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
作者:雷维, 马玥, 任嫒姝
Author:Wei Lei, Yue Ma, Aishu Ren
收稿日期:2017-06-10 年卷(期)页码:2018,36(1):23-23-28
期刊名称:华西口腔医学杂志
Journal Name:West China Journal of Stomatology
关键字:神经营养素3,人牙囊细胞,成骨分化,
Key words:neurotrophin 3,human dental follicle cells,osteogenic differentiation,
基金项目:[基金项目] 中国博士后科学基金(2013M542248);2016年重庆高校创新团队建设计划(CXTDG201602006);重庆市高校市级口腔生物医学工程重点实验室资助项目[渝教科(2014)55号]
中文摘要
目的 研究神经营养素3(NT-3)对人牙囊细胞(hDFCs)成骨分化的影响。方法 体外分离并培养hDFCs,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hDFCs增殖活性的影响,同时检测NT-3对hDFCs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)mRNA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hDFCs为间充质细胞来源。NT-3对hDFCs增殖活性无明显影响;当NT-3质量浓度为25、50、100 ng·mL-1时能明显增强ALP活性,提高BMP-2、OCN mRNA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·mL-1时BMP-2、OCN mRNA的相对表达量达到最高。当NT-3质量浓度为50、100 ng·mL-1时矿化结节数量最多。结论 适宜质量浓度的NT-3能促进hDFCs的成骨分化。
英文摘要
ObjectiveThis study aims to investigate the effect of neurotrophin 3 (NT-3) on the osteogenic differentiation of human dental follicle cells (hDFCs).MethodshDFCs were isolated and culturedin 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.ResultsVimentin 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 significantly 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-1NT-3.ConclusionAppropriate mass concentration of NT-3 can promote the osteogenic differentiation of hDFCs.
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