Abstract:

Objective: To study the effects of TiO₂ nanotubes with different diameters on the proliferation, spreading, adhesion and differentiation of mouse C2C12 myoblasts. Methods: TiO₂ nanotubes with diameters of 25 nm, 50 nm and 100 nm were prepared by anodization at 10 V, 15 V and 20 V respectively. C2C12 myoblasts were cultured on the surface of each sample. CCK-8 assay was used to examine the cell proliferation. The cell morphology was observed by scanning electron microscope (SEM). The effects of TiO₂ nanotubes with different diameters on the expression of genes related to the differentiation of mouse C2C12 myoblasts were detected by real-time quantitative polymerase chain reaction (RT-qPCR). Results: TiO₂ nanotubes with different diameters promoted the proliferation of C2C12 myoblasts, and the effect of the 50 nm group was the best, while that of the 100 nm group was weaker. The cells in the 50 nm and 100 nm nanotubes extended more pseudopodium, which exhibited promoted adhesion and spreading. The 50 nm nanotubes upregulated the expression of myogenic related genes such as Myog, Myh3 and Myod1 on the 3rd and 7th day. Conclusion: The 50 nm TiO₂ nanotubes can promote the proliferation, spreading, adhesion and differentiation of mouse C2C12 myoblasts.

Key words: TiO₂ nanotubes, different diameters, myoblast, proliferation

摘要：

目的：研究不同管径TiO₂纳米管对于小鼠C2C12成肌细胞增殖、铺展、黏附和分化的影响。方法：通过阳极氧化的方法，用10、15、20 V电压制备3组不同管径的TiO₂纳米管，扫描电镜（scanning electron microscope, SEM）测得3组管径分别为25、50、100 nm。在各组样品表面接种C2C12成肌细胞，用CCK-8法检测细胞增殖情况，SEM观察细胞形态，实时定量聚合酶链反应（real-time quantitative polymerase chain reaction，RT-qPCR）观察不同管径TiO₂纳米管对于小鼠C2C12成肌细胞分化相关基因表达的影响。结果：不同管径TiO₂纳米管均促进了C2C12成肌细胞的增殖，其中50 nm管径组效果最佳，100 nm管径组效果减弱。50 nm和100 nm管径组能伸展出更多的伪足和伪板，促进了细胞的黏附和铺展。50 nm管径组在3 d和7 d时均上调了Myog、Myh3和Myod1这类成肌相关基因的表达。结论：50 nm管径的TiO₂纳米管能促进小鼠C2C12成肌细胞的增殖、铺展、黏附和分化。

关键词: 二氧化钛纳米管, 不同孔径, 成肌细胞, 增殖