SLM打印钛合金片表面含氟纳米二氧化钛薄膜结构的生物相容性及抗菌性能评价

Evaluation of biocompatibility and antibacterial properties of fluorine-doped TiO2 nanofilm on SLM printed titanium alloy surface

目的对选择性激光熔化技术(SLM)打印的钛合金片进行表面处理获得含氟纳米二氧化钛薄膜结构,观察成骨细胞在其表面的黏附、增殖情况,评价其生物相容性;观察大肠杆菌和金黄色葡萄球菌在其表面的生长情况,评价其抗菌性能。方法SLM打印钛合金片通过前驱液浸泡和水热法表面处理,获得含氟纳米二氧化钛薄膜结构为实验组,将无表面处理的SLM打印钛合金片作为对照组。采用扫描电镜观测实验组钛合金片表面特征,并用接触角测量仪测量两组钛合金片接触角。将小鼠胚胎成骨细胞(MC3T3-E1 Subclone 14细胞)分别接种于钛合金片表面并进行体外培养,采用扫描电镜、荧光染色及CCK-8方法分析成骨细胞的黏附和增殖情况;将大肠杆菌和金黄色葡萄球菌分别与两组钛合金片共培养,采用平板活菌计数和活死菌染色实验来评价钛合金片的抗菌性能。结果细胞形态、荧光染色和黏附实验结果显示,成骨细胞在实验组钛合金片表面的黏附能力显著高于在对照组钛合金片表面(P<0.05)。增殖实验结果显示培养3、7 d后,成骨细胞在实验组钛合金片表面的增殖能力显著高于在对照组钛合金片表面(均P<0.05)。抗菌实验结果显示,实验组的含氟纳米二氧化钛薄膜结构可以抑制大肠杆菌和金黄色葡萄球菌在其表面的生长。结论通过前驱液浸泡和水热法表面处理可获得含氟纳米二氧化钛薄膜结构,该结构可促进成骨细胞在SLM打印钛合金片表面的黏附和增殖,同时可以抑制大肠杆菌和金黄色葡萄球菌的生长,具有较好的生物相容性和抗菌性能。

Objective To evaluate the biocompatibility and antibacterial properties of fluorine-doped TiO_(2) nanofilms on selective laser melting(SLM)printed titanium alloy surfaces.Methods The fluorine-doped TiO_(2) nanofilms was fabricated on the SLM printed titanium alloy surface by adjusting the formula of precursor solution and hydrothermal time.The titanium alloys with fluorine-doped TiO_(2) nanofilms were obtained as the study group,and the titanium alloy without surface treatment were used as the control group.The surface characteristics of titanium alloy samples were observed through field-emission scanning electron microscopy(FE-SEM),and the contact angle was measured through contact angle tester.Mouse embry-onic osteoblasts(MC3T3-E1 Subclone 14)were inoculated on the surface of the titanium alloy samples and cultured in vitro.The adhesion and proliferation of osteoblasts were analyzed by FE-SEM,4',6-damidino-2-phenylindole(DAPI)and fluores-cein isothiocyanate labeled phalloidin(FITC phalloidin)staining and cell counting-kit 8(CCK-8)assay.Escherichia coli(E.coli)and Staphylococcus aureus(S.aureus)were cocultured with two groups of titanium alloy samples,and the antibacterial perfor-mance of the titanium alloy samples was evaluated by plate count and the bacterial staining method.Results Cell mor-phology,DAPI and FITC phalloidin staining and cell adhesion showed that the adhesion of osteoblasts in the study group was significantly higher than that in the control group(P<0.05).Cell proliferation showed that the proliferation of osteoblasts on the study group was superior to that on the control group after 3 and 7 days(P<0.05).The antibacterial test results showed that the fluorine-doped TiO_(2) nanofilms in the study group inhibited the growth of E.coli and S.aureus on their surface.Conclusion A fluorine-doped TiO_(2) nanofilm structure has been obtained by hydrothermal surface treatment,which can promote the adhesion and proliferation of osteoblasts on the surface of titanium alloy sample,and inhibit the growth of E.coli and S.aureus,with satisfactory biocompatibility and antibacterial properties.