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

Abstract

The invention discloses a preparation method and application of a titanium-silver doped antibacterial bone electroactive material, and relates to the field of bone electroactive materials. The preparation method of the PVDF bone electroactive film comprises the following steps: cutting, cleaning and blow-drying the PVDF film, and preserving in vacuum; and depositing a titanium elementary substance and a silver elementary substance serving as magnetron sputtering target materials on the PVDF film to obtain the titanium-silver doped antibacterial bone electroactive material, the magnetron sputtering method comprises the steps that under the argon atmosphere condition, the internal air pressure of a magnetron sputtering cavity is adjusted to range from 0.5 Pa to 5 Pa, the sputtering distance ranges from 4 cm to 10 cm, the sputtering power is maintained to range from 50 W to 100 W, the substrate temperature ranges from room temperature to 300 DEG C, the argon flow speed ranges from 5 sccm to 80 sccm, and the duration time ranges from 10 min to 20 min. According to the preparation method disclosed by the invention, the surface of the PVDF bone electroactive film is effectively modified by using titanium and silver through a magnetron sputtering technology, so that the adhesion of the PVDF film and bone tissues is remarkably improved, and the PVDF film has excellent antibacterial activity and good induced osteogenesis activity.

Description

Technical Field

[0001] The invention relates to the field of bone electroactive materials, and in particular to a preparation technology of titanium-silver doped antibacterial bone electroactive materials. Background Art

[0002] Current research shows that the microscopic nanotopography and electrical signal characteristics of bone defect repair materials are crucial to bone regeneration and antibacterial environment. When subjected to external forces, polarized charges similar to the piezoelectric properties of natural bone tissue will appear on the surface of PVDF film. It has excellent piezoelectric properties and a large specific surface area, and can effectively simulate the collagen fiber structure in the natural extracellular matrix. Based on the above characteristics, PVDF film has a wide range of applications in antibacterial environment, bone repair, biosensors, drug delivery, etc.

[0003] However, conventional PVDF films have problems such as poor electrical signal stability, poor bone regeneration regulation, slow bone healing, and poor antibacterial ability. In recent years, although a large number of studies have been conducted to modify PVDF films, it is still impossible to solve all of the above problems at the same time. For the in vivo microenvironment of the human or animal body, especially the environment full of bacteria such as the oral cavity, how to use suitable bone defect repair materials to achieve bone regeneration in such a bacterial environment is an urgent problem to be solved in the field of bone electroactive materials. Summary of the invention

[0004] The present invention provides a preparation method and application of a titanium-silver doped antibacterial bone electroactive material, which has excellent biocompatibility, antibacterial properties and osteogenesis induction ability, expands its application scenarios in the oral cavity and other parts, and effectively solves the shortcomings of existing PVDF bone electroactive films in terms of poor bone integration, bone healing speed and antibacterial effectiveness. It is specifically achieved through the following technologies.

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

[0006] The PVDF film is cut, cleaned, blown dry, and stored in a vacuum; titanium and silver are used as magnetron sputtering targets and deposited on the PVDF film to obtain the titanium-silver doped antibacterial bone electroactive material;

[0007] The magnetron sputtering method is: under argon atmosphere conditions, adjust the internal gas pressure of the magnetron sputtering chamber to 0.5-5 Pa, the sputtering distance to 4-10 cm, maintain the sputtering power to 50-100 W, the substrate temperature to room temperature-300 ° C, the argon gas flow rate to 5-80sccm, and the duration is 10-20 min.

[0008] Furthermore, the internal gas pressure of the magnetron sputtering chamber was adjusted to 5 Pa, the sputtering distance was 5 cm, the sputtering power was maintained at 50-100 W, the substrate temperature was 200 °C, the argon gas flow rate was 80 sccm, and the duration was 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] The invention provides a titanium-silver doped antibacterial bone electroactive material prepared by the preparation method.

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

[0013] Compared with the prior art, the present invention is beneficial in that:

[0014] 1. The existing common PVDF bone electroactive film has a surface nanostructure that cannot establish good adhesion with bone tissue. The present invention effectively modifies the surface of the PVDF bone electroactive film with titanium and silver through magnetron sputtering technology, significantly improving the adhesion of the PVDF film to bone tissue. The titanium content is 34.36wt%, the oxygen content is 27.44wt%, and the silver content is 38.2wt%.

[0015] 2. The existing ordinary PVDF bone electroactive film has poor antibacterial performance. The titanium-silver doped antibacterial bone electroactive material prepared by the present 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 by the present invention has excellent antibacterial activity.

[0016] 3. The existing PVDF bone electroactive film has a poor effect on bone repair. The titanium-silver doped antibacterial bone electroactive material prepared by the present invention can promote osteoblasts to express RUNX family transcription factor 2 (RUNX2), and it has good inducing osteogenic activity. BRIEF DESCRIPTION OF THE DRAWINGS

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

[0018] Figure 2 This is the overall imaging of the titanium silver doped PVDF bone electroactive film prepared in Example 1 using a scanning electron microscope at a magnification of 10,000 times.

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

[0020] Figure 4 The graph is a relative mRNA expression of CD86 after culturing macrophages RAW264.7 using the titanium silver doped PVDF bone electroactive film of Example 1 and the common PVDF film of Comparative Example 1.

[0021] Figure 5 The graph is a relative mRNA expression of IL-1β after culturing macrophages RAW264.7 using the titanium silver doped PVDF bone electroactive film of Example 1 and the common PVDF film of Comparative Example 1.

[0022] Figure 6 The relative mRNA expression of IRF3 after culturing macrophages RAW264.7 using the titanium silver doped PVDF bone electroactive film of Example 1 and the common PVDF film of Comparative Example 1, respectively.

[0023] Figure 7 The relative mRNA expression of TRAF6 after culturing macrophages RAW264.7 using the titanium silver doped PVDF bone electroactive film of Example 1 and the common PVDF film of Comparative Example 1, respectively.

[0024] Figure 8 The graph is a relative mRNA expression of NF-κB after culturing macrophages RAW264.7 using the titanium silver doped PVDF bone electroactive film of Example 1 and the common PVDF film of Comparative Example 1.

[0025] Fig. 9 The relative mRNA expression of RUNX2 after culturing osteoblasts mt-3t3 using the titanium silver doped PVDF bone electroactive film of Example 1 and the common PVDF film of Comparative Example 1, respectively.

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

[0027] The technical solution of the present invention will be described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work 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 is Figure 1 The preparation method thereof is shown as follows:

[0030] 1. Cut the PVDF (polyvinylidene fluoride) film into square films of 2 cm × 2 cm, clean them in anhydrous ethanol by ultrasonic for 15 min, blow dry them with nitrogen, and store them in vacuum. The PVDF film was purchased from PolyK Company with a thickness of 28 μm.

[0031] 2. The PVDF bone electroactive film treated in step 1 is adhered to a glass slide loaded with conductive glue, and the glass slide is placed upside down in a magnetron sputtering chamber.

[0032] 3. Install titanium and silver target materials in the DC target area of ​​magnetron sputtering; close the chamber, introduce high-purity argon gas until the pressure in the chamber is maintained at 5 Pa, and deposit titanium and silver on the PVDF bone electroactive film in an argon atmosphere. The sputtering distance is 10 cm, the sputtering power is maintained at 100 W, the substrate temperature is 300 ℃, the argon 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 a magnification of 10,000 times. Figure 2 As shown in Figure 1, 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 the energy spectrum scanning module. The results are shown in Figure 1. Figure 3 As shown in Table 1 below, titanium and silver were generated with stable contents, wherein the titanium content was 34.36wt%, the oxygen content was 27.44wt%, and the silver content was 38.20wt%.

[0034] Table 1 Spectrum scanning results of PVDF bone electroactive film of Example 1

[0035]

[0036] Example 2

[0037] The titanium-silver doped antibacterial bone electroactive material provided in this embodiment has a preparation method as follows:

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

[0039] 2. The PVDF bone electroactive film treated in step 1 is adhered to a glass slide loaded with conductive glue, and the glass slide is placed upside down in a magnetron sputtering chamber.

[0040] 3. Install titanium and silver target materials in the DC target area of ​​magnetron sputtering; close the chamber, introduce high-purity argon gas until the pressure in the chamber is maintained at 5 Pa, and deposit titanium and silver on the PVDF bone electroactive film in an argon atmosphere. The sputtering distance is 10 cm, the sputtering power is maintained at 100 W, the substrate temperature is 300 ℃, the argon 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 has a preparation method as follows:

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

[0044] 2. The PVDF bone electroactive film treated in step 1 is adhered to a glass slide loaded with conductive glue, and the glass slide is placed upside down in a magnetron sputtering chamber.

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

[0046] Comparative Example 1

[0047] The processing of the PVDF bone electroactive film 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 is not modified by magnetron sputtering technology.

[0048] Comparative Example 2

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

[0050] Comparative Example 3

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

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

[0053] The PVDF film materials prepared in Example 1 and Comparative Example 1 were soaked in alcohol respectively, and sterilized by ultraviolet irradiation; they were placed in six-well cell culture plates, and macrophages RAW264.7 were inoculated at a density of 50%; two days later, when the density increased to 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), nuclear factor receptor-κB (NF-κB), etc. The software ImageJ was used for statistical analysis and drawing, and the results are shown in Figure 2. Figure 4-8 As shown, the PVDF bone electroactive film modified with titanium and silver 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 have stronger antibacterial properties.

[0054] The PVDF bone electroactive films prepared in Example 1 and Comparative Example 1 were soaked in alcohol and sterilized by ultraviolet irradiation, respectively, and placed in a six-well cell culture plate, and inoculated with 50% osteoblasts mt-3t3, respectively. Two days later, when the density increased to 80%, the cells were collected for qPCR experiments to detect the expression of RUNX family transcription factor 2 (RUNX2). Statistical analysis and drawing were performed using ImageJ software, and the results are shown in Figure 2. Fig. 9 It was shown that the PVDF bone electroactive film modified with titanium and silver in Example 1 could upregulate the expression of RUNX2 transcription factor in osteoblasts, indicating that it had better osteogenic activity.

[0055] The PVDF film materials prepared in Example 1 and Comparative Examples 2 and 3 were soaked in alcohol, and the same method as above was used to finally detect the expression of interleukin-6 (IL-6), nuclear factor receptor-κB (NF-κB), MYD88 and TNF receptor-associated factor 6 (TRAF6), and statistical analysis and drawing were performed using ImageJ software. The results are shown in FIG. Figure 10-13 As shown in the figure, 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 it enhances the anti-inflammatory activity of macrophages and makes them have stronger antibacterial properties.

[0056] The above specific embodiments describe the implementation of the present invention in detail, but the present invention is not limited to the specific details in the above embodiments. Within the scope of the claims and technical concept of the present invention, the technical solution of the present invention can be modified and changed in many simple ways, and these simple modifications all belong to 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 following steps are involved: The PVDF film is cut, cleaned, blown dry, and stored in a vacuum; titanium and silver are used as magnetron sputtering targets and deposited on the PVDF film to obtain the titanium-silver doped antibacterial bone electroactive material; The magnetron sputtering method is: under argon atmosphere conditions, adjust the internal gas pressure of the magnetron sputtering chamber to 0.5-5 Pa, the sputtering distance to 4-10 cm, maintain the sputtering power to 50-100 W, the substrate temperature to room temperature-300 ° C, the argon gas flow rate to 5-80sccm, and the duration is 10-20 min.

2. The method for preparing the titanium-silver doped antibacterial bone electroactive material according to claim 1, characterized in that: The internal gas pressure of the magnetron sputtering chamber was adjusted to 5 Pa, the sputtering distance was 5 cm, the sputtering power was maintained at 50-100 W, the substrate temperature was 200 °C, the argon gas flow rate was 80 sccm, and the duration was 10-20 min.

3. The method for preparing the titanium-silver doped antibacterial bone electroactive material according to claim 1, characterized in that: 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 %.

4. The method for preparing the titanium-silver doped antibacterial bone electroactive material according to claim 3, characterized in that: 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 %.

5. A titanium-silver doped antibacterial bone electroactive material prepared by the preparation method according to any one of claims 1 to 4.

6. An application of the titanium-silver doped antibacterial bone electroactive material according to claim 5, characterized in that: Used as bone defect repair material or antibacterial material.

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