钙离子对人成骨细胞迁移与成骨分化的影响

Effects of calcium ion on the migration and osteogenic differentiation of human osteoblasts

目的研究不同浓度Ca^(2+)对人成骨细胞迁移与成骨分化的影响,探讨促进迁移与成骨合适的Ca^(2+)浓度及相关机制。方法设置Ca^(2+)浓度,Transwell检测成骨细胞迁移;CCK-8法评估成骨细胞增殖;逆转录聚合酶链反应(RTPCR)检测细胞成骨分化相关基因的表达;茜素红染色检测成骨分化生成的矿化结节。钙敏感受体(CaSR)拮抗剂拮抗后,观察Ca^(2+)对人成骨细胞迁移与成骨分化的影响。结果在迁移实验中,2、4、6 mmol·L^(-1)的Ca^(2+)在3个时间点(8、16、24 h)都能明显地促进人成骨细胞的迁移,10 mmol·L^(-1)的Ca^(2+)在8 h时明显抑制迁移。2～10 mmol·L^(-1)Ca^(2+)能促进人成骨细胞的增殖、成骨分化与矿化,8、10 mmol·L^(-1)的Ca^(2+)诱导的矿化作用更明显。Ca SR拮抗降低Ca^(2+)诱导的人成骨细胞迁移与成骨分化作用。结论低浓度Ca^(2+)有利于人成骨细胞迁移,高浓度Ca^(2+)有利于人成骨细胞分化,4、6 mmol·L^(-1)的Ca^(2+)能较明显地同时诱导人成骨细胞迁移与成骨分化,Ca^(2+)-Ca SR通路参与相应的信号传导。

Objective 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. Methods 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. Results 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. Conclusion 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.