This study explored the role of Piezo1 in the odontogenic differentiation of dental papilla cells (DPCs) and tissue, focusing on a mechanism involving family with sequence similarity 83, member G (FAM83G). Here, we found Piezo1, a mechanosensitive cation channel, was upregulated during odontogenesis in DPCs and dental papilla tissues. Knockdown of Piezo1 impaired odontogenic differentiation, while its activation by Yoda1 enhanced the process. Using a 3D culture model and an ectopic transplantation model, we confirmed Piezo1’s role in vivo. RNA sequencing (RNA-seq) analysis revealed that FAM83G was upregulated in Piezo1-knockdown cells, and FAM83G silencing enhanced odontogenesis in DPCs. These findings indicate that Piezo1 positively regulates odontogenesis by inhibiting FAM83G in DPCs both in vitro and in vivo, with Piezo1 representing a potential target for dental tissue regeneration.

The differentiation of dental papilla cells (DPCs) into functional odontoblasts is critical for dentinogenesis, yet the role of mitochondrial dynamics remains unclear. Here, we investigated the functional role of mitochondrial fission and mitochondria-associated endoplasmic reticulum membranes (MAMs) in the odontogenic differentiation of DPCs. Using in vitro differentiation models combined with confocal microscopy, transmission electron microscopy, and gain- and loss-of-function approaches, we found that odontogenic induction triggered early mitochondrial fragmentation and increased MAM formation. Dynamin-related protein 1 (DRP1) mediated mitochondrial fission, which in turn regulated MAM architecture and promoted differentiation. Malic enzyme 2 (ME2) acted as an upstream regulator, facilitating DRP1 recruitment and organizing MAM integrity. Notably, disruption of the ME2-DRP1-MAM axis impaired dentin formation both in vitro and in vivo, either by ME2 knockdown or pharmacological inhibition of DRP1 (Mdivi-1). These findings establish the ME2-DRP1-MAM axis as a critical metabolic–organellar switch driving odontoblast differentiation, providing new mechanistic insights into dentinogenesis and identifying potential therapeutic targets for dentin–pulp complex regeneration.