Preparation method and application of fiber material with nanopore structure

Abstract

The invention provides a preparation method and application of a fiber material with a nanopore structure. The preparation method of the fiber material with the nano-pore structure comprises the following steps: preparing polyether-ether-ketone fibers, constructing a chelate coating formed by chelating phytic acid and strontium ions on the surfaces of the polyether-ether-ketone fibers, and carrying out electron bombardment treatment on the chelate coating to form the nano-pore structure on the surfaces of the chelate coating. The application is that the fiber material with the nano-pore structure is woven into the artificial ligament fabric. According to the invention, the biological activity of strontium, the chelation function of phytic acid and the physical advantages of a nano-pore structure are creatively combined to realize a synergistic effect of multiple functions, continuous and controllable strontium ion release can be provided, osteogenesis biochemical signals can be activated, bionic physical attachment points are provided for cells, cell behaviors are guided, and the bioactivity of the cells is improved. The composite material can be used for adjusting the microenvironment around the implant and relieving oxidative stress injury, provides a novel solution and material for promoting tendon-bone healing and bone regeneration, and has great popularization and application values.

Description

Technical Field

[0001] This invention relates to a method for preparing a fiber material and its application, and more particularly to a method for preparing a fiber material with a nanoporous structure and its application, belonging to the technical field of preparation and application of medical orthopedic implant materials. Background Technology

[0002] In the field of orthopedic implants, the performance of artificial ligaments is crucial, directly affecting the recovery of patients' motor function. Polyetheretherketone (PEEK) is considered a promising implant material due to its elastic modulus being similar to that of human bone and its good chemical stability. However, the inherent bioinertness and hydrophobicity of PEEK's surface result in poor integration with bone tissue, especially in the "insertion point" region where the ligament and bone unite, making it difficult to achieve firm osseointegration and thus affecting the long-term stability of the implant.

[0003] To improve the bioactivity of PEEK, common strategies include surface modification and composite bioactive materials. For example, increasing its surface hydrophilicity through plasma treatment or coating technology, or incorporating hydroxyapatite to promote osteogenic formation. However, these methods have limitations: the surface modification layer may wear off, and adding fillers may affect its excellent mechanical properties. In addition, the method of directly loading growth factors has problems such as high cost, easy inactivation, and possible side effects.

[0004] Osteogenesis is a complex biological process precisely regulated by multiple signaling pathways. Studies have shown that strontium (Sr), an essential trace element for the human body, has unique advantages in promoting bone formation. It not only promotes osteoblast differentiation and the expression of bone-forming genes (such as Runx2) by influencing the Wnt signaling pathway (e.g., reducing the adverse effects of GSK-3β), but also inhibits osteoclast activity, thus playing a dual role in promoting bone formation. On the other hand, the microstructure of the implant surface has a significant impact on cell behavior. Nanoscale topologies, especially nanopores, can mimic the physical environment of the natural bone matrix, effectively guiding osteoblast adhesion, spread, and differentiation, and enhancing the osteogenic response by activating integrin-mediated signal transduction.

[0005] Phytic acid (PA, also known as inositol hexaphosphate) is a naturally occurring organophosphorus compound containing multiple phosphate groups, giving it a strong ability to chelate metal ions and good electrical conductivity and antioxidant properties. This characteristic makes it an ideal bridge for surface modification, as it can both form stable complexes with metal ions and act as a functional coating to improve the performance of the substrate material.

[0006] Existing artificial ligament materials often focus on improving only one aspect, either by providing chemical signals (such as ion release) or physical signals (such as surface roughening), making it difficult to achieve synergistic effects and effectively promote tendon-bone healing and bone regeneration. Therefore, they have many shortcomings as medical orthopedic implant materials. Summary of the Invention

[0007] To overcome the shortcomings of existing technologies and address the key technical bottlenecks of insufficient bioactivity and weak osseointegration capacity of traditional artificial ligament materials, thereby promoting tendon-bone healing and bone regeneration and enabling patients to recover as soon as possible, the following measures are taken:

[0008] This invention first provides a method for preparing a fiber material with a nanoporous structure, comprising the following steps:

[0009] Prepare polyetheretherketone (PEEK) fibers; construct a chelate coating of phytic acid and strontium ions on the surface of the PEEK fibers; subject the chelate coating to electron bombardment treatment to form a nanoporous structure on the surface of the chelate coating.

[0010] Preferably, the chelate coating is constructed using a stacking assembly technique.

[0011] The stacked assembly technology includes the following steps:

[0012] The polyetheretherketone fiber is immersed in a phytic acid solution, so that phytic acid molecules adhere to the surface of the polyetheretherketone fiber.

[0013] Cleaning removes any unattached phytic acid molecules;

[0014] The polyetheretherketone fiber, after the phytic acid molecules have been attached, is then immersed in a strontium ion solution, so that the strontium ions in the strontium ion solution react with the phytic acid molecules attached to the surface of the polyetheretherketone fiber to form a chelate coating.

[0015] Cleaning removes unreacted strontium ions;

[0016] Repeat the above steps multiple times until the thickness of the chelate coating meets the design requirements.

[0017] Further:

[0018] The phytic acid solution has a volume percentage concentration of 3-7%, and the pH value of the phytic acid solution is adjusted to 4.5-6.5 by using an alkaline solution;

[0019] The polyetheretherketone fiber is immersed in phytic acid solution for 0.5 to 2 hours;

[0020] The strontium ion solution is a strontium nitrate (i.e., Sr(NO3)2) solution with a concentration of 0.3–0.7 M;

[0021] The polyetheretherketone fiber with phytic acid molecule attachment is immersed in strontium ion solution for 15 to 45 minutes;

[0022] The stacked assembly is repeated 5 to 9 times.

[0023] Preferred:

[0024] The phytic acid solution has a volume percentage concentration of 5% and a pH value of 5.5, while the alkaline solution for adjusting the pH value of the phytic acid solution is a 1M NaOH solution.

[0025] The polyetheretherketone fiber was immersed in the phytic acid solution for 1 hour;

[0026] The strontium nitrate solution has a concentration of 0.5 M;

[0027] The polyetheretherketone fiber, after phytic acid molecule attachment, is immersed in a strontium ion solution for 30 minutes.

[0028] Preferably, the stacked assembly is repeated 7 times.

[0029] Furthermore, the solvent used to clean and remove unattached phytic acid molecules is water, and the solvent used to clean and remove unreacted strontium ions is hydrogen peroxide.

[0030] Furthermore, the electron bombardment treatment has a bombardment voltage of 15–25 kV and a bombardment treatment duration of 5–15 seconds.

[0031] Preferably, the electron bombardment treatment has a bombardment voltage of 20 kV and a bombardment treatment duration of 10 seconds.

[0032] Secondly, the present invention also provides a fiber material with a nanoporous structure, wherein the fiber material with a nanoporous structure is prepared by any one of the preparation methods described above.

[0033] Furthermore, the present invention also provides an artificial ligament fabric, which is woven from the aforementioned fibrous material having a nanoporous structure. Compared with the prior art, the outstanding beneficial effects and significant progress of the present invention are as follows:

[0034] First, this invention utilizes the chelating ability of phytic acid (PA) to attach a PA / Sr chelate coating to the surface of polyetheretherketone (PEEK) fibers, forming a chelate coating. Then, a nanoporous structure is created on the surface of the chelate coating by electron bombardment, thus creating a novel functionalized bio-fiber material. That is, a polyetheretherketone fiber with a PA / Sr chelate coating on its surface and the chelate coating having a nanoporous structure. This fiber can be woven into medical orthopedic implants with excellent physiological and physical properties, especially for use as artificial ligaments.

[0035] This invention innovatively combines the bioactivity of strontium, the chelating function of phytic acid, and the physical advantages of nanoporous structures. Specifically, it first utilizes the chelating ability of phytic acid to construct a stable phytic acid / strontium (PA / Sr) chelate coating on the surface of PEEK fibers. Then, it uses an electron bombardment method to create a nanoporous structure on the surface of this coating to achieve a multi-functional synergistic effect. The phytic acid-strontium chelate can provide continuous and controllable strontium ion release, activating osteogenic biochemical signals; the nanoporous structure provides biomimetic physical attachment sites for cells, guiding cell behavior; and the conductivity and antioxidant properties of phytic acid itself may help regulate the microenvironment around the implant and reduce oxidative stress damage.

[0036] Therefore, by constructing this PEEK fiber with multi-layered functionalization, the present invention solves the key technical bottlenecks of insufficient bioactivity and weak bone integration capacity of traditional artificial ligament materials, and provides a new solution and material for promoting tendon-bone healing and bone regeneration.

[0037] Studies have shown that this functionalized PEEK fiber material with a nanoporous structure overcomes the shortcomings of existing technologies and solves key technical bottlenecks such as insufficient bioactivity and weak bone integration capacity of traditional medical orthopedic implants. It can promote tendon-bone healing and bone regeneration, enabling patients to recover sooner. Compared with existing technologies, it has outstanding beneficial effects and significant progress, and therefore has great value for promotion and application. Attached Figure Description

[0038] To more clearly illustrate the technical solution of the present invention and the technical effects of implementing the present invention, the accompanying drawings used in the examples of the effects of the present invention will be briefly introduced below.

[0039] It is obvious:

[0040] The accompanying drawings described below are only a portion of the drawings used in the examples of the effects of this invention. For those skilled in the art, other drawings can be obtained from these drawings without any creative effort, but these other drawings also fall within the scope of the drawings required for the embodiments of this invention, wherein:

[0041] Figure 1The SEM image set of PKPS provided in Example 1 of the present invention;

[0042] Figure 2 The SEM image set of PKPSP provided in Example 1 of the present invention;

[0043] Figure 3 The PKPS P-element distribution image provided in Example 1 of the present invention;

[0044] Figure 4 The image of Sr element distribution in PKPS provided in Example 1 of the present invention;

[0045] Figure 5 The EDS spectrum of PKPS provided in Example 1 of the present invention;

[0046] Figure 6 Example 1 of the present invention provides a P-element distribution image of PKPSP;

[0047] Figure 7 The image of Sr element distribution in PKPSP provided in Example 1 of the present invention;

[0048] Figure 8 The EDS spectrum of PKPSP provided in Example 1 of the present invention;

[0049] Figure 9 The FTIR detection spectra of PKF, PKPS, and PPKSP provided in Example 1 of the present invention;

[0050] Figure 10 The XRD patterns of PKF, PKPS, and PPKSP provided in Example 1 of the present invention;

[0051] Figure 11 A bar chart showing the contact angles of PKF, PKPS, and PKBSP in contact with diiodomethane and water, provided as an example of the effectiveness of this invention;

[0052] Figure 12 The surface energy histograms of PKF, PKPS, and PKBSP provided in Example 1 of the present invention;

[0053] Figure 13 The protein adsorption bar charts of PKF, PKPS, and PPKSP provided in Example 1 of the present invention;

[0054] Figure 14 The conductivity histograms of PKF, PKPS, and PKBSP provided in Example 1 of the present invention;

[0055] Figure 15 A bar chart showing the tensile strength of PKF, PKPS, and PKBSP provided in Example 1 of the present invention;

[0056] Figure 16 Bar chart showing the free radical scavenging capabilities of PKF, PKPS, and PPKSP as shown in Example 1 of the present invention;

[0057] Figure 17 The release curves of strontium ions after immersion in PBS for different times by PKPS and PPKSP as provided in Example 1 of the present invention;

[0058] Figure 18 SEM image sets of PKF, PKPS, and PKBSP co-cultured with BMSCs for different time periods were provided for Example 2 of the present invention.

[0059] Figure 19 The CLSM image set provided in Example 2 of the present invention, after co-culturing PKF, PKPS, and PKBSP with BMSC for different time periods;

[0060] Figure 20 The bar chart showing the adhesion performance of BMSCs after being cultured on PKF, PKPS and PPKSP surfaces for different times is provided for Example 2 of the present invention.

[0061] Figure 21 The bar chart showing the proliferation of BMSCs after being cultured on PKF, PKPS, and PPKSP surfaces for different times is provided in Example 2 of the present invention.

[0062] Figure 22 ALP staining images of BMSCs cultured on PKF, PKPS, and PPKSP surfaces for 14 days, provided as Example 2 of the present invention;

[0063] Figure 23 ARS staining images of BMSCs cultured on PKF, PKPS and PPKSP surfaces for 21 days, provided as an example of the effectiveness of this invention;

[0064] Figure 24 The bar chart for quantitative analysis of ALP activity of BMSCs after culturing on PKF, PKPS and PPKSP surfaces for different times is provided in Example 2 of the present invention.

[0065] Figure 25 A quantitative bar chart of calcium nodules formed on the surfaces of PKF, PKPS, and PPKSP after BMSCs were cultured for different times for different durations, as provided in Example 2 of the present invention;

[0066] Figure 26 The bar chart showing the expression of the osteogenic gene ALP in BMSCs after being cultured on PKF, PKPS and PPKSP surfaces for different times, as provided in Example 2 of the present invention.

[0067] Figure 27The bar chart showing the expression of the osteogenic gene OCN in BMSCs after being cultured on PKF, PKPS and PPKSP surfaces for different times, as provided in Example 2 of the present invention;

[0068] Figure 28 The bar chart showing the expression of the osteogenic gene OPN in BMSCs after being cultured on PKF, PKPS and PPKSP surfaces for different times, as provided in Example 2 of the present invention;

[0069] Figure 29 The bar chart showing the expression of the osteogenic gene Runx2 in BMSCs after being cultured on PKF, PKPS and PPKSP surfaces for different times, as provided in Example 2 of the present invention;

[0070] Figure 30 This is a set of SEM images of BMSCs cultured on PKF, PKPS and PPKSP surfaces for 3 days under electrical stimulation, as provided in Example 3 of the present invention.

[0071] Figure 31 The image set of CLSMs after culturing BMSCs on PKF, PKPS and PPKSP surfaces for 3 days under electrical stimulation, provided as Example 3 of the present invention;

[0072] Figure 32 A bar chart showing the proliferation of BMSCs under electrical stimulation after 3 days of culture on PKF, PKPS and PPKSP surfaces, as provided in Example 3 of the present invention.

[0073] Figure 33 A bar chart of ALP activity of BMSCs cultured on the surfaces of PKF, PKPS and PPKSP for 14 days under electrical stimulation, as provided in Example 3 of the present invention;

[0074] Figure 34 The image atlas of CLSMs stained with ROS fluorescence after being cultured on the surfaces of PKF, PKPS and PPKSP for 1 day under oxidative stress conditions, as provided in Example 4 of the present invention;

[0075] Figure 35 A bar chart of quantitative analysis of ROS fluorescence intensity of BMSCs cultured on the surfaces of PKF, PKPS and PPKSP for 1 day under oxidative stress conditions, provided as an example 4 of the present invention;

[0076] Figure 36 The Micro-CT three-dimensional reconstruction images of the femoral bone tunnel region after 4 weeks and 12 weeks of implantation of PKF, PKPS and PPKSP, respectively, are provided for Example 5 of the present invention. In the images, the white part is the implanted material and the yellow part is the new bone. The scale bar is 1 mm.

[0077] Figure 37The Micro-CT three-dimensional reconstruction images of the tibial bone tunnel region after 4 weeks and 12 weeks of implantation of PKF, PKPS and PPKSP are provided as an example of the effect of the present invention. The white part is the implanted material and the yellow part is the new bone. The scale bar is 1 mm.

[0078] Figure 38 Bar charts showing the BV / TV quantitative analysis of new bone formation around PKF, PKPS and PPKSP 4 and 12 weeks after implantation in the femur and tibia, as provided in Example 5 of the present invention.

[0079] Figure 39 Bar charts showing the BMD quantitative analysis of new bone formation around PKF, PKPS and PPKSP 4 and 12 weeks after implantation in the femur and tibia, as provided in Example 5 of the present invention.

[0080] Figure 40 Bar charts showing the bone tunnel area of ​​the femur and tibia 4 and 12 weeks after PKF, PKPS and PPKSP were implanted in the body, as provided in Example 5 of the present invention.

[0081] Figure 41 Bar charts showing the failure loads of PKF, PKPS, and PKBSP after 4 and 12 weeks of implantation (between the femur and tibia) in Example 5 of the present invention.

[0082] Figure 42 The set of histological HE-stained images of PKF, PKPS, and PPKSP at 4 and 12 weeks after implantation, provided as an example of the effectiveness of the present invention (Example 5);

[0083] Figure 43 This is a set of histological Masson stained images of PKF, PKPS, and PPKSP at 4 and 12 weeks after implantation, provided as an example of the effectiveness of this invention (Example 5).

[0084] In the diagram: * indicates a comparison with PKF # represents a comparison with PKPS . Detailed Implementation

[0085] To make the technical solution, beneficial effects and significant progress of the present invention clearer and more comprehensive, the technical solution provided by the present invention will be clearly and completely described below through specific embodiments and their effects. Obviously, all embodiments and their effects described below are only some embodiments and effects of the present invention, and not all of them.

[0086] Based on the embodiments and effects provided by this invention, all other embodiments and effects obtained by those skilled in the art without creative effort are within the scope of protection of this invention.

[0087] It should be noted that the terms "firstly," "secondly," etc., in the claims, specification, and examples and effects of the embodiments of this invention are only used to distinguish different objects and not to describe a specific order; in addition, the term "comprising" and any variations thereof are intended to cover non-exclusive inclusion, for example, including not only a series of listed steps or units of a process, method, system, product, or device, but also optionally steps or units not listed, or optionally other operational steps or units inherent to these processes, methods, products, or devices.

[0088] It should be understood that some basic operational terms commonly used in the art are used in the description of the embodiments of the present invention, such as "immersion" and "attachment". These terms should be interpreted broadly, that is, they can refer to conventional operations performed using various conventional equipment and instruments in the art, or they can refer to operations performed using the latest equipment, such as programmed operations and unmanned automatic operations. Unless otherwise explicitly limited, those skilled in the art should understand the specific meaning of the above terms in the present invention according to the specific circumstances and adopt specific operating methods to achieve their operating objectives.

[0089] It should also be noted that:

[0090] The following specific embodiments can be combined with each other. The same or similar concepts or processes may not be repeated in some implementation cases and comparative examples. In addition, all kinds of instruments, equipment, raw materials, reagents and standards involved in the following specific embodiments are commercially available unless otherwise specified.

[0091] The technical solution of the present invention will now be described in detail with reference to specific embodiments. Example 1

[0092] This embodiment provides a method for preparing a fiber material with a nanoporous structure.

[0093] A method for preparing a fiber material with a nanoporous structure includes the following steps:

[0094] Prepare polyetheretherketone (PEEK) fibers; construct a chelate coating of phytic acid and strontium ions on the surface of the PEEK fibers; subject the chelate coating to electron bombardment treatment to form a nanoporous structure on the surface of the chelate coating.

[0095] In the above preparation method, the chelate coating is constructed through a stacking assembly technique.

[0096] The stacked assembly technology specifically includes the following steps:

[0097] Polyetheretherketone (PEEK) fibers are immersed in a phytic acid solution, allowing phytic acid molecules to adhere to the surface of the PEEK fibers.

[0098] Cleaning removes any unattached phytic acid molecules;

[0099] The polyetheretherketone fiber with phytic acid molecules attached is then immersed in a strontium ion solution, so that the strontium ions in the strontium ion solution can chelate with the phytic acid molecules attached to the surface of the polyetheretherketone fiber to form a chelate coating.

[0100] Cleaning removes unreacted strontium ions;

[0101] Repeat the above steps multiple times until the thickness of the chelate coating meets the design requirements.

[0102] In the above process:

[0103] The volume percentage concentration of the phytic acid solution is usually 3-7%, preferably 5%, and the pH value of the phytic acid solution is adjusted to 4.5-6.5, preferably 5.5, by an alkaline solution. The alkaline solution used to adjust the pH value of the phytic acid solution is a 1M NaOH solution.

[0104] The immersion time of polyetheretherketone (PEEK) fibers in phytic acid solution is usually 0.5 to 2 hours, preferably 1 hour;

[0105] The strontium ion solution is a strontium nitrate solution with a concentration of 0.3–0.7 M, preferably 0.5 M;

[0106] The time for which the polyetheretherketone fiber with phytic acid molecules is attached to be immersed in strontium ion solution is usually 15 to 45 minutes, preferably 30 minutes;

[0107] The stacked assembly is typically repeated 5 to 9 times, preferably 7 times.

[0108] The solvent used to clean and remove unattached phytic acid molecules is water, while the solvent used to clean and remove unreacted strontium ions is hydrogen peroxide.

[0109] The bombardment voltage for electron bombardment is typically 15–25 kV, preferably 20 kV, while the bombardment duration is typically 5–15 seconds, preferably 10 seconds.

[0110] From the above description, it can be seen that:

[0111] This embodiment first utilizes the chelating ability of phytic acid to construct a stable phytic acid / strontium (PA / Sr) chelate coating on the surface of PEEK fibers. Then, a nanoporous structure is created on the surface of this coating by electron bombardment. This innovatively combines the bioactivity of strontium, the chelating function of phytic acid, and the physical advantages of the nanoporous structure, thereby achieving a synergistic effect of multiple functions. Specifically, the phytic acid-strontium chelate provides continuous and controllable strontium ion release, thereby activating osteogenic biochemical signals. The nanoporous structure provides biomimetic physical attachment points for cells, guiding cell behavior. Furthermore, the conductivity and antioxidant properties of phytic acid itself are used to regulate the microenvironment around the implant, thereby reducing oxidative stress damage. This solves the key technical bottlenecks of insufficient bioactivity and weak bone integration capacity of traditional artificial ligament materials, providing a new solution and material for promoting tendon-bone healing and bone regeneration.

[0112] Furthermore, it can be seen that the preparation method provided in this embodiment is relatively simple, the production process is stable, and it can ensure the quality of the product and achieve the corresponding technical effects.

[0113] To further aid in understanding the technical solution provided in this embodiment, as well as the specific operation process and the effects that can be obtained, the preparation method provided in this embodiment will be further explained below through specific examples.

[0114] Of course, those skilled in the art should understand that the examples described below are illustrative and not restrictive, and should not be used to limit the scope of protection claimed by the present invention. Preparation Case

[0115] First, prepare the polyetheretherketone (PEEK) fibers purchased from the market;

[0116] Then, a 5% (v / v) phytic acid (PA) solution was prepared, and the pH of the PA solution was adjusted to 5.5 using 1 M equivalent NaOH solution.

[0117] The PEEK fibers were then immersed in the PA solution for 1 hour to fix the PA molecules at the interface of the PEEK fibers. Then, the fibers were washed with H2O to remove the unattached PA molecules.

[0118] Subsequently, the fibers were immersed in a 0.5M Sr(NO3)2 solution for 30 minutes to allow the Sr... 2+ It undergoes a chelation reaction with PA molecules to form a chelate coating on the surface of PEEK fibers, and is then cleaned with H2O2 to remove excess Sr. 2+ ;

[0119] The process of immersing the PEEK fibers in PA solution and then cleaning them, followed by immersion in Sr(NO3)2 solution and cleaning them, was repeated seven times to obtain a chelate coating that meets the design requirements.

[0120] Finally, the PEEK fibers with a PA / Sr chelate coating formed on their surface were placed in an electron bombardment furnace (DZS-300, Lijie, China) and treated at 20kV for 10 seconds to obtain polyetheretherketone fibers with a PA / Sr chelate coating on their surface and the chelate coating having a nanoporous structure.

[0121] To further aid in understanding the technical effects achievable by the technical solution provided in this case, the following will further illustrate the technical effects achievable by this embodiment through specific examples of effects, particularly the polyether ether ketone fiber with a nanoporous structure obtained through the above preparation example, by conducting corresponding functional tests and comparisons.

[0122] It should be noted that, for ease of testing and comparison, the following methods are used to weave commercially available polyetheretherketone (PEEK) fibers, PEEK fibers coated with a PA / Sr chelate coating obtained in the above preparation example, and PEEK fibers coated with a PA / Sr chelate coating whose surface is formed into a nanoporous structure by electron bombardment, respectively, to form PEEK fiber artificial ligaments (PKF), PEEK fiber artificial ligaments coated with a PA / Sr chelate coating (PKPS), and PEEK fiber artificial ligaments coated with a PA / Sr chelate coating whose surface is formed into a nanoporous structure by electron bombardment (PKPSP). Example 1: Characterization of the physicochemical properties of the sample

[0123] 1.1) SEM analysis:

[0124] The surface morphology of PKF, PKPS and PPKSP was observed using a scanning electron microscope (SEM).

[0125] Through testing, we can obtain results such as Figure 1 The image shown is the SEM image set of PKPS provided in Example 1 of the present invention, and as shown in... Figure 2 The image shown is the SEM image set of PKPSP provided in Example 1 of the present invention.

[0126] from Figure 1 and Figure 2It can be seen that at low magnification, both PKPS and PPKSP exhibit smooth surfaces with fiber diameters of approximately 27 µm. At high magnification, after PA / Sr coating, the PKPS surface exhibits a rough surface, and the micro-cracks on its surface are caused by the PA / Sr composite being bombarded by the high-energy electron beam of the scanning electron microscope. After electron bombardment, a large number of nanoscale pore structures with sizes of approximately 50–200 nm can be seen on the PPKSP surface.

[0127] 1.2) EDS analysis:

[0128] The surface elemental distribution of the above-mentioned PKF, PKPS and PPKSP was analyzed by energy dispersive spectroscopy (EDS analysis).

[0129] Through testing, we can obtain results such as Figure 3 The image shown is a distribution image of phosphorus (P) in PKPS provided in Example 1 of the present invention. Figure 4 The image shown is a strontium (Sr) element distribution image obtained by PKPS according to Example 1 of the present invention. Figure 5 The image shown is the EDS spectrum of PKPS provided in Example 1 of the present invention, and... Figure 6 The image shown is a phosphorus (P) element distribution image of PKPSP provided in Example 1 of the present invention. Figure 7 The image shown is an image of the strontium (Sr) element distribution in the PKPSP provided in Example 1 of the present invention. Figure 8 The image shown is the EDS spectrum of the PPKSP provided in Example 1 of the present invention.

[0130] 1.3) FTIR analysis:

[0131] The chemical groups of PKF, PKPS, and PKPSP were analyzed using Fourier Transform Infrared Spectroscopy (FTIR), with a wavenumber range of 4000–650 cm⁻¹. -1 .

[0132] Through testing, we can obtain results such as Figure 9 The image shows the FTIR detection spectra of PKF, PKPS, and PPKSP provided in Example 1 of the present invention.

[0133] As shown in the figure, it is located at 925 cm. -1 1450 cm -1 and 1646 cm -1 The spectral peak is attributed to the characteristic peak of benzophenone in PEEK, appearing at 1589 cm⁻¹. -1The spectral peaks are attributed to the stretching vibrations of C=C in the benzene ring, while the peak at 1221 cm⁻¹ is... -1 and 1148 cm -1 The observed spectral peaks are attributed to the stretching vibrations of diaryl-COC. Furthermore, the PO4 content of PA in PKPS and PPKSP is also observed. 3- The spectral peak is located at 1100 cm⁻¹ -1 Place.

[0134] 1.4) XRD analysis:

[0135] X-ray diffraction (XRD) was used to analyze the crystal phase structure of PKF, PKPS, and PPKSP, with a degree range of 10–80°.

[0136] Through testing, we can obtain results such as Figure 10 The image shows the XRD patterns of PKF, PKPS, and PPKSP provided in Example 1 of the present invention.

[0137] Depend on Figure 4 It can be seen that the characteristic peaks of PEEK can be observed in the XRD patterns of all three samples, located at 18.6° and 24.2°, respectively. Furthermore, no characteristic peaks of PA were observed in PKPS and PPKSP, indicating that the Sr-PA composites on the surfaces of PKPS and PPKSP are in an amorphous state, and that electron bombardment treatment has no effect on the crystal phase structure of the Sr-PA composite. 1.5) Physical Property Detection

[0138] 1.5.1) Contact angle and surface energy detection: The diiodomethane and water contact angles of the PKF, PKPS and PPKSP surfaces were measured using a contact angle meter, and the surface energy of the PKF, PKPS and PPKSP was calculated using the Owen-Wendt formula.

[0139] 1.5.2) Protein adsorption detection: The adsorption capacity of PKF, PKPS and PKBSP surfaces for fibronectin (FN) and bovine serum albumin (BSA) was detected using the BCA Protein Assay Kit.

[0140] 1.5.3) Conductivity testing: The conductivity of PKF, PKPS, and PKBSP was tested using a four-probe measuring instrument;

[0141] 1.5.4) Tensile strength test: The tensile strength of PKF, PKPS and PKBSP was tested using a mechanical testing machine at a tensile rate of 5 mm / min.

[0142] 1.5.5) Antioxidant performance test: The antioxidant performance of PKF, PKPS and PPKSP was tested using a colorimetric quantitative detection kit for 1,1-diphenyl-2-trinitrophenylhydrazine organic free radical (DPPH), i.e., free radical scavenging rate test.

[0143] Through testing, we can obtain the following:

[0144] like Figure 11 The figure shown is a bar chart of the contact angles of PKF, PKPS and PKBSP provided in Example 1 of the present invention in contact with diiodomethane and water.

[0145] Figure 12 The above is a bar chart of the surface energy of PKF, PKPS, and PKBSP provided in Example 1 of the present invention;

[0146] Figure 13 The image shown is a bar chart of protein adsorption of PKF, PKPS, and PPKSP provided in Example 1 of the present invention.

[0147] Figure 14 The figure shown is a bar chart of the conductivity of PKF, PKPS and PKBSP provided in Example 1 of the present invention.

[0148] Figure 15 The figure shown is a bar chart of the tensile strength of PKF, PKPS and PKBSP provided in Example 1 of the present invention.

[0149] Figure 16 The figure shown is a bar chart of the free radical scavenging capabilities of PKF, PKPS, and PPKSP provided in Example 1 of the present invention, where * represents the free radical scavenging capability compared to PKF. # represents a comparison with PKPS .

[0150] Depend on Figure 11 It can be seen that the water contact angles of PKF, PKPS, and PPKSP are 87.0±1.8°, 33.5±1.9°, and 32±2.3°, respectively, and the diiodomethane contact angles of PKF, PKPS, and PPKSP are 57.0±1.5°, 23.5±1.6°, and 24.5±1.7°, respectively.

[0151] It can be seen that the contact angle between diiodomethane and water on the PKF surface is the largest. Compared with PKF, the contact angle between water and diiodomethane on PKPS is significantly reduced after coating with PA / Sr composite. Compared with PKPS, the contact angle between water and diiodomethane on the PKBSP surface does not change significantly, indicating that electron bombardment has no effect on the hydrophilicity of PKPSP.

[0152] Depend on Figure 12It can be seen that the surface energies of PKF, PKPS, and PKBSP are 24.6 ± 1.3 mJ / m. 2 49.9±2.4 mJ / m 2 48.2±2.8 mJ / m 2 ;

[0153] It can be seen that PKF has the lowest surface energy. Compared with PKF, the surface energy of PKPS is significantly reduced after coating with PA / Sr composite. Compared with PKPS, the surface energy of PKBSP does not change significantly, indicating that electron bombardment has no effect on the surface energy of PKBSP.

[0154] Depend on Figure 13 It can be seen that the protein adsorption rates of PKF, PKPS, and PPKSP for BSA were 12.8±2.0%, 36.5±1.7%, and 39.8±2.9%, respectively, and the protein adsorption rates for FN were 7.9±1.6%, 22.5±1.9%, and 26.4±2.4%, respectively.

[0155] It can be seen that the adsorption capacity of PKF surface for BSA and FN is the weakest. Compared with PKF, the adsorption capacity of PKPS for BSA and FN is significantly improved after coating with PA / Sr complex. Compared with PKPS, the adsorption capacity of PKBSP for BSA and FN is further improved, indicating that the nanoporous structure generated by electron bombardment promotes the protein adsorption of PKPSP.

[0156] Depend on Figure 14 It can be seen that the electrical conductivities of PKF, PKPS, and PKBSP to BSA are 0±0 S / cm, 0.015±0.002 S / cm, and 0.013±0.003 S / cm, respectively.

[0157] It can be seen that PKF is non-conductive and is an insulating material; compared with PKF, the conductivity of PKPS is significantly improved after coating with PA / Sr composite; compared with PKPS, the conductivity of PKBSP is slightly decreased, indicating that the nanoporous structure generated by electron bombardment reduces the conductivity of PKBSP.

[0158] Depend on Figure 15 It can be seen that the tensile strengths of PKF, PKPS, and PKBSP are 570.2±16.3 MPa, 572.6±19.3 MPa, and 568.3±22.1 MPa, respectively.

[0159] It can be seen that PKF, PKPS, and PPKSP all have excellent mechanical properties. Coating with PA / Sr composite or subjecting them to electron bombardment did not significantly reduce the tensile strength of PKPS and PPKSP.

[0160] Depend on Figure 16It can be seen that the scavenging rates of PKPS and PPKSP against 1,1-diphenyl-2-trinitrophenylhydrazine organic free radical (DPPH) are 81.2±5.4% and 85.1±6.3%, respectively.

[0161] 1.6) Ion release:

[0162] PKPS and PPKSP were immersed in 10 mL of PBS solution and then placed in a constant temperature shaker at 37°C to study the release behavior of strontium ions. Specifically, the concentration of strontium ions in phosphate buffered saline (PBS) solution was measured using an inductively coupled plasma atomic emission spectrometer on days 1, 3, 5, 7, 14 and 21.

[0163] Through testing, we can obtain results such as Figure 17 The figure shown is a curve of strontium ion release after PKPS and PPKSP provided in Example 1 of the present invention were soaked in PBS for different times.

[0164] Depend on Figure 17 It is known that strontium ions from PKPS and PPKSP are released rapidly in the first 3 days, and then slowly until 21 days. In addition, due to the nanoporous structure on the surface of PPKSP, the release of strontium ions is always slightly faster than that from PKPS. Example 2: Osteoblast response

[0165] 2.1) Cell morphology detection:

[0166] Rat bone marrow mesenchymal stem cells (BMSCs) were cultured on the surfaces of PKF, PKPS, and PPKSP for 1 day and 3 days, respectively, and the cell morphology on the samples was observed by scanning electron microscopy (SEM).

[0167] Rat bone marrow mesenchymal stem cells (BMSCs) were cultured on PKF, PKPS, and PPKSP surfaces for 1 and 3 days, respectively. The actin and nucleus of the cells were stained with two fluorescent dyes: fluorescein isothiocyanate (FITC), which can bind to proteins or other molecules, and 4',6-diamidinyl-2-benzimidazole (DAPI), which specifically binds to DNA. The stained cells were then observed using a confocal laser scanning microscope (CLSM).

[0168] Through testing, we can obtain results such as Figure 18 The image shown is a set of SEM images of PKF, PKPS, and PKBSP co-cultured with BMSCs for different time periods, as provided in Example 2 of the present invention. Figure 19The image shown is a set of CLSM images after PKF, PKPS, and PPKSP were co-cultured with BMSC for different time periods, as provided in Example 2 of the present invention.

[0169] Depend on Figure 18 It can be seen that with the extension of culture time, the number of BMSCs on the surface of PKF, PKPS and PPKSP all increased, indicating that none of the three materials are cytotoxic.

[0170] In addition, the cells on the surface of PKF exhibit a spherical structure, poor cell adhesion morphology, and weak affinity with PKF. Compared with PKF, the cell adhesion morphology on the surfaces of PKPS and PPKSP is significantly improved, with cells growing a large number of pseudopodia and spreading well. The number and morphology of cells on the surface of PPKSP are superior to those on PKPS.

[0171] Depend on Figure 19 It can be seen that after BMSCs were co-cultured with PKF, PKPS and PPKSP for 1 day and 3 days respectively, the number of BMSCs on the surface of PKF was small, the proliferation state was weak, and the cells were spherical and in poor condition.

[0172] In addition, compared with PKF, PKPS and PPKSP significantly promoted cell proliferation, and after 3 days of cell culture, the number of cells on the material surface was significantly increased, and cell pseudopodia and cytoskeleton spread on the material surface. Moreover, the number and morphology of cells on the PPKSP surface were superior to those on PKPS.

[0173] 2.2) Detection of cell adhesion and proliferation:

[0174] PKF, PKPS, and PPKSP were co-cultured with rat bone marrow mesenchymal stem cells (BMSCs) to evaluate the in vitro osteoblast response of the samples. Cell adhesion and proliferation were detected using a cell counting kit (CCK-8). Cell adhesion on the sample surface was detected after 6, 12, and 24 hours of co-culture with cells, and cell proliferation on the sample surface was detected after 1, 3, and 7 days of co-culture.

[0175] Through testing, we can obtain results such as Figure 20 The figure shown is a bar graph illustrating the adhesion performance of BMSCs on PKF, PKPS, and PPKSP surfaces after different cultivation times, as provided in Example 2 of the present invention. Figure 21 The bar chart shown is a graph illustrating the proliferation of BMSCs on PKF, PKPS, and PPKSP surfaces after different culturing times, as provided in Example 2 of the present invention. * indicates a comparison with PKF. # represents a comparison with PKPS .

[0176] from Figure 20 It can be seen that BMSC cells have a higher adhesion rate on PKPS and PPKSP than on PKF, and the cell adhesion rate of PPKSP is higher than that of PKPS.

[0177] from Figure 21 It can be seen that the cell proliferation rate of BMSC cells on PKPS and PPKSP is higher than that on PKF, and the cell proliferation rate of PPKSP is higher than that of PKPS.

[0178] 2.3) Osteogenic differentiation:

[0179] After co-culturing PKF, PKPS, and PPKSP with BMSC cells for 14 days, ALP in the cells was stained and quantified using an ALP staining kit (Beyotime, China) and an ALP detection kit (Beyotime) to detect alkaline phosphatase (ALP) activity. Meanwhile, after co-culturing PKF, PKPS, and PPKSP with BMSC cells for 21 days, Alizarin Red (ARS) staining was used to stain and quantify calcium nodule formation in the cells.

[0180] Through testing, we can obtain the following:

[0181] like Figure 22 The image shown is an ALP staining photograph of BMSCs cultured on the surfaces of PKF, PKPS and PPKSP for 14 days, as provided in Example 2 of the present invention. It shows that the ALP staining intensity of PKPS and PPKSP is significantly higher than that of PKF, and the ALP staining intensity of PPKSP is higher than that of PKPS.

[0182] like Figure 23 The image shown is an ARS staining photograph of BMSCs cultured on the surfaces of PKF, PKPS and PPKSP for 21 days, as provided in Example 2 of the present invention. It shows that the ARS staining intensity on the surfaces of PKPS and PPKSP is significantly higher than that on PKF. Among them, the ARS staining intensity of PPKSP is higher than that of PKPS, indicating that the amount of calcium nodules formed on the surface of PPKSP cells is the highest.

[0183] like Figure 24 The bar chart shown is a quantitative analysis of ALP activity of BMSCs after being cultured on the surfaces of PKF, PKPS and PPKSP for different times according to Example 2 of the present invention. It shows that after 7 and 14 days of culture, the ALP activity of PKPS and PPKSP is significantly higher than that of PKF, with PPKSP having the highest ALP activity. In other words, PKPS and PPKSP coated with PA / Sr complex can significantly improve the ALP activity of BMSCs, and after electron bombardment, PPKSP with nanoporous structure further improves the ALP activity of BMSCs.

[0184] like Figure 25 The figure shown is a quantitative bar graph of calcium nodule formation in BMSCs after culturing on PKF, PKPS, and PPKSP surfaces for different times, as provided in Example 2 of the present invention. It shows that after 7 and 14 days of culture, the calcium nodule formation in PKPS and PPKSP is significantly higher than that in PKF, with PPKSP showing the highest calcium nodule formation. In other words, PKPS and PPKSP coated with PA / Sr composite can significantly promote the formation of calcium nodules in BMSCs, and after electron bombardment, PKPSP with its nanoporous structure can further promote the formation of calcium nodules.

[0185] 2.4) Expression of osteogenic-related genes:

[0186] After co-culturing PKF, PKPS, and PPKSP with cells for 7 and 14 days, the expression of osteogenic-related genes (ALP, Runx2, OPN, and OCN) of differentially expressed genes (BMSCs) closely related to bone cell biology during spontaneous calcification was detected using RT-PCR technology based on reverse transcription and polymerase chain reaction.

[0187] After testing, the following can be obtained: Figure 26 The image shows a bar chart illustrating the expression of the osteogenic gene ALP in BMSCs cultured on PKF, PKPS, and PPKSP surfaces for different durations, as provided in Example 2 of this invention. Figure 27 The figure shown is a bar chart of the expression of the osteogenic gene OCN in BMSCs cultured on PKF, PKPS, and PPKSP surfaces for different times according to Example 2 of the present invention. Figure 28 The figure shown is a bar chart of the expression of the osteogenic gene OPN in BMSCs cultured on PKF, PKPS, and PPKSP surfaces for different times according to Example 2 of the present invention, and also shows... Figure 29 The image shows a bar chart of the expression of the osteogenic gene Runx2 in BMSCs after being cultured on PKF, PKPS and PPKSP surfaces for different times, as provided in Example 2 of the present invention.

[0188] from Figures 26-29 As can be seen, the expression level of osteogenic-related genes of BMSCs on the material surface increases with increasing culture time. Specifically, on day 7, the expression of osteogenic-related genes of PKPS and PPKSP is higher than that of PKF, while the expression of osteogenic-related genes of PPKSP is slightly higher than that of PKPS. On day 14, the expression of osteogenic-related genes of PKPS and PPKSP is higher than that of PKF, while the expression of osteogenic-related genes of PPKSP is slightly higher than that of PKPS. In other words, after coating with PA / Sr complex, PKPS and PPKSP significantly promote osteogenic differentiation of BMSCs, and after electron bombardment, PKPSP with nanoporous structure can further promote osteogenic differentiation of BMSCs. Example 3: Osteoblast electrical stimulation response

[0189] 3.1) Electrical stimulation response of osteoblasts:

[0190] BMSCs were co-cultured on PKF, PKPS, and PPKSP surfaces. Simultaneously, 300 mV electrical stimulation was applied to the BMSCs using an electrochemical workstation, and their cell morphology was then examined to evaluate the electrical stimulation response of osteoblasts on the samples.

[0191] After testing, the following can be obtained: Figure 30 The image shown is a set of SEM images of BMSCs cultured on PKF, PKPS, and PPKSP surfaces for 3 days under electrical stimulation, as provided in Example 3 of the present invention. Figure 31 The image shown is a set of CLSM images of BMSCs cultured on PKF, PKPS and PPKSP surfaces for 3 days under electrical stimulation, as provided in Example 3 of the present invention.

[0192] Depend on Figure 30 and 31 It can be seen that under the culture condition without electrical stimulation (0mV), the number of cells on the surface of PKF is small, the proliferation is weak, and the cells are spherical and in poor condition. However, the number of cells on the surface of PKPS and PPKSP samples is significantly greater than that of PKF. Under the culture condition of 300mV electrical stimulation, the cells on the surface of PKF did not change significantly, while the number of cells on the surface of PKPS and PPKSP increased significantly, and the cell morphology was more spread out. This indicates that electrical stimulation promotes the proliferation and spread of cells on the surface of PKPS and PPKSP.

[0193] 3.2) Cell proliferation and differentiation:

[0194] Cell proliferation was detected 3 days after culturing cells on PKF, PKPS, and PPKSP using a cell counting kit (CCK-8); ALP was quantitatively analyzed in cells on PKF, PKPS, and PPKSP using an ALP detection kit (Beyotime).

[0195] After testing, the following can be obtained: Figure 32 The figure shown is a bar chart illustrating the proliferation of BMSCs under electrical stimulation after 3 days of culture on PKF, PKPS, and PPKSP surfaces, as provided in Example 3 of the present invention. Figure 33 The image shown is a bar graph of ALP activity of BMSCs cultured on the surfaces of PKF, PKPS, and PPKSP for 14 days under electrical stimulation, as provided in Example 3 of the present invention.

[0196] The results show that:

[0197] Under culture conditions without electrical stimulation (0 mV), the cell proliferation rate and ALP activity on PKPS and PPKSP were higher than those on PKF, and the cell proliferation rate and ALP activity on PPKSP were higher than those on PKPS.

[0198] Under 300 mV electrical stimulation culture conditions, the proliferation rate and ALP activity of PKF surface cells did not change significantly, while the proliferation rate and ALP activity of PKPS and PPKSP surface cells increased significantly, indicating that electrical stimulation promoted the proliferation and osteogenic differentiation of PKPS and PPKSP surface cells. Example 4: Evaluation of Antioxidant Stress Performance

[0199] PKF, PKPS, and PPKSP were co-cultured with BMSCs, and 10 µL of hydrogen peroxide (35 v%) solution was added to the culture medium to examine the antioxidant stress performance of the samples. The experimental procedure was as follows: after co-culturing PKF, PKPS, and PPKSP with BMSCs for 1 day, the cell culture medium was removed, and the cells were fixed with 4% paraformaldehyde for 2 hours. Then, the cells were washed three times with phosphate-buffered saline (PBS). The washed samples were stained with reactive oxygen species (ROS) in the cells for 10 minutes using the DCFH-HA fluorescent probe (Beyotime, China). The stained cells were then observed using a confocal laser scanning microscope (CLSM), and the fluorescence intensity of ROS was quantitatively analyzed using ImageJ software.

[0200] After testing, the following can be obtained: Figure 34 The image shown is a collection of CLSM images stained with ROS fluorescence after being cultured on the surfaces of PKF, PKPS, and PPKSP for one day under oxidative stress conditions, as provided in Example 4 of the present invention. Figure 35 The figure shown is a bar chart of ROS fluorescence intensity quantitative analysis of BMSCs cultured on the surfaces of PKF, PKPS, and PPKSP for 1 day under oxidative stress conditions, as provided in Example 4 of the present invention. * indicates a comparison with PKF. # represents a comparison with PKPS .

[0201] from Figure 34 and Figure 35As can be seen, there is a large amount of ROS fluorescence in the cells on the surface of PKF, indicating that it does not have the ability to scavenge ROS. After coating with the PA / Sr complex, the ROS fluorescence in the BMSC cells on the surface of PKPS is significantly reduced, indicating that the PKPS surface coated with the PA / Sr complex has a strong ability to eliminate reactive oxygen free radicals. After electron bombardment, the ROS fluorescence of the BMSC cells on the surface of PKPSP is weaker than that of PKPS, indicating that the PKPSP material after electron bombardment to form a nanoporous structure of PA / Sr complex has a better ability to eliminate reactive oxygen free radicals. Example 5: In vivo tendon-bone healing test

[0202] 5.1) Micro-CT analysis:

[0203] To evaluate the in vivo tendon-bone healing performance of PKF, PKPS, and PPKSP, a rabbit anterior cruciate ligament (ACL) reconstruction model was established. This involved non-destructive three-dimensional imaging and analysis of the samples using micro-computed tomography (Micro-CT). The microstructure of the tendon-bone interface was analyzed 4 and 12 weeks after PKF, PKPS, and PPKSP implantation. The two-dimensional and three-dimensional images were then reconstructed using a CT analyzer, and the bone mineral density (BMD) and bone volume fraction (BV / TV) around the samples were quantitatively analyzed.

[0204] After testing, we can obtain the following:

[0205] like Figure 36 The image shown is a set of Micro-CT three-dimensional reconstruction images of the femoral bone tunnel area after 4 weeks and 12 weeks of implantation of PKF, PKPS and PPKSP provided in Example 5 of the present invention. The white part is the implanted material and the yellow part is the new bone. The scale bar is 1 mm.

[0206] like Figure 37 The image shown is a set of Micro-CT three-dimensional reconstruction images of the tibial bone tunnel region after 4 weeks and 12 weeks of implantation of PKF, PKPS and PPKSP provided in Example 5 of the present invention. The white part is the implanted material and the yellow part is the newly formed bone. The scale bar is 1 mm.

[0207] Depend on Figure 36 and Figure 37It can be seen that: with the increase of implantation time, the amount of new bone formation around PKF, PKPS, and PPKSP all increased; among them, the amount of new bone formation around PKF was relatively small; compared with PKF, the amount of new bone formation around PKPS increased significantly after coating with PA / Sr complex; compared with PKF, the amount of new bone formation around PPKSP increased significantly after coating with PA / Sr complex and electron bombardment, and the amount of new bone formation around PPKSP was greater than that around PKPS. After 12 weeks of implantation, the new bone tissue wrapped the surface of PPKSP and was tightly bound to the sample.

[0208] 5.2) Biomechanical analysis:

[0209] The biomechanical properties of PKF, PKPS, and PPKSP were tested using a materials testing system, including quantitative analysis of BV / TV, BMD, bone tunnel area, and failure load.

[0210] After testing and calculation, we can obtain:

[0211] like Figure 38 The figure shown is a bar chart of BV / TV quantitative analysis of new bone formation around PKF, PKPS and PPKSP 4 weeks and 12 weeks after implantation in the femur and tibia provided in Example 5 of the present invention.

[0212] like Figure 39 The bar chart shown is a quantitative analysis of BMD of the new bone formation around PKF, PKPS, and PPKSP 4 and 12 weeks after implantation into the femur and tibia provided in Example 5 of the present invention.

[0213] like Figure 40 The figure shown is a bar chart of the bone tunnel area of ​​the femur and tibia 4 weeks and 12 weeks after PKF, PKPS and PPKSP were implanted in the body, respectively, according to Example 5 of the present invention.

[0214] like Figure 41 The figure shows the failure load bars of PKF, PKPS, and PPKSP at 4 and 12 weeks after implantation (between femur and tibia) of the bone tunnel provided in Example 5 of the present invention.

[0215] Where * represents compared to PKF # represents a comparison with PKPS .

[0216] Depend on Figure 38 and 39 It can be known that:

[0217] The bone density (BMD) and bone volume (BV / TV) of the newly formed bone tissue around PKF were the lowest. Compared with PKF, the BMD and BV / TV of the newly formed bone tissue around PKPS were significantly improved after coating with PA / Sr composite. After electron bombardment, the BMD and BV / TV of the newly formed bone tissue around PKSN with nanoporous structure were further improved. At 12 weeks, the BV / TV of the femur and tibia implanted with PKF, PKPS, and PPKSP were 12.57±1.91%, 29.39±2.25%, and 36.98±1.84%, respectively. In addition, at 12 weeks, the average BMD of the femur and tibia implanted with PKF, PKPS, and PPKSP were 0.28±0.04 g / cc, 0.46±0.05 g / cc, and 0.51±0.04 g / cc, respectively.

[0218] Depend on Figure 40 It can be known that:

[0219] The femur and tibia implanted with PKF had the largest bone tunnel areas. Compared to PKF, PKPS implanted with a PA / Sr composite significantly reduced the bone tunnel areas in both the femur and tibia. Compared to PKPS, PKPSP implanted with an electron-bombarded nanoporous structure further reduced the bone tunnel areas in both the femur and tibia. After 12 weeks, the average bone tunnel areas in the femur and tibia implanted with PKF, PKPS, and PKPSP, respectively, were 6.47 ± 0.28 mm. 2 6.01±0.29 mm 2 and 5.90±0.28 mm 2 .

[0220] Depend on Figure 41 It can be known that:

[0221] Four weeks after implantation, the failure loads of PKPS and PPKSP in the femur and tibia were significantly higher than those of PKF. Furthermore, the failure load of PPKSP was higher than that of PKPS in both the femur and tibia. At 12 weeks after implantation, the failure loads of PKPS and PPKSP in the femur and tibia were again significantly higher than those of PKF, with PPKSP showing a higher failure load than PKPS in both bones. The increasing failure load of the implanted material over time indicates better tendon-bone healing between the implanted material and bone tissue. The failure loads of PKF, PKPS, and PPKSP at 12 weeks after implantation were 39.1±3.0 N, 58.5±4.1 N, and 63.6±3.5 N, respectively.

[0222] 5.3) Histological analysis:

[0223] Four and twelve weeks after implantation of PKF, PKPS, and PPKSP samples, decalcified sections were prepared from the implanted samples. The sections were then stained with hematoxylin and eosin (HE staining) and Masson staining for collagen fibers in connective tissue to analyze the tendon-bone healing status of the samples.

[0224] After testing, we can obtain the following:

[0225] like Figure 42 The image shown is a set of histological HE-stained images of PKF, PKPS, and PPKSP at 4 and 12 weeks after implantation, as provided in Example 5 of the present invention. Figure 43 The image shown is a set of histological Masson staining images of PKF, PKPS, and PPKSP 4 weeks and 12 weeks after implantation, provided in Example 5 of the present invention.

[0226] from Figure 42 and Figure 43 From this, we can see that:

[0227] Four weeks after implantation, there was a clear gap between PKF and the host bone, which was filled with fibrous tissue and had little new bone formation. Compared with PKF, there was more new bone formation around PKPS and less fibrous tissue between the material and the new bone. In contrast, the new bone formation around PPKSP was further increased and the fibrous tissue was reduced.

[0228] Twelve weeks after implantation, the fibrous tissue around PKF became dense and mature, and there was still little new bone formation; the new bone tissue around PKPS increased further, but the fibrous tissue around the sample still existed; while a large amount of new bone was formed around PPKSP; in addition, the fibrous tissue around PPKSP disappeared and was replaced by new bone, which was tightly integrated with the new bone, indicating that PPKSP and the host bone formed good tendon-bone healing.

[0229] It should be noted that:

[0230] All experiments in the above effect examples were characterized three times independently, and all experimental data were processed using Prism 8 (OriginLab, USA) software, expressed as Mean ± SD, and statistical differences in the data were analyzed. This indicates that there are significant differences between the data groups.

[0231] Based on the above examples, the following results can be obtained.

[0232] 1) The fibrous material with nanoporous structure provided in this embodiment can improve osteoblast response and promote the formation of new bone tissue.

[0233] The above examples demonstrate that:

[0234] By utilizing the chelating effect of PA on metal ions, a PA / Sr complex is coated on the surface of PEEK fibers. Through electron bombardment technology, a nanoporous structure is formed on the PA / Sr chelate coating to constitute the fiber material provided by this invention. This fiber material is woven into a fabric to serve as an artificial ligament. After implantation, the surface energy of this fiber material can directly contact the body tissue and form a biological interface. Its surface properties have a positive and effective impact on the behavior and function of cells / tissues. Its high surface energy and hydrophilic surface help with body fluid infiltration, cell spreading, migration, adhesion, proliferation and differentiation, thereby improving the formation of new bone tissue.

[0235] Compared with PKF materials, which are hydrophobic and have low surface energy, PKPS materials have significantly improved hydrophilicity and surface energy due to the PA / Sr composite coating on their surface.

[0236] Since PA is a polyhydroxy compound, the hydroxyl groups on the molecule easily form hydrogen bonds with water. Therefore, PA on the PKPS surface can form hydrogen bonds with water, which significantly improves its surface energy and hydrophilicity.

[0237] Compared with PKPS, the hydrophilicity and surface energy of PKPSP did not change significantly, indicating that the nanoporous structure generated by electron bombardment had no effect on its hydrophilicity and surface energy. However, the nanoporous structure promoted the release of strontium ions on the surface of PKPSP. Therefore, it can be seen that the fiber material with nanoporous structure provided in this embodiment is conducive to protein adsorption due to its high surface energy, hydrophilicity, specific surface area and polar groups. The adsorbed protein is conducive to subsequent cell spreading and adhesion.

[0238] The above-mentioned effect examples also show that, due to the large number of polar groups (-OH) on the PA in the surface coating of the nanoporous fiber material provided in this embodiment, the adsorption capacity of FN and BSA on the PKPS surface is significantly increased compared with PKF material. In addition, the nanopores generated by electron bombardment significantly increase the specific surface area of ​​PKPSP. Therefore, the adsorption capacity of PKPSP for BSA and FN provided in this embodiment is further enhanced, indicating that the nanoporous structure and PA / Sr chelate coating are beneficial to improving the protein adsorption capacity of PKPSP.

[0239] 2) The fiber material with nanoporous structure provided in this embodiment has good adhesion and proliferation ability to body cells, which has a positive promoting effect on the differentiation of body cells.

[0240] The above examples demonstrate that:

[0241] BMSCs showed the worst adhesion and proliferation on the PKF surface, and cell proliferation did not change significantly over time, which is due to the biological inertness of PEEK.

[0242] Compared with PKF, cell adhesion and proliferation on PKPS are significantly improved, and cell adhesion and proliferation on PKPS can be further improved due to the nanoporous structure generated by electron bombardment.

[0243] Since ALP and calcium nodules are important markers of early and late osteogenic differentiation of cells, respectively, the nanoporous fiber material provided in this embodiment has a positive effect on osteogenic differentiation of cells, as demonstrated by ALP and Alizarin Red staining. The results showed that ALP and calcium nodule formation on the surface of PKPS and PPKSP cells were higher than those on PKF, while PPKSP cells were higher than PKPS cells, indicating that PA / Sr promoted osteogenic differentiation of cells. Moreover, the nanoporous structure, together with PA / Sr, further promoted osteogenic differentiation of cells. Furthermore, the expression of Runx2, OCN, OPN, and ALP on PKPS and PPKSP cells was significantly better than that on PKF, with PPKSP cells being superior to PKPS cells. Therefore, it can be seen that the synergistic effect of PA and Strontium ions on the surface of PKPS can improve osteogenic differentiation of cells, while the synergistic effect of PA / Sr coating and nanoporous structure on the surface of PPKSP can further improve osteogenic differentiation of cells.

[0244] Furthermore, the results demonstrate that, since the nanoporous fiber material provided in this embodiment is an electroactive material, under electrical stimulation, the conductive biofiber material provided in this embodiment can enhance intercellular signal transduction, activate charged transmembrane receptors in the calcium / calmodulin-related pathway, and significantly promote osteoblast response.

[0245] 3) The results demonstrate that, under 300mV electrical stimulation, the nanoporous fiber material provided in this embodiment has a positive effect on cell response. Compared with no electrical stimulation, the proliferation and osteogenic differentiation of BMSC cells on the PKF surface did not change significantly under electrical stimulation, indicating that electrical stimulation had no effect on PKF surface cells. However, under electrical stimulation, the proliferation and osteogenic differentiation of PKPS and PPKSP surface cells were significantly enhanced, indicating that PKPS and PPKSP are electroactive and can promote cell response under electrical stimulation. During tissue repair, excessive reactive oxygen species are often generated at the injury site, causing oxidative stress in cells. Excessive reactive oxygen species directly act on the cell membranes of surrounding cells, thereby damaging cells and tissues and hindering tissue repair. Therefore, materials with antioxidant properties can provide electrons to eliminate reactive oxygen species.

[0246] 4) The co-culture effect of the materials with BMSC cells under oxidative stress was demonstrated by the example. When co-cultured with PKF, the cells still had a large amount of reactive oxygen species, indicating that PKF does not have the ability to scavenge reactive oxygen species. However, the intracellular reactive oxygen species content in cells co-cultured with PKPS decreased significantly, indicating that PA on the surface of PKPS has good antioxidant capacity and can provide electrons to reactive oxygen species in the cells, thereby eliminating reactive oxygen species. The intracellular reactive oxygen species level in cells co-cultured with PKBSP was further reduced, indicating that its ability to scavenge reactive oxygen species was further improved. This shows that the nanoporous structure of PKPSP increases its specific surface area and works synergistically with PA to enhance the antioxidant capacity of PKPSP.

[0247] 5) The effect of examining the in vivo tendon-bone healing effect using a rabbit ACL reconstruction animal model can be seen from the following examples:

[0248] Four and twelve weeks after implantation, the amount of new bone tissue around PKPS and PPKSP was significantly higher than that around PKF, and the amount of new bone tissue around PPKSP was higher than that around PKPS, indicating that PKPSP containing PA / Sr chelate coating and nanoporous structure has a good ability to promote new bone formation.

[0249] Biomechanical failure load testing showed that the failure loads of PKPS and PPKSP were significantly higher than those of PKF, with PPKSP being the highest. This indicates that PPKSP has the best tendon-bone healing with the femur and tibia, the best biomechanical stability, and can achieve long-term load.

[0250] Meanwhile, histological images showed that the amount of new bone tissue generated around PKPS and PPKSP was higher than that around PKF, and PPKSP was higher than PKPS. The gap between the new bone tissue and PKF fibers was obvious, and the implanted material was wrapped by fibrous tissue. However, the new bone was in close contact with the surface of PPKSP and there was no fibrous tissue wrapping, which showed good tendon-bone healing, which is crucial for improving the quality of tendon-bone healing.

[0251] Based on the results of the above examples, it can be shown that:

[0252] Compared with PKF, PKPS and PKBSP have the ability to release strontium ions and optimized surface properties, as well as excellent electroactivity and ROS scavenging ability;

[0253] Compared to PKPS, the nanoporous structure on the surface of PKPSP enhances protein adsorption and strontium ion release. Due to its excellent surface properties and sustained strontium ion release capability, PKPSP significantly promotes cell responses, including adhesion, proliferation, and differentiation. Furthermore, due to the conductivity and antioxidant properties of PA, PKPSP further promotes osteoblast responses under electrical stimulation and can effectively scavenge intracellular ROS, thereby providing a microenvironment with better osteogenic effects. Therefore, it can be seen that the nanoporous fiber material PKPSP provided in this embodiment has excellent osteogenic bioactivity, can promote tendon-bone healing, and has significant clinical application potential as an artificial ligament. Example 2

[0254] This embodiment provides a fiber material with a nanoporous structure.

[0255] The fiber material with a nanoporous structure provided in this embodiment is prepared by the preparation method described in Example 1 above.

[0256] Since the nanoporous fiber material provided in this embodiment is prepared by the method described in Example 1, it is known that it has optimized surface properties, excellent electroactivity and ROS scavenging ability, as well as excellent osteogenic bioactivity, which can promote tendon-bone healing and has significant clinical application potential as an artificial ligament. Example 3

[0257] This embodiment provides an artificial ligament fabric.

[0258] The artificial ligament fabric provided in this embodiment is woven from the nanoporous fiber material provided in Example 2.

[0259] Since the artificial ligament fabric provided in this embodiment is woven from the nanoporous fiber material provided in Example 2, it is known that it has optimized surface properties, excellent electroactivity and ROS scavenging ability, as well as excellent osteogenic bioactivity, which can promote tendon-bone healing. As an artificial ligament fabric, it has significant clinical application potential.

[0260] In conclusion, it can be seen that:

[0261] This invention utilizes the chelating ability of PA to coat PA / Sr chelates onto the surface of polyether ether ketone (PEEK) fibers to prepare PA / Sr chelate coated fibers. Then, an electron bombardment method is used to bombard the chelate coating surface to create a nanoporous structure, thereby constructing the fiber material with a nanoporous structure provided by this invention. This fiber material can be woven into fabric and used as an artificial ligament.

[0262] Comparative analysis of commercially available polyetheretherketone (PEEK) fiber-woven artificial ligaments (PKF), PEEK fiber-woven artificial ligaments (PKPS) coated with a PA / Sr chelate coating, and PEEK fiber-woven artificial ligaments (PKPSP) coated with a PA / Sr chelate coating whose surface is coated with a PA / Sr chelate coating and the chelate coating is subjected to electron bombardment to form a nanoporous structure, reveals the following:

[0263] Compared with PKF, PKPS and PKPSP exhibit optimized surface properties such as hydrophilicity, surface energy and protein adsorption, and also have good conductivity, antioxidant properties and sustained strontium ion release capability.

[0264] Compared with PKPS, PKPSP has further enhanced protein adsorption and strontium ion release capabilities. PKPSP also significantly promotes the cell response of BMSCs, such as adhesion, proliferation and differentiation. Under electrical stimulation, it can further promote the cell response of BMSCs. At the same time, it can remove intracellular reactive oxygen species and reduce oxidative stress. In addition, PKPSP has a significant promoting effect on bone tissue regeneration in vivo and can form tendon-bone healing.

[0265] In summary, it can be seen that due to its nanoporous structure, PKPSP optimizes its surface properties, continuously releases strontium, and exhibits good electroactivity and antioxidant properties. It can stimulate BMSC response and promote bone regeneration and tendon-bone healing in vivo. Therefore, it has broad application prospects in the field of artificial ligaments. Compared with existing technologies, it has outstanding beneficial effects and significant progress, and thus has great promotion and application value.

[0266] In the description process of the above instruction manual:

[0267] The terms “this embodiment,” “this embodiment of the invention,” “this case,” “this comparative example,” “as shown,” “further,” etc., are used to indicate that the specific features, structures, materials, or characteristics described in the embodiment or case or comparative example are included in at least one embodiment or case or comparative example of the present invention.

[0268] In this specification, the illustrative expressions of the above terms are not necessarily directed at the same embodiments, cases, or comparative examples. Moreover, the specific features, structures, materials, or characteristics described may be combined or combined in any suitable manner in one or more embodiments, cases, or comparative examples. Furthermore, without causing contradiction, those skilled in the art may combine or combine the different embodiments, cases, or comparative examples described in this specification, as well as the features in the different embodiments, cases, or comparative examples.

[0269] Finally, it should be noted that:

[0270] The above embodiments and comparative examples are only used to illustrate the technical solutions and technical effects of the present invention, and are not intended to limit them. Although the present invention has been described in detail with reference to the foregoing embodiments, examples, and comparative examples, those skilled in the art should understand that modifications or supplements can still be made to the technical solutions or technical effects described in the foregoing embodiments, examples, and comparative examples, or equivalent substitutions can be made to some or all of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions described in the embodiments of the present invention. Non-essential improvements, adjustments, or substitutions made by those skilled in the art based on the content of this specification are all within the scope of protection claimed by the present invention.

Claims

1. A method for preparing a fiber material with a nanoporous structure, characterized in that, Includes the following steps: Prepare polyetheretherketone (PEEK) fibers; A chelate coating of phytic acid and strontium ions is constructed on the surface of the polyetheretherketone fiber; The chelate coating is subjected to electron bombardment treatment to form a nanoporous structure on the surface of the chelate coating.

2. The preparation method according to claim 1, characterized in that: The chelate coating is constructed using a stacking assembly technique.

3. The preparation method according to claim 2, characterized in that: The stacked assembly technology includes the following steps: The polyetheretherketone fiber is immersed in a phytic acid solution, so that phytic acid molecules adhere to the surface of the polyetheretherketone fiber. Cleaning removes any unattached phytic acid molecules; The polyetheretherketone fiber, after the phytic acid molecules have been attached, is then immersed in a strontium ion solution, so that the strontium ions in the strontium ion solution react with the phytic acid molecules attached to the surface of the polyetheretherketone fiber to form a chelate coating. Cleaning removes unreacted strontium ions; Repeat the above steps multiple times until the thickness of the chelate coating meets the design requirements.

4. The preparation method according to claim 3, characterized in that: The phytic acid solution has a volume percentage concentration of 3-7%, and the pH value of the phytic acid solution is adjusted to 4.5-6.5 by using an alkaline solution; The polyetheretherketone fiber is immersed in phytic acid solution for 0.5 to 2 hours; The strontium ion solution is a strontium nitrate solution with a concentration of 0.3–0.7 M; The polyetheretherketone fiber with phytic acid molecule attachment is immersed in strontium ion solution for 15 to 45 minutes; The stacked assembly is repeated 5 to 9 times.

5. The preparation method according to claim 4, characterized in that: The phytic acid solution has a volume percentage concentration of 5% and a pH value of 5.5, while the alkaline solution for adjusting the pH value of the phytic acid solution is a 1M NaOH solution. The polyetheretherketone fiber was immersed in the phytic acid solution for 1 hour; The strontium nitrate solution has a concentration of 0.5 M; The polyetheretherketone fiber with phytic acid molecule attachment was immersed in a strontium ion solution for 30 minutes. The stacked assembly is repeated 7 times.

6. The preparation method according to claim 3, characterized in that: The solvent used to clean and remove unattached phytic acid molecules is water, and the solvent used to clean and remove unreacted strontium ions is hydrogen peroxide.

7. The preparation method according to claim 1, characterized in that: The electron bombardment treatment has a bombardment voltage of 15-25 kV and a bombardment treatment duration of 5-15 seconds.

8. The preparation method according to claim 7, characterized in that: The electron bombardment treatment has a bombardment voltage of 20kV and a bombardment treatment duration of 10 seconds.

9. A fiber material with a nanoporous structure, characterized in that: It is prepared by the preparation method according to any one of claims 1 to 8.

10. An artificial ligament fabric, characterized in that: The fabric is woven from the fibrous material with a nanoporous structure as described in claim 9.

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