Background
The
Conclusion
Phenytoin has the ability of promoting the differentiation of hDPSC into odontoblasts and osteoblasts. BMP4/Smad pathway, inducing odontogenic/osteogenic differentiation of hDPSC, appears a main process in this context.
Methods
Fourth-generation human hDPSC originating from healthy pulp of third molars was cultured in control as well as phenytoin-containing media (PHT) for 14 days. qPCR was applied to detect the expression of DSPP, DMP1, and ALP genes. Western blot analysis was used to confirm the findings. One-way analysis of variance (ANOVA) was used for statistical analysis (p < 0.05). Information about phenytoin was assessed from PubChem database, while targets of phenytoin were assessed from six databases. Drug targets were extracted based on the differentially expressed genes (‖logFC‖ ≥ 1, p < 0.05) in the experimental group (50 mg/L PHT, 14 days). GO BP and KEGG pathway enrichment analysis on the obtained drug targets was performed and the target protein functional network diagram was constructed.
Results
A concentration below 200 mg/L PHT had no obvious toxicity to hDPSC. The expression of DSPP, DMP1, and ALP genes in the 50 mg/L PHT concentration group increased significantly. The WB experiment showed that the protein content of BMP4, Smad1/5/9, and p-Smad1/5 was significantly increased in 50 mg/L PHT in comparison with the NC group (the group without treatment of PHT) at 14 days.