Abstract Background MiR-224-5p has been proven to play an important role in regulating cell differentiation. This study aimed to clarify the regulatory role and mechanism of miR-224-5p in the osteogenic differentiation of human dental pulp stem cells (hDPSCs), thereby laying a theoretical foundation for subsequent jaw defect repair. Methods Human dental pulp stem cells (hDPSCs) were isolated, cultured, and sorted from healthy dental pulp tissues. We performed integrated bioinformatics analysis to screen and identify the potential targets and pathways of miR-224-5p involved in the osteogenic induction of hDPSCs. Subsequently, in vitro experiments were conducted. Plasmid transfection was used to regulate the overexpression and knockdown of miR-224-5p in hDPSCs, and the expression of osteogenesis-related proteins was detected. Furthermore, luciferase reporter assays and Western blot assays were used to confirm the direct targets of miR-224-5p, and rescue experiments were performed to verify the underlying mechanism. Results The results demonstrated that overexpression of miR-224-5p inhibited the osteogenic differentiation of DPSCs, as reflected by the significantly decreased expression of osteogenic markers (OCN, Runx2, and ALP). In contrast, inhibition of miR-224-5p promoted the osteogenic differentiation of DPSCs. Bioinformatics analysis and dual-luciferase reporter gene assays indicated that miR-224-5p specifically targets the 3’ untranslated region of the PTEN gene. Rescue experiments further confirmed that miR-224-5p regulates this process by modulating the PTEN/PI3K/AKT pathway. Conclusions Inhibition of miR-224-5p promotes osteogenesis in DPSCs by targeting the PTEN/PI3K/AKT signaling axis. These findings provide reliable evidence for the fabrication of three-dimensional tissue-engineered structures and further repair of maxillofacial bone defects.
Ke et al. (Sat,) studied this question.