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目的 研究不同管径TiO2纳米管修饰的钛材料与成骨细胞在体外复合培养,观察其对成骨细胞的粘附、增殖和分化的影响,并探讨可能的机制,为纳米材料的临床应用提供理论依据.方法 将骨植入材料金属物质分成四组,其中三组经处理后分别内置有50 nm、100 nm、CDW图案的纳米管,以未经任何处理的钛片作为对照组,将MG-63成骨细胞分别接种于内置有50 nm、100 nm、CDW图案的纳米管与对照组钛片的6孔板中,采用MTT实验对四组材料复合培养MG-63的细胞活力进行评估;应用碱性磷酸酶(alklinephosphatase,ALP)试剂盒检测四组细胞碱性磷酸酶活性的改变;荧光共聚焦检测皮肤桥蛋白(DPT)的表达情况;通过Western blot实验检测DPT、黏着斑激酶,粘着斑激酶(Focal Adhesion Kinase,FAK)、骨桥蛋白(osteopontin, OPN)蛋白的表达变化.结果 MTT检测显示,三种纳米管复合培养MG-63活性比对照组钛片复合培养的MG-63活性增强,培养的第3d,5d,7d MG-63细胞活性增强显著,呈时间依赖性.与钛片复合培养组相比,50 nm、100 nm、CDW图案的纳米管复合培养组ALP、DPT、FAK及OPN表达水平增加.结论 50 nm、100 nm、CDW图案的纳米管复合培养更能促进成骨细胞的生长与铺展,具有良好的生物相容性,其机制可能与增加ALP的表达水平和激活DPT/FAK信号通路相关.“,”Objective To study the co-culture of titanium materials modified by TiO2 nanotubes with different diameters and osteoblasts in vitro,and observe its effect to the adhesion,proliferation and differentiation of osteoblasts,to explore the possible mechanism.Methods MG-63 cells were respectively implanted into 6 well culture plates of nanotubes with 50 nm,100 nm,CDW pattern and the titanium of the control group.The viability of co-cultured MG-63 cells was assessed by MTT,activity changes of cells alkaline phosphatase were detected by ALP test-kit,the expression of (Dermatopontin DPT) DPT was detected by fluorescence confocal microscope,the expressions of DPT,FAK(Focal Adhesion Kinase),OPN(osteopontin) were detected by Western blot.Results MG-63 activity in three kinds of nanotubes was increased than that in control group,especially at days of 3,5 and 7,with a time-dependent manner.The expression levels of ALP,DPT,FAK and OPN were increased in the nanotube co-cultured group(50 nm,100 nm,CDW pattern) than in titanium co-cultured group.Conclusion The nanotubes with 50 nm,100 nm,CDW pattern promoted the growth and spread of osteoblasts with good biocompatibility,which may be related to the upexpression of ALP and activating DPT/FAK signal pathway.