神经营养素3促进人牙囊细胞成骨分化

Neurotrophin 3 promotes osteogenic differentiation of human dental follicle cells

目的研究神经营养素3(NT-3)对人牙囊细胞(h DFCs)成骨分化的影响。方法体外分离并培养h DFCs,免疫细胞化学染色法鉴定细胞来源,CCK-8法检测不同质量浓度NT-3对h DFCs增殖活性的影响,同时检测NT-3对h DFCs的碱性磷酸酶(ALP)活性,以及骨形态发生蛋白-2(BMP-2)、骨钙素(OCN)m RNA相对表达量的影响,并用茜素红染色法检测矿化情况。结果波形丝蛋白和角蛋白染色结果表明h DFCs为间充质细胞来源。NT-3对h DFCs增殖活性无明显影响;当NT-3质量浓度为25、50、100 ng·m L-1时能明显增强ALP活性,提高BMP-2、OCN m RNA的相对表达量,与对照组相比差异有统计学意义(P<0.05),NT-3质量浓度为100 ng·m L-1时BMP-2、OCN m RNA的相对表达量达到最高。当NT-3质量浓度为50、100 ng·m L-1时矿化结节数量最多。结论适宜质量浓度的NT-3能促进h DFCs的成骨分化。

Objective This study aims to investigate the effect of neurotrophin 3 （NT-3） on the osteogenic differentiation of human dental follicle cells （hDFCs）. Methods hDFCs were isolated and cultured in vitro. Immunocytochemical staining was used to identify the origin of hDFCs. The effects of different NT-3 concentrations on hDFCs proliferation were detected by using CCK-8 assay. The alkaline phosphatase （ALP） activities and mRNA expression levels of bone morphogenetic protein-2 （BMP-2） and osteocalcin （OCN） were determined to investigate the effects of NT-3 on hDFCs osteogenesis. The difference in the number of mineralized nodules was detected using alizarin red staining. Results Vimentin and cytokeratin staining results showed that hDFCs originated from the mesenchymal cells. NT-3 exerted no evident effect on hDFCs proliferation. The ALP activity and the BMP-2 and OCN mRNA expression levels of hDFCs were signi？cantly improved under treatment with different NT-3 concentrations （25, 50, and 100 ng·mL^-1） compared with those in the control group. BMP-2 and OCN mRNA relative expression levels of hDFCs reached the highest when the NT-3 concentration was 100 ng·mL^-1. The number of mine-ralized nodules reached the maximum when the hDFCs were treated with 50 and 100 ng·mL^-1 NT-3. Conclusion Appropriate mass concentration of NT-3 can promote the osteogenic differentiation of hDFCs.