不同管径钛纳米管对大鼠骨髓间充质干细胞增殖及成骨分化的影响

发布时间：2018-05-17 23:35

本文选题：TiO纳米管 + 骨髓间充质干细胞　；参考：《第三军医大学学报》2014年12期

【摘要】：目的研究不同管径钛纳米管表面结构对体外培养SD大鼠骨髓间充质干细胞(rat mesenchymal stem cells,rMSCs)粘附、增殖及成骨向分化的影响。方法分离并扩增rMSCs,成骨向诱导分化及流式细胞鉴定;阳极氧化法分别制备管径30、70、100 nm的TiO2纳米管,扫描电镜观察表面形貌、接触角测试材料表面亲疏水性;MTT法检测细胞在材料表面粘附及增殖情况,激光共聚焦显微镜观察细胞在材料表面形貌。评价细胞在纯钛,管径30、70、100 nm的TiO2纳米管表面成骨向分化过程中碱性磷酸酶活性变化特征及矿化情况。Real-time PCR检测诱导14 d后Runx2及OSX基因表达情况。结果分离获得的rMSCs具有成骨向分化的能力,CD29阳性细胞为99.65%;扫描电镜观察各组材料表面管径均达到设计要求;管径为70 nm的TiO2纳米管材料表面亲水性较好(P0.05);MTT结果显示70 nm组rMSCs的粘附、增殖均较其他组明显增高(P0.05);30 nm组在成骨诱导7 d后,碱性磷酸酶活性均高于其他组(P0.05)。30 nm组在成骨诱导21 d后表面矿化高于其他组(P0.05)。30 nm组在成骨诱导14 d后Runx2及OSX基因表达高于其余组(P0.05)。结论与光滑面纯钛相比,不同管径的TiO2纳米管对rMSCs的作用各不相同,rMSCs在70nm表面增殖能力和粘附性较好,但在30 nm表面的成骨向分化能力较强。
[Abstract]:Objective to study the effect of different diameter titanium nanotubes on the adhesion, proliferation and osteogenic differentiation of SD rat bone marrow mesenchymal stem cells (BMSCs) in vitro. Methods rMSCs were isolated and amplified, osteoblast induced differentiation and flow cytometry were identified. TiO2 nanotubes with the diameter of 30 ~ 70100nm were prepared by anodizing method, and the surface morphology was observed by scanning electron microscope (SEM). The cell adhesion and proliferation were detected by MTT method and the morphology of cells on the surface was observed by laser confocal microscopy. To evaluate the changes of alkaline phosphatase (ALP) activity during osteogenesis and differentiation of TiO2 nanotubes with pure titanium and 30 ~ 70100nm diameter. Real-time PCR was used to detect the expression of Runx2 and OSX genes 14 days after induction. Results the CD29 positive cells of rMSCs with osteogenic differentiation ability were 99.65 and the diameter of surface tubes of each group were all up to the design requirements. The surface hydrophilicity of TiO2 nanotubes with diameter of 70 nm was better than that of other groups. The results showed that the adhesion of rMSCs in 70 nm group was significantly higher than that in other groups, and the proliferation of 70 nm group was significantly higher than that of other groups after 7 days of osteogenic induction. The activity of alkaline phosphatase was higher than that of other groups after 21 days of osteogenesis. The expression of Runx2 and OSX genes in other groups was higher than that in other groups at 14 days after osteogenesis induction. Conclusion compared with pure titanium with smooth surface, TiO2 nanotubes with different diameters have different effects on the proliferation and adhesion of rMSCs on the surface of 70nm, but the osteogenic differentiation ability on the surface of 70nm is stronger than that on the surface of 30 nm.
【作者单位】：重庆医科大学口腔医学院口腔疾病与生物医学重庆市重点实验室;
【基金】：重庆医科大学口腔医学院博士科研启动基金(2011) 重庆市卫生局面上项目(2012-2-121) 重庆市科委自然科学基金(CSTC2012jjA0178) 高等学校博士学科点专项科研基金新教师类资助课题(20125503120009)~~
【分类号】：R783.1

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