Preparation method and application of titanium-silver doped antibacterial bone electroactive material

Abstract

This invention discloses a method for preparing and applying a titanium-silver doped antibacterial bone electroactive material, relating to the field of bone electroactive materials. The method for preparing the PVDF bone electroactive film of this invention involves: cutting, cleaning, and drying the PVDF film, and storing it under vacuum; depositing elemental titanium and silver as magnetron sputtering targets onto the PVDF film to obtain the titanium-silver doped antibacterial bone electroactive material; the magnetron sputtering method is as follows: under an argon atmosphere, the internal pressure of the magnetron sputtering chamber is adjusted to 0.5-5 Pa, the sputtering distance is 4-10 cm, the sputtering power is maintained at 50-100 W, the substrate temperature is room temperature to 300℃, the argon flow rate is 5-80 sccm, and the duration is 10-20 min. This invention effectively modifies the surface of the PVDF bone electroactive film with titanium and silver using magnetron sputtering technology, significantly improving the adhesion of the PVDF film to bone tissue, and exhibiting excellent antibacterial activity and good osteogenic induction activity.

Description

Technical Field

[0001] This invention relates to the field of bone electroactive materials, and particularly to the preparation technology of titanium-silver doped antibacterial bone electroactive materials. Background Technology

[0002] Current research indicates that the microscopic nano-topography and electrical signal properties of bone defect repair materials are crucial for bone regeneration and antibacterial environments. When subjected to external forces, PVDF films exhibit polarized charges on their surface, mimicking the piezoelectric properties of natural bone tissue. They possess excellent piezoelectric performance and a large specific surface area, thus effectively mimicking the collagen fiber structure in the natural extracellular matrix. Based on these characteristics, PVDF films have wide applications in antibacterial environments, bone repair, biosensors, and drug delivery.

[0003] However, conventional PVDF films suffer from poor electrical signal stability, poor bone regeneration regulation, slow bone healing, and inadequate antibacterial properties. Although numerous studies have been conducted in recent years to modify PVDF films, it remains impossible to simultaneously resolve all of these issues. For the in vivo microenvironment of the human or animal body, especially environments teeming with bacteria such as the oral cavity, how to utilize suitable bone defect repair materials to achieve bone regeneration in this bacterial environment is a pressing problem in the field of bone electroactive materials. Summary of the Invention

[0004] This invention provides a method for preparing and applying a titanium-silver-doped antibacterial osteoactive material. This titanium-silver-doped antibacterial osteoactive material exhibits superior biocompatibility, antibacterial properties, and osteogenic induction ability, expanding its application scenarios in the oral cavity and other sites. It effectively addresses the shortcomings of existing PVDF osteoactive films in terms of poor bone integration, bone healing speed, and antibacterial effectiveness. Specifically, this is achieved through the following techniques.

[0005] A method for preparing a titanium-silver doped antibacterial bone electroactive material includes the following steps:

[0006] The PVDF film was cut, cleaned and dried, and then stored in a vacuum. Titanium and silver were used as magnetron sputtering targets to deposit onto the PVDF film to obtain the titanium-silver doped antibacterial bone electroactive material.

[0007] The magnetron sputtering method is as follows: under an argon atmosphere, the internal gas pressure of the magnetron sputtering cavity is adjusted to 0.5-5 Pa, the sputtering distance is 4-10 cm, the sputtering power is maintained at 50-100 W, the substrate temperature is room temperature to 300 ℃, the argon flow rate is 5-80 sccm, and the duration is 10-20 min.

[0008] Furthermore, the internal gas pressure of the magnetron sputtering cavity was adjusted to 5 Pa, the sputtering distance to 5 cm, the sputtering power to 50-100 W, the substrate temperature to 200 ℃, and the argon flow rate to 80 sccm for 10-20 min.

[0009] Furthermore, in the titanium-silver doped antibacterial bone electroactive material, the titanium content is 34.26-34.46 wt%, the silver content is 38.10-38.30 wt%, and the oxygen content is 27.34-27.54 wt%.

[0010] Furthermore, in the titanium-silver doped antibacterial bone electroactive material, the titanium content is 34.36 wt%, the silver content is 38.2 wt%, and the oxygen content is 27.44 wt%.

[0011] This invention provides a titanium-silver doped antibacterial bone electroactive material prepared by the above-described preparation method.

[0012] The present invention also provides an application of the above-mentioned titanium-silver doped antibacterial bone electroactive material, which is used as a bone defect repair material or an antibacterial material.

[0013] Compared with the prior art, the advantages of the present invention are:

[0014] 1. Existing conventional PVDF bone electroactive films have surface nanostructures that cannot establish good adhesion with bone tissue. This invention effectively modifies the surface of the PVDF bone electroactive film with titanium and silver using magnetron sputtering technology, significantly improving the adhesion between the PVDF film and bone tissue. The titanium content is 34.36 wt%, the oxygen content is 27.44 wt%, and the silver content is 38.2 wt%.

[0015] 2. Existing ordinary PVDF bone electroactive films have poor antibacterial properties. The titanium-silver doped antibacterial bone electroactive material prepared in this invention can effectively stimulate macrophages to express T lymphocyte activation antigen (CD86), interleukin-1β (IL-1β), interferon regulatory factor 3 (IRF3), TNF receptor-associated factor 6 (TRAF6), and nuclear factor receptor-κB (NF-κB), indicating that the titanium-silver doped antibacterial bone electroactive material prepared in this invention has excellent antibacterial activity.

[0016] 3. Existing PVDF osteoactive films have poor effects on bone repair. The titanium-silver doped antibacterial osteoactive material prepared in this invention can promote osteoblasts to express RUNX family transcription factor 2 (RUNX2), and its surface shows good osteogenic induction activity. Attached Figure Description

[0017] Figure 1 This is a schematic diagram of the titanium-silver-doped PVDF bone electroactive film modified by magnetron sputtering technology in this invention.

[0018] Figure 2 The image shows the overall imaging of the prepared titanium-silver doped PVDF bone electroactive film at 10,000x magnification using a scanning electron microscope, as described in Example 1.

[0019] Figure 3 This is the energy spectrum generated by scanning the surface elements of the titanium-silver-doped PVDF bone electroactive film using an energy spectrum scanning module in Example 1.

[0020] Figure 4 The images show the relative mRNA expression of CD86 in RAW264.7 macrophages after culturing the titanium-silver-doped PVDF osteoactive membrane of Example 1 and the ordinary PVDF membrane of Comparative Example 1.

[0021] Figure 5 The graph shows the relative mRNA expression of IL-1β after culturing RAW264.7 macrophages using the titanium-silver doped PVDF osteoactive membrane of Example 1 and the ordinary PVDF membrane of Comparative Example 1.

[0022] Figure 6 The images show the relative mRNA expression of IRF3 in RAW264.7 macrophages after culturing the titanium-silver-doped PVDF osteoactive membrane of Example 1 and the ordinary PVDF membrane of Comparative Example 1.

[0023] Figure 7 The images show the relative mRNA expression of TRAF6 in RAW264.7 macrophages after culturing the titanium-silver-doped PVDF osteoactive membrane of Example 1 and the ordinary PVDF membrane of Comparative Example 1.

[0024] Figure 8 The images show the relative mRNA expression of NF-κB in RAW264.7 macrophages after culturing the titanium-silver-doped PVDF osteoactive membrane of Example 1 and the ordinary PVDF membrane of Comparative Example 1.

[0025] Figure 9 The images show the relative mRNA expression of RUNX2 after osteoblasts (mt-3t3) were cultured using the titanium-silver doped PVDF osteoactive membrane of Example 1 and the ordinary PVDF membrane of Comparative Example 1.

[0026] Figure 10-13 The images show the relative mRNA expression of IL-6, NF-κB, MYD88, and TRAF6 in RAW264.7 macrophages after culturing the PVDF film materials of Example 1 and Comparative Examples 2 and 3, respectively. Detailed Implementation

[0027] The technical solution of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0028] Example 1

[0029] The titanium-silver doped antibacterial bone electroactive material provided in this embodiment, such as Figure 1 The preparation method shown is as follows:

[0030] 1. After cutting the PVDF (polyvinylidene fluoride) film into 2 cm × 2 cm square films, ultrasonically clean them in anhydrous ethanol for 15 min, then dry them with nitrogen and store them under vacuum. The PVDF films were purchased from PolyK and were 28 μm thick.

[0031] 2. Adhere the PVDF bone electroactive film processed in step 1 onto a glass slide loaded with conductive adhesive, and invert the glass slide into the magnetron sputtering chamber.

[0032] 3. Install titanium and silver elemental targets in the DC target area of ​​magnetron sputtering; close the chamber and introduce high-purity argon gas until the internal pressure is maintained at 5 Pa. Under the argon atmosphere, titanium and silver are deposited on the PVDF bone electroactive film. The sputtering distance is 10 cm, the sputtering power is maintained at 100 W, the substrate temperature is 300 ℃, the argon gas flow rate is 5 sccm, and the duration is 20 min. Finally, titanium and silver doped PVDF bone electroactive film is obtained.

[0033] The PVDF bone electroactive film prepared in Example 1 was imaged as a whole using a scanning electron microscope at 10,000x magnification. The results are as follows: Figure 2 As shown, the surface of the modified PVDF bone electroactive film is relatively smooth. Subsequently, the surface elements of the modified PVDF bone electroactive film were scanned using an energy dispersive spectroscopy (EDS) module, and the results are as follows. Figure 3 As shown in Table 1 below, titanium and silver with stable content were generated, with titanium content of 34.36 wt%, oxygen content of 27.44 wt%, and silver content of 38.20 wt%.

[0034] Table 1. Energy dispersive spectroscopy (EDS) results of PVDF bone electroactive thin film in Example 1

[0035]

[0036] Example 2

[0037] The titanium-silver doped antibacterial bone electroactive material provided in this embodiment is prepared by the following method:

[0038] 1. After cutting the PVDF film into square films of appropriate size, ultrasonically clean it in anhydrous ethanol for 15 min, then dry it with nitrogen and store it under vacuum.

[0039] 2. Adhere the PVDF bone electroactive film processed in step 1 onto a glass slide loaded with conductive adhesive, and invert the glass slide into the magnetron sputtering chamber.

[0040] 3. Install titanium and silver elemental targets in the DC target area of ​​magnetron sputtering; close the chamber and introduce high-purity argon gas until the internal pressure is maintained at 5 Pa. Under the argon atmosphere, titanium and silver are deposited on the PVDF bone electroactive film. The sputtering distance is 10 cm, the sputtering power is maintained at 100 W, the substrate temperature is 300 ℃, the argon gas flow rate is 5 sccm, and the duration is 15 min. Finally, titanium and silver doped PVDF bone electroactive film is obtained.

[0041] Example 3

[0042] The titanium-silver doped antibacterial bone electroactive material provided in this embodiment is prepared by the following method:

[0043] 1. After cutting the PVDF film into square films of appropriate size, ultrasonically clean it in anhydrous ethanol for 15 min, then dry it with nitrogen and store it under vacuum.

[0044] 2. Adhere the PVDF bone electroactive film processed in step 1 onto a glass slide loaded with conductive adhesive, and invert the glass slide into the magnetron sputtering chamber.

[0045] 3. Install titanium and silver elemental targets in the DC target area of ​​magnetron sputtering; close the chamber and introduce high-purity argon gas until the internal pressure is maintained at 5 Pa. Under the argon atmosphere, titanium and silver are deposited on the PVDF bone electroactive film. The sputtering distance is 5 cm, the sputtering power is maintained at 50 W, the substrate temperature is 200 ℃, the argon gas flow rate is 80 sccm, and the duration is 20 min. Finally, titanium and silver doped PVDF bone electroactive film is obtained.

[0046] Comparative Example 1

[0047] The PVDF bone electroactive film treated in this comparative example is the same as the S1 process in Example 1, except that the PVDF electroactive film obtained in this comparative example 1 has not been modified by magnetron sputtering technology.

[0048] Comparative Example 2

[0049] The difference between the preparation method of the thin film material in this comparative example and that in Example 1 is that only elemental titanium is installed as the target material in the DC target region of magnetron sputtering, and elemental titanium is doped onto the PVDF thin film; the titanium content on the surface of the thin film material is approximately 72.56%, which is the sum of the titanium and silver content in Example 1. No elemental silver or any other metal / non-metal materials are doped.

[0050] Comparative Example 3

[0051] The difference between the preparation method of the thin film material in this comparative example and that in Example 1 is that only elemental silver is installed as the target material in the DC target region of magnetron sputtering, and elemental silver is doped onto the PVDF thin film; the silver content on the surface of the thin film material is approximately 72.56%, which is the sum of the titanium and silver content in Example 1. No elemental titanium or any other metal / non-metal materials are doped.

[0052] Experimental Example: Performance Study of PVDF Thin Film Materials in Example 1 and Comparative Examples 1-3

[0053] The PVDF film materials prepared in Example 1 and Comparative Example 1 were soaked in alcohol and then sterilized by UV irradiation. They were then placed in six-well cell culture plates and seeded with RAW264.7 macrophages at a density of 50%. After two days, when the density reached 80%, the cells were collected for qPCR experiments to detect the expression of T lymphocyte activation antigen (CD86), interleukin-1β (IL-1β), interferon regulatory factor 3 (IRF3), TNF receptor-associated factor 6 (TRAF6), and nuclear factor receptor-κB (NF-κB). Statistical analysis and plotting were performed using ImageJ software. The results are shown below. Figure 4-8 As shown, the titanium and silver modified PVDF osteoactive membrane can promote the expression of inflammatory factors such as CD86, IL-1β, IRF3, TRAF6 and NF-κB in macrophages, indicating that it enhances the anti-inflammatory activity of macrophages and makes them more effective in antibacterial activity.

[0054] The PVDF osteoactive films prepared in Example 1 and Comparative Example 1 were soaked in alcohol and then sterilized by UV irradiation. They were then placed in six-well cell culture plates and seeded with 50% confluence of osteoblasts (mt-3t3). After two days, when the confluence reached 80%, the cells were collected for qPCR to detect the expression of RUNX family transcription factor 2 (RUNX2). Statistical analysis and plotting were performed using ImageJ software. The results are shown below. Figure 9 As shown in Example 1, the PVDF osteogenic membrane modified with titanium and silver can upregulate the expression of osteoblast RUNX2 transcription factor, suggesting that it has better osteogenic activity.

[0055] The PVDF film materials prepared in Examples 1 and Comparative Examples 2 and 3 were soaked in alcohol using the same method as described above. The expression levels of interleukin-6 (IL-6), nuclear factor receptor-κB (NF-κB), MYD88, and TNF receptor-associated factor 6 (TRAF6) were then detected, and statistical analysis and plotting were performed using ImageJ software. The results are as follows: Figure 10-13 As shown, PVDF films doped with elemental titanium and silver can promote the expression of inflammatory factors such as IL-6, NF-κB, MYD88, and TRAF6 in macrophages, indicating that they enhance the anti-inflammatory activity of macrophages and give them stronger antibacterial properties.

[0056] The above detailed embodiments describe the implementation of the present invention; however, the present invention is not limited to the specific details described in the above embodiments. Within the scope of the claims and technical concept of the present invention, various simple modifications and changes can be made to the technical solution of the present invention, and these simple modifications all fall within the protection scope of the present invention.

Claims

1. A method for preparing a titanium-silver doped antibacterial bone electroactive material, characterized in that, The titanium-silver doped antibacterial osteoactive material is used as a bone defect repair material or an antibacterial material, and includes the following steps: The PVDF film was cut, cleaned, dried, and vacuum-preserved. Titanium and silver were deposited onto the PVDF film using magnetron sputtering targets to obtain the titanium-silver doped antibacterial bone electroactive material. The titanium-silver doped antibacterial bone electroactive material contained 34.36 wt% titanium, 38.2 wt% silver, and 27.44 wt% oxygen.

2. A titanium-silver doped antibacterial bone electroactive material prepared by the preparation method described in claim 1.

3. An application of the titanium-silver doped antibacterial bone electroactive material according to claim 2, characterized in that, It is used as a material for repairing bone defects or as an antibacterial material.

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