diagram of an end-to-end adhesion experiment of the bone adhesive of the present invention; Figure 5 This is a photo of the bone adhesive of the present invention used to bond rabbit ulna and radius fractures; Figure 6 This is a diagram showing the antibacterial effect of the bone adhesive of the present invention on Staphylococcus aureus; Figure 7 Graph showing the cytotoxicity test results of the bone adhesive of the present invention on rabbit bone marrow mesenchymal stem cells; Figure 8 The figure is a schematic diagram of the process for preparing the antibacterial bone adhesive of the present invention. DETAILED DESCRIPTION

[0021] The following will clearly and completely describe the technical solutions of the present invention in conjunction with the accompanying drawings. Obviously, the embodiments described are only some of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making any creative efforts are within the scope of protection of the present invention.

[0022] Please see the attached Figure 1 - Attachment Figure 8 An embodiment of the present invention provides a moisture-resistant antibacterial bone adhesive based on porcine collagen, comprising porcine collagen, tannic acid and nano-hydroxyapatite, 0.01 to 100 grams of porcine collagen, 0.01 to 100 grams of tannic acid, and 0.01 to 100 grams of nano-hydroxyapatite.

[0023] Specifically, porcine collagen, tannic acid and nano-hydroxyapatite are rapidly combined through hydrogen bonds and coordination bonds to form a bone adhesive that has wet bone tissue adhesion and promotes the generation of new bone tissue. This invention is suitable for various clinical fracture types, including long bone fractures, weight-bearing bone fractures, comminuted fractures and irregular bone defects.

[0024] Please see the attached Figure 1 - Attachment Figure 8 A method for preparing a moisture-resistant antibacterial bone adhesive based on porcine collagen, for use in a moisture-resistant antibacterial bone adhesive based on porcine collagen according to claim 1, the method comprising the following steps: S1. Preparation of porcine collagen aqueous solution: dissolving porcine collagen in deionized water to obtain a porcine collagen aqueous solution; S2. Preparation of tannic acid aqueous solution: dissolving tannic acid powder in deionized water to obtain a tannic acid aqueous solution; S3, preparation of tannic acid-nanohydroxyapatite mixed solution: dissolving hydroxyapatite powder in deionized water and adding the solution to the tannic acid aqueous solution obtained in S2 to obtain a tannic acid-nanohydroxyapatite mixed solution; S4, mixing and oscillating molding: the porcine collagen aqueous solution obtained in S1 and the tannic acid-nanohydroxyapatite mixed solution obtained in S3 are mixed to obtain a moisture-resistant antibacterial bone adhesive based on porcine collagen.

[0025] Specifically, in step S1, porcine collagen is one of the main components of bone matrix. Its aqueous solution can provide a biomimetic microenvironment for bone cells, promote cell adhesion, proliferation and new bone tissue formation. The preparation method is simple and low-cost. Dissolving porcine collagen in deionized water in the form of an aqueous solution avoids the use of organic solvents, reduces the risk of cytotoxicity, and meets the biosafety requirements of medical devices. In step S2, tannic acid releases polyphenol active ingredients, destroying bacterial cell membranes and inhibiting their metabolism. This results in an inhibition rate of ≥90% against common orthopedic pathogens such as Staphylococcus aureus, significantly reducing the risk of postoperative infection. The phenolic hydroxyl groups of tannic acid combine with the amino groups of porcine collagen and the calcium ions of hydroxyapatite through hydrogen bonds and coordination bonds, forming a three-dimensional network structure that significantly enhances the adhesive's immediate adhesion to moist bone tissue surfaces. In step S3, nano-hydroxyapatite, as the main inorganic component of bone tissue, has high hardness and rigidity that enhances the performance of the adhesive, meeting the mechanical requirements of load-bearing bone fractures. The phenolic hydroxyl groups of tannic acid and the calcium ions of hydroxyapatite combine through coordination bonds to form a stable network structure, reducing the slippage of molecular chains in a wet environment and improving the long-term stability of the adhesive. In step S4, porcine collagen simulates the organic matrix of natural bone, and nanohydroxyapatite provides an inorganic mineral phase to form an "organic-inorganic hybrid network" similar to natural bone, promoting the attachment and differentiation of osteoblasts. The adhesive can be injected or preformed and is suitable for complex scenarios such as comminuted fractures and irregular bone defects.

[0026] After step S4, an ethylene oxide sterilization step is also included, with an ethylene oxide concentration of 600-1000 mg / L, a sterilization temperature of 45-55°C, and a sterilization time of 2-5 hours.

[0027] Ethylene oxide can penetrate the porous adhesive structure and effectively kill bacteria, fungi, and spore-forming microorganisms. The antibacterial rate is maintained at ≥90%, avoiding the risk of postoperative infection. Ethylene oxide sterilization is a gas-phase reaction and will not dissolve or destroy the hydrogen bond / coordination bond structure of collagen-tannic acid-hydroxyapatite. The adhesion strength remains ≥0.5MPa. Ethylene oxide sterilization can process large quantities of products at the same time, with high efficiency and suitable for industrial production.

[0028] In step S1, the amount of porcine collagen is 0.01 to 100 g, the amount of deionized water is 0.01 to 100 g, and the stirring time during dissolution is 20 to 60 min.

[0029] Specifically, 0.01 to 100 grams of porcine collagen is taken, covering laboratory research to industrial production needs. It is stirred for 20 to 60 minutes to prevent metal ions from interfering with the collagen conformation and maintain its natural triple helix structure. The fully dissolved collagen aqueous solution can quickly cross-link with tannic acid through hydrogen bonds and quickly cross-link with hydroxyapatite calcium ions through electrostatic effects, thereby improving the curing speed and wet adhesion strength of the adhesive.

[0030] In step S2, the amount of tannic acid is 0.01 to 100 g, the amount of deionized water is 0.01 to 100 g, and the stirring time during dissolution is 10 to 30 min.

[0031] Specifically, by taking 0.01 to 100 grams of tannic acid and 0.01 to 100 grams of deionized water, tannic acid can destroy bacterial cell membranes to reduce the risk of postoperative infection. By stirring for 10 to 30 minutes, metal ions are prevented from interfering with the activity of tannic acid phenolic hydroxyl groups, thereby maintaining its antioxidant and cross-linking capabilities.

[0032] In step S3, the amount of hydroxyapatite powder is 0.01 to 100 grams, the amount of deionized water is 0.01 to 100 grams, and the stirring time during dissolution is 10 to 30 minutes to form a nanohydroxyapatite suspension. The volume ratio of the tannic acid aqueous solution obtained in step S2 to the nanohydroxyapatite suspension is 1:1, and the stirring time during mixing is 15 to 30 minutes.

[0033] Specifically, stirring for 10 to 30 minutes ensures that the hydroxyapatite powder nanoparticles are fully dispersed in deionized water to avoid agglomeration, providing a uniform inorganic phase for subsequent composite preparation. The volume ratio of tannic acid to hydroxyapatite is 1:1, so that the phenolic hydroxyl groups of tannic acid can completely cover the surface of the nanoparticles, forming a stable coordination bond network. The rigidity of nano-hydroxyapatite enhances the adhesion strength of the adhesive, meeting the mechanical requirements of load-bearing bones. Mixing and stirring for 15 to 30 minutes allows the tannic acid and hydroxyapatite to be fully combined through the Ca²⁺-phenolic hydroxyl coordination bond, and the wet adhesion strength is ≥0.5MPa.

[0034] In step S4, the volume ratio of the porcine collagen aqueous solution obtained in step S1 to the tannic acid-nanohydroxyapatite mixed solution obtained in step S3 is 1:1, the oscillation time during mixing is 5 to 15 minutes, and the standing time after oscillation is 10 to 30 minutes.

[0035] Specifically, a volume ratio of 1:1 is used to ensure that the amino groups of collagen, the phenolic hydroxyl groups of tannic acid, and the calcium ions of hydroxyapatite are quickly cross-linked through hydrogen bonds and coordination bonds, so that the adhesive can form a firm adhesion on the wet bone surface within 1 minute, with an adhesion strength of ≥0.5MPa. Vibration is used to promote uniform mixing of the components, avoid the generation of bubbles, and prevent excessive shearing from destroying the collagen fiber structure. The hydrophobic-hydrophilic balance structure formed after standing can resist the erosion of physiological saline.

[0036] The concentration of the porcine collagen aqueous solution obtained in step S1 is 5 to 50 wt %, and the concentration of the tannic acid aqueous solution obtained in step S2 is 5 to 50 wt %.

[0037] Specifically, the concentration of porcine collagen aqueous solution is controlled at 5-20wt% to form a porous structure to promote cell migration and blood vessel growth, the concentration is controlled at 30-50wt% to form a high-density collagen network to simulate natural bone matrix and directly induce osteogenic differentiation, and the concentration of tannic acid aqueous solution is controlled at 5-50wt%, while the free radical scavenging ability is positively correlated with the concentration, ensuring the antibacterial rate.

[0038] The concentration of the nano-hydroxyapatite suspension obtained in step S3 is 5-50 wt %, and the particle size of the nano-hydroxyapatite powder is 20-100 nm.

[0039] Specifically, the weak alkalinity of nanohydroxyapatite destroys the bacterial membrane potential, and the combined antibacterial rate with tannic acid is ≥95%. The 20-100 nm particle size optimizes the ion release kinetics and continuously activates osteoblasts. Combined with 10-30 minutes of stirring, it ensures that the nanohydroxyapatite is evenly dispersed and there is no precipitation after standing for 24 hours.

[0040] The bone adhesive forms adhesion on the wet bone tissue surface within 1 minute, with an adhesion strength of not less than 0.5 MPa, and an inhibition rate against Staphylococcus aureus of ≥90%.

[0041] Specifically, traditional bone cement takes 5 to 10 minutes to cure, and the adhesion strength decays to <0.3 MPa in a humid environment. The present invention cures in 1 minute and maintains a strength of ≥0.5 MPa, shortening the operation time, destroying the Staphylococcus aureus biofilm, inhibiting bacterial metabolism, and reducing the infection rate.

[0042] The experimental methods used below are conventional methods unless otherwise specified. The materials, reagents, methods, and instruments used, unless otherwise specified, are conventional in the art and can be obtained commercially or prepared according to literature by those skilled in the art.

[0043] Example 1 A method for preparing an antibacterial bone adhesive based on porcine collagen comprises the following steps: Select 0.01g of porcine collagen, 0.01g of tannic acid, 0.01g of nanohydroxyapatite and 0.01g of deionized water. S1, dissolving 0.01 g of porcine collagen in 0.01 g of deionized water and stirring for 20 minutes to obtain a 5 wt% collagen aqueous solution; S2. Dissolve 0.01 g of tannic acid in 0.01 g of deionized water and stir for 10 minutes to obtain a 5 wt% tannic acid aqueous solution.

[0044] S3, 0.01g of nano-hydroxyapatite with a particle size of 20nm was dissolved in 0.01g of deionized water and stirred for 10 minutes to form a 5wt% suspension, which was then mixed with tannic acid aqueous solution at a volume ratio of 1:1 and stirred for 15 minutes. S4. The solutions obtained in S1 and S3 were mixed in a volume ratio of 1:1, shaken for 5 minutes, and then allowed to stand for 10 minutes to obtain a moisture-resistant antibacterial bone adhesive based on porcine collagen.

[0045] Example 2 A method for preparing an antibacterial bone adhesive based on porcine collagen comprises the following steps: Select 50g of porcine collagen, 30g of tannic acid, 40g of nanohydroxyapatite and 50g of deionized water. S1, dissolving 50g of porcine collagen in 50g of deionized water and stirring for 40 minutes to obtain a 50wt% collagen aqueous solution; S2. Dissolve 30 g of tannic acid in 50 g of deionized water and stir for 20 minutes to obtain a 37.5 wt % tannic acid aqueous solution.

[0046] S3, 40 nanometers of hydroxyapatite with a particle size of 50nm was dissolved in 50g of deionized water and stirred for 20 minutes to form a 44.4wt% suspension, which was then mixed with a tannic acid aqueous solution at a volume ratio of 1:1 and stirred for 20 minutes. S4. The solutions obtained in S1 and S3 were mixed in a volume ratio of 1:1, shaken for 10 minutes, and then allowed to stand for 20 minutes to obtain a moisture-resistant antibacterial bone adhesive based on porcine collagen.

[0047] Example 3 A method for preparing an antibacterial bone adhesive based on porcine collagen comprises the following steps: Select 100g of porcine collagen, 100g of tannic acid, 100g of nanohydroxyapatite and 100g of deionized water. S1, dissolving 100g of porcine collagen in 100g of deionized water and stirring for 60 minutes to obtain a 50wt% collagen aqueous solution; S2. Dissolve 100 g of tannic acid in 100 g of deionized water and stir for 30 minutes to obtain a 50 wt % tannic acid aqueous solution.

[0048] S3, 100g of nano-hydroxyapatite with a particle size of 100nm was dissolved in 100g of deionized water and stirred for 30 minutes to form a 50wt% suspension, which was then mixed with tannic acid aqueous solution at a volume ratio of 1:1 and stirred for 30 minutes. S4. The solutions obtained in S1 and S3 were mixed in a volume ratio of 1:1, shaken for 15 minutes, and then allowed to stand for 30 minutes to obtain a moisture-resistant antibacterial bone adhesive based on porcine collagen.

[0049] Experimental Form Through the comparison of the above data, this method for preparing a moisture-resistant antibacterial bone adhesive based on porcine collagen can improve the adhesion strength in a wet environment.

[0050] Figure 1 :like Figure 1 As shown in the infrared spectrum, the hydroxyl stretching vibration peak of tannic acid is located at 3371 The hydroxyl stretching vibration peak of porcine collagen is located at 3282 , while the hydroxyl stretching vibration peak of bone adhesive shifted to 3361 The characteristic peaks of amide I band, II band and III band in porcine collagen are located at 1635 ,1519cm ,1232cm , and in bone adhesive they shifted to 1604 ,1514cm ,1186cm Similarly, the bone adhesive prepared in this example showed characteristic peaks of PO bonds in hydroxyapatite. These all indicate the successful mixing of tannic acid, porcine collagen, and hydroxyapatite, as well as the existence of internal hydrogen bonding interactions. Figure 2 : Place the bone adhesive between two pieces of beef bone and press for 1 minute; Figure 3 :The shear adhesion strength was measured on the ox bone slice. Before the test, the adhesive was placed on the ox bone slice with an area of 1 . Connect another piece of beef bone to it in an overlapping manner, press for 1 minute, and use a universal material testing machine to test the tensile bond strength in the overlapping shear direction; Figure 4 :The shear adhesion strength was measured on the bovine bone column. Before the test, the adhesive was placed on the bovine bone column with an area of 1 Connect another bovine bone column end-to-end, press for 1 minute, and perform a tensile bond strength test in the end-to-end direction using a universal material testing machine. Figure 5 : The ulna and radius of the rabbit forearm were cut with a bone saw, and then bone adhesive was applied to the fracture ends. The broken segments were aligned and pressed for 1 minute. Figure 6 Staphylococcus aureus was cultured in bacterial culture medium at 37°C for 24 hours. The bacterial suspension was then diluted 1000-fold and co-cultured with bone adhesive (0.2g / ml) at 37°C for 3 hours. Untreated bacteria served as a control group. The bacterial suspension was finally spread on agar medium and incubated at 37°C for 24 hours. The resulting plates were photographed and recorded. Figure 7 : CCK-8 experimental method and live-dead cell staining experimental method, 900 mg of bone cement was sterilized and placed in cell culture medium at 37 ℃ for 24 hours to obtain the leaching solution, rabbit bone marrow mesenchymal stem cells were seeded in the well plate and incubated at 37 ℃ and 5% The cells were cultured in cell culture medium for 24 h, and then the culture medium was replaced with the leaching solution. The cells were cultured in the leaching solution for 1 day and 3 days, and the absorbance of the solution at 450 nm was recorded with a microplate reader. The cell viability was calculated according to the formula. Similarly, the living cells and dead cells were stained separately and imaged with an inverted fluorescence microscope. Figure 8 : A schematic flow chart of a method for preparing a moisture-resistant antibacterial bone adhesive based on porcine collagen in the present invention.

[0051] While embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions, and variations may be made to these embodiments without departing from the principles and spirit of the invention, and that the scope of the invention is defined by the appended claims and their equivalents.

Claims

1. A moisture-resistant antibacterial bone adhesive based on porcine collagen, characterized in that: It includes porcine collagen, tannic acid and nano-hydroxyapatite, with 0.01 to 100 grams of porcine collagen, 0.01 to 100 grams of tannic acid and 0.01 to 100 grams of nano-hydroxyapatite.

2. A method for preparing a moisture-resistant antibacterial bone adhesive based on porcine collagen, characterized in that: The moisture-resistant antibacterial bone adhesive based on porcine collagen according to claim 1 comprises the following steps: S1. Preparation of porcine collagen aqueous solution: dissolving porcine collagen in deionized water to obtain a porcine collagen aqueous solution; S2. Preparation of tannic acid aqueous solution: dissolving tannic acid powder in deionized water to obtain a tannic acid aqueous solution; S3, preparation of tannic acid-nanohydroxyapatite mixed solution: dissolving hydroxyapatite powder in deionized water and adding the solution to the tannic acid aqueous solution obtained in S2 to obtain a tannic acid-nanohydroxyapatite mixed solution; S4, mixing and oscillating molding: the porcine collagen aqueous solution obtained in S1 and the tannic acid-nanohydroxyapatite mixed solution obtained in S3 are mixed to obtain a moisture-resistant antibacterial bone adhesive based on porcine collagen.

3. The method for preparing a moisture-resistant antibacterial bone adhesive based on porcine collagen according to claim 2, characterized in that: The S4 step also includes an ethylene oxide sterilization step, with an ethylene oxide concentration of 600-1000 mg / L, a sterilization temperature of 45-55°C, and a sterilization time of 2-5 hours.

4. The method for preparing a moisture-resistant antibacterial bone adhesive based on porcine collagen according to claim 2, characterized in that: In step S1, the amount of porcine collagen is 0.01 to 100 grams, the amount of deionized water is 0.01 to 100 grams, and the stirring time during dissolution is 20 to 60 minutes.

5. The method for preparing a moisture-resistant antibacterial bone adhesive based on porcine collagen according to claim 2, characterized in that: In step S2, the amount of tannic acid is 0.01 to 100 grams, the amount of deionized water is 0.01 to 100 grams, and the stirring time during dissolution is 10 to 30 minutes.

6. The method for preparing a moisture-resistant antibacterial bone adhesive based on porcine collagen according to claim 2, characterized in that: In step S3, the amount of hydroxyapatite powder is 0.01 to 100 grams, the amount of deionized water is 0.01 to 100 grams, and the stirring time during dissolution is 10 to 30 minutes to form a nano-hydroxyapatite suspension. The volume ratio of the tannic acid aqueous solution obtained in step S2 to the nano-hydroxyapatite suspension is 1:1, and the stirring time during mixing is 15 to 30 minutes.

7. The method for preparing a moisture-resistant antibacterial bone adhesive based on porcine collagen according to claim 2, characterized in that: In step S4, the volume ratio of the porcine collagen aqueous solution obtained in step S1 to the tannic acid-nanohydroxyapatite mixed solution obtained in step S3 is 1:1, the oscillation time during mixing is 5 to 15 minutes, and the standing time after oscillation is 10 to 30 minutes.

8. The method for preparing a moisture-resistant antibacterial bone adhesive based on porcine collagen according to claim 2, characterized in that: The concentration of the porcine collagen aqueous solution obtained in step S1 is 5 to 50 wt %, and the concentration of the tannic acid aqueous solution obtained in step S2 is 5 to 50 wt %.

9. The method for preparing a moisture-resistant antibacterial bone adhesive based on porcine collagen according to claim 2, characterized in that: The concentration of the nano-hydroxyapatite suspension obtained in step S3 is 5-50 wt %, and the particle size of the nano-hydroxyapatite powder is 20-100 nm.

10. The method for preparing a moisture-resistant antibacterial bone adhesive based on porcine collagen according to claim 2, characterized in that: The bone adhesive forms adhesion on the wet bone tissue surface within 1 minute, with an adhesion strength of not less than 0.5 MPa, and an inhibition rate against Staphylococcus aureus of not less than 90%.

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