Effect of Vitamin D3 on the Modulation of Differentiation in Periodontal Ligament Mesenchymal Cells with Low Osteogenic

Abstract

The regeneration of periodontal structures depends on the activation and differentiation ofmesenchymal cells from the periodontal ligament (PDLMSCs), which are responsible for formingnew cementum, alveolar bone, and periodontal ligament. However, these cells areheterogeneous, with approximately 80% displaying low osteogenic potential (LOP). Previousstudies identified the Asporin (ASPN) gene as an inhibitor of osteogenic/cementogenicdifferentiation by blocking BMP2 signaling and suggested that its suppression could enhancemineralized tissue formation. Vitamin D3 has emerged as a promising modulator, capable ofinhibiting ASPN expression and promoting mineralization. This study aims to investigate theeffects of vitamin D3 on the osteo/cementogenic differentiation of human LOP cells.Five LOP cell populations from the Periocells Biobank (B041) at FOP-UNICAMP will be culturedin standard medium, osteogenic medium (OM), and osteogenic medium supplemented withvitamin D3 (VitD) at different concentrations. The effects will be evaluated through assays ofcellular metabolism and apoptosis, mineralized matrix production (Alizarin Red), geneexpression of osteogenic markers (RUNX2, ALP, OC), and vitamin D3-modulated genes (VDR,ASPN, BMP2) via RT-qPCR, as well as protein analysis of ASPN and BMP2. This study aims toelucidate the role of vitamin D3 in promoting osteogenesis in LOP cells, suggesting newstrategies for periodontal regeneration. (AU)