Cytotoxic Activity of Gambier Leave (Uncaria gambir) Ethyl Acetate Extract on Mouse Embryonic Fibroblast Cell (NIH-3T3) using MTT Assay

Abstract

Background Uncaria gambir or Gambier is one of plant that widely found in Indonesia. Gambier is locally known as an antioxidant and antibacterial agent because it has high catechin content. Ethyl acetate extract of gambier leaves has been investigated to contain the highest catechin content than other extraction solvents. Fibroblasts are often used in biomaterial viability and toxicity tests because they have a highly reproducible growth rate and biological response. NIH-3T3 is commonly used as a substitute for human gingival fibroblasts. However, no study has been evaluated conducted to the cytotoxic activity of gambier extract on fibroblast cells. Objective The aim in this study is to evaluate whether the cytotoxic activity of gambier ethyl acetate extract (GEE) exerts on NIH-3T3 cell lines with MTT assay. Methods The group of cytotoxic activity of gambier extract was evaluated in three incubation periods. The cytotoxicity test was conducted using an ethyl acetate extract of gambier (Uncaria gambir Roxb.) leaves. The NIH-3T3 cell was treated by GEE in ten concentrations (0, 2.5, 5, 10, 25, 50, 100, 250, 500, and 1000 ppm) for 24-, 48, and 72-hours incubation periods. Cell viability was determined with MTT (3-4,5-dimethylthiazol-2yl -2,5-diphenyltetrazolium bromide) assay. The data was analyzed statistically using SPSS based on ANOVA followed by Tukey HSD post hoc with p<0.05, ANOVA paired sample T test with p<0.05, and the CD50 value was measured by Sigma Plot software. Result GEE 2.5, 5, 10, 25, 50, 100, 250 ppm have viability cell% >80%, and increase the viability cell% based on incubation period. GEE 1000 ppm significantly decreased the viability cell % from GEE 0 ppm in 24-, 48-, and 72- hours incubation periods (23.83%, 30.14%, and 19.02%, respectively). Moreover, GEE 500 ppm becomes toxic by significantly decreasing the viability cell % in 48- and 72-hours (40.43% and 23.03%, respectively). The CD50 value of GEE at 24-, 48-, and 72-hours incubation are 578.03 ppm, 488.63 ppm, and 470.70 ppm, respectively. Conclusion GEE 2.5, 5, 10, 25, 50, 100, 250 ppm has not toxic to NIH-3T3 cells for 24-, 48-, and 72-hours incubation periods.