摘要：

目的： 通过体外细胞实验探讨纳米钽颗粒对成骨细胞的增殖能力，对碱性磷酸酶（alkaline phosphatase，ALP）、骨形成蛋白-2（bone morphogenetic protein 2，BMP-2）、骨钙素（osteocalcin，OCN）基因表达的影响，分析纳米钽的作用机制，为临床选择种植体材料提供依据。方法： 通过超声乳化制备3种质量浓度（10 ng/mL、10 μg/mL和100 μg/mL）的纳米钽母液，采用透射电镜和扫描电镜对钽颗粒进行表征；3组纳米钽母液与成骨细胞系MC3T3-E1共培养6、12、24 h，用CCK8法检测不同培养时间点的细胞增殖率，PCR法检测细胞内ALP、BMP-2及OCN mRNAs的表达水平。结果： 纳米钽母液颗粒表面电镜表征显示，纳米钽颗粒形状较规则，圆球形多见，粒径约8~15 nm，平均水合粒径为（286.5±13.6） nm。培养12 h后， 3组细胞增殖率，ALP、BMP-2及OCN 的mRNAs水平明显高于培养6 h后，培养24 h后下降，但仍高于培养6 h后；组间比较发现，10 μg/mL组不同时间点的细胞增殖率，ALP、BMP-2及OCN 的mRNAs水平均明显高于其他2组。结论： 纳米钽与成骨细胞接触有较好的增殖诱导作用，并与其颗粒浓度和大小有关。但结果尚需进一步验证。

关键词: 纳米钽, 种植体, 成骨细胞, 增殖

Abstract:

Objective: To investigate the effects of nano-tantalum particles on cell proliferation, and gene expression of alkaline phosphatase (ALP), bone morphogenetic protein 2(BMP-2) and osteocalcin (OCN) of osteoblasts in vitro.The mechanism of nano tantalum was analyzed to provide a basis for clinical selection of implant materials. Methods: The mother liquor of nano-tantalum was prepared by phacoemulsification with three concentrations, namely 10 ng/mL, 10 μg/mL and 100 μg/mL respectively. Tantalum particles were characterized by transmission electron microscope(TEM) and scanning electron microscope(SEM). Three groups of nano-tantalum particles with different concentrations were cultured with osteoblasts MC3T3-E1 cell line for 6, 12 and 24 hours. CCK8 assay was used to detect the cell proliferation rate at different time points. The mRNAs levels of ALP, BMP-2 and OCN were detected by PCR. Results: The shape of nano-tantalum particles is regular, and the spherical shape is more common, the particle size is about 8-15 nm, and the average hydrated particle size is (286.5±13.6) nm. At 12 h, the cell proliferation rate, ALP, BMP-2 and OCN mRNAs expression levels of the three groups were significantly higher than those at 6 h and decreased at 24 h, but still higher than those at 6 h. Comparison between groups found at all time points, the cell proliferation rate, ALP, BMP-2 and OCN mRNAs expression levels of 10 μg/mL group were significantly higher than those of the other two groups. Conclusion: The results show that nano-tantalum still has a good induction effect of proliferation in contact with osteoblasts, which was related to particle concentration and size. The results need to be further verified.

Key words: nano-tantalum, implants, osteoblasts, proliferation