Flavonoid Diosmetin Prevents Periodontitis via Synergistic Anti‐Inflammatory and Osteogenic Effects: Dual In Vitro/In Vivo Validation

OBJECTIVE: Periodontitis is a chronic inflammatory disorder affecting periodontal tissues; diosmetin, exhibiting anti-inflammatory and anti-osteoporotic properties, emerges as a promising novel agent for periodontitis prevention. METHODS: In this study, we utilized in vitro primary hPDLCs cultures and in vivo ligature-induced periodontitis models in rats to demonstrate the dual anti-inflammatory and osteogenic effects of diosmetin, alongside its protective role against alveolar bone resorption in periodontal disease. RESULTS: CCK-8 assays demonstrated that diosmetin exhibited no cytotoxicity in hPDLCs at concentrations up to 100 μM. Diosmetin significantly suppressed LPS-induced mRNA and protein expression of pro-inflammatory mediators, including COX-2, IL-1β, and IL-6. The MAPK and NF-κB signaling pathways may represent the underlying mechanisms for its anti-inflammatory effects. Quantitative analysis further revealed upregulated mRNA and protein levels of osteogenic markers-RUNX-2, BMP-2, and BSP, which may be associated with Wnt/β-catenin signaling activation. Alkaline phosphatase (ALP) and alizarin red S (ARS) staining validate the osteogenic differentiation potential of diosmetin in hPDLCs. In the rat ligature-induced periodontitis model, administration of diosmetin significantly reduced the RANKL/OPG ratio, potentially contributing to its inhibition of alveolar bone resorption. CONCLUSION: This study demonstrated the dual roles of diosmetin's anti-inflammatory and osteogenic differentiation effects in preventing periodontitis, along with a preliminary exploration of its underlying mechanisms.