A polyetheretherketone surface nanoporous microcolumn structure for enhancing osteogenicity and antibacterial properties and its preparation method

Abstract

The present invention discloses a poly(ether ether ketone) surface nanoporous micro-pillar structure for enhancing osteogenic and antibacterial properties and its preparation method. The structure consists of a plurality of rhombus-shaped pillars arranged in a staggered array. The rhombuses have a side length of 15-20 μm, a height of 10-15 μm, and a gap of 8-12 μm between adjacent rhombuses. The top surfaces of the rhombuses are provided with nanoscale meshes. Nanosilver particles are adhered to the surfaces of the nanoscale meshes and the rhombuses, and zinc ions are chelated. Sulfonation, femtosecond, and chemical treatments are used to synergistically regulate the poly(ether ether ketone) surface, significantly improving its osteogenic activity and antibacterial properties.

Description

Technical Field

[0001] The present invention belongs to the technical field of bone implant preparation, and specifically relates to a polyetheretherketone surface structure with a nanoporous microcolumn structure and zinc-silver ions for enhancing osteogenicity and antibacterial properties, and further provides a preparation method of the structure. Background Art

[0002] The repair of bone defects caused by trauma and disease remains a clinical challenge, and bone implant materials such as autologous bone, ceramics, metals, and polymers are widely used in clinical applications. Due to the shortage of autologous bone sources, the current mainstream implants are metal implant materials. However, such materials are prone to release harmful excess metal ions that endanger human health, ceramic materials have poor ductility, and polymers have weak cell adhesion ability, which has led to a sharp increase in demand for bone implants. Compared with metals and ceramics, biomedical polymer materials have the advantages of high safety and good stability. Polyetheretherketone, as a polymer material with an elastic modulus close to that of natural cancellous bone, has a special molecular structure that gives it high mechanical strength, biocompatibility, and biostability, and is considered to be a substitute for titanium and its alloys. However, due to the insufficient surface hydrophilicity of polyetheretherketone materials, they cannot absorb proteins, inhibiting cell adhesion, resulting in poor bone integration and insufficient antibacterial properties, which greatly hinders its clinical application.

[0003] There is existing technology for surface processing of polyetheretherketone materials in order to achieve excellent hydrophilic properties. For example, application number 2020114742498 proposes the use of femtosecond laser to form a groove structure on the surface of the scanned material. After testing, it shows good hydrophilicity and osteogenic properties.

[0004] However, the above scheme does not further disclose the surface structure, its specific structure is unclear, and there is no theoretical basis for explanation and description. In addition, the preparation method is relatively simple and the corresponding surface structure cannot be prepared therefrom.

[0005] At the same time, the above scheme has not been studied in combination with antibacterial properties. Although the structure constructed by femtosecond laser design has a certain inhibitory effect on bacterial adhesion in terms of scale and arrangement, when the microstructure is filled with body fluids that can provide energy for the colony, it is still difficult to prevent the massive reproduction of bacteria, which ultimately leads to implant failure.

[0006] In addition, how to specifically set up this microstructure in terms of scale and arrangement to play a better barrier role has not yet been specifically addressed.

[0007] Therefore, as a person working in a related field, you should explore relevant issues. Summary of the Invention

[0008] In order to solve the problem that the existing technology has not proposed a surface structure based on polyetheretherketone material that can simultaneously enhance osteogenicity and antibacterial properties, the present invention provides a polyetheretherketone surface nanoporous microcolumn structure for enhancing osteogenicity and antibacterial properties, which has staggered diamond columns and specifically stipulates the size of the diamond columns and the distance between them, thereby constructing an active surface with excellent osteogenic and antibacterial properties. Furthermore, the present invention also provides a preparation method of the structure, so that the structure can be industrialized, providing the possibility for wide clinical application.

[0009] The purpose of the present invention is achieved through the following technical solutions:

[0010] The first aspect of the present invention discloses a polyetheretherketone surface nanoporous microcolumn structure for enhancing osteogenicity and antibacterial properties:

[0011] It consists of multiple diamond-shaped columns in a staggered array, each with a side length of 15-20 μm and a height of 10-15 μm. The gap between adjacent diamond-shaped columns is 8-12 μm. The top surface of the diamond-shaped columns is provided with nano-scale meshes. Nano-silver particles are adhered to the surface of the nano-scale meshes and the surface of the diamond-shaped columns, and are chelated with zinc ions.

[0012] Furthermore, the rhombus angle of the rhombus column is 50-85°.

[0013] Furthermore, the diamond-shaped columns are made of polyetheretherketone material.

[0014] A second aspect of the present invention discloses a method for preparing a nanoporous microcolumn structure on the surface of polyetheretherketone for enhancing osteogenicity and antibacterial properties, comprising the following steps:

[0015] S1: Sulfonating the surface of polyetheretherketone to obtain a nano-scale mesh surface structure;

[0016] S2: Performing femtosecond laser treatment on the nano-scale mesh surface structure to obtain a staggered diamond column array surface structure;

[0017] S3: Surface treatment is performed on the surface structure of the dislocated rhombus column array to obtain a surface structure of nanosilver particles and zinc ions.

[0018] Furthermore, in step S2, the power of the femtosecond laser treatment is 2-5 mW, the scanning speed is 0.005-0.015 m / s, and the scanning line spacing is 23-32 μm.

[0019] Furthermore, in step S3, the nanosilver particles and the zinc ion surface structure are immersed in a mixed solution of polydopamine and zinc chloride for 20 hours; and then immersed in a silver ammonia solution for 3 hours.

[0020] Furthermore, the polydopamine and zinc chloride mixed solution includes a 2 mg / mL polydopamine solution and a 2000 μmol / L zinc chloride solution.

[0021] Furthermore, in step S1, the sulfonation treatment includes ultrasonic cleaning, drying, soaking in concentrated sulfuric acid, soaking in deionized water, rinsing with acetone and distilled water, boiling in water, and vacuum drying.

[0022] Furthermore, the femtosecond laser treatment is deepened layer by layer, and each layer is scanned twice.

[0023] Furthermore, in each layer scan, one direction is scanned first and then the other direction is scanned.

[0024] Compared with the prior art, the present invention has at least the following advantages and beneficial effects:

[0025] 1. The main structure of the surface structure of the present invention is a rhombus column. The special design of the parameters related to the rhombus column makes the polyetheretherketone surface gain structural antibacterial properties:

[0026] On the one hand, the side length of the rhombus column is 15-20 μm, and its top area is obviously less than 400 μm 2 , thus helping to prevent bacteria from forming biofilms on top;

[0027] On the other hand, the gap between the diamond columns of 8–12 μm can effectively prevent bacteria from entering the interior of the groove;

[0028] Furthermore, its height is 10-15 μm, and the high aspect ratio structure is easy to tear bacteria;

[0029] Furthermore, experiments have shown that the staggered rhombus structure exhibits a dense structure, which reduces the air gap and reduces the contact angle. This structure is similar to the hexagonal structure of a natural honeycomb, with extremely high space utilization and good stability.

[0030] 2. The present invention also uses sulfonation treatment to make the surface of the structure nanoporous, which is conducive to more biological anchoring and improves implant stability. In combination with the staggered diamond column structure, it can promote the adhesion, proliferation and differentiation of osteoblasts, thereby improving osteogenic activity;

[0031] 3. The present invention also uses a chemical immersion treatment to form a chemical coating on the surface. The coating contains nanosilver particles and zinc ions. The nanosilver particles kill bacteria by releasing corresponding ions, further enhancing antibacterial properties. The zinc ions not only exhibit antibacterial properties against Gram-negative bacteria, but also stimulate bone formation by activating the proliferation and differentiation of osteoblasts, and also exhibit antibacterial properties against Gram-negative bacteria. Through polydopamine modification, the zinc ions can be combined with the coating to achieve an antibacterial effect.

[0032] 4. The present invention innovatively proposes a process scheme of first sulfonation and then femtosecond treatment, forming a porous mesh microstructure array composited with the microstructure, and the microstructure has a very regular shape, a dense arrangement and no defects, which is conducive to retaining the results of the two processes.

[0033] In summary, the present invention significantly improves the osteogenic activity and antibacterial properties by simultaneously regulating the structure and coating of the polyetheretherketone surface. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0035] Figure 1 This is a flow chart of the method for preparing the nanoporous microcolumn structure on the surface of polyetheretherketone of the present invention;

[0036] Figure 2 Schematic diagram of the sample of the present invention;

[0037] Figure 3 This is a schematic diagram of a local enlargement after femtosecond laser treatment;

[0038] Figure 4 The microscopic morphology of the sample and the samples obtained in each step;

[0039] Figure 5 Schematic diagram of the microscopic surface structure of the present invention;

[0040] Figure 6 is the microscopic morphology and contact angle of each structure;

[0041] Figure 7 Cell viability after 1, 3, 5, and 7 days of culture for PEEK, PP-Zn-Ag, SPP-Zn-Ag, and SLPP-Zn-Ag samples;

[0042] Figure 8 Live / dead staining of cells after 7 days of culture for PEEK, PP-Zn-Ag, SPP-Zn-Ag, and SLPP-Zn-Ag samples;

[0043] Figure 9 The cell morphology on the surface of PEEK, PP-Zn-Ag, SPP-Zn-Ag, and SLPP-Zn-Ag samples;

[0044] Figure 10The cell adhesion status on the surface of PEEK, PP-Zn-Ag, SPP-Zn-Ag and SLPP-Zn-Ag samples;

[0045] Figure 11 Semi-quantitative analysis of surface cell Col lagen Ⅰ staining of PEEK, PP-Zn-Ag, SPP-Zn-Ag and SLPP-Zn-Ag samples;

[0046] Figure 12 Semi-quantitative analysis of Runx2 staining of surface cells of PEEK, PP-Zn-Ag, SPP-Zn-Ag, and SLPP-Zn-Ag samples;

[0047] Figure 13 Semi-quantitative analysis of surface alizarin red (ARS) staining of PEEK, PP-Zn-Ag, SPP-Zn-Ag and SLPP-Zn-Ag samples;

[0048] Figure 14 The inhibition zones of PEEK samples and samples from different steps against Escherichia coli and Staphylococcus aureus are shown;

[0049] Figure 15 The diagram shows the effects of sulfonation and laser treatment in different sequences. DETAILED DESCRIPTION

[0050] To make the objectives, technical solutions, and advantages of the embodiments of the present invention more clear, the technical solutions of the embodiments of the present invention will be clearly and completely described below in conjunction with the accompanying drawings of the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Generally, the components of the embodiments of the present invention described and shown in the drawings herein can be arranged and designed in various different configurations.

[0051] Therefore, the following detailed description of the embodiments of the present invention provided in the accompanying drawings is not intended to limit the scope of the invention as claimed, but rather merely represents selected embodiments of the present invention. All other embodiments derived by persons of ordinary skill in the art based on the embodiments of the present invention without creative effort are also within the scope of protection of the present invention.

[0052] It should be noted that, in the absence of conflict, the embodiments and features of the embodiments of the present invention can be combined with each other.

[0053] It should be noted that similar reference numerals and letters denote similar items in the following drawings, and therefore, once an item is defined in one drawing, it does not need to be further defined or explained in subsequent drawings.

[0054] In the description of the present invention, it should be noted that the terms "center," "upper," "lower," "left," "right," "vertical," "horizontal," "inner," "outer," etc., indicating orientations or positional relationships, are based on the orientations or positional relationships shown in the accompanying drawings, or are the orientations or positional relationships in which the inventive product is typically placed when in use, or are the orientations or positional relationships commonly understood by those skilled in the art. These terms are intended only to facilitate the description of the present invention and to simplify the description, and are not intended to indicate or imply that the device or element referred to must have a specific orientation, be constructed, or operate in a specific orientation. Therefore, they should not be construed as limiting the present invention. Furthermore, the terms "first," "second," etc., are used only to distinguish descriptions and should not be construed as indicating or implying relative importance.

[0055] In the description of the present invention, it should also be noted that, unless otherwise expressly specified or limited, the terms "disposed," "installed," "connected," and "connected" should be understood in a broad sense. For example, they may refer to fixed connections, detachable connections, or integral connections; they may refer to mechanical connections or electrical connections; they may refer to direct connections or indirect connections through an intermediate medium; and they may refer to internal communication between two components. Those skilled in the art will understand the specific meanings of the above terms in the present invention based on the specific circumstances.

[0056] The present invention discloses a method for manufacturing a nanoporous microcolumn structure on the surface of polyetheretherketone for enhancing osteogenicity and antibacterial properties, and the equipment involved includes:

[0057] (1) A femtosecond laser integrated system (Spectra-Physics, USA, with a wavelength of 800 nm (adjustable), a pulse width of 120 fs, and a repetition rate of 1 kHz) that can be used to prepare micro-nano composite structures on the surface of polyetheretherketone;

[0058] (2) An independently equipped optical system and a set of high-speed scanning galvanometers (HurryScan II 10 from Germany); capable of focusing the light spot below 2 μm and moving the laser at high speed along a specific trajectory;

[0059] (3) A set of high-precision three-dimensional moving stages (Newport IMS-LM, USA, sub-micron repeatability) that can accurately control the multi-axis motion of the object being processed in the X, Y, and Z directions to achieve complex processing;

[0060] (4) Magnetic stirrer (S10-3 model), used to control the temperature and flow rate of concentrated sulfuric acid during the sulfonation reaction of polyetheretherketone to ensure a full and uniform reaction;

[0061] (5) A reactor that can remove residual sulfur ions on the surface of polyetheretherketone after the sulfonation reaction;

[0062] (6) Contact angle measurement system (Kruss, Germany, DSA30), which can observe the contact angle state and analyze the surface properties;

[0063] (7) Field emission scanning electron microscope (Zeiss, Ultra 55, Germany) for characterization of sample surface morphology; Energy dispersive X-ray spectrometer (EDX) for analysis and characterization of sample surface chemical composition;

[0064] (8) Optical profilometer (Bruker Countor GT K 3D), characterizing the 3D morphology and roughness of the sample;

[0065] (9) Inductively coupled plasma emission spectrometer (ICP, Thermo iCAP6500) was used to detect the concentration of ions released by the sample in PBS solution;

[0066] (10) X-ray photoelectron spectroscopy (XPS, Thermo Scientific K-Alpha), used for qualitative elemental analysis of the sample surface;

[0067] (11) reflectance-Fourier transform infrared spectroscopy (ATR-FTIR), used to verify the presence of polydopamine;

[0068] (12) Cell culture incubator (Sanyo, Japan), used for constant temperature simulation culture, heating reactor and drying samples;

[0069] (13) α-MEM basal medium (Gibco, USA), used to support the growth and reproduction of bone marrow mesenchymal stem cells;

[0070] (14) 24-well cell culture plates (Corning, USA), used for small-scale cell culture experiments;

[0071] (15)25 / 75cm 2 Cell culture flasks (Corning, USA) were used for large-scale cell culture experiments;

[0072] (16) Microplate spectrophotometer (Synergy H1), used to measure the optical density (OD) of the working solution;

[0073] (17) Two-photon confocal microscope (Nikon), used to observe the status of cells;

[0074] (18) Ultrasonic cleaning apparatus, used to remove residual concentrated sulfuric acid after sulfonation and sample residues after laser processing.

[0075] The materials used are:

[0076] Medical grade polyetheretherketone (10×10×1 mm) was purchased from Dongguan Senyue Changhong Trading Co., Ltd.; concentrated sulfuric acid (concentration 95-98 wt%); dopamine hydrochloride (Beijing Biolab Technology Co., Ltd.); acetone (Chengdu Kelong Chemical Co., Ltd.); Tri s-HCl buffer (10 mM, pH = 8.5, Aladdin); zinc chloride solution (2000 μM, Aladdin); silver nitrate solution (0.05 M, Aladdin); ammonia water (concentration 25%, Aladdin); phosphate-buffered saline (PBS, pH = 7.2-7.4); Escherichia coli (ATCC25922, Science Compass); Staphylococcus aureus (ATCC29213, Science Compass); CCK-8 kit (Dojindo Chemical Research Institute, Japan); third-generation bone marrow mesenchymal stem cells (BMSCs) (Shanghai Cell Bank, Chinese Academy of Sciences); penicillin-streptomycin (Gibco, USA); phalloidin-rhodamine backbone detection kit (Beijing Thermo Fisher Scientific Co., Ltd.); 6-diamindo-2-phenylindole (DAPI) (I Invitrogen, USA), calcein (Sigma, USA); fetal bovine serum (FBS) (Gibco, USA); ascorbic acid (Sigma, USA); dexamethasone (Sigma, USA); β-glycerophosphate (Sigma, USA); alizarin red (1% concentration, Sigma, USA); cetylpyridinium chloride (10% concentration, Sigma, USA).

[0077] The integrated femtosecond laser system used consists of three main components: a femtosecond laser system, a processing optical system, and a laser texturing system. First, the femtosecond laser system emits a femtosecond laser, which is then transmitted to the laser texturing system via a self-built processing optical system. Finally, the laser texturing system controls the movement of the femtosecond laser and the sample being processed, achieving surface microstructuring.

[0078] (1) Femtosecond laser system: This system consists of an oscillator, a stretcher, an amplifier, and a compressor. The principle of this system to generate ultrafast pulses with high peak power and short pulse width is that the femtosecond pulse is first generated by the oscillator, then stretched by the stretcher. The pulse then passes through the amplifier to increase the pulse energy, and finally compressed back to the femtosecond time scale by the compressor.

[0079] (2) Processing optical path system: This system is mainly used to control femtosecond laser energy, adjust laser energy density, and conduct light.

[0080] (3) Laser texturing system: This system mainly consists of a high-speed scanning galvanometer and a high-precision three-dimensional motion platform. The high-speed scanning galvanometer is used to control the ultra-high-speed and precise movement of the femtosecond laser, and the high-precision three-dimensional motion platform is used to adjust the relative position between the processed sample and the femtosecond laser. Both are controlled by computer software to control the specific movement speed, trajectory, position and distance. The coordinated use of the two motion control systems is conducive to achieving the diversity and precision of surface microstructures.

[0081] like Figure 1 As shown, the preparation process mainly includes the following steps:

[0082] S1: Sulfonating the surface of polyetheretherketone to obtain a nano-scale mesh surface structure;

[0083] S2: Performing femtosecond laser treatment on the nano-scale mesh surface structure to obtain a staggered diamond column array surface structure;

[0084] S3: Surface treatment is performed on the surface structure of the dislocated rhombus column array to obtain a surface structure of nanosilver particles and zinc ions.

[0085] The following specific embodiments are proposed below:

[0086] Example 1:

[0087] Step 1: Make samples

[0088] like Figure 2 As shown, a 10x10x1 mm PEEK plate was ultrasonically cleaned with ethanol and ultrapure water. After 5 minutes of cleaning, the sample was placed in a vacuum drying oven and dried at 60°C for 20 minutes. The PEEK sample was then immersed in 95-98 wt% concentrated sulfuric acid at 60°C and stirred for 5 minutes. After removal, it was immersed in deionized water to stop the reaction and further rinsed with acetone and distilled water. Subsequently, the sample was boiled at 120°C for 4 hours and then vacuum dried to obtain Sample SP.

[0089] Step 2: Femtosecond laser treatment

[0090] The process is carried out in a layer-by-layer deepening manner, with each layer progressing by 2-3 μm, ultimately reaching a height or depth of 10-15 μm. Each layer is scanned twice, once for deepening and twice for finishing, to make the surface neat and the outline clear. The scanning is performed in a linear manner, first scanning in one direction, then rotating the corresponding angle according to the rhombus angle and scanning in another direction. This allows a single scan in a certain direction, reducing the time for repeated adjustment of the scanning angle and improving operational efficiency. The rhombus angle of this embodiment is not limited and can be any value between 50-85°. When the side length of the rhombus column is less than 20 μm, its surface area is always less than 400 μm. 2, which helps prevent bacteria from forming biofilm on the top. In this embodiment, the laser power is 2mW, the scanning speed is 0.005-0.015m / s, and the line spacing is 23-32μm. Rinse with ultrapure water and dry to obtain sample SLP, such as Figure 3 shown.

[0091] Step 3: Dissolve dopamine hydrochloride in 10mM Tris-HCl (pH=8.5) buffer to prepare a 2mg / mL polydopamine solution. Then immerse the sample in a solution containing polydopamine and 2000μM zinc chloride, protect from light, and soak at 60°C for 20h. Gradually add ammonia water to the 0.05M AgNO3 solution and stir continuously to prepare the Tol Lens reagent. Then soak the sample in the Tol Lens reagent at room temperature for 3 hours. Finally, remove the sample, rinse with ultrapure water, and air-dry at room temperature to obtain the sample SLPP-Zn-Ag.

[0092] Through the above steps, we conducted microscopic morphology analysis on the bare PEEK sample and the samples obtained in each step, such as Figure 4 .

[0093] The surface microstructure prepared by the above scheme is as follows Figure 5 As shown, it is composed of multiple diamond columns in a staggered array, with a side length of 15-20 μm, a height of 10-15 μm, and a gap of 8-12 μm between adjacent diamond columns. Nanoscale meshes are provided on the top surface of the diamond columns, and nanosilver particles are adhered to the top surface of the diamond columns and the surface of the nanoscale meshes, and zinc ions are chelated.

[0094] Compared with other structures: the surface of periodic groove structure, the surface of pit array structure, the surface of micro protrusion array structure, etc. Figure 6 As shown, the size of each structure is between 15-25 μm, and the height or depth is greater than 10 μm.

[0095] Among them, (a) the surface and contact angle of the periodic groove structure; (b) the surface and contact angle of the pit array structure; (c) the surface and contact angle of the micro-protrusion array structure; (d) the surface and contact angle of the staggered diamond structure.

[0096] In order to reflect the roughness and hydrophilicity of the surface, the contact angle of each surface was measured. The results show that, affected by femtosecond laser processing, the contact angles of the surfaces of each structure increased compared to the original smooth surface CA = 75°, and were all close to the hydrophobic state. Among them, the micro-protrusion array structure surface showed the maximum contact angle CA = 99°, while the staggered diamond structure surface showed the minimum contact angle A = 81°, and the other two structures were in the middle. The main reason is due to the relatively water-repellent properties of the material and the different structural shapes and arrangements. According to the Cass ie-Baxter contact angle model state analysis, due to the large gaps between the periodic groove structure, the pit array structure, and the micro-protrusion array structure, a large air cushion will be formed, and the supported droplet cannot collapse, thus showing a larger contact angle. Compared with other structures, the staggered diamond structure surface shows the density between the structures, which reduces the air cavity gap and the contact angle is low. This structure is similar to the hexagonal structure of a honeycomb in nature, with extremely high space utilization and good stability.

[0097] In addition, in the case of equal-length dimensions obtained under the same precision, the top area of ​​the rhombus structure is much smaller than that of the hexagonal structure, which is consistent with the fact that the smaller the top structure area is and the smaller it is, the better. 2 The conclusion is that it helps prevent bacteria from forming biofilms on the top. At the same time, structures of specific sizes affect the initial attachment and biofilm formation of E. coli. For example, extremely small gaps prevent bacteria from entering the grooves, while high aspect ratio structures with a depth greater than 10 μm easily tear bacteria apart. Therefore, Figure 5 As shown, the inventors have optimized the design of the staggered diamond structure array and constructed a staggered diamond structure with a side length of about 20 μm (area less than 400 μm) under the maximum precision of laser processing. 2 ), with a height of about 10-15μm (>10μm) and a gap of about 8-12μm, thereby making the polyetheretherketone surface gain in structural antibacterial properties.

[0098] The above scheme stipulates the process route of sulfonation first and then femtosecond laser, such as Figure 15 As shown in the figure, when the laser is first used and then sulfonated, that is, from c to d, the micron-scale array constructed by the femtosecond laser is very fragile and cannot withstand the sulfonation effect, and cannot maintain the clear array of the femtosecond laser. However, when the solution from a to b is adopted, that is, sulfonation first and then femtosecond laser, a very regular micron-scale array structure can be formed, which is densely arranged and has no defects.

[0099] Example 2:

[0100] Step 1: Ultrasonic cleaning of the polyetheretherketone (PEEK) sample was performed using ethanol and ultrapure water. After 5 minutes of cleaning, the sample was placed in a vacuum drying oven and dried at 60°C for 20 minutes. The PEEK sample was then immersed in concentrated sulfuric acid (95-98 wt%) at 60°C and stirred for 5 minutes. After removal, the sample was immersed in deionized water to stop the reaction and further rinsed with acetone and distilled water. Subsequently, the sample was boiled at 120°C for 4 hours and then vacuum dried to obtain Sample SP.

[0101] Step 2: Construct a micron-scale columnar structure by femtosecond laser. The process is carried out in a layer-by-layer deepening manner, with each layer increasing by 1.6-2.2μm, and finally reaching a height of 10-15μm. Each layer is scanned twice, once for deepening and twice for finishing, to make the surface neat and the outline clear. The scanning is done in a linear manner, first scanning in one direction, then rotating the corresponding angle according to the rhombus angle, and scanning in the other direction. The rhombus angle of this embodiment is not limited and can be any value between 50-85°. When the side length of the rhombus column is less than 20μm, its surface area is always less than 400μm. 2 , thereby helping to prevent bacteria from forming biofilms on the top, the laser power in this embodiment is 5 mW, the scanning speed is 0.005-0.015 m / s, and the line spacing is 23-32 μm. The sample SLP is rinsed with ultrapure water and dried.

[0102] Step 3: Dissolve dopamine hydrochloride in 10mM Tris-HCl (pH=8.5) buffer to prepare a 2mg / mL polydopamine solution. Then immerse the sample in a solution containing polydopamine and 2000μM zinc chloride, protect from light, and soak at 60°C for 20h. Gradually add ammonia water to the 0.05M AgNO3 solution and stir continuously to prepare the Tol Lens reagent. Then soak the sample in the Tol Lens reagent at room temperature for 3 hours. Finally, remove the sample, rinse with ultrapure water, and air-dry at room temperature to obtain the sample SLPP-Zn-Ag.

[0103] Example 3:

[0104] Step 1: First, ultrasonically clean the PEEK sample with ethanol and ultrapure water. After 5 minutes of cleaning, the sample was placed in a vacuum drying oven and dried at 60°C for 20 minutes. Next, the PEEK sample was immersed in 60°C concentrated sulfuric acid (95-98 wt%) and stirred for 5 minutes. After removal, the sample was immersed in deionized water to stop the reaction and further rinsed with acetone and distilled water. Subsequently, the sample was boiled at 120°C for 4 hours and then vacuum dried to obtain Sample SP.

[0105] Step 2: Construct a micron-scale columnar structure by femtosecond laser. The process is carried out in a layer-by-layer deepening manner, with each layer increasing by 1.6-2.2μm, and finally reaching a height of 10-15μm. Each layer is scanned twice, once for deepening and twice for finishing, to make the surface neat and the outline clear. The scanning is done in a linear manner, first scanning in one direction, then rotating the corresponding angle according to the rhombus angle, and scanning in the other direction. The rhombus angle of this embodiment is not limited and can be any value between 50-85°. When the side length of the rhombus column is less than 20μm, its surface area is always less than 400μm. 2 , thereby helping to prevent bacteria from forming biofilms on the top, the laser power in this embodiment is 3 mW, the scanning speed is 0.01 m / s, and the line spacing is 28 μm. The sample SLP is rinsed with ultrapure water and dried.

[0106] Step 3: Dissolve dopamine hydrochloride in 10mM Tris-HCl (pH=8.5) buffer to prepare a 2mg / mL polydopamine solution. Then immerse the sample in a solution containing polydopamine and 2000μM zinc chloride, protect from light, and soak at 60°C for 20h. Gradually add ammonia water to the 0.05M AgNO3 solution and stir continuously to prepare the Tol Lens reagent. Then soak the sample in the Tol Lens reagent at room temperature for 3 hours. Finally, remove the sample, rinse with ultrapure water, and air-dry at room temperature to obtain the sample SLPP-Zn-Ag.

[0107] Example 4:

[0108] Step 1: Ultrasonic clean the sample with ethanol and ultrapure water. After rinsing for 5 minutes, place the sample in a vacuum drying oven and dry at 60°C for 20 minutes. Subsequently, boil the sample at 120°C for 4 hours and then vacuum dry it. Dissolve dopamine hydrochloride in 10 mM Tris-HCl (pH = 8.5) buffer to prepare a 2 mg / mL polydopamine solution. Then, immerse the sample in a solution containing polydopamine and 2000 μM zinc chloride at 60°C for 20 hours in the dark. Tollens reagent is prepared by gradually adding ammonia to a 0.05 M AgNO3 solution with constant stirring. The sample is then immersed in Tollens reagent at room temperature for 3 hours. Finally, remove the sample, rinse with ultrapure water, and air-dry at room temperature to obtain the PP-Zn-Ag sample.

[0109] Step 2: Ultrasonic clean the sample with ethanol and ultrapure water. After 5 minutes of cleaning, place the sample in a vacuum drying oven and dry it at 60°C for 20 minutes. Then, immerse the polyetheretherketone sample in 60°C concentrated sulfuric acid (95-98 wt%) and stir for 5 minutes. Remove the sample and immerse it in deionized water to stop the reaction. Rinse it further with acetone and distilled water. Then, boil the sample at 120°C for 4 hours and vacuum dry it. Dissolve dopamine hydrochloride in 10 mM Tris-HCl (pH = 8.5) buffer to prepare a 2 mg / mL polydopamine solution. Then, immerse the sample in a solution containing polydopamine and 2000 μM zinc chloride at 60°C for 20 hours, protected from light. Tollens reagent is prepared by gradually adding ammonia to a 0.05 M AgNO3 solution with constant stirring. The sample is then immersed in Tollens reagent at room temperature for 3 hours. Finally, the sample was fished out, rinsed with ultrapure water, and air-dried at room temperature to obtain the sample SPP-Zn-Ag.

[0110] Step 3: Ultrasonic cleaning of the sample with ethanol and ultrapure water. After 5 minutes of cleaning, the sample was placed in a vacuum drying oven and dried at 60°C for 20 minutes. The polyetheretherketone sample was then immersed in 60°C concentrated sulfuric acid (95-98 wt%) and stirred for 5 minutes. After removal, it was immersed in deionized water to stop the reaction and further rinsed with acetone and distilled water. Subsequently, micrometer-scale columnar structures were constructed using a femtosecond laser. In this experiment, a linear scanning method was used. The laser power was 3 mW, and the line spacing was 28 μm. The sample was rinsed with ultrapure water and dried. Dopamine hydrochloride was dissolved in 10 mM Tris-HCl (pH = 8.5) buffer to prepare a 2 mg / mL polydopamine solution. The sample was then immersed in a solution containing polydopamine and 2000 μM zinc chloride, protected from light, and soaked at 60°C for 20 hours. Ammonia water was gradually added to a 0.05 M AgNO3 solution with constant stirring to prepare the Tol lens reagent. The sample was then immersed in Tol lens reagent at room temperature for 3 hours. Finally, the sample was fished out, rinsed with ultrapure water, and air-dried at room temperature to obtain the SLPP-Zn-Ag sample.

[0111] The following is an analysis of the proliferation capacity of the bare PEEK sample and the samples obtained in each step:

[0112] (1) CCK-8 proliferation assay

[0113] This experiment was conducted with cell culture for 7 days. The results showed that in the first three days, there was no significant difference in cell proliferation between PEEK, PP-Zn-Ag, SPP-Zn-Ag, and SLPP-Zn-Ag. However, by the fifth day, all processed samples showed higher cell viability than the original PEEK (see Figure 7By day seven, the difference was even more pronounced, with SLPP-Zn-Ag exhibiting the highest cell viability. These results demonstrate that both femtosecond laser processing and the introduction of active metal ions positively impact cell viability. This suggests that these processing and functionalization methods have the potential to enhance the cytocompatibility of polyetheretherketone materials for biomedical applications.

[0114] (2) Live / dead cell staining experiment

[0115] In order to improve the antibacterial properties of the samples, silver ions were introduced into this experiment. However, when the concentration of silver ions released by the implant material is too high, it may cause cytotoxicity and have a negative impact on surrounding tissues and cells. In order to comprehensively analyze the effect of silver ions released by the samples on cells, we conducted a live / dead cell staining experiment. After statistically analyzing the cell live / dead rates, we found that the values ​​were within the normal range, such as Figure 8 This phenomenon indicates that the concentration of silver ions released from all samples was lower than its toxic concentration in the body. Furthermore, the sulfonation treatment employed an appropriate cleaning method to effectively remove any sulfuric acid that may have remained on the implant material, and the cleaned implant material was not toxic to cells.

[0116] (3) Cytoskeleton staining experiment

[0117] Implant surface morphology can affect the behavior of cells on the implant surface, thereby affecting the tissue integration and biocompatibility of the implant. In order to gain a deeper understanding of the effects of different surface morphologies on the cytoskeleton, we performed cytoskeleton staining and observed and analyzed the Figure 9 The following conclusions were drawn. At day 7, there was little difference in cell arrangement between the polyetheretherketone, PP-Zn-Ag, and SPP-Zn-Ag samples. However, on the SLPP-Zn-Ag, cells were regularly arranged and exhibited a unique weaving effect, forming a distinct cellular braided structure. This weaving structure was consistent with the diamond-shaped structure formed on the implant surface using femtosecond laser processing.

[0118] (4) Scanning electron microscopy observation

[0119] In addition, scanning electron microscopy revealed that the cell shape on the surface of the SLPP-Zn-Ag sample was significantly different from that of other samples, showing a bulging state. It is worth noting that some cells' pseudopodia even extended into the nanopores produced by the sulfonation treatment (see Figure 10 This suggests that the microstructure of the implant surface may directly influence cell attachment and morphology. Although the underlying mechanisms underlying these cell morphologies and arrangements require further investigation, it is presumed that there is a close correlation with the sample surface structure.

[0120] This discovery not only highlights the regulatory role of implant surface morphology on cell behavior, but also reminds us to pay attention to the influence of microstructure on the interaction between cells and implant surfaces. These observations provide important clues for further revealing the mechanism of interaction between implant surfaces and cells. In addition, the formation of the aforementioned woven structure may provide cells with a more ordered and stable support matrix, which has potential benefits for improving tissue integration and biocompatibility. This comprehensive study helps to deepen the understanding of the interaction between implants and biological tissues and provides inspiration for the design of more effective biomaterials.

[0121] In addition, the inventors also performed Col agen Ⅰ staining, Runx2 staining and Alizarin red (ARS) staining analysis on polyetheretherketone, PP-Zn-Ag, SPP-Zn-Ag and SLPP-Zn-Ag samples, respectively. Figure 11 、 12 , as shown in 13.

[0122] Furthermore, the inventors analyzed the antibacterial ability of the bare PEEK sample and the samples obtained in each step, as shown in FIG. Figure 14 Figure 2 shows the inhibition zones of samples prepared at different steps against Escherichia coli and Staphylococcus aureus (a: E. coli, b: S. aureus). As can be seen, the surface structures of polyetheretherketone and PP-Zn-Ag did not exhibit significant antibacterial effects. However, SPP-Zn-Ag and SLPP-Zn-Ag, which possess a porous micropillar structure, exhibited significant antibacterial properties. Further analysis revealed that SLPP-Zn-Ag exhibited superior inhibition against both bacteria compared to SPP-Zn-Ag. This difference can be attributed to the significantly higher Zn and Ag contents in SLPP-Zn-Ag compared to SPP-Zn-Ag, which possesses only a porous structure. Therefore, the combination of the porous micropillar structure and the enhanced release of Zn and Ag ions enables SLPP-Zn-Ag to exhibit superior antibacterial activity against E. coli and S. aureus.

[0123] Although the present invention has been described in detail with reference to the aforementioned embodiments, it is still possible for those skilled in the art to modify the technical solutions described in the aforementioned embodiments, or to make equivalent substitutions for some of the technical features therein. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A polyetheretherketone capable of enhancing osteogenic and antibacterial properties, characterized in that: The surface of the polyetheretherketone is prepared with a nanoporous microcolumn structure, which is composed of a plurality of diamond columns in a staggered array. The diamond columns have a side length of 15-20 μm, a height of 10-15 μm, and a gap of 8-12 μm between adjacent diamond columns. The top surface of the diamond columns is provided with nanoscale meshes. Nanosilver particles are adhered to the surface of the nanoscale meshes and the surface of the diamond columns, and zinc ions are chelated. A porous mesh microstructure array composited with the microstructure is formed by first sulfonating the polyetheretherketone surface and then performing a femtosecond treatment. Then, a chemical immersion treatment is performed to form a chemical coating on the surface, forming a coating containing nanosilver particles and zinc ions.

2. The polyetheretherketone capable of enhancing osteogenic and antibacterial properties according to claim 1, characterized in that: The rhombus angle of the rhombus column is 50-85°.

3. The polyetheretherketone capable of enhancing osteogenic and antibacterial properties according to claim 1, characterized in that: The diamond-shaped columns are made of polyetheretherketone material.

4. A method for producing a polyetheretherketone capable of enhancing osteogenic and antibacterial properties according to any one of claims 1 to 3, characterized in that: The following steps are involved: S1: Sulfonating the surface of polyetheretherketone to obtain a nano-scale mesh surface structure; S2: Performing femtosecond laser treatment on the nano-scale mesh surface structure to obtain a staggered diamond column array surface structure; S3: Surface treatment is performed on the surface structure of the dislocated rhombus column array to obtain a surface structure of nanosilver particles and zinc ions.

5. The method for producing polyetheretherketone capable of enhancing osteogenic and antibacterial properties according to claim 4, characterized in that: In step S2 , the power of the femtosecond laser treatment is 2-5 mW, the scanning speed is 0.005-0.015 m / s, and the scanning line spacing is 23-32 μm.

6. The method for producing polyetheretherketone capable of enhancing osteogenic and antibacterial properties according to claim 4, characterized in that: In step S3, the preparation method of the nanosilver particles and the zinc ion surface structure is to soak them in a mixed solution of polydopamine and zinc chloride for 20 hours; Then soak in silver ammonia solution for 3 hours.

7. The method for producing polyetheretherketone capable of enhancing osteogenic and antibacterial properties according to claim 6, characterized in that: The polydopamine and zinc chloride mixed solution includes a 2 mg / mL polydopamine solution and a 2000 μmol / L zinc chloride solution.

8. The method for producing polyetheretherketone capable of enhancing osteogenic and antibacterial properties according to claim 4, characterized in that: In step S1, the sulfonation treatment includes ultrasonic cleaning, drying, soaking in concentrated sulfuric acid, soaking in deionized water, rinsing with acetone and distilled water, boiling in water, and vacuum drying.

9. A method for producing polyetheretherketone capable of enhancing osteogenic and antibacterial properties according to any one of claims 4 to 8, characterized in that: Femtosecond laser treatment deepens layer by layer, and each layer is scanned twice.

10. The method for producing polyetheretherketone capable of enhancing osteogenic and antibacterial properties according to claim 9, characterized in that: In each layer scan, scan one direction first and then the other direction.

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