Stem Cell-Derived Vesicles For Periodontal Regeneration

To investigate the biological functions of intracellular and extracellular vesicles derived from Mo-doped bioactive glass ceramic (Mo-BGC) - pretreated human periodontal ligament stem cells (hPDLSCs). We will evaluate their anti-inflammatory effects and their ability to promote osteogenic differentiation of hPDLSCs under inflammatory conditions, and explore the underlying molecular mechanisms involved. Intracellular and extracellular vesicles (EVs, Mo-BGC/EVs, IVs, Mo-BGC/IVs) were successfully isolated from hPDLSCs—both pretreated with Mo-BGC and untreated controls, followed by comprehensive characterization. In vitro, the four types of vesicles were individually introduced into the E. coli-derived lipopolysaccharide (LPS)-treated cultures to assess their effects on cell viability, inflammatory cytokine release, and osteogenic differentiation capacity. In vivo, they were applied to the defect sites to evaluate their regenerative efficacy on periodontal bone repair under inflammatory conditions. Finally, RNA-seq was performed to compare the effects of EVs vs. IVs and IVs vs. Mo-BGC/IVs on the gene expression profiles of hPDLSCs. In vitro and in vivo experiments demonstrated that all four types of vesicles inhibited inflammation while promoting the proliferation and osteogenic differentiation of hPDLSCs, with IVs exhibiting stronger effects than EVs. Following Mo-BGC pretreating, the vesicles showed enhanced anti-inflammatory capacity and osteogenic differentiation potential. IVs significantly upregulated the YAP/TAZ signaling pathway compared to EVs, whereas Mo-BGC/IVs further activated the PI3K/AKT signaling pathway relative to IVs. These findings establish a critical molecular biology foundation for elucidating the functional mechanisms of intracellular and extracellular vesicles.