Lipus Enhances Osteogenic Differentiation Of Pdlscs Via Wnt7b/Β-Catenin Pathway

To evaluate LIPUS effects on proliferation, migration, and osteogenic differentiation of periodontal ligament stem cells (PDLSCs) and elucidate its regulatory mechanism through the Wnt7B/β-catenin pathway. PDLSCs were isolated via enzymatic digestion, characterized by flow cytometry, and treated with LIPUS at 30–120 mW/cm². Optimal intensity was determined by ALP staining and RT-PCR after osteogenic induction. Proliferation/migration were assessed using CCK-8, colony formation, scratch, and Transwell assays. Osteogenesis was evaluated via ALP/ARS staining, RT-PCR, and Western blot. RNA-seq identified differentially expressed genes (DEGs), with KEGG pathway analysis highlighting Wnt/β-catenin involvement. Wnt7B was overexpressed or silenced via lentiviral transfection to assess its role in LIPUS-mediated osteogenesis. LIPUS at 80 mW/cm² maximized ALP activity and osteogenic gene expression (Runx2, OCN). It enhanced proliferation (CCK-8/colony formation) and migration (scratch/Transwell) while promoting mineralization (ARS). RNA-seq revealed 117 DEGs, with Wnt/β-catenin pathway enrichment. LIPUS upregulated Wnt7B, Wnt11, and β-catenin. Overexpression of Wnt7B synergized with LIPUS to amplify osteogenesis, whereas silencing Wnt7B attenuated differentiation, which LIPUS partially rescued. β-catenin/TCF3 expression correlated with Wnt7B modulation. LIPUS at 80 mW/cm² enhances PDLSC osteogenesis via the Wnt7B/β-catenin axis, positioning Wnt7B as a critical mediator. These findings provide insights into LIPUS as a therapeutic stimulus for periodontal regeneration.