Role of the Extracellular Signal-regulated Kinase 1/2 Pathway in Driving Tricalcium Silicate-induced Proliferation and Biomineralization of Human Dental Pulp Cells In Vitro
Role of the Extracellular Signal-regulated Kinase 1/2 Pathway in Driving Tricalcium Silicate-induced Proliferation and Biomineralization of Human Dental Pulp Cells In Vitro
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DOI：
10.1016/j.joen.2013.03.002
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发表时间：
2013-08-01
影响因子：
4.2
通讯作者：
Zhu, Yaqin
中科院分区：
文献类型：
--
作者：
Du, Rong;Wu, Tiantian;Zhu, Yaqin
简介：本研究旨在探讨细胞外信号调节激酶1/2（ERK 1/2）通路在硅酸三钙（C3 S）诱导的人牙髓细胞（hDPC）体外增殖和生物矿化中的作用。方法：人DPC在含C3 S的培养基中培养，并与未处理的对照组进行比较。通过甲基-噻唑-四唑测定来测量细胞活力。通过用von Kossa和茜素红S染色对细胞外基质上的钙沉积进行染色来评估生物矿化。通过免疫印迹法评价磷酸化ERK 1/2。分别采用流式细胞术和实时荧光定量聚合酶链反应（real-time polymerase chain reaction，RT-PCR）检测ERK 1/2抑制剂U 0126对hDPC细胞周期和矿化相关基因表达的影响。通过方差分析，然后进行Student-Newman-Keuls事后检验来分析数据，显著性设定为P <0.05。结果：C3 S提取物能促进hDPC的活力和生物矿化（P <0.05）。在C3 S提取物中培养30分钟后，磷酸化ERK 1/2强烈出现。此外，在C3 S处理的hDPC中抑制ERK 1/2通路降低了增殖和矿化依赖性基因的表达，包括I型胶原、牙本质唾液磷蛋白、骨桥蛋白和骨钙素（P <0.05）。结论：C3 S在体外刺激hDPC的增殖和生物矿化，ERK 1/2通路在这些效应的调节中起关键作用。
Introduction: The aim of this study was to investigate the role of the extracellular signal-regulated kinase 1/2 (ERK1/2) pathway in regulating tricalcium silicate (C3S)-driven proliferation and biomineralization of human dental pulp cells (hDPCs) in vitro. Methods: Human DPCs were cultured in C3S-containing medium and compared with untreated controls. Cell viability was measured by the methyl-thiazol-tetrazolium assay. Biomineralization was assessed by staining calcium deposits on the extracellular matrix with von Kossa and alizarin red S stains. Phosphorylated ERK1/2 was evaluated by immunoblotting. The ERK1/2 inhibitor U0126 was used to assess the role of this pathway on stage of the cell cycle and mineralization-dependent gene expressions of hDPCs by using flow cytometry and real-time polymerase chain reaction, respectively. Data were analyzed by analysis of variance followed by the Student-Newman-Keuls post hoc test, with significance set at P < .05. Results: The viability and biomineralization of hDPCs were promoted by C3S extracts (P < .05). Phosphorylated ERK1/2 strongly appeared after hDPCs were cultured in the C3S extracts for 30 minutes. Moreover, inhibition of the ERK1/2 pathway in C3S-treated hDPCs decreased proliferation and the expression of mineralization-dependent genes, including collagen type I, dentin sialophosphoprotein, osteopontin, and osteocalcin (P < .05). Conclusions: C3S stimulated the proliferation and biomineralization of hDPCs in vitro, with the ERK1/2 pathway playing a key role in the regulation of these effects.