The roles of calcium-sensing receptor and calcium channel in osteogenic differentiation of undifferentiated periodontal ligament cells
The roles of calcium-sensing receptor and calcium channel in osteogenic differentiation of undifferentiated periodontal ligament cells
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DOI：
10.1007/s00441-014-1918-5
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发表时间：
2014-09-01
影响因子：
3.6
通讯作者：
Akamine, Akifumi
中科院分区：
文献类型：
--
作者：
Koori, Katsuaki;Maeda, Hidefumi;Akamine, Akifumi
升高的细胞外钙已被证明可以促进成骨细胞的分化。然而，钙影响人牙周韧带干细胞/祖细胞（PDLSCs）成骨分化的方式尚不清楚。我们的目的是研究我们最近建立的钙暴露的人PDLSC系（细胞系1-17）的增殖和成骨分化，并阐明钙敏感受体（CaSR）和l型电压依赖性钙通道（L-VDCC）在这一过程中的作用。WST-1法检测细胞增殖活性，定量逆转录酶加聚合酶链反应和免疫染色法检测基因和蛋白表达。采用von Kossa法和茜素红染色进行钙化试验。5mm CaCl2显著诱导细胞系1-17的增殖、骨相关基因表达和钙化。在5 mM CaCl2培养过程中，该细胞系的CaSR基因表达量上调，7天后表达量降低。与单独使用CaCl2相比，同时使用NPS2143（一种CaSR抑制剂）和钙治疗可进一步显著增加骨相关基因表达和钙化。与单独钙处理相比，L-VDCC抑制剂硝苯地平显著抑制5 mM CaCl2处理的细胞系1-17的成骨分化，促进CaSR的表达。因此，细胞外钙的升高促进了PDLSC细胞系的增殖和成骨分化。拮抗CaSR进一步增强了钙对成骨分化的作用，细胞外钙作用下，CaSR的表达受L-VDCC调控。因此，细胞外钙可能通过相互调节CaSR和L-VDCC来调节PDLSCs的成骨分化。
Elevated extracellular calcium has been shown to promote the differentiation of osteoblasts. However, the way that calcium affects the osteogenic differentiation of human periodontal ligament stem/progenitor cells (PDLSCs) remains unclear. Our aim has been to investigate the proliferation and osteogenic differentiation of a calcium-exposed human PDLSC line (cell line 1-17) that we have recently established and to elucidate the roles of the calcium-sensing receptor (CaSR) and L-type voltage-dependent calcium channel (L-VDCC) in this process. Proliferation activity was investigated by WST-1 assay, and gene and protein expression was examined by quantitative reverse transcriptase plus the polymerase chain reaction and immunostaining, respectively. Calcification assay was performed by von Kossa and Alizarin red staining. Treatment with 5 mM CaCl2 significantly induced proliferation, bone-related gene expression, and calcification in cell line 1-17. During culture with 5 mM CaCl2, this cell line up-regulated the gene expression of CaSR, which was reduced after 7 days. Simultaneous treatment with NPS2143, a CaSR inhibitor, and calcium significantly further increased bone-related gene expression and calcification as compared with CaCl2 exposure alone. The L-VDCC inhibitor, nifedipine, significantly suppressed osteogenic differentiation of cell line 1-17 treated with 5 mM CaCl2 and promoted the expression of CaSR, as compared with calcium treatment alone. Thus, elevated extracellular calcium promotes the proliferation and osteogenic differentiation of a PDLSC line. Antagonizing CaSR further enhances the effect of calcium on osteogenic differentiation, with CaSR expression being regulated by L-VDCC under extracellular calcium. Extracellular calcium might therefore modulate the osteogenic differentiation of PDLSCs through reciprocal adjustments of CaSR and L-VDCC.