应用钛表面二氧化钛纳米管诱导牙周膜干细胞成骨分化的实验研究

Experimental study on the osteogenic differentiation of hPDLSCs induced by titanium dioxide nanotubes on titanium surface

目的:研究应用钛表面二氧化钛纳米管诱导人牙周膜干细胞(hPDLSCs)成骨分化的效果。方法:将人hPDLSCs分为空白对照组(DMEM培养基)、阳性对照组(成骨诱导培养基)、实验组(接种二氧化钛纳米管-羟基二咪唑-rhBMP-2复合生物层置于DMEM培养基);SEM观察细胞黏附情况;碱性磷酸酶(ALP)染色、茜素红染色检测细胞成骨;逆转录聚合酶链反应法(RT-PCR)、免疫印迹法(WB)测定hPDLSCs中DMP-1、Runx-2、Osterix、OCN基因和蛋白表达。结果:SEM显示细胞粘附于复合生物层表面,细胞突起互相连接。阳性对照组和实验组ALP阳性细胞、钙结节数量增多,DMP-1、Runx-2、Osterix、OCN表达均升高(P<0.05)。结论:二氧化钛纳米管-羟基二咪唑-rhBMP-2复合生物层可诱导hPDLSCs细胞成骨分化。

Objective: To investigate the osteogenic differentiation of human periodontal ligment stem cells(hPDLSCs) on titanium im plants with titanium dioxide nanotubes. Methods: Human hPDLSCs were in vitro cultured and divided into the blank control(DMEM) group, positive control(osteogenesis inducing medium) group and experimental group (the titanium with dioxide nanotube-hydroxyl di imidazole- rhBMP-2 composite biological layer in DMEM). The adhesion of the cells was observed by SEM. Alkaline phosphatase (ALP) staining and alizarin red staining were used to detect the cell osteogenesis. RT-PCR and Western blotting were used to detect DMP-1 , Runx-2, Osterix, OCN gene and protein expression of hPDLSCs. Results: SEM observation showed that cells adhered to the surface of the composite biological layer, and the cells were connected to each another by protrusions. In positive control group and ex perimental group, the number of ALP positive cells and calcium nodules increased, the expression of DMP-1, Runx-2, Osterix and OCN increased(P <0. 05). Conclusion: TiO2 nanotube hydroxy diimidazole -rhBMP-2 complex biological layer can promote the osteo genic differentiation of hPDLSCs.