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

Abstract

AbstractPeriodontitis is a chronic inflammatory disease caused byPorphyromonas gingivalisand 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 1α,25-dihydroxyvitamin D3[1,25(OH)2VitD3] on osteogenic differentiation of hPDLSCs in anin vitroperiodontitis model and whether it can improve inflammatory status. hPDLSCs werein vitroisolated and identified. After treatment with 1,25(OH)2VitD3and ultrapure purePorphyromonas gingivalislipopolysaccharide (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 inflammatory genes in cells using immunofluorescence assay. It was found that 1,25(OH)2VitD3reversed 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)2VitD3. Meanwhile, LPS-G upregulated the expressions of inflammatory genes IL-1β and Casp1, whereas 1,25(OH)2VitD3antagonized such an effect and improved the inflammatory status. In conclusion, 1,25(OH)2VitD3can reverse the inhibitory effect of LPS-G on hPDLSCs proliferation and osteogenic differentiation and suppress LPS-G-induced upregulation of inflammatory gene expressions.