钙离子对人牙囊细胞增殖、迁移和成骨分化的影响

Effect of calcium on proliferation,migration and osteogenic differentiation of human dental follicle cells

PURPOSE: To determine the role of Ca²⁺ in proliferation,migration and osteogenic differentiation of human dental follicle cells(hDFCs). METHODS: hDFCs were isolated and cultured. The source of hDFCs was detected by immunofluorescence staining. Osteogenesis and adipogenic differentiation of hDFCs was detected by alizarin red staining and oil red O staining, to identify its multi-directional differentiation ability. A series of Ca²⁺ solutions with different concentrations was prepared, CCK8 assay was used to detect the proliferative abilities at 1, 3, 5, and 7 d;migratory ability of 24 h was detected by Transwell assay. Calcium nodules were detected by semiquantitative analysis of alizarin red staining. mRNA expression of osteogenic differentiation related genes was examined by real-time quantitative polymerase chain reaction (RT-qPCR).Statistical analysis was performed using SPSS 17.0 software package. RESULTS: Compared with the control group, 3,4 and 5 mmol/L Ca²⁺ significantly promoted proliferation of hDFCs at 3, 5 and 7 d (P<0.05). 3, 4, 5 and 6 mmol/L Ca²⁺ significantly promoted the migration of hDFCs at 24 h(P<0.01). High concentration of Ca²⁺ had no significant effect on its proliferation and migration. The results of alizarin red staining showed that when Ca²⁺ concentration reached 4 mmol/L, formation of mineralized nodules were increased(P<0.01), and Ca²⁺ concentration-dependent. RT-qPCR results showed that Ca²⁺ up-regulated the expression of RUNX2 and OCN in osteogenic differentiation genes (P<0.01). CONCLUSIONS: Low Ca²⁺ concentration is beneficial to proliferation and migration, and high Ca²⁺ concentration is beneficial to osteogenic differentiation of human dental follicle cells.