The purpose of the present study was to compare the cytotoxicity of mineral trioxide aggregate (MTA) to other commonly used retrofilling materials, Super-EBA and amalgam. This was accomplished using a cell viability assay for mitochondrial dehydrogenase activity in human periodontal ligament fibroblasts after 24-hr exposure to extracts of varying concentrations of the test materials, in both freshly mixed and 24-hr set states. Methyl methacrylate 2% (vol/vol) served as the positive control, and complete culture medium served as the negative control. Differences in mean cell viability values were assessed by ANOVA (p < 0.05). In the freshly mixed state, the sequence of toxicity was amalgam > Super-EBA > MTA. In the 24-hr set state the sequence of toxicity at a low extract concentration was Super-EBA > MTA, amalgam, and Super-EBA > amalgam > MTA at a higher extract concentration. This study supports the use of MTA in the root-end environment.