TiO_2纳米管对DPT在成骨细胞中表达的影响及机制研究  

The effect and mechanism of TiO_2 nanotubes on theexpression of DPT in osteoblast

作　　者：王欣[1] 王一维[1] 臧晋[1] 孙振祥[1] 王效杰[1] WANG Xin WANG Yi-wei ZANG Jin SUN Zhen-xiang WANG Xiao-jie(Department of Anatony, Shenyang Medial College, Shenyang 110034, China)

机构地区：[1]沈阳医学院基础医学院解剖学教研室,辽宁沈阳110034

出　　处：《解剖科学进展》2017年第3期231-234,238,共5页Progress of Anatomical Sciences

基　　金：沈阳市科技局计划项目(F-13-221-9-33)

摘　　要：目的研究不同管径TiO_2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(alkline phosphatase,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 cell＇ s 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.

关 键 词：生物材料 TIO2纳米管 成骨细胞 皮肤桥蛋白 

分 类 号：R318.17[医药卫生—生物医学工程]