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The attachment and morphology of human periodontal ligament fibroblasts to mineral trioxide aggregate (MTA) was evaluated using a scanning electron microscope. The material was placed at an apical cavity of 30 single-rooted slices of extracted human teeth. The specimens were divided into two groups of 15 root slices each (freshly mixed and set state). For each experimental group, five root slices were used per observation period (4, 8, and 24 h). A set of two glass slides was used per observation period for the control group. The experiments were performed in tissue-culture cluster 96-well plates in which 1 ml of human periodontal ligament fibroblast cell suspension was placed over the MTA filling and the control glass slides. For the positive-control group, 0.5 ml of methyl methacrylate 2% (vol/vol) was added to the cell suspensions before being dispensed into the wells. Results showed the normal cell morphology in the negative controls. Few round cells with less smooth surfaces and many rough blebs were seen in the positive control, and most of these cells did not show any attachment to the substratum. Similar observations were seen with the freshly prepared-MTA group. In the set-MTA group, cells were round and flattened, displayed smooth surfaces, and appeared to be tightly attached to MTA. It was concluded that the quality and quantity of cell attachment to the retrofilling material could be used as a criterion to evaluate material's toxicity.