A bone promoting antibacterial biological functional material and a preparation method thereof

Abstract

The present application relates to a kind of promote bone antibacterial biological function material and its preparation method, belong to laser application technical field.The present application is after polishing nickel-titanium alloy material ultrasonic and dry, using femtosecond laser direct writing technology, through the laser flux of synergic control processing process, processing translation table moving speed and scanning pitch and other processing parameters, prepare the self-organizing composite periodic micro-wave groove structure;It is again after being handled by anodic oxidation, form rich nanometer hole on the basis of the structure, prepare to get groove-micro-wave groove-micropore three-level micro-nano composite structure;Subsequently, the three-level micro-nano composite structure prepared is placed in the alkaline Tris buffer solution containing hydrochloric acid dopamine, and the polydopamine coating is attached;Subsequently, the groove-micro-wave groove-micropore three-level micro-nano composite structure with polydopamine coating is placed in silver nitrate solution, to load nano-silver on its surface, finally, the biological function material with good bone bonding ability, promote bone growth ability and antibacterial ability is prepared.

Description

TECHNICAL FIELD

[0001] The present application relates to a kind of promoting bone antibacterial biological function material and its preparation method, belong to laser application technical field. BACKGROUND

[0002] Bone implants play a crucial role in bone defect repair and bone disease treatment. However, the long-term stability and therapeutic effect of bone implants are severely affected by poor bone integration and bacterial infection. Poor bone integration makes the bonding force between the implant and the surrounding bone tissue insufficient, leading to implant loosening, displacement and even surgical failure. On the other hand, bacterial infection is one of the common complications in the clinical application of bone implants, which can cause inflammatory reaction and bone resorption in the surrounding tissue of the implant, seriously affecting the long-term stability of the implant and the therapeutic effect of the patient. Therefore, solving the problems of poor bone integration and bacterial infection is crucial for improving the biocompatibility of bone implants, promoting bone tissue regeneration and improving the clinical treatment effect.

[0003] The surface morphology and chemical properties of bone implants are crucial for their combination with surrounding bone tissue. Femtosecond laser technology uses ultrashort pulse laser to control the microstructure and nanoscale structure of the implant surface. It can increase the surface roughness and specific surface area, thereby improving the biocompatibility and cell adhesion. In addition, femtosecond laser can also induce surface chemical reactions such as oxidation, carbonization, etc., further improving the chemical properties of the implant, which is beneficial to the growth of bone cells and the regeneration of bone tissue. Anodic oxidation technology increases the surface roughness, bioactivity and biocompatibility of the implant by forming a dense oxide layer on the surface, which is beneficial to the attachment and growth of bone cells and can promote the formation and regeneration of bone tissue. Femtosecond laser technology and anodic oxidation technology as surface treatment methods can effectively improve the surface morphology and surface properties of the implant, thereby promoting the attachment and growth of bone cells, improving the bone integration performance of the implant, and have attracted more and more attention.

[0004] In current research, for example, Mengxia Peng et al. achieved the improvement of antibacterial performance of biological metal materials through double surface modification treatment of anodic oxidation and preparation of nano-silver coating, but they focused on the antibacterial performance of the material and did not pay attention to the osteogenic performance of the material. Su Wang et al. used femtosecond laser to modify the surface of titanium alloy bone scaffold, making the surface morphology of the bone scaffold present micro-nano morphology, increasing the surface roughness, and significantly enhancing the surface activity and integration ability with surrounding bone tissue, but they focused on the osteogenic performance of the material and did not pay attention to the antibacterial function of the material. SUMMARY

[0005] The application aims to solve the problems of poor bone integration and bacterial infection in the application of bone implants in bone defect repair and bone disease treatment, and provides a bone-promoting antibacterial biological functional material and a preparation method thereof; the method is characterized in that: the polished nickel-titanium alloy material is ultrasonically treated and dried, a self-organized composite periodic micro-ripple groove structure is prepared by using a femtosecond laser direct writing technology, through synergistically regulating processing parameters such as laser flux, processing translation table moving speed and scanning spacing, the nickel-titanium alloy material is anodized, rich nanopores are formed on the basis of the structure, and a three-level micro-nano composite structure of groove-micro-ripple-micro-pore is prepared; then, the prepared three-level micro-nano composite structure is coated with a polydopamine coating in an alkaline Tris buffer solution containing dopamine hydrochloride; then, the three-level micro-nano composite structure with the polydopamine coating is placed in a silver nitrate solution to load nano-silver on the surface, and finally, the biological functional material with good bone combination ability, bone growth promoting ability and antibacterial ability is prepared.

[0006] To achieve the above application purposes, the application provides the following technical solutions.

[0007] A preparation method of a bone-promoting antibacterial biological functional material, comprising the following steps:

[0008] Step one, ultrasonically treating and drying the polished nickel-titanium alloy material;

[0009] Step two, vertically focusing the femtosecond laser to the surface of the nickel-titanium alloy material;

[0010] Step three, generating relative motion between the femtosecond laser and the nickel-titanium alloy, and processing along the same motion path in a cycle to obtain a self-organized composite periodic micro-ripple groove structure;

[0011] Step four, placing the groove structure prepared in step three into an electrolytic cell to perform an anodization treatment, forming rich nanopores on the groove structure to obtain a three-level micro-nano composite structure of groove-micro-ripple-micro-pore;

[0012] Step five, placing the three-level micro-nano composite structure prepared in step four into an alkaline Tris buffer solution containing dopamine hydrochloride to form a polydopamine coating on the three-level micro-nano composite structure, and obtaining a composite structure;

[0013] Step six, placing the composite structure obtained in step five into a silver nitrate solution to obtain the bone-promoting antibacterial biological functional material.

[0014] The specific implementation steps of step three are as follows:

[0015] (1) fixing the clean nickel-titanium alloy material on a glass slide, and then fixing the glass slide on a high-precision six-degree-of-freedom translation table;

[0016] (2) Set the femtosecond laser repetition rate to 1000Hz, use a 5x objective lens to focus the femtosecond laser on the material surface; control the laser flux to be 1J / cm 2 ~10J / cm 2 , the machining speed is 500um / s-2000um / s, and the machining pitch is 8um-20um; a high-precision six-degree-of-freedom translation table is controlled by a computer control system to make the sample move relative to the laser; and a groove structure is obtained.

[0017] The anodizing treatment method in step four is as follows: an electrolyte composed of sodium chloride, deionized water and ethylene glycol is configured, the composite periodic micro-ridge groove structure prepared in step three is connected with the working electrode of the electrochemical workstation, in the constant current mode, the voltage of the electrochemical workstation is set to 5V-10V, and the anodizing time is set to 180s-600s.

[0018] The specific implementation steps of step five are as follows: a dopamine hydrochloride solution with a concentration of 1mg / ml-3mg / ml is configured, the solvent is an alkaline Tris buffer solution with a concentration of 10mM-50mM and a pH value of 8-10; and the tertiary micro-nano composite structure prepared in step four is immersed in the dopamine hydrochloride solution, the soaking time is 12h-24h, and then ultrasonic cleaning and vacuum drying are performed.

[0019] The specific implementation steps of step six are as follows: a silver nitrate solution with a concentration of 10mM-50mM is configured, the composite structure prepared in step five is placed in the silver nitrate solution, and the reaction time is 1h-12h; and then ultrasonic cleaning and drying are performed.

[0020] The obtained bone-promoting antibacterial biological functional material is used for bone defect repair and postoperative bacterial infection; the obtained bone-promoting antibacterial biological functional material has the functions of resisting bacterial infection and promoting bone defect repair.

[0021] Advantages:

[0022] 1. The bone-promoting antibacterial biological functional material and the preparation method thereof have the ability of patterned processing by using the relative motion of femtosecond laser and nickel-titanium alloy material, and can be used for nickel-titanium alloy materials of various sizes and different shapes.

[0023] 2. The bone-promoting antibacterial biological functional material and the preparation method thereof can effectively inhibit the formation of biofilm, have excellent antibacterial infection ability by the certain antibacterial adhesion ability of the multi-level micro-nano composite structure and the significant killing of various bacteria adhering to the surface through the surface silver element. At the same time, the biological functional material has excellent cell compatibility in vitro, has no cytotoxicity and can significantly induce bone regeneration. It can be used to prevent bacterial infection, promote bone defect repair and improve the problem of poor osseointegration of bone implants.

[0024] 3. The bone-promoting antibacterial biological functional material and the preparation method thereof, the prepared three-level micro-nano composite structure of groove-microripples-micropores has excellent drug loading capacity, and the three-level micro-nano composite structure of groove-microripples-micropores with high specific surface area can provide rich drug loading space and thus improve the drug loading capacity.

[0025] 4. The bone-promoting antibacterial biological functional material and the preparation method thereof, the preparation method of the three-level micro-nano composite structure of groove-microripples-micropores is flexible and simple, the process parameters are easy to control, and the laser technology field application can be easily realized. BRIEF DESCRIPTION OF DRAWINGS

[0026] Figure 1 is the optical path diagram of the processing method of the application.

[0027] Figure 2 is the SEM image of the material prepared in the embodiment of the application, wherein (a) is the SEM image of the groove-microripples composite structure prepared by polishing nickel-titanium alloy through femtosecond laser processing; (b) is the SEM image of the three-level micro-nano composite structure of groove-microripples-micropores prepared by anodizing the sample after femtosecond laser processing; and (c) is the SEM image of the three-level micro-nano composite structure of groove-microripples-micropores loaded with antibacterial nano-silver.

[0028] Figure 3 is the surface contact angle test result of each type of material prepared in the embodiment of the application.

[0029] Figure 4 is the cytotoxicity of the bone-promoting antibacterial biological functional material prepared in the embodiment of the application.

[0030] Figure 5 is the live and dead staining fluorescence microscopic image of the osteoblasts (MC3T3-E1, co-cultured for 24 hours) of the bone-promoting antibacterial biological functional material prepared in the embodiment of the application, wherein (a) is the survival of the osteoblasts on the surface of the clean polished nickel-titanium alloy; (b) is the survival of the osteoblasts on the surface of the groove-microripples composite structure prepared by polishing nickel-titanium alloy through femtosecond laser processing; (c) is the survival of the osteoblasts on the surface of the three-level micro-nano composite structure of groove-microripples-micropores prepared by anodizing the sample after femtosecond laser processing; and (d) is the survival of the osteoblasts on the surface of the three-level micro-nano composite structure of groove-microripples-micropores loaded with antibacterial nano-silver.

[0031] Figure 6Figure 1 is a fluorescence microscopic image of osteoblasts (MC3T3-E1, co-cultured for 24 hours) adhered to the antibacterial bone-promoting biological functional material prepared in the embodiments of the present application. Among them, (a) is the adhesion of osteoblasts on the surface of clean polished nickel-titanium alloy; (b) is the adhesion of osteoblasts on the surface of the groove-micro-ridge composite structure prepared by polishing nickel-titanium alloy with femtosecond laser processing; (c) is the adhesion of osteoblasts on the groove-micro-ridge-micro-pore three-level micro-nano composite structure prepared by anodizing the sample after femtosecond laser processing; (d) is the adhesion of osteoblasts on the surface of the groove-micro-ridge-micro-pore three-level micro-nano composite structure after loading of antibacterial nano-silver.

[0032] Figure 7 Figure 2 is a fluorescence microscopic image of Staphylococcus aureus colonization on the antibacterial bone-promoting biological functional material prepared in the embodiments of the present application. (a) is the colonization of Staphylococcus aureus on the surface of clean polished nickel-titanium alloy; (b) is the colonization of Staphylococcus aureus on the surface of the groove-micro-ridge composite structure prepared by polishing nickel-titanium alloy with femtosecond laser processing; (c) is the colonization of Staphylococcus aureus on the groove-micro-ridge-micro-pore three-level micro-nano composite structure prepared by anodizing the sample after femtosecond laser processing;

[0033] (d) is the colonization of Staphylococcus aureus on the surface of the groove-micro-ridge-micro-pore three-level micro-nano composite structure after loading of antibacterial nano-silver.

[0034] Figure 8 Figure 3 is a fluorescence microscopic image of Escherichia coli colonization on the antibacterial bone-promoting biological functional material prepared in the embodiments of the present application. Among them, (a) is the colonization of Escherichia coli on the surface of clean polished nickel-titanium alloy; (b) is the colonization of Escherichia coli on the surface of the groove-micro-ridge composite structure prepared by polishing nickel-titanium alloy with femtosecond laser processing; (c) is the colonization of Escherichia coli on the groove-micro-ridge-micro-pore three-level micro-nano composite structure prepared by anodizing the sample after femtosecond laser processing; (d) is the colonization of Escherichia coli on the surface of the groove-micro-ridge-micro-pore three-level micro-nano composite structure after loading of antibacterial nano-silver.

[0035] Among them, 1 is a femtosecond laser; 2 is a continuous attenuation sheet; 3 is a first reflecting mirror; 4 is a second reflecting mirror; 5 is a light shutter; 6 is a third reflecting mirror; 7 is a focusing lens; 8 is a nickel-titanium alloy material to be processed; 9 is a six-degree-of-freedom translation stage; 10 is a dichroic mirror; 11 is a white light source; 12 is a CCD camera; and 13 is a computer. DETAILED DESCRIPTION

[0036] The present application will be described in detail below in conjunction with the accompanying drawings and examples.

[0037] The experimental methods used in the following examples are conventional methods unless otherwise specified.

[0038] Embodiment 1

[0039] The application discloses a kind of promoting bone antibacterial biological function materials and preparation method thereof, comprising the following specific steps:

[0040] Step one: polished nickel-titanium alloy material with surface roughness less than 5 angstrom is cleaned using ultrasonic cleaner, immerse sample in deionized water and anhydrous ethanol solution at room temperature respectively, and the ultrasonic frequency of ultrasonic cleaner is set to 80kHZ, and cleaning is performed for 5min.

[0041] Step two: the cleaned sample of step one is dried using ear bulb, to obtain clean nickel-titanium alloy sample.

[0042] Step three: using femtosecond laser processing system as shown in Figure 1 , laser generated by femtosecond laser passes through continuous attenuation sheet, first mirror, second mirror, light shutter, third mirror and focusing lens in sequence, and is focused on the surface of nickel-titanium alloy material to be processed; the opening and closing of light shutter and the movement of six-degree-of-freedom translation table are controlled by computer; white light generated by white light source passes through dichroic mirror, third mirror and focusing lens in sequence, and is irradiated to the surface of nickel-titanium alloy material to be processed, and then is reflected to CCD camera along focusing lens, third mirror and dichroic mirror in sequence to generate real-time imaging of processing area. Groove-micro-ripple micro-nano composite structure is prepared on the surface of nickel-titanium alloy, and the laser processing parameters are specifically set as follows: the wavelength of femtosecond laser is 800nm, the repetition frequency is 1kHz, the processing objective lens is selected as 5 times objective lens, the focal point is processed, the laser flux is 5.43J / cm 2 , the scanning speed is 600μm / s, the scanning interval is 10um, and high-pressure nitrogen is used to blow away chips during the whole processing process to process large-area and consistent groove-micro-ripple micro-nano composite structure.

[0043] Step four: the sample prepared in step three is subjected to anodic oxidation using electrochemical workstation, platinum sheet and the sample prepared in step three are connected to the counter electrode and working electrode respectively, a saturated calomel reference electrode is selected, the anodic oxidation electrolyte is a glycol solution containing 5.0vol.%H2O and 0.3M NaCl, the electrolysis voltage is 10V, and the anodic oxidation time is 5min, to prepare groove-micro-ripple-micropore three-level micro-nano composite structure.

[0044] Step five: Put the groove-microrough-micropore three-level micro-nano composite structure prepared in step four into a basic Tris buffer solution containing dopamine hydrochloride, wherein the concentration of dopamine hydrochloride (DA) is 2 mg / mL, the pH of the Tris buffer solution is 8.5, and the solution is left to stand at room temperature for 24 h.

[0045] Step six: Put the groove-microrough-micropore three-level micro-nano composite structure with a polydopamine coating prepared in step five into a silver nitrate solution, and form nano-silver particles on the surface of the groove-microrough-micropore three-level micro-nano composite structure by using the reducing property of polydopamine, wherein the concentration of the silver nitrate solution is 50 mM, and the reaction time is 6 h.

[0046] Step seven: Rinse the groove-microrough-micropore three-level micro-nano composite structure loaded with nano-silver particles prepared in step six with deionized water, and vacuum dry (37℃, 4 h) in a vacuum drying box to obtain a nickel-titanium alloy antibacterial and bone integration promoting functional material.

[0047] The product prepared by the method has excellent antibacterial infection function, bone integration promoting function and bone defect repair function, and can be used for bone implants and other medical devices to repair bone defects and treat postoperative bacterial infections.

[0048] As shown in Figure 1 The optical path diagram of the processing method of the application is shown.

[0049] As shown in Figure 2 The SEM images of the surfaces of various types of materials prepared in the embodiment of the application are shown

[0050] As shown in Figure 3 The static contact angles of the surfaces of various types of materials prepared in the embodiment of the application are shown. As can be seen from the figure, the contact angles of the original polished surface of the nickel-titanium alloy (denoted as P-NiTi), the groove-microrough structure surface (denoted as Fs-NiTi), the groove-microrough-micropore three-level micro-nano composite structure surface (denoted as Fs-NPs) and the groove-microrough-micropore three-level micro-nano composite structure loaded with nano-silver (denoted as Fs-NPs-PDA-Ag) are 72.6°±5.4°, 74°±10.1°, 36°±4.8° and 41.5°±2.6°, respectively. After the sample surface is processed by femtosecond laser and anodized, and then loaded with nano-silver after being coated with a polydopamine coating, the sample surface has a strong ability to adsorb water molecules, so that the surface contact angle is reduced to 41.5°±2.6°, and the sample surface has good hydrophilicity.

[0051] As shown in Figure 4The diagram shows the cell activity and proliferation tests of the bone-promoting and antibacterial biomaterial prepared according to an embodiment of the present invention. At culture times of 1 day, 3 days, and 5 days, the relative activity of cells in each group showed almost no significant difference on the first day, maintaining nearly 100% cell activity. With increasing incubation time, the cell proliferation rate of each treated sample group significantly increased compared to the original surface, indicating that femtosecond laser processing, anodizing, and loading with antibacterial silver nanoparticles did not significantly inhibit cell proliferation. The bone-promoting and antibacterial biomaterial exhibits excellent biocompatibility and can promote cell proliferation to a certain extent.

[0052] like Figure 5 The image shows the in vitro cell viability test of the bone-promoting and antibacterial biofunctional material prepared according to an embodiment of the present invention. As can be seen from the fluorescence micrograph, the surface of each group of samples in the embodiment of the present invention shows a large area of ​​green fluorescence, with almost no red fluorescence. The cells in each group are basically alive, indicating that the prepared materials are basically non-cytotoxic.

[0053] like Figure 6 The image shows the bone-promoting and antibacterial biomaterials prepared according to embodiments of the present invention, which exhibit bone regeneration characteristics. Fluorescence microscopy images show that, compared to the original polished surface ( Figure 6 (a)) Osteoblasts with abundant pseudopodia almost completely cover the surface of the groove-microwave-micropore three-level micro-nano composite structure. Figure 6 (c) and a three-level micro / nano composite structure of groove-microwave texture-micropore loaded on a nano-silver surface ( Figure 6 (d) The number of cells also increased significantly compared to the original polished surface, indicating that the osteogenic and antibacterial biofunctional material prepared in the embodiments of the present invention can significantly induce the adhesion and proliferation of osteoblasts and has excellent osteogenic properties.

[0054] like Figure 7 The image shown is a fluorescence micrograph of Staphylococcus aureus colonization on the bone-promoting and antibacterial biomaterial prepared according to an embodiment of the present invention. Original polished surface ( Figure 7 (a) The coverage of live bacteria (green) on the surface was 38.9%, and the coverage of dead bacteria (red) was 0.01%. Staphylococcus aureus was almost colonized throughout the entire sample surface, with almost no dead bacteria. Groove-microwave textured surface ( Figure 7 (b) The coverage rate of live bacteria (green) was 31.4%, and the coverage rate of dead bacteria (red) was 0.1%. The number of Staphylococcus aureus decreased. The surface of the groove-microwave texture-micropore three-level micro-nano composite structure ( Figure 7(c) The live bacteria (green) coverage on the surface is 15.8%, the dead bacteria (red) coverage is 1.2%, and the number of S. aureus is reduced compared with the original surface, indicating that the three-level micro-nano composite structure of groove-microripples-microporous greatly reduces the colonization sites of S. aureus. The groove-microripples-microporous three-level micro-nano composite structure loaded with nano-silver surface Figure 7 (d) The live bacteria (green) coverage on the surface is 0.1%, and the dead bacteria (red) coverage is 2.3%, and there is almost no live S. aureus colonization.

[0055] As shown in Figure 8 The fluorescence micrograph of E. coli colonization on the bone-promoting antibacterial biological functional material prepared in the embodiment of the application is shown. The original polished surface Figure 8 (a) The live bacteria (green) coverage on the surface is 49.9%, and the dead bacteria (red) coverage is 0.1%, and the surface is uniformly covered with E. coli. The groove-microripples structure surface Figure 8 (b) The live bacteria (green) coverage on the surface is 38.9%, and the dead bacteria (red) coverage is 0.4%, and the number of E. coli colonization is reduced. The groove-microripples-microporous three-level micro-nano composite structure surface Figure 8 (c) The live bacteria (green) coverage on the surface is 20.6%, and the dead bacteria (red) coverage is 0.3%, and the groove-microripples-microporous three-level micro-nano composite structure greatly reduces the number of E. coli colonization, indicating that the structure has the effect of resisting E. coli colonization. The groove-microripples-microporous three-level micro-nano composite structure loaded with nano-silver surface Figure 8 (d) The live bacteria (green) coverage on the surface is 0.1%, and the dead bacteria (red) coverage is 2.1%, and the number of E. coli on the surface is very small, and there is almost no live E. coli colonization.

[0056] The above specific description further details the purpose, technical solutions and beneficial effects of the application, and it should be understood that the above description is only a specific embodiment of the application, and is not limited to the protection scope of the application, and any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the application should be included in the protection scope of the application.

Claims

1. A method for preparing a bone-promoting and antibacterial biomaterial, characterized in that: Includes the following steps: Step 1: Ultrasonically clean and dry the polished nickel-titanium alloy material; Step 2: The femtosecond laser is vertically focused onto the surface of the nickel-titanium alloy material; Step 3: The femtosecond laser and the nickel-titanium alloy generate relative motion and are processed repeatedly along the same motion path to obtain a self-organized composite periodic microwave groove structure. The specific implementation steps for step three are as follows: (1) Fix the clean nickel-titanium alloy material onto the glass slide, and then fix the glass slide onto the high-precision six-degree-of-freedom translation stage; (2) Set the femtosecond laser repetition rate to 1000Hz, and focus the femtosecond laser onto the material surface using a 5x objective lens; control the laser flux to 1J / cm. 2 ~10J / cm 2 The processing speed is 500um / s-2000um / s, and the processing spacing is 8um-20um; a high-precision six-degree-of-freedom translation stage is controlled by a computer control system to make the sample move relative to the laser. A groove structure with composite periodic microwave ripples was obtained; Step 4: Place the trench structure prepared in Step 3 into an electrolytic cell for anodic oxidation treatment to form abundant nanopores on the trench structure, thus obtaining a three-level micro-nano composite structure of trench-microwave texture-micropore. The anodizing process described in step four is as follows: an electrolyte composed of sodium chloride, deionized water and ethylene glycol is prepared, the groove structure of the composite periodic microwave texture prepared in step three is connected to the working electrode of the electrochemical workstation, and the voltage of the electrochemical workstation is set to 5V-10V in constant current mode, and the anodizing time is set to 180s-600s. Step 5: Place the tertiary micro-nano composite structure prepared in Step 4 into an alkaline Tris buffer solution containing dopamine hydrochloride to form a polydopamine coating on the tertiary micro-nano composite structure, thus obtaining the composite structure. Step 6: Place the composite structure obtained in Step 5 into a silver nitrate solution to obtain a bone-promoting and antibacterial biofunctional material.

2. The preparation method of the bone-promoting and antibacterial biofunctional material as described in claim 1, characterized in that: The specific implementation steps of step five are as follows: Prepare a dopamine hydrochloride solution with a concentration of 1 mg / ml to 3 mg / ml, using alkaline Tris buffer solution with a concentration of 10 mM to 50 mM and a pH value of 8 to 10; immerse the tertiary micro-nano composite structure prepared in step four into the dopamine hydrochloride solution for 12 h to 24 h, followed by ultrasonic cleaning and vacuum drying.

3. The method for preparing a bone-promoting and antibacterial biomaterial as described in claim 1, characterized in that: The specific implementation steps of step six are as follows: prepare a silver nitrate solution with a concentration of 10mM-50mM, place the composite structure prepared in step five into the silver nitrate solution, and react for 1h-12h; then ultrasonically clean and dry.

4. The preparation method of the bone-promoting and antibacterial biofunctional material as described in claim 1, characterized in that: The obtained bone-promoting and antibacterial biofunctional material is used for bone defect repair and postoperative bacterial infection; the bone-promoting and antibacterial biofunctional material has the functions of antibacterial infection and promoting bone defect repair.

5. An apparatus for implementing the method of any one of claims 1 to 4, characterized in that: include: Femtosecond laser; Continuous attenuation plate; First reflecting mirror; Second reflecting mirror; Light shutter; Third reflecting mirror; Focusing lens; Nickel-titanium alloy material to be processed; Six-DOF translation stage; dichroic mirror ； A white light source; a CCD camera and a computer; a laser generated by a femtosecond laser passes sequentially through a continuous attenuator, a first reflecting mirror, a second reflecting mirror, an optical shutter, a third reflecting mirror, and a focusing lens, and is focused onto the surface of the nickel-titanium alloy material to be processed; the computer controls the opening and closing of the optical shutter and the movement of the six-degree-of-freedom translation stage; the white light generated by the white light source passes sequentially through a dichroic mirror, a third reflecting mirror, and a focusing lens to illuminate the surface of the nickel-titanium alloy material to be processed, and is then reflected sequentially along the focusing lens, the third reflecting mirror, and the dichroic mirror to the CCD camera to generate real-time imaging of the processing area.

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