H19 promotes odontogenic differentiation of human dental pulp cells via miR-103a-3p-mediated PIK3R1/AKT and KLF4 pathways

Background Functional regeneration of the dentin–pulp complex is essential for restoring tooth integrity after injury. Odontoblastic differentiation of dental pulp stem cells (DPSCs) plays a central role in reparative dentinogenesis. Although lncRNA H19 is known to regulate biomineralization, its downstream network remains unclear. This study identified miR-103a-3p as a novel downstream effector of H19 and investigated its regulatory network in DPSCs odontoblastic differentiation. Methods Interactions between H19 and miR-103a-3p as well as miR-103a-3p and targets mRNAs (PIK3R1 and KLF4), were validated via bioinformatic and dual luciferase reporter assays. Quantitative Real-Time PCR (qRT-PCR) and western blots were used to investigate the expression pattern of H19 and its potential signal axis and odontogenic markers. Alkaline phosphatase (ALP) and alizarin red S (ARS) staining were used to evaluated odontogenic ability. Finally, a heterotopic pulp regeneration model was established to reveal the regulating effects of H19. Results H19 acted as a sponge for miR-103a-3p, which otherwise inhibited the expression of odontogenic markers. PIK3R1 and KLF4 were identified as direct targets of miR-103a-3p. Overexpression of either PIK3R1 or KLF4 rescued the odontogenic differentiation capacity of DPSCs suppressed by miR-103a-3p. Furthermore, PIK3R1 promoted odontogenesis by activating the PI3K/AKT signaling pathway, while KLF4 functioned as an independent transcriptional regulator. Finally, in a heterotopic pulp regeneration model, H19 overexpression enhanced the expression of PIK3R1 and KLF4 and promoted odontoblastic differentiation of DPSCs in vivo. Conclusion Our results suggest that the H19-mediated miR-103a-3p/PIK3R1/AKT and miR-103a-3p/KLF4 axes promote the odontogenic differentiation of DPSCs and are expected to serve as therapeutic targets for pulp regeneration.