The effects of leptin on osteogenesis/odontogenic related gene expression of human apical papillary stem cells

Abstract

Objective To investigate the effects of leptin on the proliferation of stem cells from human stem cells from the apical papilla (hSCAPs) and the expression of osteogenic/dentinogenic genes in vitro to provide an experimen⁃ tal basis for the sustainable development of young permanent teeth. Methods The tissue block method was used to iso⁃ late and culture hSCAPs from the apical papilla of the immature third permanent molar. The expression of leptin and OBRb in hSCAPs was detected using immunocytofluorescence staining, western blotting and reverse transcription poly⁃ merase chain reaction (RT⁃PCR) analysis. The hSCAPs was treated with 0.1 μg/mL of leptin (0.1 μg/mL group) or 1.5 μg/mL of leptin (1.5 μg/mL group) at different time points. The control group was treated with alpha⁃MEM medium. Cell proliferation was measured using the CCK8 assay and cell cycle analysis. QRT⁃PCR was used to detect the expres⁃ sion of related osteoblast/odontogenic genes for alkaline phosphatase (ALP), dentin matrix protein ⁃1 (DMP⁃1), dentin si⁃ alophosphoprotein (DSPP), and osteocalcin (OCN) mRNA. The differences between the treatment groups and the control group were analyzed statistically using one⁃way ANOVA followed by Bonferroni analysis. Results The expression of both leptin and OBRb were found in hSCAPs. Compared with the control group, the cell proliferation capacity and S phase cells in the treatment groups were higher than those in the control group, with the 1.5 μg /mL group displaying higher levels than 0.1 μg /mL group, and the treated hSCAPs demonstrated a higher proliferation rate and a higher ex⁃ pression of ALP, DSPP, and DMP⁃1 from day 3 to day 7, with the 1.5 μg /mL group displaying higher levels than 0.1 μg /mL group , and the difference was statistically significant (P < 0.05), at day 7. The treated hSCAPs demonstrated a lower expression of ALP, DSPP, and DMP⁃1. Compared with the control group, the treated hSCAPs demonstrated a high⁃ er expression of OCN from day 7 to day 14, with significantly higher expression in the 1.5 μg /mL group compared to the 0.1 μg /mL group. Conclusion Leptin may promote cell proliferation and upregulate the expression of relative os⁃ teogenic/dentinogenic genes.

Keywords

• Odontogenic，
• Apical papilla stem cells，
• Leptin，
• Osteogenic/dentinogenic genes，
• Differentiation，
• Dentin matrix protein⁃1，
• Dentin sialophosphoprotein，
• Osteocalcin，