Calcium Hydroxide-induced Proliferation, Migration, Osteogenic Differentiation, and Mineralization via the Mitogen-activated Protein Kinase Pathway in Human Dental Pulp Stem Cells
Calcium Hydroxide-induced Proliferation, Migration, Osteogenic Differentiation, and Mineralization via the Mitogen-activated Protein Kinase Pathway in Human Dental Pulp Stem Cells
复制标题
氢氧化钙通过丝裂原激活蛋白激酶途径诱导人牙髓干细胞的增殖、迁移、成骨分化和矿化
DOI：
10.1016/j.joen.2016.04.025
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发表时间：
2016-09-01
影响因子：
4.2
通讯作者：
Fan, Yubo
中科院分区：
文献类型：
--
作者：
Chen, Luoping;Zheng, Lisha;Fan, Yubo
前言：氢氧化钙已被广泛用作临床牙科直接盖髓的金标准。它诱导牙髓干细胞（DPSCs）增殖、迁移和矿化，但其潜在机制尚不清楚。本研究的目的是探讨有丝分裂原活化蛋白（MAP）激酶通路在氢氧化钙诱导的人DPSCs增殖、迁移、成骨分化和矿化中的作用。方法：使用第3代和第6代之间的人DPSC。DPSC与MAP激酶抑制剂预孵育，并与氢氧化钙一起培养。Western blot检测磷酸化MAP激酶。通过甲基噻唑四唑测定分析细胞活力。使用伤口愈合测定来估计细胞迁移。使用碱性磷酸酶（ALP）染色测定分析ALP表达。茜素红染色法研究矿化作用。结果如下：氢氧化钙显著促进c-Jun N-末端激酶（JNK）、p38和细胞外信号调节激酶的磷酸化。抑制JNK和p38信号通路可抑制氢氧化钙诱导的DPSC增殖。抑制JNK、p38和细胞外信号调节激酶信号传导抑制DPSC的迁移、ALP表达和矿化。结论：我们的研究表明，MAP激酶通路参与了氢氧化钙诱导的人DPSCs的增殖、迁移、成骨分化和矿化。
Introduction: Calcium hydroxide has been extensively used as the gold standard for direct pulp capping in clinical dentistry. It induces proliferation, migration, and mineralization in dental pulp stem cells (DPSCs), but the underlying mechanisms are still unclear. The aim of this study was to investigate the role of the mitogen-activated protein (MAP) kinase pathway in calcium hydroxide induced proliferation, migration, osteogenic differentiation, and mineralization in human DPSCs. Methods: Human DPSCs between passages 3 and 6 were used. DPSCs were preincubated with inhibitors of MAP kinases and cultured with calcium hydroxide. The phosphorylated MAP kinases were detected by Western blot analysis. Cell viability was analyzed via the methylthiazol tetrazolium assay. Cell migration was estimated using the wound healing assay. Alkaline phosphatase (ALP) expression was analyzed using the ALP staining assay. Mineralization was studied by alizarin red staining analysis. Results: Calcium hydroxide significantly promoted the phosphorylation of the c-Jun N-terminal kinase (JNK), p38, and extracellular signal regulated kinase. The inhibition of JNK and p38 signaling abolished calcium hydroxide induced proliferation of DPSCs. The inhibition of JNK, p38, and extracellular signal regulated kinase signaling suppressed the migration, ALP expression, and mineralization of DPSCs. Conclusions: Our study showed that the MAP kinase pathway was involved in calcium hydroxide induced proliferation, migration, osteogenic differentiation, and mineralization in human DPSCs.