2023, Vol. 27 ›› Issue (19): 3005-3010

Different concentrations of calcium ions interfere with the proliferation and osteogenic differentiation of human periodontal ligament stem cells

Li Yinghui1, 2 , Qi Fangfang2, 3, Han Xing1, 2 , Zhao Liru1, 2 , Ma Wensheng1, 2   

Abstract: BACKGROUND: As a second messenger, Ca2+ regulates many physiological processes, but its biological effect on the osteogenic differentiation of periodontal ligament stem cells is still unclear.  
OBJECTIVE: To investigate the effect of Ca2+ on the osteogenic differentiation of human periodontal ligament stem cells.
METHODS: The proliferation ability and osteogenic differentiation of the third generation human periodontal ligament stem cells were detected by the complete medium containing Ca2+ at the concentrations of 0 (control group), 1, 2, 5, 10, and 15 mmol/L and osteogenic induction solution.   CCK-8 assay was used to detect cell proliferation at 1, 3, 5, and 7 days. Real-time PCR and western blot assay were used to detect the expression levels of genes and proteins related to osteogenic differentiation factors RUNX2, OPN and OCN on days 3 and 7 of osteogenic induction. Alizarin red staining was performed on day 14 to analyze the osteogenic mineralization of human periodontal ligament stem cells.  
RESULTS AND CONCLUSION: (1) CCK-8 assay showed that the proliferation ability of human periodontal ligament stem cells in 2 and 5 mmol/L groups was better than that in control group on days 5 and day 7 (P < 0.05).   (2) On day 7 after in vitro osteogenic induction, the expression levels of osteogenic genes RUNX2, OPN and OCN in human periodontal ligament stem cells of 5 and 10 mmol/L groups were higher than those in control group (P < 0.05). (3) After 14 days of in vitro osteogenic induction, the numbers of mineralization nodules of human periodontal ligament stem cells in Ca2+ groups were higher than that in control group (P < 0.05). (4) These results suggest that Ca2+ can promote the proliferation and osteogenic differentiation of human periodontal ligament stem cells in a certain concentration range.

Key words: Ca2+, human periodontal ligament stem cell, proliferation, osteogenic differentiation