Abstract

BACKGROUND Employing a comprehensive strategy that integrated network pharmacology, molecular docking, and in vitro experimental techniques, this study examined the effect of neohesperidin on the proliferation and osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) upon lipopolysaccharide (LPS)-induced inflammatory conditions. METHODS Core genes were screened using molecular docking and network pharmacology. hPDLSCs were isolated and cultured, and their surface markers were identified by flow cytometry. The effects of neohesperidin on hPDLSC proliferation and osteogenic differentiation upon LPS stimulation were investigated using ELISA, CCK-8, alkaline phosphatase, alizarin red staining, qRT-PCR, and Western blot analyses. RESULTS hPDLSCs highly expressed CD90 but did not express CD34 and CD45, which is consistent with the characteristics of stem cells. Network pharmacology and molecular docking analyses revealed that inflammatory and osteogenesis-related signaling pathways were significantly enriched. Treatment with 50 μM neohesperidin promoted proliferation, alkaline phosphatase activity, and mineralized nodule formation while suppressing the inflammatory response in LPS-stimulated hPDLSCs (P < 0.05). It also upregulated the expression of osteogenic markers, such as Runx family transcription factor 2 (RUNX2), osteocalcin (OCN), and osterix (OSX) (P < 0.05). Furthermore, neohesperidin activated the PI3K/AKT signaling pathway and reversed the inhibitory effect of LY294002 (P < 0.05). CONCLUSION Through PI3K/AKT activation, neohesperidin stimulated the osteogenic transformation of hPDLSCs under inflammatory conditions, suggesting that it may be a pharmacologically active compound with the potential for use as an adjuvant in periodontitis treatment.