Mir-23A-3P Regulates Osteogenesis Via Lrp5 In Hpdlscs

Aim or purpose: This study investigates the regulatory role of miR-23a-3p in the osteogenic differentiation of human periodontal ligament stem cells (HPDLSCs) by targeting LRP5, using lentivirus-mediated inhibition/overexpression models. Understanding this pathway may provide insights into periodontal bone remodeling and therapeutic strategies. Materials and methods: HPDLSCs were isolated from extracted premolars of orthodontic patients (12–20 years old) and characterized via morphological analysis, colony formation assays, and immunofluorescence staining. Lentiviral vectors were designed for miR-23a-3p overexpression/inhibition and LRP5 silencing. Transfection efficiency was determined by fluorescence microscopy and RT-qPCR. Osteogenic differentiation was induced, and the expression of LRP5, ALP, and RUNX2 was assessed by RT-qPCR and Western blot at multiple time points. Statistical significance was analyzed (p<0.05). Results: 1. LRP5 expression peaked on day 7 of osteogenic induction and declined thereafter (p<0.01). 2. miR-23a-3p overexpression suppressed LRP5, ALP, and RUNX2 expression, while its inhibition promoted osteogenic differentiation (p<0.01). LRP5 silencing reduced osteogenic differentiation, confirming its role in the process. 3. The ''miR-23a-3p inhibition + LRP5 silencing'' model demonstrated that miR-23a-3p negatively regulates osteogenesis by targeting LRP5. Conclusions: This study successfully established lentiviral gene modulation models and demonstrated that miR-23a-3p inhibits osteogenic differentiation of HPDLSCs via LRP5 suppression. These findings provide a potential molecular target for periodontal bone regeneration strategies.