Lei Qun, Lin Dong, Huang Wen-Xiu, Wu Dong, Chen Jiang
Stomatological Hospital of Fujian Medical University, Fuzhou 350000, China.
Hua Xi Kou Qiang Yi Xue Za Zhi. 2018 Dec 1;36(6):602-608. doi: 10.7518/hxkq.2018.06.004.
This study aimed to investigate the effect of calcium ion (Ca²⁺) on the migration and osteogenic differentiation of human osteoblasts and explore the proper concentration and correlation mechanism.
A series of Ca²⁺ solutions with different concentrations was prepared. Osteoblast migration was assessed by Transwell assay, and proliferation was studied via the CCK-8 colorimetric assay. The mRNA expression of osteogenic genes was examined via reverse transcription-polymerase chain reaction (RT-PCR), and the mineralized nodule was examined by alizarin red-S method. After calcium sensitive receptor (CaSR) antagonism, Ca²⁺-induced migration and osteogenic differentiation were analyzed.
In the migration experiment, 2, 4, and 6 mmol·L⁻¹ Ca²⁺ could promoted osteoblast migration at three timepoints (8, 16, and 24 h), whereas 10 mmol·L⁻¹ Ca²⁺ considerably inhibited migration at 8 h. The Ca²⁺ concentration range of 2-10 mmol·L⁻¹ could promote proliferation, osteogenic differentiation, and mineralization of human osteoblasts. Moreover, mineralization was predominantly induced by 8 and 10 mmol·L⁻¹ Ca²⁺. CaSR antagonism could reduce Ca²⁺-induced migration and osteogenic differentiation of human osteoblasts.
Low Ca²⁺ concentration favored osteoblast migration, whereas high Ca²⁺ concentration favored osteogenic differentiation. The Ca²⁺ concentrations of 4 and 6 mmol·L⁻¹ could substantially induce osteoblast migration and osteogenic differentiation, and the Ca²⁺-CaSR pathway participated in signal transduction.
本研究旨在探讨钙离子（Ca²⁺）对人成骨细胞迁移和成骨分化的影响，并探索合适的浓度及相关作用机制。
制备一系列不同浓度的Ca²⁺溶液。通过Transwell实验评估成骨细胞迁移，采用CCK-8比色法研究细胞增殖。通过逆转录-聚合酶链反应（RT-PCR）检测成骨基因的mRNA表达，并用茜素红-S法检测矿化结节。在拮抗钙敏感受体（CaSR）后，分析Ca²⁺诱导的迁移和成骨分化情况。
在迁移实验中，2、4和6 mmol·L⁻¹的Ca²⁺在三个时间点（8、16和24小时）均可促进成骨细胞迁移，而10 mmol·L⁻¹的Ca²⁺在8小时时显著抑制迁移。2-10 mmol·L⁻¹的Ca²⁺浓度范围可促进人成骨细胞的增殖、成骨分化和矿化。此外，8和10 mmol·L⁻¹的Ca²⁺主要诱导矿化。CaSR拮抗可降低Ca²⁺诱导的人成骨细胞迁移和成骨分化。
低Ca²⁺浓度有利于成骨细胞迁移，而高Ca²⁺浓度有利于成骨分化。4和6 mmol·L⁻¹的Ca²⁺浓度可显著诱导成骨细胞迁移和成骨分化，且Ca²⁺-CaSR途径参与信号转导。
