Berberine Promotes Hpdlsc Osteogenic Differentiation Through Egfr

To further investigate the affinity of Berberine (BER) with epidermal growth factor receptors (EGFR) and to clarify that BER promotes osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) through activation of ERK-fos pathway by EGFR. Periodontal ligament tissues from orthodontic teeth were used to culture hPDLSCs, EGFR-overexpressing hPDLSCs were constructed by lentiviral infection. The target sites and related signaling pathways of BER binding to hPDLSCs were predicted by network pharmacology and bioinformatics, the affinity properties of BER and EGFR were examined using cell membrane chromatography (CMC), surface plasmon resonance biomolecular interaction analyzer (Biacore), molecular docking and dynamics. BER (0.1 mg/L), EGFR inhibitor Gefitinib (GEF, 2μg/L), and ERK inhibitor PD98059 (50μM) were applied to hPDLSCs, EGFR-hPDLSCs, and EGFR NC-hPDLSCs cells. Expression levels of pathway factors (ERK), intracellular target genes (EGR1, fos) and osteogenic factors (ALP, OPN, OCN, COL1) were analyzed; osteomineralization was observed by alkaline phosphatase staining and alizarin red staining. BER promoted osteogenic differentiation of hPDLSCs through the EGFR-ERK-fos pathway. CMC, Biacore, molecular docking and dynamics all showed BER and EGFR have good affinity performance. BER elevated intracellular levels of target genes and osteogenic factors and promoted osteogenic mineralization in hPDLSCs. EGFR overexpression cells enhanced the above effects of BER, but GEF and PD98059 decreased the effects. This study demonstrated that BER can further regulate the EGFR-ERK-fos pathway to promote osteogenic differentiation of hPDLSCs through affinity for EGFR targets on the cell membrane of hPDLSCs.