【摘要】 目的研究不同浓度Ca²⁺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Ca²⁺浓度及相关机制。方法设置Ca²⁺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Ca²⁺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Ca²⁺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Ca²⁺在8 h时明显抑制迁移。2～10 mmol·L^-1Ca²⁺能促进人成骨细胞的增殖、成骨分化与矿化,8、10 mmol·L^-1的Ca²⁺诱导的矿化作用更明显。Ca SR拮抗降低Ca²⁺诱导的人成骨细胞迁移与成骨分化作用。结论低浓度Ca²⁺有利于人成骨细胞迁移,高浓度Ca²⁺有利于人成骨细胞分化,4、6 mmol·L^-1的Ca²⁺能较明显地同时诱导人成骨细胞迁移与成骨分化,Ca²⁺-Ca SR通路参与相应的信号传导。更多还原

【Abstract】 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.更多还原