PEEK-ZrO2-FeNC surface modified material as well as preparation method and application thereof

Abstract

The invention discloses a PEEK-Zr / ZrO2-FeNC surface modified material as well as a preparation method and application thereof. The surface modified material comprises a PEEK base material, Zr / ZrO2 deposited on the surface of the PEEK base material, and FeNC nano-enzyme particles uniformly distributed on the surface of the PEEK base material subjected to Zr / ZrO2 deposition. According to the invention, Zr / ZrO2 is deposited on the surface of the PEEK base material, so that the formation of a biological membrane on the surface of the material can be reduced, and the risk of occurrence of peri-implantitis is reduced; in addition, the introduction of zirconium can increase the toughness of the special appearance of the PEEK surface, and reduce the surface damage when the implant material is beaten; the FeNC nano-enzyme particles incarcerated in the Zr / ZrO2 layer can remove ROS in the early stage of osseointegration, macrophages are polarized to an M2 type, an anti-inflammatory microenvironment is formed, finally, adhesion of osteoblasts is enhanced by using a zirconium oxide columnar structure, and the cells are anchored on the surface of PEEK to promote osseointegration.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedical materials, and in particular to a PEEK-Zr / ZrO 2 -FeNC surface modified material and its preparation method and application. Background Art

[0002] Degenerative spinal diseases are common and frequent diseases in the middle-aged and elderly, mainly including herniated disc, spinal stenosis and spondylolisthesis. Patients who do not respond to conservative treatment need surgical treatment to achieve the purpose of spinal sequence restoration, spinal cord and nerve decompression and spinal stability reconstruction. Intervertebral fusion cage (cage) is one of the most commonly used spinal implants in surgical treatment. However, failure of bone formation, connection or shaping will lead to non-fusion of intervertebral bone grafts. The most direct manifestation of intervertebral fusion cage failure is loosening and displacement of the fusion cage, as well as collapse of the fusion cage and formation of false joints. Therefore, how to reduce the occurrence of cage failure after bone graft fusion surgery has become an important research topic in spinal surgery.

[0003] The cage made of polyetheretherketone (PEEK) is still the most commonly used implant in clinical practice because of its good transmittance, elastic modulus and biocompatibility, and its clinical effect has also been confirmed. In order to improve the fusion rate, the bone graft space is increased as much as possible when designing the fusion cage. Filling the intervertebral fusion cage with small granular blocks of autologous bone, allogeneic bone, artificial bone and bone morphogenetic protein (BMP) and other materials can improve the bone fusion rate and cell adhesion. However, the contact area between the granular filler in the cage and the upper and lower end plates of the vertebral body is limited, and the bone healing is slow. In order to improve the problem of difficulty in bone ingrowth between the bony end plates and the surface of the PEEK fusion cage caused by the poor osteogenic efficiency of the PEEK material, and to increase the stability of the contact surface between the upper and lower surfaces of the cage and the vertebral end plates, modifying the surface of the PEEK fusion cage is a direct and effective method. So far, material coatings with osteogenic activity have been widely used to change the surface properties of PEEK. However, there is currently no recognized optimal coating material. Studying which material or how to mix it can provide the best biomechanical stability and osteogenesis has important clinical value and research significance. Summary of the invention

[0004] The purpose of the present invention is to provide a PEEK-Zr / ZrO 2 -FeNC surface modified material and its preparation method and application.

[0005] To achieve the above object, the technical solution adopted by the present invention is:

[0006] The first aspect is to provide a PEEK-Zr / ZrO 2-FeNC surface modified material, comprising a PEEK substrate, and Zr / ZrO deposited on the surface of the PEEK substrate 2 , and uniformly distributed on the deposited Zr / ZrO 2 FeNC nanozyme particles on the surface of PEEK substrate.

[0007] Furthermore, the Zr / ZrO deposited on the surface of the PEEK substrate 2 It presents a uniform nanoscale three-dimensional columnar structure.

[0008] Furthermore, the FeNC nanozyme particles are embedded in the gaps of the three-dimensional columnar structure.

[0009] The second aspect is to provide the above-mentioned PEEK-Zr / ZrO 2 -The preparation method of FeNC surface modified material comprises the following steps:

[0010] Step 1: After polishing and cleaning the medical PEEK substrate, PBHS deposition was performed using a radio frequency ionization plasma pulse bias injection system to obtain PEEK-Zr / ZrO 2 ;

[0011] Step 2: weigh ferric chloride hexahydrate and dissolve it in formamide solution, stir at room temperature to obtain a precursor solution; 2 Place in the precursor solution, react at 180-200°C for 4-8h, wash and dry after the reaction to obtain the PEEK-Zr / ZrO 2 -FeNC surface modified materials.

[0012] Furthermore, the specific method of PBHS deposition using the RF ionization plasma pulse bias injection system is as follows: equip a Zr metal target, place the target on a turntable connected to a bias power supply, and then place the PEEK substrate on the target surface; use an RF plasma source to generate H 2 Plasma, using BDP-5A to apply unipolar voltage pulses to the target to provide a controlled constant bias pulse to ensure stability during the deposition process.

[0013] Furthermore, when the PEEK substrate is a cylinder, Zr / ZrO 2 Deposited on the upper and lower surfaces of the cylinder.

[0014] Furthermore, the concentration of ferric chloride hexahydrate in the precursor solution is 0.6-1.0 mg / mL.

[0015] The third aspect is to provide the above-mentioned PEEK-Zr / ZrO 2 -Application of FeNC surface modified materials in the preparation of intervertebral fusion products.

[0016] The present invention adopts the above technical solution, and has the following technical effects compared with the prior art:

[0017] PEEK-Zr / ZrO 2 -FeNC surface modification material, depositing Zr / ZrO on the surface of PEEK substrate 2 , can reduce the formation of biofilm on the material surface and reduce the risk of peri-implantitis; in addition, the introduction of zirconium can increase the toughness of the special morphology of the PEEK surface and reduce surface damage when knocking the implant material; embedded in Zr / ZrO 2 The FeNC nanozyme particles with a layer can remove ROS in the early stage of bone integration, polarize macrophages to M2 type, form an anti-inflammatory microenvironment, and finally use the zirconium oxide columnar structure to enhance osteoblast adhesion and anchor the cells on the PEEK surface to promote bone integration. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 The general view of each group of samples in Example 1, where (A) PEEK; (B) PEEK-Zr / ZrO 2 ;(C)PEEK-FeNC;(D)PEEK-Zr / ZrO 2 -FeNC.

[0019] Figure 2 The surface SEM morphology of each group of samples in Example 1.

[0020] Figure 3 X-ray photoelectron spectroscopy spectra of each group of samples in Example 1, including: (A) high-precision C1s spectrum; (B) high-precision O1s spectrum; (C) high-precision Zr3d spectrum; (D) high-precision N1s spectrum.

[0021] Figure 4 The water contact angle and surface energy of each group of samples in Example 1, where (A) the contact angle of water droplets on the surface of each group of samples; (B) the water contact angle value of each group of samples; (C) the calculated surface energy of each group of samples. Data are expressed as mean ± standard deviation; ns, p>0.05; *, p<0.05; **, p<0.01; ***, p<0.001; ****, p<0.0001; n=4.

[0022] Figure 5 is the H at different times in each group of samples in Example 1 2 O 2 concentration; ns, p>0.05; *, p<0.05; **, p<0.01; ***, p<0.001; ****, p<0.0001; n=3.

[0023] Figure 6 OD values ​​of each group of samples and cells co-cultured on the 1st, 3rd and 7th day in Example 1; *, p<0.05; **, p<0.01; n=5.

[0024] Figure 7 Live / dead staining of cells of each group of samples in Example 1 (Day 7).

[0025] Figure 8 The SEM images of cells adhering and spreading on the material surface on the third day in Example 1; (A) PEEK group, (B) PEEK-Zr / ZrO 2 group, (C) PEEK-FeNC group, (D) PEEK-Zr / ZrO 2 -FeNC group; a1, b1, c1, d1 and a2, b2, c2, d2 are enlarged images of cells in A, B, C, and D, respectively.

[0026] Fig. 9 The results of alkaline phosphatase staining in Example 2 are shown; wherein, (A) alkaline phosphatase staining of BMSCs after co-culture with different groups of materials in osteogenic induction medium for 7 days; (B) quantification of alkaline phosphatase activity of BMSCs after co-culture with different groups of materials in osteogenic induction medium for 7 days; (C) and (D) alkaline phosphatase staining and activity quantification of BMSCs after co-culture with different groups of materials in osteogenic induction medium for 7 days after oxidative stress.

[0027] Fig.10 The results of Alizarin Red staining in Example 2 are shown; wherein, (A) Alizarin Red staining of mineralized nodules on the 21st day of culture; (B) Alizarin Red calcium nodule quantification after BMSCs were co-cultured with osteogenic induction medium containing different groups of materials for 21 days; (C) and (D) Alizarin Red staining and calcium nodule quantification after oxidative stress after BMSCs were co-cultured with osteogenic induction medium and different groups of materials for 21 days.

[0028] Fig.11 The mRNA expression of BMSCs co-cultured with different groups of materials in osteogenic induction medium in Example 2 (day 7) is shown; ns, p>0.05; *, p<0.05; **, p<0.01; ***, p<0.001; ****, p<0.0001; n=3.

[0029] Fig.12 The effects of each group of materials in Example 2 on the expression of ALP, RUNX2, COL1 and OCN proteins are shown.

[0030] Fig.13 The results of immunological marker staining of each group in Example 2 are shown, wherein: (A) immunological marker staining of each group without oxidative stress treatment; (B) immunological marker staining of each group after H2 O 2 Immunomarker staining of each group after oxidative stress treatment.

[0031] Fig.14 The flow cytometry experiment in Example 2 shows the effect of the conditioned medium of each group on macrophage polarization after 3 days of culture.

[0032] Fig.15 The results of the expression of inflammation-related factors in Example 2 are shown, wherein (A) the expression of inflammation-related factors in each group without oxidative stress treatment; (B) the expression of inflammation-related factors in each group after H 2 O 2 Expression of inflammatory-related factors in each group after oxidative stress treatment; ns, p>0.05; *, p<0.05; **, p<0.01; ***, p<0.001; ****, p<0.0001; n=3.

[0033] Fig.16 This is a general picture of the specimens after 12 weeks of implantation of various materials in Example 3, and the implanted materials of each group have been marked.

[0034] Fig.17 This is a Micro-CT plain scan sagittal image of each material in Example 3 12 weeks after implantation.

[0035] Fig.18 This is a Micro-CT three-dimensional reconstruction image of each material in Example 3 after 12 weeks of implantation.

[0036] Fig.19 The thickness, number and separation of trabeculae in the specimens of each group 12 weeks after surgery in Example 3; ns, p>0.05; *, p<0.05; **, p<0.01; n=3.

[0037] Fig. 20 The HE staining results at 12 weeks after surgery in Example 3 are shown.

[0038] Fig.21 The Masson staining results at 12 weeks after surgery in Example 3 are shown.

[0039] Fig. 22 The Trap staining results in Example 3 at 12 weeks after surgery are shown.

[0040] Fig.23 The results of the three-point compression fracture mechanical test at 12 years after surgery in Example 3 are shown, including: (A) mechanical curves of each group; (B) comparison of the maximum force of each group at three-point compression fracture; ns, p>0.05; *, p<0.05; **, p<0.01; n=3. DETAILED DESCRIPTION

[0041] The present invention is further described below in conjunction with the accompanying drawings and specific embodiments, but is not intended to be limiting of the present invention. It should be noted that the embodiments and features in the embodiments of the present invention may be combined with each other without conflict.

[0042] Example 1

[0043] The main reagents are shown in Table 1 below:

[0044] Table 1

[0045]

[0046] 1. This embodiment provides a PEEK-Zr / ZrO 2 -The preparation method of FeNC surface modified material specifically comprises the following steps:

[0047] 1) PEEK surface plasma bias homopolar sputtering (PBHS)

[0048] Biomedical PEEK was polished with 2000 grit sandpaper and then cleaned three times with acetone, alcohol and ultrapure water, each for 30 minutes. PEEK samples included round pieces (10 mm × 1 mm for cell culture and performance characterization) and cylindrical pieces (3 mm × 3 mm for in vivo implantation experiments). PEEK-Zr / ZrO was prepared by PBHS method at -800 V bias voltage with zirconium (Zr) as the target for 15 min. 2 .

[0049] PBHS ​​deposition was performed using a radio frequency ionized plasma pulse bias injection system. A rotating circular oil-cooled turntable was equipped with a Zr metal target with a diameter of 160 mm and a thickness of 2 mm. The substrate consisted of PEEK with a diameter of 10 mm and a thickness of 1 mm. The oil-cooled target stage was installed in a vacuum chamber and pumped to 5.0 × 10 -3 Pa pressure, pure argon (Ar) with a minimum purity of 99.9% was introduced into the chamber, and the pressure in the chamber was maintained at 0.8 Pa during sample preparation. The target was placed on a turntable connected to the bias power supply, and then PEEK was placed on the target surface. All depositions were performed at room temperature, and at least three samples were deposited under each process condition, with the upper and lower circular surfaces of the in vivo implanted cylinder deposited. In the case of PBHS, the RF plasma source generated H 2 Plasma, and a unipolar voltage pulse is applied to the target by a pulse unit (BDP-5A, CNNC Cochrane) to provide a controlled constant bias pulse. In this embodiment, the average RF power is kept constant at 300W, the pulse bias frequency is 40kHz, and the duty cycle is kept at 30%.

[0050] 2) Preparation of FeNC nanozymes and different groups of polyetheretherketone samples

[0051] Weigh 30 mg of ferric chloride hexahydrate and dissolve it in 50 mL of formamide solution. Stir magnetically for 30 min at room temperature to obtain a precursor solution. 2 Place in a 150mL reactor, with 8 samples placed in each reactor. Then take 50mL of the precursor solution and place it in the reactor lining, react at 180℃ for 4h, and cool naturally. The obtained samples are ultrasonically cleaned with ultrapure water and anhydrous ethanol for 3min respectively, and dried at 60℃ for later use.

[0052] The four groups of samples were named PEEK, PEEK-Zr / ZrO 2 、PEEK-FeNC、PEEK-Zr / ZrO 2 -FeNC.

[0053] 2. The above-prepared materials (PEEK, PEEK-Zr / ZrO 2 、PEEK-FeNC、PEEK-Zr / ZrO 2 -FeNC) for characterization and experiments

[0054] 1) Field Emission Scanning Electron Microscopy (SEM)

[0055] Each group of samples with a size of 10 mm × 1 mm were adhered to the surface of the conductive adhesive and subjected to scanning electron microscopy inspection 30 seconds after gold spraying.

[0056] 2) X-ray photoelectron spectroscopy (XPS) spectrum analysis

[0057] Using an X-ray photoelectron spectrometer, the surface elements of each group of samples were qualitatively and quantitatively analyzed in an ultra-high vacuum environment, the element binding energy was calculated, and a high-resolution map was drawn.

[0058] 3) Water contact angle (WCA) measurement and surface energy calculation

[0059] The water contact angles of different groups of samples were measured using a contact angle measurement (OCA), and the surface energy of the samples was calculated using the Owen-Wendt two-liquid formula based on the results.

[0060] 4) Detection of nanozyme activity on the surface of each group of materials

[0061] Catalase activity test: The colorimetric method was used to determine the activity of each group of samples on H 2 O 2 The reaction system was PBS solution (pH = 7.4), with a total volume of 1 mL, H2 O 2 The final concentration was 1 mM. The samples were placed in the reaction solution. After mixing evenly, the mixture was incubated at 37°C for 2 h, 6 h, and 12 h. 200 μl of the reaction mixture was placed in a 96-well plate and Ti(SO 4 ) 2 20 μL of the solution was mixed and reacted for 10 min, and the absorbance of the mixture at 405 nm was measured using a multifunctional microplate reader.

[0062] 5) In vitro biosafety study (CCK-8 experiment)

[0063] Each group of samples was placed in a 6-well plate, soaked in 75% alcohol for 30 minutes, and then naturally air-dried on a clean bench and sterilized by ultraviolet radiation (1 hour per side). The samples were then placed in a 15 mL centrifuge tube and 6 mL of culture medium was added to each group to soak for 48 hours.

[0064] Observe the density of cells and make them adhere to the culture dish under the microscope. Carefully discard the culture medium, rinse twice with PBS, add trypsin to digest for about 1-2 minutes. After the cells are seen floating under the microscope, add complete culture medium to terminate the digestion.

[0065] Transfer the cells into a 15 mL centrifuge tube and centrifuge (1000 rpm, 5 min), then remove the culture medium containing trypsin and resuspend the cells with new complete culture medium;

[0066] Count the cells according to the experimental needs and dilute the extract to make up the culture medium. 3 Cells (about 100 μL) were inoculated into a 96-well plate and placed in a 37°C, 5% CO 2 Culture in an incubator and change the medium every other day;

[0067] After the cells were cultured in the extract for 1, 3, and 7 days, CCK-8 working solution was added and placed in the incubator for incubation;

[0068] After incubation for 2 h, the supernatant of each well was transferred to another well plate, and the absorbance (OD value) of each well at 450 nm was measured using an enzyme-labeled instrument to further determine the cell activity of each group, with 5 samples tested each time.

[0069] 6) Cell dead / alive fluorescence staining

[0070] Add material extract and cell culture, change the solution every 3 days, and culture for 5 days. Add 20μL PI (2mM) stock solution and 5μL Calcein-AM stock solution (4mM) to 10mL PBS to prepare the stain. On the 5th day, after washing with PBS twice, add 200μL staining solution to each well of the 24-well plate and incubate in a cell culture incubator at 37°C for 20 minutes. Observe under a fluorescence microscope, first excite with a wavelength of 490±10nm, green is live cells, and dead cells are red.

[0071] 7) In vitro cell adhesion and spreading assay

[0072] After disinfection, each group of samples was placed in a 6-well plate. 2 mL of BMSCs was added to each well at a density of 1×10 5 / mL of cell suspension. The sample was then placed in an incubator and cultured for 72 hours. Take out the inoculated cells and observe the cell adhesion under a microscope. Rinse thoroughly with PBS three times and fix the sample with electron microscope fixative. Rinse again with PBS three times, 2 minutes each time. The treated sample was frozen at -20℃ for 6 hours, and then dried in a freeze dryer for 24 hours. Finally, the sample surface was sprayed with gold, and the morphology of the cells and their attachment and expansion on the material surface were observed by scanning electron microscopy (SEM).

[0073] 8) Statistical analysis: The experimental data were expressed as mean ± standard deviation. The inter-group comparison was performed using analysis of variance. P < 0.05 was considered statistically significant. SPSS 26.0 and GraphPad Prism 10.0 were used for analysis of the results.

[0074] 3. Experimental Results

[0075] 1) Macroscopic photos of each group of samples are as follows Figure 1 As shown in the figure, it can be seen that after sputtering coating, PEEK-Zr / ZrO 2 Group, PEEK-Zr / ZrO 2 The surface of the -FeNC group showed metallic luster, while the PEEK discs in each group did not undergo obvious deformation.

[0076] 2)Reference Figure 2 Through SEM observation, it can be seen that the surface of untreated PEEK is generally smooth, without special structure, and the grinding marks are clearly visible, while the surface of ZrO2 deposited by PBHS is smooth. 2 The PEEK surface presents a uniform nanoscale three-dimensional columnar structure. The diameter of these micro-cylinders is about 200nm, and the distance between the micro-cylinders is between 40 and 500nm. 2After being placed in the reactor with FeNC precursor solution for 4 hours, it can be seen that FeNC particles are evenly distributed on the surface of the material. The difference is that the nanozymes in the PEEK-FeNC group are attached to the PEEK surface, while the PEEK-Zr / ZrO 2 -FeNC group nanozyme particles are embedded in the gaps of the columnar structure.

[0077] 3) X-ray photoelectron spectroscopy (XPS) spectrum analysis (reference Figure 3 ):

[0078] XPS shows high-resolution spectra of C, O, Zr and Fe on the sample surface. The outermost surface of PEEK and PEEK-FeNC is mainly CC and CO bonds, with a small amount of C=O bonds. PEEK was ion-etched to a depth of 400nm and tested. It can be seen from the figure that the C1s peaks on PEEK are 284.8eV, 286.4eV, and 291.5eV, corresponding to CC, CO, and C=O. The Ols peaks are 533.3eV and 531.3eV, corresponding to OC and O=C. After Ar&Zr-PIII treatment, the CC binding energy did not change significantly, the CO bond was reduced to almost disappear, and C=O still existed, but the binding energy was reduced to 288.5eV. This shows that PEEK is completely carbonized in a certain depth during ion implantation, and amorphous carbon may be formed. The OC binding energy dropped to 529.95eV, and O=C rose to 531.45eV without significant changes. The Zr3d peaks are at 184.35 eV and 181.95 eV, corresponding to single-element Zr and ZrO, respectively. 2 After Fe nanozyme coating, it can be seen that it does not affect the CC, CO, and C=O binding energies of the corresponding treated PEEK, and FeNC nanozyme is successfully loaded on the sample surface. 2 -FeNC has a higher surface Fe content, indicating that the columnar nanostructure formed after PEEK treated with Ar&Zr-PIII can embed more nanozymes.

[0079] 4)Reference Figure 4 , PEEK, PEEK-Zr / ZrO 2 、PEEK-FeNC、PEEK-Zr / ZrO 2 The water contact angles of the samples in the -FeNC group were 94.17±2.04°, 148.0±3.06°, 96.41±2.67° and 95.35±2.48°, respectively. The surface energy of each group was calculated based on the water contact angle. Compared with the untreated PEEK sample, the surface energy of PEEK-Zr / ZrO 2The apparent contact angle is significantly improved, showing a hydrophobic / superhydrophobic state. After plasma deposition treatment, a nanoscale columnar structure appears on the PEEK surface, and the contact surface is transformed from a Wenzel state to a Cassie state.

[0080] PEEK-FeNC and PEEK-Zr / ZrO 2 The -FeNC group has a similar contact angle with the untreated PEEK water. This indicates that the FeNC nanozyme particles are insufficient to change the surface morphology of PEEK, but can be embedded in the gaps of the columnar structure after plasma deposition, reducing the incomplete contact effect caused by the air chamber between the liquid and the solid in the Cassie state.

[0081] 5) The iron nanozyme has catalase-like activity. By detecting the ability to remove hydrogen peroxide at different time periods, the enzyme activity of the iron nanozyme at different time periods can be measured. Figure 5 It can be seen that at 37°C, 2h, PEEK-Zr / ZrO 2 -FeNC group showed CAT-like activity, which was significantly different from other groups (p<0.0001). At 6h, PEEK-FeNC group showed CAT-like activity, which was significantly different from the two groups without nanozyme coating (p<0.01), but not as good as PEEK-Zr / ZrO 2 -FeNC group has high enzyme activity. At 12h, PEEK-Zr / ZrO 2 The -FeNC group still maintained a high CAT-like activity, and the enzyme activity of the PEEK-FeNC group increased further, but was still not as good as that of the PEEK-Zr / ZrO 2 -FeNC group.

[0082] 6) The OD values ​​of each group ( Figure 6 ) found that after 1d and 3d of co-culture of cells and material extracts, there was no statistical difference in absorbance among the groups (p>0.05); at the 7th day, the PEEK group, PEEK-Zr / ZrO 2 There was no significant difference in OD values ​​between the PEEK-FeNC group and the PEEK-Zr / ZrO 2 The OD value of the -FeNC group was greater than that of the other three groups, and the difference was statistically significant (p<0.05).

[0083] 7)Reference Figure 7 The cells in each group survived well, with the majority of live cells showing green fluorescence and only a few dead cells showing red fluorescence. 2 Compared with the PEEK group and PEEK-Zr / ZrO 2 The number of cell colonies in the PEEK-FeNC group and the PEEK-FeNC group increased, which was consistent with the results of cytocompatibility analysis.

[0084] 8) After 3 days of co-culture of cells and materials, it can be seen that BMSCs cells can adhere to the surface of the material ( Figure 8 ). In terms of cell number, after rinsing with PBS, the PEEK group had the least cells on the surface, and the PEEK-Zr / ZrO 2 Group and PEEK-Zr / ZrO 2 -FeNC group had the same number of cells and the largest number. Long spindle cells were observed adhering to the surface of PEEK group, and the cell bodies were stretched; 2 The number of cells adhering to the surface of the PEEK-FeNC group was large, the cell spreading area was good, the cells grew evenly, and the cells extended pseudopodia to adhere to the material. A mesh structure was visible between adjacent cells. The cells in the PEEK-FeNC group spread well, the cells were dispersed, and they were spindle-shaped. 2 In the -FeNC group, the number of cells adhering to the surface was greater, the extension area was further increased, the cells grew evenly, and extended a large number of pseudopodia to connect with adjacent cells and materials.

[0085] In summary, this embodiment constructs a columnar zirconium oxide coating on the PEEK surface through plasma treatment to enhance cell adhesion, and covers the iron nanozyme coating on its surface. The zirconium oxide columnar structure strengthens osteoblast adhesion and anchors the cells on the PEEK surface to promote bone integration; in the early stage of bone integration, the material can remove ROS, polarize macrophages to M2 type, form an anti-inflammatory microenvironment, and promote bone formation on the surface of the material. Material characterization and experimental analysis prove that it has Zr / ZrO 2 The modified polyetheretherketone has a rough surface, but the hydrophilicity has not changed significantly. Cell compatibility experiments were conducted on each group of PEEK materials, and it was found that all four groups of samples showed good biocompatibility, and there was no significant difference in the number of dead cells stained with red fluorescence, indicating that these samples have minimal cytotoxicity. In particular, Fe nanozymes and Zr / ZrO 2 The surface-modified polyetheretherketone showed cell proliferation activity on the 7th day. This may be because under neutral conditions in vitro, the Fe nanozyme was slowly released and played a role similar to CAT, effectively removing ROS generated by cells during proliferation. This ability to remove ROS helps maintain the stability of the intracellular environment, thereby promoting cell proliferation and survival.

[0086] Example 2 PEEK-Zr / ZrO 2 -FeNC in vitro experimental study

[0087] 1. Experimental Methods

[0088] 1. Evaluation of the materials for inducing osteogenic differentiation of BMSCs in vitro

[0089] 1) Alkaline phosphatase staining and activity detection.

[0090] 2) Alizarin red staining and calcium content determination.

[0091] 2. Osteogenic gene expression assay

[0092] 1) Total RNA extraction;

[0093] 2) Primer design;

[0094] 3) Reverse transcription to obtain cDNA;

[0095] 4) RT-PCR amplification.

[0096] 3. Detection of osteogenic protein expression

[0097] 1) Total protein extraction;

[0098] 2) Glue making;

[0099] 3) Electrophoresis;

[0100] 4) Transfer and block;

[0101] 5) Antibody incubation;

[0102] 6) Chemiluminescence imaging and analysis.

[0103] 4. Evaluation of the Immunomodulatory Effect of the Materials in Vitro

[0104] 1) Cultivation of macrophages;

[0105] 2) Immunofluorescence assessment of macrophage polarization;

[0106] 3) flow cytometry to assess macrophage polarization;

[0107] 4) PCR was used to detect the expression levels of inflammation-related genes.

[0108] The above experimental methods can be obtained by conventional technical means in the art, and the present invention will not elaborate on them in detail.

[0109] 5. Statistical analysis: The experimental data were expressed as mean ± standard deviation. Paired t-test and analysis of variance were used for comparison between groups. Statistical differences were considered when p < 0.05. SPSS26.0 and GraphPad Prism 10.0 were used for analysis of the results.

[0110] 2. Experimental Results

[0111] 1. Alkaline Phosphatase Staining

[0112] Alkaline phosphatase is an important marker for early osteogenic differentiation of BMSCs. BMSCs were co-cultured with different groups of materials in osteogenic induction medium. Alkaline phosphatase staining and activity detection were performed 7 days later to evaluate the effects of each group of materials on early osteogenic differentiation of BMSCs. Alkaline phosphatase was stained into blue-black particles by alkaline phosphatase staining agent, and the density of blue-black particles was used to reflect the expression level of alkaline phosphatase. The results showed that ( Fig. 9 ), in H 2 O 2 In the (-) group, PEEK-Zr / ZrO 2 、PEEK-FeNC、PEEK-Zr / ZrO 2 The blue-dyed particle density of the three groups of -FeNC was higher than that of the PEEK group, especially the PEEK-Zr / ZrO 2 -FeNC had the deepest staining, and PEEK-FeNC group was darker than PEEK-Zr / ZrO 2 The granule staining density of the group was high. The quantification of alkaline phosphatase activity showed the same results, PEEK-Zr / ZrO 2 The alkaline phosphatase activity was the highest in the -FeNC group, and that in the PEEK-FeNC group was higher than that in the PEEK-Zr / ZrO 2 There were statistically significant differences among the four groups (p<0.05). 2 O 2 Compared with the (-) group, H 2 O 2 The density of alkaline phosphatase blue-stained particles in the (+) group decreased, and the PEEK-Zr / ZrO 2 、PEEK-FeNC、PEEK-Zr / ZrO 2 The blue-dyed particle density of the -FeNC group was significantly higher than that of the PEEK group, and the blue-dyed particle density of the PEEK-Zr / ZrO 2 The -FeNC group was the most obvious. The quantitative results of alkaline phosphatase activity were consistent with the staining.

[0113] 2. Alizarin Red Staining

[0114] Calcium nodules are an important marker for the late osteogenic differentiation of BMSCs. Alizarin red S can dye calcium nodules red, which can quantify the amount of calcium deposition during bone formation. Alizarin red staining is often used to evaluate the level of late osteogenic differentiation of BMSCs. BMSCs were co-cultured with each group of materials in osteogenic induction medium for 21 days and then stained with alizarin red calcium nodules to evaluate the effects of each group of materials on the late osteogenic differentiation of BMSCs. The results showed that ( Fig.10 ), in H 2 O 2 In the (-) group, scattered red nodules were found in the PEEK group; more red nodules were found in the PEEK-FeNC group, which were larger in size and some of the nodules were translucent.2 The red mineralized nodules were more visible in the PEEK-ZrO group, most of which were oval, obvious, and stained darker. 2 -FeNC group showed more red mineralized nodules deposited to form clumps, with strong red positive reaction. 2 -FeNC group>PEEK-Zr / ZrO 2 group>PEEK-FeNC group>PEEK group. Quantitative analysis showed that PEEK-Zr / ZrO 2 The difference between the -FeNC group and the other three groups was statistically significant (p<0.0001). 2 O 2 (+) group and H 2 O 2 Compared with the (-) group, the staining of calcium nodules became lighter, the number of calcium nodules was relatively sparse, and the amount of calcium deposition decreased significantly. 2 -FeNC group, PEEK-Zr / ZrO 2 Compared with the PEEK group, the calcium deposition in the PEEK-Zr / ZrO 2 The -FeNC group was the most significant.

[0115] 3. Expression of Osteogenesis-related Genes

[0116] On the 7th day, the expression levels of ALP, Bglap, Sp7, Runx2, and Col1a1 were different in each group (see Fig.11 ). The expression of each target gene in the PEEK group was significantly lower than that in other groups, while the expression of each target gene in the PEEK-Zr / ZrO 2 -FeNC group had higher expression of osteogenic differentiation-related genes than other groups, and the difference was statistically significant (p<0.05). 2 There was no significant difference between the PEEK-FeNC group and the PEEK-Zr / ZrO2 group (p>0.05). 2 -FeNC group was significantly higher than PEEK group (p<0.0001), and higher than PEEK-FeNC group. 2 There was no statistically significant difference between the groups (p>0.05). 2 -FeNC group had no statistically significant difference (p>0.05). 2 O 2 The expression of osteogenic differentiation-related genes in BMSCs of the (+) group was higher than that in the H2 O 2 Compared with the PEEK group, the PEEK-Zr / ZrO 2 group, PEEK-FeNC group and PEEK-Zr / ZrO 2 -FeNC group showed different degrees of increase in the expression of osteogenic differentiation-related genes, among which PEEK-Zr / ZrO 2 The -FeNC group had the most significant increase.

[0117] 4. Expression of Osteogenesis-related Proteins

[0118] The target protein bands obtained by Western blot experiment are as follows Fig.12 As can be seen from the figure, each group of coating materials has different expressions of osteogenic differentiation-related genes. 2 -FeNC group showed the deepest expression of each protein, PEEK-ZrO 2 The expression of ALP and RUNX2 proteins in the PEEK-FeNC group was deeper than that in the PEEK-Zr / ZrO 2 After oxidative stress, the expression of osteogenic differentiation-related proteins in BMSCs decreased to varying degrees, but compared with the PEEK group and PEEK-Zr / ZrO 2 group and PEEK-FeNC group, PEEK-Zr / ZrO 2 -FeNC composition significantly increased the expression of bone differentiation-related proteins. Combined with gene expression, ALP staining and Alizarin red staining, the results showed that iron nanozymes and zirconium oxide nanostructured coatings have a promoting effect in the osteogenic differentiation process and can reduce the adverse effects of oxidative stress on the osteogenic differentiation of BMSCs.

[0119] 5. Immunomarker Staining

[0120] In immunology, the M1 and M2 types of macrophages represent two different activation states: M1 macrophages: usually associated with inflammatory responses, they produce pro-inflammatory cytokines such as tumor necrosis factor α (TNF-α) and interleukin 6 (IL-6), which play an important role in clearing pathogens and promoting inflammatory responses. M2 macrophages: associated with anti-inflammation and tissue repair, they produce anti-inflammatory cytokines such as interleukin 10 (IL-10) and transforming growth factor β (TGF-β), which help reduce inflammation and promote tissue repair. Staining was performed using antibodies against specific markers for M1 and M2 macrophages. M1 macrophages use CD86 as a marker, while M2 macrophages may use CD206 as a marker. The results are as follows: Fig.13 As shown, due to the PEEK group and PEEK-Zr / ZrO 2The PEEK-FeNC group had no CAT-like enzyme activity and could not promote the transformation of macrophages from M1 to M2. The number of M1 markers (CD86) after staining was large and the expression was high. However, the number of M2 markers (CD206) was small and the expression was low. The PEEK-FeNC group had the effect of inhibiting inflammation and could promote the transformation of macrophages from M1 to M2. The number of fluorescent spots of M1 markers decreased, while the fluorescence expression of M2 markers increased. Compared with the PEEK-FeNC group, the PEEK-Zr / ZrO 2 The -FeNC group had a more obvious effect in inhibiting inflammation and promoting the transformation of macrophages from M1 to M2. The number of CD86 staining was similar to that of the PEEK-FeNC group, and the number of CD206 marker staining was greater. After oxidative stress treatment, the number of CD86 staining in the two groups without iron nanozyme coating increased further, while the number of CD206 staining decreased further. 2 -FeNC group had less CD86 staining and more CD206 staining, which was 2 The difference was more obvious in -FeNC group.

[0121] 6. Flow cytometry analysis of macrophage phenotypes

[0122] Flow cytometry was used to determine the proportion of M1 and M2 phenotypes of macrophages in each group. The results showed the same trend as immunofluorescence. Fig.14 As shown in the figure, after induction with CM-3 conditioned medium, the proportion of CD86(+) macrophages in each group was 26.46%, 20.32%, 17.04%, and 15.57%, respectively, while the proportion of CD206(+) macrophages in each group was 36.41%, 36.95%, 45.54%, and 57.46%, respectively. The flow cytometry results of each group after oxidative stress are shown in the figure. The proportion of CD86(+) macrophages in each group was 32.54%, 30.68%, 14.84%, and 8.05%, respectively; the proportion of CD206(+) macrophages was 15.65%, 13.86%, 26.31%, and 36.40%, respectively. That is, in the early stage, iron nanozymes can induce macrophages to polarize to the M2 phenotype, and after oxidative stress, they can still promote the induced cells to the M2 phenotype.

[0123] 7. Analysis of Inflammation-related Gene Expression

[0124] By detecting the expression of inflammation-related genes (iNOS, TNF, CD86) and anti-inflammatory genes (Arg1, mIL10, CD206), it can be found that ( Fig.15)Under normal conditions, the two groups without Fe nanozymes showed higher expression of genes related to inflammatory factors. The expression of genes related to inflammation inhibition was very low, and the difference between the two groups was not statistically significant. The two groups containing Fe nanozymes showed lower expression of genes related to inflammatory factors and higher expression of genes that inhibited inflammation. The difference between the two groups was not statistically significant, but the difference was significant from the first two groups without Fe nanozymes. Under oxidative stress conditions, the expression of genes related to inflammatory factors in the two groups without Fe nanozymes was further upregulated, and the difference in expression of anti-inflammatory genes was more obvious, but there was no significant difference in the expression of each gene between the two groups. PEEK-FeNC, PEEK-Zr / ZrO 2 Compared with the two groups without Fe nanozymes, the -FeNC group showed lower expression of inflammatory factor-related genes and higher expression of anti-inflammatory genes, but there was no significant difference in the expression of inflammatory factor-related genes between the two groups. In the expression of mIL10 and mCD206, PEEK-Zr / ZrO 2 -FeNC group was higher than PEEK-FeNC group.

[0125] Example 3 PEEK-Zr / ZrO 2 -FeNC in vivo experimental study

[0126] 1. Experimental Methods

[0127] The present invention uses 21 clean-grade New Zealand white rabbits, all male, 6 months old, weighing between 2.5-3.0 kg, provided by Shengwang Animal Co., Ltd. They are raised according to the standard conditions of GB14925-2010 and are adaptively raised 1 week before surgery.

[0128] 1. Implant Preparation

[0129] The upper and lower bottom surfaces of the medical grade cylindrical PEEK (3 mm × 3 mm) were treated according to the method described in Example 1. 20 New Zealand rabbits were randomly divided into groups A, B, C, and D, with 5 rabbits in each group. Group A: implanted with PEEK; Group B: implanted with PEEK-Zr / ZrO 2 Coated PEEK; Group C: PEEK-FeNC coated PEEK implanted; Group D: PEEK-Zr / ZrO implanted 2 -FeNC coated PEEK.

[0130] 2. Retroperitoneal anterior vertebral fusion experiment

[0131] 1) Establishment of critical lumbar bone defect model and material implantation: Experimental animals were fasted for 12 hours and water deprived for 4 hours before surgery. After weighing the animals, they were anesthetized by injection of sudamine through the ear vein. The skin of the lateral abdomen was prepared, and the rabbit was placed in the lateral position, and the incision was marked on the body surface. First, the highest point of the rabbit iliac crest was located, and the head-tail axis of the rabbit was made 1 cm below this point. This line was used for the horizontal positioning of the incision. The line connecting the posterior superior iliac crest of the rabbit was level with the spinous process of the 7th lumbar vertebra. After touching the three spinous processes upward, the spinous processes of L4, L5, and L6 were located, and vertical lines were made respectively. The position between the three vertical lines was the target L4-5 and L5-6 intervertebral disc positions, which was the incision range. Routine disinfection and draping were performed. The skin was cut at the marked position, about 3.0 cm long, and the subcutaneous tissue was separated layer by layer to expose the junction of the white abdominal muscle aponeurosis and the back muscle. In order to facilitate the layer-by-layer suture after the operation, an incision was made 0.5 cm above the red and white dividing line. Continue to separate deeper along the muscle gap, and pay attention to protecting the nerves during exposure until the edge of the transverse process is touched. Continue to peel along the ventral side of the transverse process until the bulge at the front edge of the vertebral body is touched, which is the intervertebral disc. Use retractors on both sides to open the field of vision, and bluntly separate the muscle tissue at the front edge of the intervertebral disc to expose the intervertebral disc. Use a sharp knife to remove the anterior fiber ring, drill a hole to a depth of 4mm with a 2.5mm drill, and expand the hole to 3mm in the head-to-tail direction. Rinse with 0.9% saline, and press the two bottom surfaces of the cylindrical PEEK experimental comparison into the grooved position in the head-to-tail direction. Rinse again with 0.9% saline. Suture the surgical incision layer by layer. Each rabbit was randomly implanted with 2 groups of materials.

[0132] 2) Postoperative treatment: After surgery, all experimental rabbits were placed in clean cages for resuscitation. They were forbidden to eat or drink on the day of surgery, and their diet and water intake were resumed from the second day after surgery. Within 3 days after surgery, 800,000 units of veterinary penicillin sodium were injected intramuscularly every day. In addition, the rabbits' mental state, appetite, excretion, lower limb movement, wound healing, and survival status were observed daily.

[0133] 3. Gross specimen collection and observation

[0134] At 12 weeks after surgery, all 20 rabbits were killed by air embolism. After a midline incision was made on the back, the skin was cut open and the muscles on both sides of the spine were peeled off. The lumbar spine specimen was removed to observe the position of the implant, and to evaluate whether the implant was wrapped by tissue and whether there were signs of inflammation or abscess around it. At the same time, the hardness of the surrounding tissue after implantation was evaluated, and the degree of curvature of the fusion segment was compared with that of the adjacent normal intervertebral disc.

[0135] 4. Micro-CT examination

[0136] After gross observation of the specimens, the specimens were scanned using Micro-CT to measure and analyze bone tissue parameters: trabecular thickness, trabecular number, trabecular separation, bone volume fraction, and bone density.

[0137] 5. Histological Analysis

[0138] HE staining, Masson staining and Trap staining were used to evaluate the formation, maturation and osteoclastogenesis of new bone.

[0139] 6. Mechanical analysis

[0140] 12 weeks after surgery, the redundant structures such as the transverse process were cut off, and the surface muscles and connective tissue were removed, leaving only the vertebral bony structure. The vertebra was placed in a specific fixture for a compression fracture test. The cones at both ends and the middle intervertebral disc were a unit. The narrowest part of the cones at both ends was clamped on the two supports of the fixture, fixed with rubber bands, and crushed by a mechanical testing machine. The compression fracture curve and the maximum bearing capacity peak were analyzed.

[0141] 7. Statistical methods

[0142] The measurement data were expressed as mean ± standard deviation, and the comparison among multiple groups was performed using analysis of variance. When the p value was less than 0.05, the results were considered statistically significant. Data analysis was completed using SPSS26.0 and GraphPad Prism10.0 software.

[0143] 2. Experimental Results

[0144] 1. General appearance of the specimen

[0145] At 12 weeks after surgery, the lumbar specimens in each group were intact, but two groups of materials in the PEEK group fell off from the intervertebral space, and the PEEK-Zr / ZrO 2 、PEEK-FeNC、PEEK-Zr / ZrO 2 -FeNC, one group of materials fell out of the intervertebral space, and all other materials were in place. Considering that during the material implantation, due to the limitation of the use of non-professional tools, the surgeon could not use the drill to open up the ideal material implantation slot. The left and right sides of the slot were too wide or the depth was too shallow, which was the main reason for the material to fall out. Compared with the normal intervertebral disc on the head and tail sides, each group of implanted materials was covered with a thick layer of tough fibrous tissue, which was the same color and texture as the surrounding host bone surface, and no inflammatory reaction was observed. The surface of the PEEK group was covered with tough fibrous tissue, which was slightly white in color. The material was not visible from the outside, and it was slightly deformed after the bending force was applied. PEEK-Zr / ZrO 2 The appearance of the group showed that the new bone callus completely covered the material. The center of the callus was dark in color, similar to the color of the coating, and the surrounding color was the same as that of the normal lumbar spine. The implant site was slightly enlarged, and bone tissue hyperplasia and coverage were seen. Hyperplastic fibrous tissue was seen around the new bone wrapping, but the connection between the head and tail vertebrae was firm. 2 -The implanted part of the FeNC group was covered with tough fibrous tissue, which was thick and hard, closely integrated with the surrounding bone tissue, and had a smooth surface ( Fig.16 ).

[0146] 2. Micro-CT scan results

[0147] At 12 weeks after surgery, the Micro-CT scan results of the specimens in each group were shown in Fig.17 All samples in each group were successfully implanted into the intervertebral space. The upper and lower bottom surfaces of the cylindrical material corresponded to the head and tail of the rabbit, and the bone structure of the two vertebrae fit the surface of the material. A small amount of bone structure grew on the edges of the upper and lower bottom surfaces of the PEEK group material. A few trabeculae were discontinuous, with low thickness and large spacing between trabeculae. The interface between the bone and the material surface was rough and blurred. PEEK-Zr / ZrO 2 Compared with the PEEK group, the PEEK-FeNC group had continuous bone growth at the edges of the upper and lower bottom surfaces, and the trabeculae were continuously formed with average thickness. The interface between the bone and the material surface was relatively smooth, without obvious cavities and defects. 2 The thickness, density, and continuity of trabecular bone in the -FeNC group were similar and better than those in the first two groups, but the surface of the PEEK-FeNC group was rough and uneven. 2 The area of ​​new bone tissue around the -FeNC group was larger than that of the PEEK-FeNC group, and the surface was smoother.

[0148] 12 weeks after surgery, Micro-CT three-dimensional reconstruction images are shown in Fig.18 , the PEEK material was successfully implanted into the intervertebral space, and the outer surface was covered by a layer of callus. The normal intervertebral disc was no longer visible in the ventral side, new bone formation was seen at the ventral edge of the intervertebral space, the space became narrower, and the upper and lower vertebral edges were blurred. 2 The callus of the FeNC group was relatively smooth, and the PEEK and PEEK-Zr / ZrO 2 The callus formed in the group is rough and irregular in shape. In intervertebral fusion surgery, callus formation is an important indicator for evaluating the fusion effect. Round callus is usually regarded as a good sign of fusion because it indicates that the bone around the intervertebral implant is healing and forming a stable bone bridge. The formation of this callus usually means that the operation is successful, the fusion between the vertebrae is ongoing, and the implant is well integrated with the surrounding bone. In contrast, irregular callus may mean that there are some problems in the fusion process. These problems may include: poor implant position, which may affect the uniform formation of callus and cause irregular callus shape. Ideally, the fusion device should be located in the middle of the intervertebral space to ensure the mechanical stability of the anterior column and promote uniform callus formation; poor bone healing ability, slow callus formation and inflammatory response may form irregular callus; surgical techniques can also affect the fusion effect, insufficient intervertebral preparation, improper endplate processing, etc. However, combined with sagittal CT observation, the function of implant material surface modification has an important influence on bone integration.

[0149] 12 weeks after surgery, the parameters of new bone tissue around the two bottom surfaces of the implants in each group were shown in Fig.19 .PEEK-Zr / ZrO 2 -FeNC group had the best parameters, which were better than PEEK and PEEK-Zr / ZrO 2 In the trabecular thickness, there was no statistical significance in the pairwise comparison (p>0.05), but the two groups coated with Fe nanozymes were superior to the PEEK group (p<0.05). The number of trabeculae was inversely proportional to the separation of trabeculae. 2 -FeNC has no statistically significant difference with PEEK-FeNC (p>0.05), but has significant advantages over the two groups without nanozyme coating, and the difference is statistically significant (p<0.05). 2 -FeNC implant surface has excellent bone integration effect.

[0150] 3. Histological Evaluation

[0151] 1) HE staining

[0152] See also Fig. 20 , 12 weeks after surgery, the surface trabeculae of the PEEK group were sparse, no obvious bone integration was observed, and there was a defect at the contact point between the plant surface and the bone. 2 New bone tissue can be seen at the upper and lower interfaces of the group materials. The number of trabeculae is abundant, the texture is full, and they are interconnected. Osteoblasts can be seen in a single layer around the trabeculae, and the connective tissue is slightly hyperplastic. In the PEEK-FeNC group, more new bone tissue can be observed, the density of trabeculae is higher, the bone matrix is ​​lighter in color, the trabeculae are interconnected, and osteoblasts can be seen in a single layer around some trabeculae. A small amount of new blood vessels can be seen, and the connective tissue on the surface of the cortical bone is slightly hyperplastic. PEEK-Zr / ZrO 2 In the -FeNC group, a large area of ​​new bone tissue was observed at the bone contact surface, with a large number of trabeculae that were interconnected, and osteoblasts arranged in a single layer around the trabeculae. The trabeculae were not limited to the adjacent interface of the implant, but were larger in range and greater in number than in other groups.

[0153] 2) Masson staining

[0154] At 12 weeks after surgery, a small amount of new woven bone was observed in the PEEK group, which did not completely surround or cover the material. 2More new bone tissue can be observed in the PEEK group than in the PEEK group. These bone tissues surround the upper and lower interfaces of the material. At the same time, there is a slight proliferation of collagen fibers. A large amount of new bone tissue can be observed at the upper and lower interfaces of the PEEK-FeNC group, and some red and blue mature lamellar bones appear. At the same time, collagen fibers also show proliferation. PEEK-Zr / ZrO 2 The -FeNC group had the best bone integration effect, and the material was almost completely surrounded by dense lamellar bone. These bone tissues were not only dense, but also had regular arrangement of osteocytes, dense and numerous trabeculae, and mature lamellar bone ( Fig.21 ).

[0155] 3) Trap staining

[0156] 12 weeks after surgery, PEEK, PEEK-Zr / ZrO 2 Mature osteoclasts were observed around the trabeculae in the PEEK-FeNC and PEEK-Zr / ZrO 2 -FeNC group, the surface of the material and the larger area around the material were not stained with osteoclasts, indicating that the CAT enzyme reduction activity of the nanozyme may have a certain inhibitory effect on osteoclasts ( Fig. 22 ).

[0157] 4. Three-point compression test

[0158] 12 weeks after surgery, a three-point compression test was used to quantitatively evaluate intervertebral fusion. The experimental results showed that ( Fig.23 ), PEEK-Zr / ZrO 2 The intervertebral space implanted in the -FeNC group performed best in terms of force and deformation capacity, reaching a maximum peak of approximately 750N. 2 The intervertebral space implanted in the PEEK-FeNC group also showed higher strength, but the peak force was slightly lower than that of PEEK-Zr / ZrO 2 -FeNC. The mechanical properties of the intervertebral space implanted by PEEK were poor, and the peak force was significantly lower than that of other materials. 2 The peak pressure of the implants with surface structure treatment was significantly lower than that of the two groups with three-dimensional columnar structures on the surface, and the difference was statistically significant (p<0.05). 2 The results of the -FeNC group were statistically significant compared with the PEEK group (p<0.01), indicating that surface structure modification and nanozyme coating can improve the integration of PEEK plant material surface with bone.

[0159] The above description is only a preferred embodiment of the present invention, and does not limit the implementation mode and protection scope of the present invention. For those skilled in the art, it should be aware that all solutions obtained by equivalent substitutions and obvious changes made using the contents of the specification and illustrations of the present invention should be included in the protection scope of the present invention.

Claims

1. A PEEK-Zr / ZrO2-FeNC surface modified material, characterized in that: The invention comprises a PEEK substrate, Zr / ZrO2 deposited on the surface of the PEEK substrate, and FeNC nanozyme particles uniformly distributed on the outer surface after the Zr / ZrO2 is deposited.

2. The PEEK-Zr / ZrO2-FeNC surface modified material according to claim 1, characterized in that: The Zr / ZrO2 deposited on the surface of the PEEK substrate presents a uniform nanoscale three-dimensional columnar structure.

3. The PEEK-Zr / ZrO2-FeNC surface modified material according to claim 2, characterized in that: The FeNC nanozyme particles are embedded in the gaps of the three-dimensional columnar structure.

4. A method for preparing a PEEK-Zr / ZrO2-FeNC surface modified material according to any one of claims 1 to 3, characterized in that: The steps include: Step 1: After polishing and cleaning the medical PEEK substrate, PBHS deposition is performed using a radio frequency ionization plasma pulse bias injection system to obtain PEEK-Zr / ZrO2; Step 2, weighing ferric chloride hexahydrate and dissolving it in formamide solution, stirring at room temperature to obtain a precursor solution; placing the PEEK-Zr / ZrO2 obtained in step 1 in the precursor solution, reacting at 180-200°C for 4-8h, washing and drying after the reaction to obtain the PEEK-Zr / ZrO2-FeNC surface modified material.

5. The preparation method according to claim 4, characterized in that: The specific method of PBHS deposition using the RF ionization plasma pulse bias injection system is as follows: equip a Zr metal target, place the target on a turntable connected to a bias power supply, and then place the PEEK substrate on the target surface; use an RF plasma source to generate H2 plasma, and use BDP-5A to apply a unipolar voltage pulse to the target to provide a controlled constant bias pulse to ensure stability during the deposition process.

6. The preparation method according to claim 5, characterized in that: The PEEK substrate can be of any shape, and Zr / ZrO2 can be deposited on the upper surface of the substrate in a single preparation. When the PEEK substrate is a cylinder, Zr / ZrO2 is deposited on the upper and lower surfaces of the cylinder in two steps.

7. The preparation method according to claim 4, characterized in that: The concentration of ferric chloride hexahydrate in the precursor solution is 0.6-1.0 mg / mL.

8. Use of the PEEK-Zr / ZrO2-FeNC surface modified material according to any one of claims 1 to 3 in the preparation of an intervertebral fusion device product.

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