Cellular responses of stem cells isolated from human exfoliated deciduous teeth towards different concentration of calcium ion

Abstract

Objectives: The purpose of this study was to investigate the effect of Ca2+ on proliferation, osteogenic differentiation, and migration of stem cells from human exfoliated teeth (SHEDs) in vitro Materials and methods: SHEDs were seeded in culture media and osteogenic induction media containing 1.8-16.2 mM of Ca2+. SHEDs proliferation was determined using MTT assay and colony forming unit assay. Osteogenic differentiation was evaluated using mineralization assay and osteogenic marker gene expression and cell migration was evaluated using wound healing assay. Values were expressed as mean + S.D. Statistical analysis of MTT assay and wound healing assay were performed using two-way ANOVA followed by Tukey’s range test. Other assays were analyzed using one-way ANOVA. Results: Different Ca2+ concentration did not affect cell proliferation and colony forming unit. While, osteocalcin, an osteogenic marker gene, was significantly increased at day 7 in 9.0 and 12.6 mM group (p<0.05). The result was also consistent with alizarin red and Von kossa staining, which had the most staining in 9.0 mM group. However, higher Ca2+ concentration inhibited SHEDs migration at 24 hours compared to control (1.8 mM). Conclusion: Ca2+ concentration between 1.8 and 16.2 mM did not have any effects on proliferation and colony forming unit. On the other hand, higher Ca2+ seemed to induce late stage osteogenic differentiation and mineralization, but migration was inhibited.