YTHDC1 Modulates the Osteogenic Capacity of hPDLSCs via Wnt/β-Catenin Signalling Pathway for the Treatment of Bone Defects in Osteoporosis Rats

Abstract

Human periodontal ligament stem cells (hPDLSCs) have emerged as promising candidates for the treatment of osteoporotic bone defects. Previous studies have indicated that m⁶A plays a crucial role in regulating the osteogenic differentiation of hPDLSCs. However, research on the relationship between YTHDC1, as a reading protein, and the osteogenic differentiation of hPDLSCs remains unexplored. This study aimed to investigate the biological roles of YTHDC1 in the osteogenic differentiation of hPDLSCs and to explore underlying mechanisms. Dot blot analysis revealed a progressive increase in m⁶A methylation during osteogenic differentiation, accompanied by significant upregulation of YTHDC1 expression, as evidenced by qPCR and Western blot. Functional assays utilising siRNA-mediated knockdown and lentiviral-mediated overexpression demonstrated that YTHDC1 positively regulated the osteogenic differentiation potential of hPDLSCs. Mechanistically, mRNA-seq analysis implicated the Wnt/β-catenin signalling pathway, which was further validated through rescue experiments with the Wnt inhibitor DKK1. Notably, in vivo experiments showed that hPDLSCs overexpressing YTHDC1 exhibited enhanced bone formation capacity in the osteoporotic rats. In conclusion, our findings suggested that YTHDC1 modulated the osteogenic capacity of hPDLSCs through the Wnt/β-catenin signalling pathway, highlighting its therapeutic potential for treating bone defects in osteoporotic conditions.

Keywords

bone regeneration / hPDLSCs / N6-methyladenosine / Wnt/β-catenin signalling pathway / YTHDC1

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