钛种植体表面GO-CG涂层的制备及性能研究

Research of GO-CG Coating Construction and Properties on Titanium Implant Surface

目的在纯钛(Ti)种植体表面构建由氧化石墨烯(GO)负载葡萄糖酸氯己定(CG)的表面涂层,研究该涂层材料的抗菌性能。方法利用碱热处理、硅烷化处理和浸渍法在纯钛表面制备GO-CG涂层,通过扫描电子显微镜(SEM)、能谱(EDS)分析、接触角测试、体外药物缓释实验对该涂层材料进行表征。对MC3T3-E1细胞进行DAPI染色实验、CCK-8实验、Live/Dead染色实验检测材料的细胞毒性;使用金黄色葡萄球菌和大肠杆菌通过抑菌环法、比浊法、平板计数法以及SEM观察检测该涂层材料的抗菌性能。结果SEM下Ti组表面布满喷砂酸蚀后的微小凹陷,Ti-GO组及Ti-GO-CG组样本表面可见一层均匀的薄膜状物质覆盖在基底的凹陷形貌上,且出现堆叠的褶皱。EDS分析显示Ti-GO组较Ti组出现Si波峰,且C、O含量增加,Ti含量降低;在GO涂层的基础上,Ti-GO-CG组出现了CG特征元素Cl的波峰。Ti组、Ti-GO组和Ti-GO-CG组表面的接触角依次降低(P<0.05)。Ti-GO-CG组材料体外药物释放时间长达5~7 d。Ti-GO组和Ti-GO-CG组样本表面黏附的细胞数多于Ti组。Ti-GO-CG-es组有抑制细胞增殖的现象,与对照组比较,差异有统计学意义(P<0.05),但抑制效果不明显。Live/Dead染色结果显示死亡细胞数量无明显增加。Ti-GO-CG组金黄色葡萄球菌和大肠杆菌出现抑菌环。比浊法显示Ti-GO-CG能抑制细菌的增殖(P<0.05)。平板计数法显示Ti-GO组呈现出较弱的抗菌性,Ti-GO-CG组呈现出较强的抗菌性(P<0.05)。SEM下Ti-GO-CG组表面黏附的细菌少于Ti组和Ti-GO组。结论利用GO负载CG可以在钛种植体表面成功构建抗菌涂层,制备的Ti-GO-CG涂层材料无细胞毒性,同时具备良好的抗菌性能。

Objective To study the antibacterial properties of chlorhexidine gluconate(CG)coated with graphene oxide(GO)on the surface of pure titanium(Ti)implants.Methods The GO-CG coating was prepared on the surface of pure titanium by alkali heat treatment,silanization treatment and impregnation.The coating material was characterized by scanning electron microscope(SEM),energy dispersive spectrum(EDS),contact angle test and drug sustained release test in vitro.MC3T3-E1 cells were used to detect the cytotoxicity of the materials by DAPI staining,CCK-8 test and Live/Dead staining,and Staphylococcus aureus and Escherichia coli were used to detect the antibacterial properties of the coating material by bacteriostatic ring method,turbidimetric method,plate counting method and scanning electron microscope.Results SEM showed the surface of the Ti group was covered with small depressions after sandblasting and acid etching.A uniform thin film like substance can be seen on the surface of the Ti-GO group and Ti GO-CG group samples,covering the depression morphology of the substrate,with stacked folds appearing.EDS analysis showed that the Ti-GO group exhibited Si peaks compared to the Ti group,with an increase in C and O content and a decrease in Ti content.On the basis of GO coating,the Ti-GO-CG group exhibited a wave peak of CG characteristic element Cl.The contact angle on the surface of the Ti group,Ti-GO group,and Ti-GO-CG group decreased sequedtially(P<0.05).The drug release time of Ti-GO-CG group was 5 to 7 days in vitro.The cells adhered to the surface of samples in the Ti-GO group and Ti-GO-CG group more than that in the Ti group.The Ti-GO-CG-es group showed an inhibitory effect on cell proliferation,with a statistically difference compared to the control group(P<0.05),but the inhibitory effect was not significant.The results of Live/Dead staining showed that the number of dead cells did not increase significantly.In the Ti-GO-CG group,Staphylococcus aureus and Escherichia coli showed inhibitory rings.Turbidimetric analysis showed that Ti-GO-CG can inhibit bacterial proliferation(P<0.05).The plate counting method showed that the Ti-GO group showed weak antibacterial activity,while the Ti-GO-CG group showed strong antibacterial activity(P<0.05).SEM showed the number of bacteria adhering to the surface of the Ti-GO-CG group was less than that of the Ti group and Ti-GO group.Conclusion The antibacterial coating on the surface of titanium implant can be successfully constructed by GO loaded CG.The prepared Ti-GO-CG coating material has no cytotoxicity and good antibacterial properties.