Mechanisms Of Helns Modulating Periodontal Inflammation And Osteo-Angiogenic Coupling

This study aimed to explore the therapeutic potential of Houttuynia cordata-derived exosome-like nanoparticles (HELNs) in periodontal regeneration by supressing inflammation and promoting osteogenesis-angiogenesis coupling. Network pharmacology identified multi-target mechanisms of Houttuynia cordata in periodontitis treatment. HELNs were isolated via differential centrifugation and sucrose density gradient ultracentrifugation, characterized structurally and compositionally. The osteogenic differentiation of hPDLSCs enhanced by HELNs was validated through ALP staining, ARS mineralization assay, qPCR and Western blot. Similarly, the pro-angiogenic effects of HELNs on HUVECs were confirmed by Matrigel-based tubule formation assay and qPCR-mediated quantification of angiogenesis-related gene expression. Herein, HELNs isolated via differential centrifugation and sucrose density gradient ultracentrifugation demonstrated characteristic bilayer structures and contained flavonoids, amino acids, and organic acids. Functionally, HELNs dose-dependently enhanced osteogenesis in human periodontal ligament stem cells (hPDLSCs), evidenced by elevated alkaline phosphatase activity, mineralization, and osteogenic gene expression. Mechanistic studies revealed dual regulation of bone metabolism through Wnt/β-catenin activation and RANKL-mediated osteoclast suppression. Concurrently, HELNs stimulated angiogenesis in endothelial cells by improving migration, tubulogenesis, and angiogenic gene expression, establishing osteogenesis-angiogenesis coupling. HELNs regulate the periodontal microenvironment via a multi-target synergistic mechanism and promote tissue regeneration through osteogenesis-angiogenesis coupling. Future directions include developing HELNs-loaded hydrogels for localized delivery, validating regenerative efficacy in preclinical models, and employing spatial metabolomics/single-cell sequencing to decipher cellular network modulation. This study provides an innovative strategy for periodontal regeneration and establishes a novel paradigm for inflammatory bone defect repair.