Methods: Phase 1: hESCs were differentiated into hESCs-MSCs. hESCs-MSCs were derived into odontoblast like cells (hESCs-Odont) under the stimulation of FGF-8 and BMP-4. Phase 2: hESCs-Odont were seeded on 1 mm thick, 17 % EDTA treated coronal dentin disc in the presence of FGF-8 and BMP-4 for terminal differentiation.
Results: Phase 1: Our RT-PCR and Real Time PCR results confirmed the expression of odontontoblast specific markers (DSPP, DMP, ALP and Collagen I) at mRNA by hESCs-Odont. Alternatively we confirmed the expression of DSP by hESCs-Odont at protein level by immunofluorescence staining. Phase 2: hESCs-Odont on 17% EDTA treated dentin surface in the presence of FGF-8 and BMP-4 showed change in morphology from fibroblast-like shape into more spherical shape with well-formed cytoplasmic extension into the dentinal tubules. Results were confirmed by SEM. Alternatively, histological sectioning results showed the polarization of the hESCs-Odont under the same conditions.
Conclusions: In this study we demonstrated the hESCs as a possible novel cell source for odontoblastic differentiation. Alternatively, we demonstrated a novel effect of FGF-8 and BMP-4 growth factors in odontontoblastic differentiation from hESCs. Concurrently, this study showed the significant synergetic effect of different surface treatments of dentin scaffolds on terminal odontoblastic differentiation.