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An important requirement for dental materials placed in direct contact with living tissues is biocompatibility. The purpose of this study was to evaluate the antiproliferative activity of three dental materials (mineral trioxide aggregate, zinc oxide-eugenol cement, and glass-ionomer cement) against a panel of established fibroblastic cell lines (L929, BHK21/C13, and RPC-C2A). The materials were prepared according to the manufacturer's instructions and were tested in insert wells for 12, 24, and 48 h. Cell number fraction was estimated by the sulforhodamine-B assay, in reference to controls. The degree of antiproliferative effect in ascending order was mineral trioxide aggregate, glass-ionomer cement, and zinc oxide-eugenol cement in all cell lines tested.