A method for preparing a biofunctionalized polyether ether ketone implant and uses thereof

Abstract

The application provides a preparation method and application of a biofunctionalized polyether ether ketone implant, wherein polyether ether ketone is subjected to surface modification through strong acid treatment, and is combined with strontium-doped bioactive glass by using a sol-gel method, so as to endow the polyether ether ketone with the ability of promoting osteogenic differentiation of bone marrow mesenchymal stem cells and inhibiting osteoclastic differentiation of mononuclear macrophages. Performance test results show that the contact angle of the acidized and surface-modified PEEK is reduced to different levels, which indicates that the hydrophilicity of the PEEK is significantly improved. At the same time, the proliferation and growth activity of bone marrow mesenchymal stem cells on the surface of the Sr-SPK are also better than before the surface modification. Differentiation test results show that, compared with the PEEK, the Sr-SPK can obviously promote the osteogenic differentiation of rat bone marrow mesenchymal stem cells, can obviously inhibit the differentiation of rat mononuclear macrophages into osteoclasts, and can obviously inhibit the formation of osteoclasts, and the biofunctionality is better.

Description

Technical Field

[0001] This invention belongs to the field of medical materials technology, and relates to an orthopedic implant, specifically to a method for preparing and applying a biofunctionalized polyether ether ketone implant. Background Technology

[0002] A perfect orthopedic implant should avoid complications, and the key to effectively avoiding complications lies in the osseointegration capacity of the implant. Goodman (Goodman SB, Biomaterials 2013, 34(13):3174-3183) proposed that the future focus of orthopedic bio-coated implants is to improve osseointegration; Han (Biomaterials, 2010, 31(13):p.3465-70.) believes that excellent orthopedic prostheses should not only have the potential to promote bone tissue growth, but also have good biocompatibility. In summary, the functional modification of the surface of orthopedic implants needs to improve the osseointegration capacity of the implants.

[0003] Currently, most in vitro experiments on orthopedic prostheses are limited to studies on promoting osteogenic properties, with very few studies on implants that also inhibit osteoclastogenesis. Some modified orthopedic prostheses show good osteogenic effects in vitro, but their in vivo osseointegration is less than ideal, mainly because it is difficult to simulate an in vivo environment in vitro. Current studies evaluating the osseointegration of implant materials primarily use single-factor culture models of materials and bone marrow mesenchymal stem cells or osteoblasts stimulated by osteogenic induction fluid, evaluating them based on osteogenic differentiation indicators (expression of osteogenic-related genes such as alkaline phosphatase, alizarin red, and bone morphogenetic protein-2), without considering the ability to inhibit osteoclastogenesis. The performance of implants in the host is largely influenced by both osteogenic and osteoclastogenesis-inhibiting effects, namely, osteogenic differentiation of bone marrow mesenchymal stem cells and osteoclastogenesis of mononuclear macrophages.

[0004] Polyetheretherketone (PEEK) is a semi-crystalline linear polycyclic aromatic thermoplastic, first developed by British scientists in 1978 (Ma, Int J Mol Sci. 2014; 15(4):5426-5445.). The molecular structure of PEEK is based on aromatic molecules with ketone and ester functional groups linked between the aromatic rings. This unique chemical structure gives PEEK stable physical and chemical properties, such as high temperature resistance, wear resistance, flame resistance, hydrolysis resistance, and chemical resistance (Luo, J Mech Behav Biomed Mater. 2014; 29:103-113, 12-15; Kurtz SM, Biomaterials. 2007; 28(32):4845-4869.). PEEK has good in vitro and in vivo biocompatibility, is non-toxic, non-teratogenic, non-mutagenic, and non-carcinogenic, and is radiolucent without producing artifacts on magnetic resonance imaging. Currently, PEEK and its composites have become a research hotspot for materials scientists and orthopedic experts, and have been applied clinically in interbody fusion devices and artificial joint prostheses with good short-term follow-up results. However, PEEK is bioinert and hydrophobic, lacks bioactivity, and cannot integrate well with the host bone after implantation, which greatly limits its widespread clinical application.

[0005] Therefore, surface modification of PEEK biomaterials should aim to enhance their osteointegration capabilities. However, most functional materials currently used in orthopedics only promote osteogenic differentiation, with few considering their crucial role in inhibiting osteoclastogenesis. Strontium ions have a positive promoting effect on osteogenesis and are frequently used clinically in the form of strontium ranitidine. Bioactive glass, due to its excellent biocompatibility, is currently used as an implant material for treating various orthopedic and other diseases. This invention prepared strontium-doped bioactive glass using the sol-gel method and immobilized it on a PEEK surface to evaluate the effect of biofunctionalized polyetheretherketone implants on osteointegration. Summary of the Invention

[0006] This invention addresses the aforementioned problems by providing a method for preparing biofunctionalized polyether ether ketone implants to solve the bioinertness issue of PEEK materials and to endow them with various biological functions through surface modification.

[0007] To achieve the above objectives, the technical solution adopted by the present invention is summarized as follows: Polyetheretherketone is surface modified by strong acid treatment, and then combined with strontium-doped bioactive glass using the sol-gel method, thereby endowing it with the ability to promote osteogenic differentiation of bone marrow mesenchymal stem cells while inhibiting osteoclast differentiation of monocytes and macrophages.

[0008] Specifically, the preparation method of the biofunctionalized polyether ether ketone implant provided by the present invention includes the following steps:

[0009] A. Concentrated sulfuric acid sulfonated PEEK

[0010] After cleaning and pretreatment, PEEK was immersed in concentrated sulfuric acid solution, ultrasonically irradiated at 25°C for a few minutes, and then washed with deionized water until sulfuric acid residue was removed. All materials were subjected to hydrothermal treatment at 120°C in an autoclave lined with inert material, and then cooled to 25°C to obtain PEEK sulfonated with concentrated sulfuric acid.

[0011] B. Preparation of Strontium Ion-Containing Mesoporous Bioglass Nanospheres

[0012] A modified microemulsion-assisted sol-gel method was used to synthesize CATB: Cetylammonium bromide was dissolved in deionized water and rapidly stirred in a water bath at 55-60°C until completely dissolved; then ethyl acetate was added, and the mixture was stirred again at 55-60°C for a certain period. After stirring, 1 mol / L ammonia was added, followed by stirring again, and then tetraethyl orthosilicate (TEOS) was added and stirred. TEOS and strontium nitrate tetrahydrate were added sequentially every 30 minutes. The resulting colloid was centrifuged at 8000 g and then washed sequentially with deionized water and anhydrous ethanol. The washed precipitate was placed in a dry environment at 65°C overnight and then calcined at 750°C to obtain strontium-containing mesoporous bioglass nanospheres (Sr-MBG).

[0013] C. Preparation of biofunctionalized polyetheretherketone Sr-SPK

[0014] The Sr-MBG from step B was placed in a chitosan solution, ultrasonicated in a water bath, and then coated onto the surface of PEEK sulfonated with concentrated sulfuric acid. The mixture was then dried overnight at 60°C to obtain biofunctionalized polyether ether ketone Sr-SPK.

[0015] The specific process conditions for the above three steps are as follows:

[0016] In step A, the PEEK cleaning pretreatment steps are as follows: First, the sample is ultrasonically cleaned with ethanol for 30 minutes, then cleaned two to three times with deionized water, and dried overnight at 37°C; after cleaning pretreatment, the PEEK is immersed in a concentrated sulfuric acid solution with a concentration of 98% to 99%, ultrasonically irradiated at 25°C for 5 minutes, and then washed with deionized water at least three times until sulfuric acid residue is removed; all materials are subjected to hydrothermal treatment at 120°C for 12 hours in an autoclave lined with polytetrafluoroethylene, and then cooled to 25°C to obtain concentrated sulfuric acid sulfonated PEEK.

[0017] In step B, the concentration of hexadecylammonium bromide is 0.05–0.06 mol / L, the volume ratio of deionized water, ethyl acetate, ammonia, and TEOS is 165:50:35:18, and the mass ratio of strontium nitrate tetrahydrate to hexadecylammonium bromide is 9:14. The stirring time after adding ethyl acetate is 30 min, the stirring time after adding ammonia is 15 min, and the stirring time after adding TEOS is 30 min. The colloid is centrifuged at 8000 g for 10 min.

[0018] In step C, the chitosan solution is prepared as follows: add 2 times the volume of acetic acid to distilled water and adjust to an acidic environment, then add chitosan powder, and stir and mix with a magnetic stirrer to obtain a chitosan solution. The weight ratio of chitosan to the volume of distilled water is 2:1.

[0019] In another aspect, the present invention provides a biofunctionalized polyether ether ketone implant, which is prepared by any of the methods described above.

[0020] By adjusting the amount of Sr-MBG added to the chitosan solution, different concentrations of Sr-SPK can be obtained. This invention prepared Sr-SPK at a concentration of 0.1 and 0.5 times the concentration.

[0021] Performance test results showed that ordinary PEEK had the smoothest surface. The contact angle of acidified and surface-modified PEEK decreased to different levels, with the 0.5 times concentration of Sr-SPK showing the best contact angle, indicating that the treatment significantly improved the hydrophilicity of PEEK. At the same time, the proliferation and growth activity of bone marrow mesenchymal stem cells on the Sr-SPK surface were also better than before surface modification.

[0022] Differentiation assay results showed that, compared with PEEK, Sr-SPK significantly promoted osteogenic differentiation of rat bone marrow mesenchymal stem cells, while significantly inhibiting the differentiation of rat monocytes and macrophages into osteoclasts and significantly suppressing osteoclast formation, demonstrating better biological functionality.

[0023] Therefore, in a third aspect, the present invention provides the application of the aforementioned biofunctionalized polyetheretherketone implant in the preparation of orthopedic implants. Orthopedic implants prepared from this material possess both the ability to promote osteogenic differentiation and the ability to inhibit osteoclast differentiation.

[0024] The role and effect of invention

[0025] This invention provides a bioactive modification for PEEK materials. First, its surface morphology is altered through strong acidification. Then, it is combined with strontium-containing mesoporous bioglass nanospheres via a sol-gel method, exhibiting the ability to promote osteogenic differentiation of bone marrow mesenchymal stem cells and inhibit osteoclast differentiation of monocytes and macrophages. This method functionalizes bioinert PEEK materials, potentially promoting osseointegration in PEEK orthopedic implants, significantly improving the success rate of PEEK implants in orthopedic surgery, and substantially reducing the probability of postoperative complications. Attached Figure Description

[0026] Figure 1 For the surface morphology analysis and water contact angle measurement of each group of PEEK materials;

[0027] Figure 2 Cell proliferation experiments at 1d, 3d, and 5d showed that Sr-SPK promoted cell proliferation at all time points.

[0028] Figure 3 CLSM observation of the extension of rat bone marrow mesenchymal stem cells on different material surfaces;

[0029] Figure 4 The effects of acidified and surface-modified PEEK at 7D and 14D on osteogenic differentiation of rat bone marrow mesenchymal stem cells;

[0030] Figure 5 Alizarin Red staining of extracellular matrix on the surface of different materials at 14D and 21D;

[0031] Figure 6 Comparison of expression levels of osteogenic differentiation-related genes (ALP, BMP2, OCN, OPN, RUNX2, COL-1) on the surface of each group of materials;

[0032] Figure 7 Comparison of expression levels of osteoclast differentiation-related genes (TRAP, c-FOS, CTSK, VATPs-d2, VATPs-a3, NFATc1) on the surface of materials from different groups;

[0033] Figure 8 Comparison of tartrate-resistant acid phosphatase staining results on the surfaces of materials from different groups. Detailed Implementation

[0034] The present invention will now be described in detail with reference to embodiments and accompanying drawings. However, the following embodiments should not be construed as limiting the scope of the present invention.

[0035] I. Preparation of Biofunctionalized Polyetheretherketone Implants (Sr-SPK)

[0036] A. Concentrated sulfuric acid sulfonated PEEK

[0037] Before using PEEK, the samples were ultrasonically cleaned with ethanol for 30 minutes, then rinsed two to three times with deionized water and dried overnight at 37°C (similar to body temperature). To form a three-dimensional porous structure on the surface, pure PEEK was immersed in a concentrated sulfuric acid (98%-99%) solution, ultrasonically irradiated at 25°C for 5 minutes, and then washed with deionized water at least three times. To remove sulfuric acid residue, all materials were hydrothermally treated in a 100 mL PTFE-lined autoclave at 120°C for 12 hours, and then cooled to 25°C to obtain sample SPK.

[0038] B. Synthesis of Strontium-containing mesoporous bioglass nanospheres (Sr-MBG)

[0039] A modified microemulsion-assisted sol-gel method was used for synthesis. The specific experimental method is as follows: First, 1.4 g of cetylammonium bromide (CTAB) was dissolved in 66 mL of deionized water and rapidly stirred in a water bath at 55-60 °C until CATB was completely dissolved. Then, 20 mL of ethyl acetate was added, and the mixture was stirred again at approximately 60 °C. After stirring for about 30 minutes, 14 mL of 1 mol / L ammonia solution was added, and the mixture was stirred again for 15 minutes. Then, 7.2 mL of tetraethyl orthosilicate (TEOS) was added and stirred for 30 minutes. TEOS and strontium nitrate tetrahydrate were added sequentially every 30 minutes. The resulting colloid was centrifuged at 8000 g for about 10 minutes, and then washed sequentially with deionized water and anhydrous ethanol (99%). The washed precipitate was placed in a dry environment at 65 °C overnight, and then calcined at 750 °C to obtain the sample, which is Sr-MBG.

[0040] C. Preparation of Sr-SPK

[0041] To synthesize Sr-MBG coated with chitosan, a chitosan solution was first prepared. The specific steps were as follows: 100 μL of acetic acid was added to 50 mL of distilled water to adjust the pH to acidic. Then, 25 mg of chitosan powder was added, and the mixture was stirred with a magnetic stirrer for approximately 15 minutes to obtain a chitosan solution. 5 mg of Sr-MBG was added to the chitosan solution, and after sonication in a water bath for 20 minutes, it was coated onto the surface of PEEK sulfonated with concentrated sulfuric acid. The resulting Sr-SPK was dried overnight in a desiccator at 60 °C to obtain a 0.1-fold concentration of Sr-SPK. A 0.5-fold concentration of Sr-SPK was prepared using the same method.

[0042] II. Performance Testing

[0043] 1. Morphological characterization and hydrophilicity measurement

[0044] Polyether ether ketone (PEEK) was biofunctionalized by strong acid treatment and sol-gel method, and then characterized by scanning electron microscopy.

[0045] Characterization of the surface morphology of PEEK after biofunctionalization. The experiment was divided into 4 groups: PEEK (ordinary PEEK), SPK (sulfonated biological PEEK with concentrated sulfuric acid), 0.1-Sr-SPK (0.1-fold concentration of strontium-doped bioactive glass surface modified PEEK), and 0.5-Sr-SPK (0.5-fold concentration of strontium-doped bioactive glass surface modified PEEK).

[0046] Each group of PEEK samples was sterilized and placed in 24-well plates with three replicates per group. For SEM characterization, the PEEK samples were fixed overnight with 2.5% glutaraldehyde. Each PEEK sample was dehydrated using a series of fractionated ethanol solutions (30%, 50%, 70%, 80%, 90%, 100%, and 100%). The PEEK samples were then freeze-dried, coated with platinum by sputtering, and observed using a scanning electron microscope (JEOL JSM-6700F, Japan).

[0047] Measurement of water contact angle of PEEK after biofunctionalization treatment. The static contact angle was measured on PEEK and biofunctionalized PEEK by titration with 2 mL of double-distilled water at room temperature.

[0048] The results are as follows Figure 1 As shown, comparing Bare PEEK (ordinary PEEK), SPK (sulfonated PEEK), 0.1-Sr-SPK (0.1 times concentration of strontium-doped bioactive glass surface-modified PEEK), and 0.5-Sr-SPK (0.5 times concentration of strontium-doped bioactive glass surface-modified PEEK), the surface morphologies of all PEEK materials are slightly different. Ordinary PEEK has the smoothest surface and the largest contact angle. The roughness of acidified and surface-modified PEEK gradually increases, while the contact angle gradually decreases to different levels, indicating that the treatment significantly improves the hydrophilicity of PEEK.

[0049] 2. Study on adhesion and proliferation of bone marrow mesenchymal stem cells

[0050] The experiment was divided into four groups: PEEK (ordinary PEEK), SPK (concentrated sulfuric acid sulfonated bio-PEEK), 0.1-Sr-SPK (0.1 times concentration of strontium-doped bioactive glass surface modified PEEK), and 0.5-Sr-SPK (0.5 times concentration of strontium-doped bioactive glass surface modified PEEK).

[0051] After UV sterilization, each group of PEEK cells was placed in 24-well plates with three replicates per group. Third-generation rat bone marrow mesenchymal stem cells (rBMSCs) were seeded onto the PEEK cells in each group, with cell density controlled at 5 × 10⁻⁶ cells / well. 4 / ml), 1ml per well. Each PEEK sample was washed with PBS at 1, 3, and 5 days post-inoculation. Cell adhesion and proliferation were studied using the CCK-8 assay at predetermined time points.

[0052] The results of proliferation are as follows Figure 2 As shown, SPK has no promoting effect on cell proliferation and even has a slight inhibitory effect. Sr-SPK promotes cell proliferation at all time points, but only on day 1, 0.5-Sr-SPK has a better promoting effect on cell proliferation than 0.1-Sr-SPK. On day 3 and day 5, there is no significant difference in the cell proliferation promoting effect between the two.

[0053] Figure 3 CLSM observations show the extension of rat bone marrow mesenchymal stem cells on different material surfaces. Compared to PEEK, cells on Sr-SPK showed more active growth and greater spread.

[0054] 3. Research on osteogenic differentiation of bone marrow mesenchymal stem cells

[0055] Biological properties of PEEK cells before and after biofunctionalization (promoting osteogenic differentiation and inhibiting osteoclast differentiation) were investigated. Rat bone marrow mesenchymal stem cells were used in the osteogenic differentiation promotion experiment, but other rat adult stem cells with osteogenic differentiation potential were also suitable for this experiment. Rat mononuclear macrophages were used in the osteoclast differentiation inhibition experiment.

[0056] The experiment was divided into four groups: PEEK (ordinary PEEK), SPK (concentrated sulfuric acid sulfonated biological PEEK), 0.1-Sr-SPK (0.1 times concentration of strontium-doped bioactive glass surface modified PEEK), and 0.5-Sr-SPK (0.5 times concentration of strontium-doped bioactive glass surface modified PEEK).

[0057] After sterilization, each group of PEEK cells was placed in a 24-well plate with three replicates per group. Third-generation rat bone marrow mesenchymal stem cells (rBMMSCs) were seeded onto the PEEK cells in each group, with cell density controlled at 5 × 10⁻⁶ cells / well. 4 / ml), 1ml per well. Cells were cultured at 37℃ and 5% CO2 for 24 hours until adherence. The culture medium was then replaced with α-MEM medium containing osteogenic induction solution (osteogenic induction solution: 10% fetal bovine serum culture medium with 50μM vitamin C, 10mM β-glycerophosphate sodium, and 100nM dexamethasone). The medium was changed every two days thereafter. Alkaline phosphatase staining was performed at 7 and 14 days to evaluate early differentiation of rat bone marrow mesenchymal stem cells into osteoblasts. RNA was extracted on day 7 and reverse transcribed for qPCR quantitative detection of osteogenic-related gene expression. Alizarin red staining was performed at 14 and 21 days to evaluate extracellular matrix mineralization, i.e., markers of late osteogenic differentiation.

[0058] Alkaline phosphatase staining: After co-culturing PEEK materials with rBMMSCs for 7 days, osteogenic medium was removed, and PEEK materials from each group were taken out. ALP staining was performed according to the ALP staining kit (Beyotime) instructions: After washing three times with PBS, fixation was performed at 4°C for 30 seconds; after washing three times with PBS again, working solution was added and the mixture was placed in a 37°C water bath for 45 minutes; finally, the working solution was removed, the mixture was rinsed three times with PBS, dried, and photographed.

[0059] Alizarin Red Staining: The experiment was also divided into four groups and placed in 24-well plates, with three replicates per well. Alizarin Red staining was used to observe the mineralization of the extracellular matrix of osteoblasts. Culture was terminated at 14 and 21 days. The culture medium was aspirated, and the cells were gently washed three times with PBS, fixed with 95% ethanol for 10 min, and then washed three more times. 0.1% Alizarin Red solution was added to 48-well plates and incubated at 37°C for 45 min. Excess staining was removed by repeated gentle washing with double-distilled water, and the plates were finally air-dried and photographed. The formation of mineralized nodules in the acidified PEEK groups at each time point was higher than that in the non-acidified PEEK groups.

[0060] Figure 4 The effects of 7D and 14D acidified and surface-modified PEEK on alkaline phosphatase (ALP) production and activity during osteogenic differentiation of rat bone marrow mesenchymal stem cells were demonstrated. Results showed that Sr-SPK produced a deeper ALP staining compared to PEEK.

[0061] Figure 5 Alizarin Red staining of the extracellular matrix was shown for 14D and 21D cells. Results showed that Sr-SPK stained more deeply with ARS compared to PEEK.

[0062] Figure 6 The expression levels of osteogenic differentiation-related genes (ALP, BMP2, OCN, OPN, RUNX2, COL-1) in different groups of materials were shown. The results showed that Sr-SPK significantly promoted osteogenic differentiation of rat bone marrow mesenchymal stem cells compared with PEEK.

[0063] 4. Study on osteoclast differentiation of bone marrow macrophages

[0064] The experiment was divided into four groups: PEEK (ordinary PEEK), SPK (concentrated sulfuric acid sulfonated bio-PEEK), 0.1-Sr-SPK (0.1 times concentration of strontium-doped bioactive glass surface modified PEEK), and 0.5-Sr-SPK (0.5 times concentration of strontium-doped bioactive glass surface modified PEEK).

[0065] After sterilization, each group of PEEK cells was placed in 24-well plates with three replicates per group. Bone marrow macrophages (BMMs) were seeded onto the PEEK cells and cultured at 37°C and 5% CO2 for 24 hours until cell adhesion was achieved. The culture medium was then replaced with conditioned medium supplemented with M-CSF (50 ng / mL) and RANKL (50 ng / mL), and the medium was changed every 48 hours. On day 5, cellular RNA was extracted and reverse transcribed for qPCR to quantitatively detect the expression of osteoclast-related genes. Tartrate-resistant acid phosphatase staining was performed after multinucleated osteoclasts appeared.

[0066] Tartrate-resistant acid phosphatase (TRAP) staining: After bone marrow macrophages (BMMs) adhered to the culture medium overnight, the medium was replaced with conditioned medium supplemented with M-CSF (50 ng / mL) and RANKL (50 ng / mL). The medium was changed every 48 h until multinucleated osteoclasts (three or more nuclei) appeared. Cells were washed in PBS, fixed in PBS with 4% paraformaldehyde for 15 min, and then stained for tartrate-resistant acid phosphatase (TRAP).

[0067] Figure 7 The expression levels of osteoclast differentiation-related genes (TRAP, c-FOS, CTSK, VATPs-d2, VATPs-a3, NFATc1) on different material surfaces were displayed. The results showed that Sr-SPK significantly inhibited the differentiation of rat monocytes / macrophages into osteoclasts compared to PEEK. Tartrate-resistant acid phosphatase staining results showed that Sr-SPK significantly inhibited osteoclast formation compared to PEEK. Figure 8 ).

[0068] The foregoing has shown and described the basic principles, main features, and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited to the above embodiments. The embodiments and descriptions in the specification are merely illustrative of the principles of the invention. Various changes and modifications can be made to the invention without departing from its spirit and scope, and all such changes and modifications fall within the scope of the present invention as claimed. The scope of protection of this invention is defined by the appended claims and their equivalents.

Claims

1. A method for preparing a biofunctionalized polyetheretherketone implant, characterized in that, Includes the following steps: A. Concentrated sulfuric acid sulfonated PEEK After cleaning and pretreatment, PEEK was immersed in concentrated sulfuric acid solution, ultrasonically irradiated at 25°C for a few minutes, and then washed with deionized water until sulfuric acid residue was removed. All materials were subjected to hydrothermal treatment at 120°C in an autoclave lined with inert material, and then cooled to 25°C to obtain PEEK sulfonated with concentrated sulfuric acid. B. Preparation of Strontium Ion-Containing Mesoporous Bioglass Nanospheres A modified microemulsion-assisted sol-gel method was used to synthesize CATB: Cetylammonium bromide was dissolved in deionized water and rapidly stirred in a water bath at 55-60°C until completely dissolved; then ethyl acetate was added, and the mixture was stirred again at 55-60°C for a certain period. After stirring, 1 mol / L ammonia was added, followed by stirring again, and then tetraethyl orthosilicate (TEOS) was added and stirred. TEOS and strontium nitrate tetrahydrate were added sequentially every 30 minutes. The resulting colloid was centrifuged at 8000 g and then washed sequentially with deionized water and anhydrous ethanol. The washed precipitate was placed in a dry environment at 65°C overnight, and then calcined at 750°C to obtain strontium-containing mesoporous bioglass nanospheres (Sr-MBG). The concentration of hexadecylammonium bromide was 0.05~0.06 mol / L, the volume ratio of deionized water, ethyl acetate, ammonia, and TEOS was 165:50:35:18, and the mass ratio of strontium nitrate tetrahydrate to hexadecylammonium bromide was 9:

14. The stirring time was 30 min after adding ethyl acetate, 15 min after adding ammonia, and 30 min after adding TEOS. The colloid was centrifuged at 8000g for 10 minutes; C. Preparation of biofunctionalized polyetheretherketone Sr-SPK The Sr-MBG from step B was placed in a chitosan solution, ultrasonicated in a water bath, coated onto the surface of PEEK sulfonated with concentrated sulfuric acid, and dried overnight at 60°C to obtain biofunctionalized polyether ether ketone Sr-SPK. In step A, the PEEK cleaning pretreatment steps are as follows: first, the sample is ultrasonically cleaned with ethanol for 30 minutes, then cleaned with deionized water two to three times, and dried overnight at 37°C. After cleaning and pretreatment, PEEK was immersed in a concentrated sulfuric acid solution with a concentration of 98%~99%, ultrasonically irradiated at 25°C for 5 minutes, and then washed with deionized water at least three times until sulfuric acid residue was removed. All materials were subjected to hydrothermal treatment at 120°C for 12 hours in an autoclave lined with polytetrafluoroethylene, and then cooled to 25°C to obtain concentrated sulfuric acid sulfonated PEEK. In step C, the chitosan solution is prepared as follows: acetic acid with a volume of 2 times is added to distilled water and the mixture is adjusted to an acidic environment. Then, chitosan powder is added, and the mixture is stirred with a magnetic stirrer to obtain a chitosan solution. The weight ratio of chitosan to the volume of distilled water is 2:

1.

2. A biofunctionalized polyetheretherketone implant, characterized in that, It is prepared by the method described in claim 1.

3. The application of the biofunctionalized polyether ether ketone implant according to claim 2 in the preparation of orthopedic implants.

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