Abstract:
Human dental pulp cells (hDPCs) have shown their plasticity to differentiate into odontoblast-like cell lineages under the treatment of two-members of hydroxamates HDAC inhibitors (HDACis), Trichostatin A (TSA) and Suberoylanilide hydroxamic acid (SAHA). However, a comparison of the potency for stimulating odontoblast-like differentiation and mineralization process among these two HDACis has not been reported. Therefore, we aimed to confirm and compare the stimulatory effect of TSA and SAHA in inducing odontoblast-like differentiation and promoting mineralized-nodule formation. The primary cultured hDPCs was used for MTT assay, ALP activity assay, and alizarin red staining in the presence and absence of TSA and SAHA, with various concentration. Odontoblast-related gene expression was observed by RT-qPCR. Scratch wound healing assay was performed to observe the TSA and SAHA effects on cell migration. Either TSA or SAHA treatment have no effect on hDPCs viability as observed by MTT assay. The presence of TSA and SAHA induced odontoblast-like differentiation, confirmed by; the significant increase of ALP activity and mineral deposition during TSA 400 nM or SAHA 1µM treatment, the significant acceleration pattern of NFI-C, KLF4, DMP1, DSPP, COL1, ALP, BSP, OC, VEGF, and p21 gene expressions analyzed by RT-qPCR, at 24h, 72h, 7d and 5d. Furthermore, Scratch wound healing assay displayed the enhanced cell migration at 72h after TSA or SAHA treatment. Our findings showed that TSA and SAHA might have similar stimulatory effect in inducing odontogenic differentiation and mineralization process of hDPCs and proposed another potential application of TSA and SAHA to promote dentin regeneration.
