Downregulating microRNA-152-3p promotes the viability and osteogenic differentiation of periodontal ligament stem cells via targeting integrin alpha 5

Abstract

ObjectiveThe aim of this study was to investigate the role of microRNA-152-3p (miR-152-3p) in the osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs). DesignHPDLSCs were isolated and identified using immunofluorescence staining, and their osteogenic differentiation capability was evaluated by alkaline phosphatase staining and Alizarin Red staining. HPDLSC viability was measured using cell counting kit-8. alkaline phosphatase level in hPDLSCs was measured by enzyme-linked immunosorbent assay. Target gene and potential binding sites between miR-152-3p and integrin alpha 5 (ITGA5) were predicted using TargetScan and confirmed by dual-luciferase reporter assay. Relative expressions of miR-152-3p and factors related to hPDLSC osteogenic differentiation were measured by quantitative real-time polymerase chain reaction and Western blot as needed. ResultsCollected cells were observed and identified as hPDLSCs. MiR-152-3p expression was downregulated during hPDLSC osteogenic differentiation in a time-dependent manner, and downregulating miR-152-3p promoted cell viability, enhanced alkaline phosphatase level, and increased the expressions of genes related to hPDLSC osteogenic differentiation. ITGA5 was the target gene of miR-152-3p and ITGA5 expression was upregulated during osteogenic differentiation in a time-dependent manner. Silencing ITGA5 partially reversed the effects of downregulating miR-152-3p on hPDLSCs. ConclusionDownregulating miR-152-3p may promote hPDLSC viability and osteogenic differentiation via targeting ITGA5, and have potential effects on periodontal and alveolar bone regeneration.