Aim: To evaluate the cytotoxicity of 15% citric acid, 5% phosphoric acid and 2.5% NaOCl on cultured fibroblasts using MTT colorimetric assay. Methodology: Irrigating solutions of 5% phosphoric acid, 15% citric acid, and 2.5% NaOCl, diluted at 0.1% and 0.5%, were applied to cell cultures of 3T3L1 fibroblasts. The cell viability was determined by means of MTT colorimetric assay after a period of 1, 6 and 24 hours. Percentages of cell viability were analyzed using the Kruskal-Wallis test for global comparisons and the Mann-Whitney U-test for pairwise comparisons. Results: The percentage of cell viability diminished progressively over a 24 hour period in all solutions at both dilutions. At 0.1% dilution, 2.5% NaOCl (63.39%) and 15% citric acid (53.91%) showed the highest percentage of cell viability (p=0.083). At 0.5% dilution, 2.5% NaOCl again showed the highest cell viability value (48.51%). Conclusions: The irrigating solution with the highest percentage of cell viability was 2.5% NaOCl at both 0.1% and 0.5% dilutions. A very low percentage of cell viability was obtained with 15% citric acid and 5% phosphoric acid at 0.5% dilution.