Expression of Oct-4, SOX-2, and MYC in dental papilla cells and dental follicle cells during in-vivo tooth development and in-vitro co-culture.

Abstract

During tooth development, the special structure of dental follicle and dental papilla enables dental papilla cells (DPCs) and dental follicle cells (DFCs) to make contact with each other. Octamer-binding transcription factor 4 (Oct-4), sex determining region Y box-2 (SOX-2), and cellular homologue of avian myelocytomatosis virus oncogene (MYC) (OSM) are associated with reprogramming and pluripotency. However, whether the expression of OSM could be activated through cell-cell communication is not known. In this study, the distribution of OSM in rat tooth germ was investigated by immunohistochemical staining. An in-vitro co-culture system of DPCs and DFCs was established. Cell proliferation, cell apoptosis, cell cycle stages, and expression of OSM were investigated by Cell Counting Kit 8 (CCK8) analysis, flow cytometry, real-time PCR, and immunohistochemical staining. We found that Oct-4 and SOX-2 were strongly expressed in tooth germ on days 7 and 9 after birth, whereas MYC was expressed only on day 9. Cell proliferation and apoptosis were inhibited, the cell cycle was arrested in the G0/G1 phase, and the propidium iodide (PI) value was downregulated. Expression of Oct-4 and SOX-2 was significantly elevated in both cell types after 3 d of co-culture, whereas expression of MYC was not significantly elevated until day 5. These results indicate that the optimized microenvironment with cell-cell communication enhanced the expression of reprogramming markers associated with reprogramming capacity in DPCs and DFCs, both in vivo and in vitro.