Evaluation of Cytotoxicity of 4-Hydroxycinnamic Acid Using Tetrazolium Bromide Assay and Zebrafish Embryotoxicity: An In-Vitro Study

Abstract

Aim This study aimed to evaluate the cytotoxicity of a novel compound, 4-hydroxycinnamic acid (4-HCA), with the help of a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and zebrafish embryotoxicity. Materials and methods In this in vitro study, MTT fibroblast assays using dental pulp stem cells, which were cultured in Modified Eagle’s Medium or Dulbecco's Modified Eagle Medium, and zebrafish cytotoxicity and embryotoxicity were done to evaluate the cytotoxicity of the novel compound 4-HCA. The data was analyzed by plotting cell number versus absorbance, allowing quantitation of changes in cell proliferation. Results 4-HCA (40 μl) showed acceptable levels of cell viability according to the American Society for Testing and Materials standards. Cell viability is reduced with increased exposure time and concentrations of 4-HCA. Similarly, the cytotoxicity assessment in zebrafish ( Danio rerio ) showed an acceptable range of toxicity levels in embryonic stages used to evaluate the mortality rate of zebrafish embryos. Conclusion Considering the constraints of this research, it can be deduced that hydroxycinnamic acid at a concentration of 40 μl was non-toxic. The findings from the MTT assay indicated a correlation between the concentration and the toxicity of the compound. Likewise, the zebrafish test demonstrated minimal toxicological effects.