摘要
目的初步探讨Wnt/β-连环蛋白(β-catenin)信号通路在人牙髓干细胞(hDPSCs)成骨向分化中的作用。方法采用激光扫描共聚焦技术检测氢氧化钙作用14d 后hDPSCs的细胞骨架变化和经典Wnt信号通路关键启动因子β-catenin 的表达及细胞定位。分别上调和抑制Wnt信号通路信号,通过蛋白质印迹(Western Blot)实验和茜素红染色实验检测氢氧化钙作用14d 后hDPSCs 的成骨向分化和矿化情况。结果氢氧化钙作用效应下hDPSCs的细胞骨架出现结构重排,β-catenin 自细胞核内向核外转移。Dickkopf相关蛋白1(DKK-1)阻断Wnt信号通路后,hDPSCs的钙结节数少于单纯氢氧化钙效应细胞;氢氧化钙作用下的hDPSCs 表达牙本质涎磷蛋白(DSPP)。结论氢氧化钙可下调经典Wnt信号通路的表达,促进hDPSCs的成骨向分化和矿化以及成牙本质向分化。
Objective To investigate the role of Wnt/β-catenin signaling pathway in the osteogenic differentiation of human dental pulp stem cells (hDPSCs). Methods After 14 days of the calcium hydroxide training, the cytoskeletal changes of hDPSCs,the expression of β-catenin,i.e. the key promoter of in the Wnt signaling pathway,and the cell localization were detected by laser scanning confocal technique. The Wnt signaling pathways were up-regulated and inhibited,and the osteogenic differentiation and mineralization of hDPSCs were detected by Western Blot and alizarin red staining after 14 days of the training. Results The cytoskeleton of hDPSCs was rearranged by the effect of calcium hydroxide,and the β-catenin migration from nucleus to cytoplasm were observed. The number of calcium nodules in hDPSCs was decreased after blocking Wnt signaling pathway by Dickkopf-related protein 1 (DKK-1). The calcium hydroxide treatment can promoted dentin sialophosphoprotein (DSPP) expression in hDPSCs. Conclusions Calcium hydroxide can down-regulate the expression of canonical Wnt signaling pathway and promote osteogenic differentiation and mineralization and odontogenetic differentiation of hDPSCs.
作者
陈罗萍
郑丽沙
樊瑜波
Chen Luoping;Zheng Lisha;Fan Yubo(Key Laboratory for Biomechanics and Mechanobiology of Ministry of Education,School of Biological Science and Medical Engineering,Beihang University,Beijing 100191,China)
出处
《国际生物医学工程杂志》
CAS
2019年第2期120-124,共5页
International Journal of Biomedical Engineering
