[Effects of Calcium Ion on the Migration and Osteogenic Differentiation of Human Osteoblasts]

Hua Xi Kou Qiang Yi Xue Za Zhi. 2018 Dec 1;36(6):602-608. doi: 10.7518/hxkq.2018.06.004.
[Article in Chinese]

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.

Conclusions: 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.

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(RT-PCR)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⁻¹ Ca²⁺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。CaSR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⁺-CaSR通路参与相应的信号传导。.

Keywords: bone substitute; calcium ion; calcium sensitive receptor; human osteoblast; migration.

MeSH terms

  • Calcium* / physiology
  • Cell Differentiation*
  • Cell Movement*
  • Cell Proliferation
  • Humans
  • Osteoblasts*
  • Osteogenesis* / physiology
  • Signal Transduction

Substances

  • Calcium

Grant support

[基金项目] 国家科技支撑计划(2012BAI17B03-6);福建省卫计委青年基金(2015-1-66);福建省口腔医院博士科研基金(2015-KQ-YY-BS-6)