H19 Facilitates Hdpscs Dentinogenesis Via Mir-103A3p/Pik3r1/Klf4 Pathway

Functional reconstruction of the dentin-pulp complex is critical for repairing damaged teeth, with odontogenic differentiation of human dental pulp stem cells (hDPSCs) serving as a key process of reparative dentin formation. This study aimed to elucidate the role and mechanism of H19-mediated miR-103a-3p in regulating hDPSC odontogenic differentiation, providing molecular targets to enhance reparative dentin regeneration. Bioinformatics tools and dual-luciferase reporter assays were utilized to validate miR-103a-3p as a downstream target of H19 and its interaction with PIK3R1/KLF4. Plasmid vectors were constructed to over/downexpress miR-103a-3p, PIK3R1, and KLF4. Functional effects were assessed using qRT-PCR, western blotting, alkaline phosphatase staining, and alizarin red S staining. Western blotting was performed to detect the activation of the PI3K/AKT pathway. Subcutaneous ectopic pulp regeneration models in nude mice were established to validate the role of the H19 in vivo. (1) Dual-luciferase assays confirmed that miR-103a-3p is a direct target of H19. Inhibition of miR-103a-3p significantly promoted hDPSCs odontogenic differentiation with concurrent upregulation of PIK3R1 and KLF4. (2) Direct binding between miR-103a-3p and PIK3R1/KLF4 was verified. KLF4 overexpression promoted odontogenic differentiation by upregulating mineralization markers, while PIK3R1 overexpression activated the PI3K/AKT pathway, synergistically amplifying dentinogenesis. (3) Stable overexpression of H19 in hDPSCs stimulated ectopic dentin-like structure formation in vivo. H19-mediated miR-103a-3p promotes odontoblastic differentiation of hDPSCs by activating PIK3R1-dependent PI3K/AKT signaling and enhancing KLF4-mediated mineralization.