Abstract.Objective: This study aimed to evaluate cytotoxicity of three commercial brands of orthodontic elastomeric ligatures. Materials and Methods: The three commercial orthodontic elastomeric ligature brands were C-brand, T-brand and U-brand from different manufacturing countries. The cytotoxic assay was performed using cell cultures (L929) which were subjected to the cell viability test with methyl-tetrazolium (MTT). The L929 cell line was grown in 96-well tissue culture plates (1x10 5 cells/cm 3 ). Elastomeric ligatures (0.2 g) from each brand was placed in 1 ml of Minimum Essential Medium (MEM) at 37°C with 5% CO2 at 100% humidity in an incubator for 24 hours. After incubation, the cell mitochondrial activity was evaluated by the MTT test for detecting viable cells from optical density according to ISO 10993-5: 2009 (E) test procedure. Cytotoxic effects were determined by the percentage of cell viability. Results: Microscopic investigation showed that all commercial elastomeric ligatures caused alterations in the fibroblast cell morphology from an elongated to a spherical form. The cell viability of C-brand, T-brand and U-brand were 51.10, 73.04 and 93.04%, respectively. Conclusion: T-brand and U-brand orthodontic elastomeric ligatures were non cytotoxic potential. U-brand had the greatest cell viability and C-brand had the lowest cell viability at 24 hours.