Roles of human periodontal ligament stem cells in osteogenesis and inflammation in periodontitis models: Effect of 1 alpha,25-dihydroxyvitamin D-3

Periodontitis is a chronic inflammatory disease caused by Porphyromonas gingivalis and other bacteria, and human periodontal ligament stem cells (hPDLSCs) are a promising candidate for the treatment of periodontal supporting tissue defects. This study aimed to investigate the effect of 1 alpha,25-dihydroxyvitamin D-3 [1,25 (OH)(2)VitD(3)] on osteogenic differentiation of hPDLSCs in an in vitro periodontitis model and whether it can improve inflammatory status. hPDLSCs were in vitro isolated and identified. After treatment with 1,25(OH)(2)VitD(3) and ultrapure pure Porphyromonas gingivalis lipopolysaccharide (LPS-G), the viability of hPDLSCs was detected using Cell Counting Kit-8, the expressions of osteogenic markers and inflammatory genes using Western blotting and quantitative reverse transcription PCR (qRT-PCR), the levels of inflammatory factors in cells using enzyme linked immunosorbent assay (ELISA), and the fluorescence signal intensity of osteoblastic markers and inflam-matory genes in cells using immunofluorescence assay. It was found that 1,25(OH)(2)VitD(3) reversed the inhibition of hPDLSCs proliferation by LPS-G; LPS-G exhibited inhibitory effect on ALP, Runx2, and OPN expressions, and such inhibitory effect was significantly weakened when co-acting with 1,25(OH)(2)VitD(3). Meanwhile, LPS-G upregulated the expressions of inflammatory genes IL-1 beta and Casp1, whereas 1,25(OH)(2)VitD(3) antagonized such an effect and improved the inflammatory status. In conclusion, 1,25(OH)(2)VitD(3) can reverse the inhibitory effect of LPS-G on hPDLSCs proliferation and osteogenic differentiation and suppress LPS-G-induced upregulation of inflammatory gene expressions.

Automated summary

This study investigated the effect of 1,25(OH)(2)VitD(3) on osteogenic differentiation of hPDLSCs in an in vitro periodontitis model and its impact on inflammatory status. The results showed that 1,25(OH)(2)VitD(3) can reverse the inhibitory effect of LPS-G on hPDLSCs proliferation and osteogenic differentiation, and also suppress LPS-G-induced upregulation of inflammatory gene expressions, improving the inflammatory status.