Differentiation and biomineralization induction potential of caffeic acid and caffeic acid phenethyl ester on pulp cells.

Abstract

Studies are searching for compounds capable of inducing stem cell differentiation that can be incorporated into scaffolds for application in endodontic regeneration techniques for the treatment of young permanent teeth. Bioactive compounds derived from plants, such as caffeic acid (CA) and its derivative caffeic acid phenethyl ester (CAPE), have shown an effect on the differentiation and induction of mineralization markers in osteoblastic cells, but studies have not yet been found in pulpal cells. Thus, this research aims to evaluate the differentiation-inducing effect and the biomineralizing potential of CA and CAPE on human dental pulp stem cells(hDPC). hDPC will be cultured in alpha-MEM medium and exposed to serial concentrations of CA, CAPE and calcium hydroxide solution (control) and cytotoxicity determined by the Alamar blue® assay for 48 and 72h. Once the non-cytotoxic concentrations have been determined, the hDPCs will be cultured in osteogenic alpha-MEM or alpha-MEM for 7, 14 and 28 days and cell viability, alkaline phosphatase (ALP) activity and deposition of mineralization nodules (NM) by theAlamar blue, thymophthalein and alizarin red S methods. hDPC will be exposed to the concentrations of the compounds with the best effect on ALP and NM and the expression of the dentin matrix protein genes - 1 (DMP-1) and dentin sialophosphoprotein (DSPP) will be evaluated.by quantitative PCR. Data will be statistically analyzed considering p<0.05. If the data values present normal distribution (homogeneity), the variables will be submitted to analysis of variance and Bonferroni, if they present heterogeneity, they will be submitted to the Kruskal-Wallis and Student-Newman-Keuls tests.