Ltcc Channel Enhances Odontogenic Differentiation Via Erk Signaling In Dpscs

This study aimed to determine whether L-type calcium channel (LTCC) agonist BayK8644 promotes odontogenic differentiation in human dental pulp stem cells (hDPSCs), addressing a gap in understanding the role of calcium signaling in dental regeneration. hDPSCs were isolated from healthy donors (ethics approval: #2021-045) and cultured with BayK8644 (0.1∼10 μM). Odontogenic differentiation was assessed via alkaline phosphatase (ALP) activity (day 7), Alizarin Red staining (day 21), and RT-qPCR for ALP, dentin sialophosphoprotein (DSPP), and osteocalcin (OCN) (day 14). A subcutaneous pulp regeneration model in immunodeficient mice (n=8/group) evaluated mineralized tissue formation after 8 weeks. Mechanistic analysis included immunofluorescence for Cav1.2, Western blotting for ERK phosphorylation, and calcium imaging. Data were analyzed using ANOVA test (α = 0.05). BayK8644 significantly increased ALP activity, mineralization, and odontogenic gene/protein expression. In vivo, BayK8644-treated groups exhibited more mineralized tissue and elevated DSPP expression. ERK phosphorylation and calcium oscillations were enhanced, while ERK inhibition abolished differentiation. BayK8644 drives hDPSC odontogenic differentiation via ERK signaling, identifying a novel therapeutic target for dentin-pulp regeneration. This study provides the first mechanistic evidence linking LTCC activation to ERK-mediated dental tissue engineering.