OBJECTIVES: This study aimed to investigate the effects of aspirin on the early stages of odontogenic differentiation. The roles of FOXC1, RUNX2, and MCAM gene expression in the mechanism of odontogenic differentiation were evaluated by examining the effects of downregulated FOXC1 or RUNX2 expression using small interfering RNAs (siRNAs). METHODS: Dental pulp cells were treated with aspirin (0, 2.5, 50, 100 μg/ml) to assess its impact on mineralization. The gene expression levels of FOXC1, RUNX2, and MCAM were measured using digital polymerase chain reaction, and the effects of siRNA-mediated knockdown of FOXC1 and RUNX2 were analyzed. The mineralization potential was quantitatively assessed using Alizarin Red S staining and a calcium assay. RESULTS: Analysis of cell growth curves and doubling times indicated that aspirin did not affect cell proliferation at 2.5 μg/ml and 50 μg/ml
however, 50 μg/ml aspirin promoted mineralization. In the FOXC1 and RUNX2 knockdown experiments, fluctuations in FOXC1, RUNX2, and MCAM gene expression were observed in the aspirin-treated group, suggesting the involvement of these genes in mineralization. Alizarin red S staining and calcium assays further demonstrated that aspirin enhanced mineralization. CONCLUSIONS: These findings indicate that aspirin promotes odontogenic differentiation and regulates the expression of FOXC1, RUNX2, and MCAM. This suggests that aspirin may serve as a promising new therapeutic agent in dental pulp regenerative medicine.