微弧氧化钛表面对2型糖尿病大鼠骨髓基质干细胞成骨分化与种植体骨结合的影响

Effect of micro arc oxidation treated Ti surface on osteogenic differentiation of bone marrow mesenchymal stem cells and osseointegration of induced type 2 diabetes mellitus rats

目的研究微弧氧化钛表面对2型糖尿病大鼠骨髓基质干细胞成骨分化与种植体骨结合的影响。方法对TC4表面微弧氧化处理获得微纳米表面,高糖高脂饮食结合腹腔内注射链脲佐菌素溶液建立2型糖尿病大鼠模型,分离大鼠骨髓基质干细胞。接种细胞到微弧氧化钛表面,观察细胞附着形态、增殖活性、碱性磷酸酶(ALP)活性和成骨分化相关基因。植入微弧氧化种植体到2型糖尿病患者股骨内,荧光素标记新骨形成。采用激光共聚焦显微镜(EDS)观察新骨形成速度,苦味酸品红染色观察骨结合率。采用配对t检验(SPSS 2.0软件)分析实验组和对照组之间细胞增殖、成骨分化相关基因表达、碱性磷酸酶半定量活性、新骨形成量和骨结合率差异(α=0.05)。结果 TC4经酸蚀处理后形成粗糙表面,经微弧氧化处理后可以观察到微纳米结构的孔隙。EDS检测到微弧氧化后的钛表面有钠、硅和磷等元素。微弧氧化钛表面的细胞伸展形态由于对照组,ALP活性增加,成骨分化基因表达上调。动物实验显示微弧氧化钛表面种植体新骨形成速度在前两个月快于对照组,骨结合率高于对照组。结论对TC4表面微弧氧化处理后可以促进2型糖尿病大鼠骨髓基质干细胞成骨分化与种植体骨结合。

Objective To investigate the effect of micro arc oxidation treated Ti surface on osteogenic differentiation of bone marrow mesenchymal stem cells and osseointegration of induced type 2 diabetes mellitus rats. Methods TC4 surface was treated by micro arc oxidation to obtain a hierarchy structure. Type 2 diabetic rat model was established by feeding with high-fat diet and injection of streptozotocin. The BMSCs of diabetic rat were cultured in vitro. The cells were seeded on the treated surface to observe the adhesion morphology, proliferative activity, alkaline phosphatase(ALP) activity and osteogenic differentiation related genes. The implant with the surface was inserted in the diabetic rats’ femurs and fluorescein was used to mark the new bone formation. EDS was applied to observe the new bone formation speed and picric acid fuchsia solution dyeing was used to observe the osseointegration rate. Pair t test was applied to analyze the cell differentiation difference, the expression difference of osteogenic differentiation related genes, ALP semi-quantitative activity, and the difference of new bone formation amount and osseointegration rate with SPSS 2.0 software. Results TC4 formed rough surface by acid etching. The pore of micro-nano-structure could be observed by micro arc oxidation treatment. EDS detected that there was sodium(Na), silicon, and phosphorus on the titanium surface after micro arc oxidation. The cell stretching morphology on the treated surface was better than the control group, ALP activity increased and the osteogenic differentiation gene expression up-regulated. The animal experiment revealed that in the first two months the new bone formation speed of the implant with micro arc oxidation surface was faster, and the osseointegration rate was higher than the control group. Conclusion Micro arc oxidation treated TC4 surface can improve osteogenic differentiation of bone marrow mesenchymal stem cells of type 2 diabetes mellitus rats and osseointegration in diabetic environment.