An oral implant having a surface with a bone-promoting and antibacterial coating and a method of preparation

Abstract

The application discloses an oral implant with a surface provided with an osteogenesis-promoting and antibacterial coating and a preparation method of the oral implant, wherein the coating comprises chitosan and an antibacterial osteogenesis-promoting fusion polypeptide, and the amino acid sequence of the chitosan and the antibacterial osteogenesis-promoting fusion polypeptide is shown as SEQ ID NO. 1 or SEQ ID NO. 2. The application further provides a method for preparing the oral implant by using the coating of the implant. The oral implant has obvious bacteriostatic and osteogenesis-promoting effects, has good biocompatibility, and has a wide application prospect in the field of oral implantation.

Description

Technical Field

[0001] This invention relates to oral implants, and more specifically to an oral implant with a surface coating that promotes bone growth and has an antibacterial coating, and a method for preparing the implant. Background Technology

[0002] In the field of dental implantology, good integration of the implant with the surrounding bone tissue and its ability to resist bacterial infection are key factors in ensuring implant success. Traditional implant surface treatment techniques often focus on improving a single function, making it difficult to simultaneously meet the dual needs of promoting osseointegration and antibacterial properties. Therefore, developing a composite coating with highly efficient osseointegration-promoting and antibacterial properties has significant clinical implications.

[0003] CN119015492A discloses an implant with a composite coating on its surface that promotes bone growth and has antibacterial properties. However, this implant achieves these effects through a nano-Ce-Ta composite material. CN118767213A discloses an implant with a hydrophilic, antibacterial surface coated with a nano-graphene oxide-chitosan-antimicrobial peptide melittin composite coating. The antimicrobial peptide melittin enhances the antimicrobial properties of the implant material, while graphene oxide strengthens the hydrophilicity of chitosan, promoting osteoblast attachment, proliferation, and migration to the implant surface, and promoting osteoblast differentiation, thereby accelerating the bone integration process and enhancing its antimicrobial effect. This invention provides a new approach to exerting antimicrobial and bone-promoting effects through proteins, but its bone-promoting effect is relatively poor, and the antimicrobial peptide melittin is an exogenous protein, which may trigger allergic reactions.

[0004] Currently, there are methods to modify antimicrobial peptides to possess both antibacterial and osteogenic properties (Li Jiamin, Li Yuchen, Ge Zhangjie, et al. Research progress of antimicrobial peptides in dental titanium implant coatings [J]. International Journal of Stomatology, 2024, 51(05): 572-584.). This provides a new approach for manufacturing implants with composite coatings that have good biocompatibility and simultaneously promote osteogenic and antibacterial properties. Summary of the Invention

[0005] Human β-defensin 1 (HBD1), a naturally occurring antimicrobial peptide in the human body, possesses excellent biocompatibility and antimicrobial properties, effectively inhibiting the growth of common oral pathogens. Osteogenic growth peptide (OGP) is a natural polypeptide, a YGFGG pentapeptide, and the derivative sequence that minimizes OGP activity. OGP and OGP10-14 play important roles in osteoblast differentiation, alkaline phosphatase (ALP) expression, and matrix mineralization.

[0006] This invention utilizes structural analysis of naturally occurring antimicrobial peptides in the human body to screen for a series of peptides with antimicrobial activity. These antimicrobial peptides are then linked with existing osteogenic peptides to form an antimicrobial-osteogenic fusion peptide (hereinafter referred to as a fusion peptide) that simultaneously possesses antimicrobial and osteogenic effects. This fusion peptide is used to construct a composite coating, achieving a synergistic effect of antimicrobial and osteogenic integration. On one hand, the antimicrobial peptide in the fusion peptide can rapidly exert its antimicrobial effect in the early stages of implant placement, reducing the risk of bacterial infection and creating a favorable local microenvironment for osseointegration. On the other hand, the osteogenic peptide can promote osteoblast activity, accelerate the formation and repair of peri-implant bone tissue, and improve implant stability. Furthermore, the fusion peptide includes a partial sequence of the naturally occurring antimicrobial peptide β-defensin 1 (HBD1), exhibiting good biocompatibility and lower sensitization.

[0007] This invention provides a composite coating loaded with fusion proteins, which uses micro-arc oxidation and chitosan gel loading technology to fix bioactive substances on the implant surface, providing a new option for the field of dental implants.

[0008] In a first aspect, the present invention provides a dental implant coating, wherein the coating material comprises a 1%-3% by mass chitosan solution and 0.05-0.15 mg / mL of an antibacterial bone-fusion promoting polypeptide, wherein the antibacterial bone-fusion promoting polypeptide comprises at least one polypeptide as shown in SEQ ID NO.1 or as shown in SEQ ID NO.2. Preferably, the mass ratio of the antibacterial bone-fusion promoting polypeptide to the chitosan is 1:15 to 1:25. In a specific embodiment of the present invention, the mass ratio of the antibacterial bone-fusion promoting polypeptide to the chitosan is 1:20.

[0009] Further, the concentration ratio of the antimicrobial bone fusion-promoting peptide shown in SEQ ID NO.1 to the antimicrobial bone fusion-promoting peptide shown in SEQ ID NO.2 is 0:1 to 1:0. Preferably, the concentration ratio of the antimicrobial bone fusion-promoting peptide shown in SEQ ID NO.1 to the antimicrobial bone fusion-promoting peptide shown in SEQ ID NO.2 is ≥1:1.

[0010] In a second aspect, the present invention provides an oral implant comprising the oral implant coating and a metal substrate.

[0011] Furthermore, the metal matrix includes at least one selected from pure titanium matrix, titanium alloy matrix, and zirconium oxide matrix. In a specific embodiment of the present invention, the metal matrix is ​​a pure titanium matrix.

[0012] In a third aspect, the present invention provides a method for preparing the oral implant, the method comprising the following steps:

[0013] S1. Place the surface-activated pure titanium in a silicate electrolyte solution and perform micro-arc oxidation with stainless steel as the cathode and pure titanium substrate as the anode to obtain a titanium substrate;

[0014] S2. Prepare an antibacterial osteofusion-promoting polypeptide solution of 0.05-0.15 mg / mL, and add the antibacterial osteofusion-promoting polypeptide solution dropwise to a 1%-3% chitosan gel solution to obtain the oral implant coating gel.

[0015] S3. The titanium matrix obtained in S1 after micro-arc oxidation pretreatment is immersed in the oral implant coating gel obtained in S2, so that the antibacterial and bone fusion-promoting peptides are loaded onto the surface of the titanium matrix to obtain an oral implant.

[0016] Furthermore, the titanium surface activation step described in step S1 includes at least one of the following operations: cleaning and degreasing, mechanical treatment, and acid pickling activation.

[0017] Furthermore, the cleaning and degreasing process includes the steps of ultrasonic cleaning with acetone and anhydrous ethanol in sequence, or the step of neutralizing residual acidic impurities with an alkaline solution.

[0018] Furthermore, the mechanical treatment includes a step-by-step sanding process using silicon carbide sandpaper.

[0019] Furthermore, the pickling activation includes the step of cleaning pure titanium with a mixed acidic solution, preferably, the mixed acidic solution includes 2% HF (v / v) and 10% HNO3 (v / v).

[0020] Furthermore, the silicate electrolyte solution in step S1 comprises at least 8-12 g / L Na2SiO3, 2-3 g / L NaOH, 5-8 g / L Na3PO4, 3-5 g / L Ca(CH3COO)2, and 1-2 g / L H2O2.

[0021] Furthermore, the micro-arc oxidation parameters obtained in step S1 are a voltage of 350-450V, an oxidation time of 30-60min, and a temperature below 35℃.

[0022] Furthermore, step S2 also includes the step of adding a crosslinking agent to crosslink the antibacterial bone fusion peptide and chitosan. Preferably, the crosslinking agent includes at least one of sodium tripolyphosphate (TPP), genipin, or glutaraldehyde.

[0023] Furthermore, in step S2, the final mass ratio of the antibacterial osteofusion-promoting peptide to chitosan is 1:15-1:25.

[0024] Furthermore, step S3 also includes cross-linking curing, sterilization and preservation steps.

[0025] The beneficial effects of the present invention include, but are not limited to:

[0026] This invention modifies the human β-defensin 1 (HBD1) antimicrobial peptide to form a fusion polypeptide with both antibacterial and osteopromoting effects. Then, the antibacterial and osteopromoting fusion polypeptide is enhanced through synergistic action with chitosan. Because the antibacterial and osteopromoting fusion polypeptide of this invention contains a portion of the naturally occurring human β-defensin 1 (HBD1) sequence, it exhibits good biocompatibility and lower sensitization potential.

[0027] This invention also increases the stability of the antibacterial bone-fusion-promoting peptide by cross-linking it with chitosan, which is beneficial for long-lasting antibacterial and bone-promoting effects.

[0028] The preparation method of this invention is environmentally friendly, the resulting implants integrate quickly with bone and exhibit good antibacterial properties, and have promising prospects for industrial production and clinical application. Attached Figure Description

[0029] The accompanying drawings, which are included to provide a further understanding of the invention and form part of this invention, illustrate exemplary embodiments of the invention and are used to explain the invention, but do not constitute an undue limitation of the invention. In the drawings:

[0030] Figure 1 This is a schematic diagram of the standard curve results for detecting alkaline phosphatase (ALP) in Example 7 of the present invention;

[0031] Figure 2 This is a schematic diagram of the in vitro release results of the fusion polypeptide in the oral implant in Example 8 of the present invention. Detailed Implementation

[0032] The present invention is described in detail below with reference to the embodiments, but the present invention is not limited to these embodiments. Unless otherwise specified, the raw materials and catalysts in the embodiments of the present invention are all purchased through commercial channels.

[0033] Example 1: Preparation of an antibacterial osteogenic fusion polypeptide (hereinafter referred to as fusion polypeptide)

[0034] Using bioinformatics methods, we analyzed the surface charge and other characteristics of the antimicrobial peptide HBD1 protein, screened out a series of antimicrobial peptides, and then added the osteogenic growth peptide YGFGG (studies have shown that YGFGG pentapeptide can play an important role in osteogenic differentiation of osteoblasts, alkaline phosphatase (ALP) expression and matrix mineralization) to form a fusion peptide.

[0035] The sequence of the constructed fusion protein is shown in Table 1.

[0036] Table 1

[0037] name amino acid sequence BH1 SGAICHPVFCPRRYKQIGTYGFGG(SEQ ID NO.1) BH2 YGFGGSGAICHPVFCPRRYKQIGT(SEQ ID NO.2)

[0038] We commissioned a biotechnology company to chemically synthesize fusion peptides BH1 and BH2 for subsequent experiments.

[0039] Example 2: Preparation of Titanium Matrix

[0040] Micro-arc oxidation pretreatment: Dental columnar implants made from commercial medical-grade TA2 pure titanium (Φ3.3mm*6mm) were ultrasonically cleaned sequentially with acetone and anhydrous ethanol for 15 minutes to remove surface grease and organic contaminants. They were then immersed in a 60-80℃ NaOH (50g / L) solution for 10 minutes to further remove grease and activate the surface. The surface was then progressively sanded using 400# to 1200# silicon carbide sandpaper to eliminate scratches and achieve a mirror finish (Ra < 0.1μm). Next, Al2O3 abrasive (50μm particle size) was used for blasting (Ra ≈ 1-2μm) to increase surface roughness and improve film adhesion. Finally, the implants were immersed in a mixed acid solution of HF (2%) and HNO3 (10%) for 30 seconds to remove the oxide layer and form a uniformly activated surface. The implants were immediately rinsed with deionized water and dried with nitrogen. The substrate was then placed in a silicate electrolyte solution containing 10 g / L Na₂SiO₃, 2.5 g / L NaOH, 6 g / L Na₃PO₄, 4 g / L Ca(CH₃COO)₂, and 1.5 g / L H₂O₂. Using stainless steel as the cathode and a pure titanium substrate as the anode, a working voltage of 450 V and a current density of 10 A / dm² were applied. 2 Constant-voltage micro-arc oxidation was performed at a frequency (pulse mode) of 750 Hz for 45 min, with the oxidation temperature controlled below 35℃. Immediately afterwards, the surface was ultrasonically cleaned three times with deionized water to remove residual electrolyte. The surface was then dried at 60℃ for 2 h to obtain a titanium substrate with a porous structure.

[0041] Example 3: Preparation of dental implant coating

[0042] The fusion peptide BH1 obtained in Example 1 was prepared into a 0.1 mg / mL solution. 2% chitosan (degree of deacetylation ≥90%) was dissolved in 1% acetic acid, and the pH was adjusted to 6.5 with PBS. TPP was dissolved in deionized water (concentration 0.5-1 mg / mL). Under magnetic stirring (500 rpm), the TPP solution was slowly added dropwise to the chitosan solution (volume ratio 2:1 to 5:1), and stirring was continued for 30 min to form a milky white suspension. Alternatively, free chitosan was removed by dialysis (deionized water, 24 hours). The fusion peptide BH1 solution was added dropwise to the chitosan gel, and the mixture was slowly stirred for 30 min under ice bath conditions until fully homogeneous. The gel was then allowed to stand at 37°C for 1 hour to form. Free peptides or TPP were removed by dialysis with PBS (pH 6.5) for 48 h. The final mass ratio of peptide to chitosan was 1:20, yielding the BH1 gel for dental implant coating.

[0043] The above experiment was repeated by replacing the actual fusion peptide BH1 with antimicrobial peptide HBD1, fusion peptide BH2, fusion peptides BH1 and BH2 (1:1), and YGFGG pentapeptide to obtain coatings HBD1, BH2, BH1+BH2, and YGFGG respectively.

[0044] Example 4: Preparation of Dental Implants

[0045] The titanium matrix pretreated with micro-arc oxidation in Example 2 was immersed in the dental implant coating gel obtained in Example 3 at a pull-up speed of 1 mm / s, repeated 3 times, and cured by ultraviolet crosslinking (365 nm, 5 min) to load the corresponding peptides onto the matrix surface. The titanium matrix loaded with the corresponding peptides was removed and then cured at 37°C for 1 h to enhance the coating stability. Sterilization was performed by gamma rays (25 kGy). Finally, composite coated implants loaded with fusion peptide BH1, composite coated implants loaded with antimicrobial peptide HBD1, composite coated implants loaded with fusion peptide BH2, composite coated implants loaded with fusion peptides BH1+BH2, and composite coated implants loaded with YGFGG pentapeptide were obtained.

[0046] Example 5 Cytotoxicity Experiment

[0047] After sterilization of the implants by irradiation with 254 nm ultraviolet light, they were dissolved in DMEM culture medium (DMEM medium, 10% fetal bovine serum, 1% penicillin + streptomycin, 50 μg / ml ascorbic acid, 10 mM β-glycerophosphate sodium, 10 nM dexamethasone) at a material / medium ratio of 1 mg / mL. The solutions were filtered through a 0.22 μm filter to obtain implant extracts with concentrations of 1.25, 1, 0.75, 0.5, 0.25, and 0.1 mg / mL, which were stored at 4°C for later use. The viability of human gingival fibroblasts (HGFs) after co-culturing with the implant extracts for 24 h was assessed using the CCK-8 assay. 1 × 10⁶ cells were seeded per well in a 96-well plate. 4 Cells (100 μL / well) were cultured for 24 h until adherence. The experimental group was then cultured in DMEM medium containing different concentrations of extract, while the control group was cultured in DMEM medium without extract. The 96-well plates were incubated at 37°C with 5% (v / v) CO2 for 24 h. 10 μL of CCK-8 reagent was added to each well, and the plates were placed in a cell culture incubator for 1–1.5 h. Cell viability was measured using a microplate reader at 450 nm.

[0048] Calculation formula:

[0049] The results are shown in Table 2. The test results show that when the mass concentration of the implant extract is 1.25 mg / mL, the cell viability of human gingival fibroblasts (HGFs) after co-incubation with the implant extract for 24 h is maintained above 80%, which meets the requirement of no cytotoxicity (≥70%).

[0050] Table 2

[0051] Concentration (mg / mL) 0.1 0.25 0.5 0.75 1 1.25 BH1 implant group (%) 109 115 106 98 90 84 BH2 implant group (%) 112 119 109 102 92 87 BH1+BH2 implant group (%) 110 117 108 99 90 85

[0052] Example 6: In vitro antibacterial performance evaluation

[0053] The experimental groups were set up as follows: blank control group (blank drug sensitivity tablet), HBD1 implant group, BH1 implant group, and BH2 implant group. All implants in the three groups were cut into small round pieces with a diameter of 5 mm, the same size as the drug sensitivity tablets, and then sterilized.

[0054] Staphylococcus aureus: In a clean bench, single-clone Staphylococcus aureus colonies were picked from colonies isolated from agar plates and inoculated into nutrient agar medium. The culture conditions were 37°C for 16 hours, after which a bacterial suspension was prepared. The optical density (OD) of the suspension was measured at 600 nm using a UV spectrophotometer, and the suspension was diluted to the appropriate OD value using nutrient agar medium. 600 The value was approximately 0.6. Next, 100 μL of bacterial suspension was evenly spread in the center of an agar plate. Then, each group of supports was placed in the center of the agar plate and incubated at 37°C for 48 hours. Finally, the diameter of the inhibition zone formed around the sample was measured and recorded to evaluate its antibacterial effect.

[0055] Candida albicans: The culture medium is Sabouraud agar; the culture conditions are to place the petri dishes in a constant temperature incubator at 28°C for 16 hours, and other steps are the same as for Staphylococcus aureus.

[0056] Porphyromonas gingivalis: The culture medium was pre-reduced BHI blood agar (containing 5% defibrinated sheep blood + heme chloride / vitamin K1); the culture conditions were to place the culture dish in an anaerobic incubator at 37°C for 80 hours, and the other steps were the same as for Staphylococcus aureus.

[0057] The results for each group are shown in Table 3.

[0058] Table 3

[0059]

[0060] As shown in Table 3, compared with the single chitosan implant group, the antibacterial effects of BH1 implant, BH2 implant, and BH1+BH2 implant groups were significantly increased. The antibacterial effects of the fusion peptides BH1 and BH2 were not much different from those of HBD1, but the antibacterial effect of the fusion peptide BH1 was better.

[0061] Example 7 promotes osteoporosis detection

[0062] Alkaline phosphatase (ALP) activity is an important marker of osteogenic differentiation, and the osteogenic capacity of MC3T3-E1 cells can be assessed by measuring ALP activity.

[0063] Preparation of extracts from implants in each group: 1cm 2 Plant surface area / 1ml culture medium, extracted at 37℃ and 5% CO2 for 24h, and sterilized by filtration through a 0.22μm filter membrane.

[0064] The MC3T3-E1 subclonal 4 pre-osteoblast cell line was seeded into 96-well plates at a density of 4 × 10⁶ cells per well. 4 When the cells reached 70% confluence, the culture medium was changed to osteogenic induction medium (DMEM medium, 10% fetal bovine serum, 1% penicillin + streptomycin, 50 μg / ml ascorbic acid, 10 mM β-glycerophosphate sodium, 10 nM dexamethasone). The prepared extract was co-incubated with the cells for 21 days (replacing with fresh induction medium every 48 h). Osteoblast activity was assessed using a CCK-8 assay, and absorbance at 450 nm was measured using a microplate reader. The calculation formula is as follows:

[0065]

[0066] The results are shown in Table 4.

[0067] Table 4

[0068] Group Cell viability (%) Blank group (culture medium + blank antimicrobial susceptibility testing tablets only) —— Negative control group (normal cells + culture medium + blank antimicrobial susceptibility test strip) 100％ YGFGG implant group (positive control) 148％ Chitosan implants 106％ BH1 implant group 131％ BH2 implant group 141％ BH1+BH2 implant group 138％

[0069] The alkaline phosphatase quantitative kit (enzyme-linked immunosorbent assay) (purchased from Shanghai Beyotime Biotechnology Co., Ltd.) was used for detection. The absorbance was measured at 405 nm using an enzyme-linked immunosorbent assay reader according to the product instructions. The absorbance values ​​were then converted to ALP activity. The standard curve parameters are as follows: Figure 1 As shown in Table 5, the results are as follows.

[0070] Table 5

[0071] Grouping ALP activity (mU / mg) Blank drug sensitivity test strip (negative control) 50.48±4.14 YGFGG implant group (positive control) 215±6.98 Chitosan implants 116±3.37 BH1 implant group 168.87±5.47 BH2 implant group 183±7.29 BH1+BH2 implant group 176±6.73

[0072] As shown in Tables 4 and 5, compared with the chitosan group, BH1 implant, BH2 implant, and BH1+BH2 implant can significantly promote osteoblast proliferation and effectively increase ALP activity, proving that the fusion peptides BH1 and BH2 have osteogenic effects, with the fusion peptide BH2 showing better osteogenic effects.

[0073] Example 8: In vitro release experiment of fusion peptide

[0074] Oral implants BH1, BH2, and BH1+BH2 were placed in dialysis bags (MWCO 14kDa), sealed, and immersed in 50mL of PBS solution (pH 7.4). The solution was incubated at a constant temperature with shaking (37℃, 100rpm) to prevent photolysis of the peptides. Alternatively, samples were directly immersed in PBS solution (pH 7.4), and 1mL samples were collected at time points of 0.5, 1, 2, 4, 8, 12, 16, 20, 24, 36, 48, 60, 72, 84, 96, 120, 144, and 168 hours. The samples were centrifuged (12,000rpm, 5min), and the supernatant was collected, with 1mL of PBS solution added simultaneously. The content of released peptides was detected using UV spectrophotometry, and the release rate of the fusion peptides was calculated.

[0075] Ultraviolet method: Detection of the absorption peak of tyrosine at 280 nm of the fusion peptide.

[0076] Test results as follows Figure 2 As shown, by Figure 2 It is known that the fusion peptides can be rapidly released in a short period of time (up to 40% within 24 hours), which is conducive to the rapid exertion of antibacterial and bone-promoting effects. The stable release of about 80% within 7 days is conducive to long-term antibacterial and bone-promoting effects, providing a good antibacterial and bone-promoting environment for dental implants.

[0077] The above description is merely an embodiment of the present invention, and the scope of protection of the present invention is not limited to these specific embodiments, but is determined by the claims of the present invention. For those skilled in the art, the present invention can have various modifications and variations. Any modifications, equivalent substitutions, improvements, etc., made within the technical concept and principle of the present invention should be included within the scope of protection of the present invention.

Claims

1. A coating for dental implants, characterized in that, The coating material comprises 1%-3% by mass chitosan solution and 0.05-0.15 mg / mL antibacterial osteofusion-promoting peptide, wherein the antibacterial osteofusion-promoting peptide comprises at least one peptide as shown in SEQ ID NO.1 or as shown in SEQ ID NO.2, and the mass ratio of the antibacterial osteofusion-promoting peptide to the chitosan is 1:15 to 1:

25.

2. The dental implant coating according to claim 1, characterized in that, The concentration ratio of the antibacterial osteofusion-promoting peptide shown in SEQ ID NO.1 to the antibacterial osteofusion-promoting peptide shown in SEQ ID NO.2 is 0:1 to 1:

0.

3. The dental implant coating according to claim 1, characterized in that, The concentration ratio of the antibacterial osteofusion-promoting peptide shown in SEQ ID NO.1 to the antibacterial osteofusion-promoting peptide shown in SEQ ID NO.2 is ≥1:

1.

4. A dental implant, characterized in that, The dental implant comprises the dental implant coating and metal substrate as described in any one of claims 1-3.

5. The dental implant according to claim 4, characterized in that, The metal matrix includes at least one of pure titanium matrix, titanium alloy matrix, and zirconium oxide matrix.

6. A method for preparing the oral implant according to any one of claims 4 or 5, characterized in that, The method includes the following steps: S1. Place the surface-activated pure titanium in a silicate electrolyte solution and perform micro-arc oxidation with stainless steel as the cathode and pure titanium substrate as the anode to obtain a titanium substrate; S2. Prepare an antibacterial osteofusion-promoting polypeptide solution of 0.05~0.15mg / mL, and add the antibacterial osteofusion-promoting polypeptide solution dropwise to a 1%~3% chitosan gel solution to obtain the oral implant coating gel according to any one of claims 1-3. S3. The titanium matrix obtained in S1 after micro-arc oxidation pretreatment is immersed in the oral implant coating gel obtained in S2, so that the antibacterial and bone fusion-promoting peptides are loaded onto the surface of the titanium matrix to obtain an oral implant.

7. The method according to claim 6, characterized in that, The surface activation step described in step S1 includes at least the steps of cleaning and degreasing, mechanical treatment, and acid pickling activation.

8. The method according to claim 6, characterized in that, The silicate electrolyte solution in step S1 comprises at least 8-12 g / L of Na2SiO3, 2-3 g / L of NaOH, 5-8 g / L of Na3PO4, 3-5 g / L of Ca(CH3COO)2, and 1-2 g / L of H2O2.

9. The method according to claim 6, characterized in that, The micro-arc oxidation parameters described in step S1 are: voltage 350-450V, oxidation time 30-60min, and temperature below 35℃.

10. The method according to claim 6, characterized in that, Step S2 also includes the step of adding a cross-linking agent to cross-link the antibacterial bone fusion peptide and chitosan.

11. The method according to claim 10, characterized in that, The crosslinking agent includes at least one of sodium tripolyphosphate (TPP), genipin, or glutaraldehyde.

12. The method according to claim 6, characterized in that, In step S2, the final mass ratio of the antibacterial osteofusion-promoting peptide to chitosan is 1:15-1:

25.

13. The method according to claim 6, characterized in that, Step S3 also includes cross-linking curing, sterilization and preservation steps.

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