Cytotoxic and mutagenic effects of root canal filling cements of various chemical composition were determined in vitro. Materials set for 24 h and 1 wk were eluted for 24 h in cell culture medium (cytotoxicity testing) and dimethyl sulfoxide or physiological saline (mutagenicity testing). The differences between cytotoxic potencies of eluates of the endodontic materials on L-929 cells were quantified colorimetrically (MTT test). Eluates of Traitment SPAD were about 5- to 30-fold more toxic than silver-free AH26, TubliSeal, CRCS, and Endomethsone N. The rank order of the toxic effects depended on the setting time of mixed materials. Dimethyl sulfoxide and saline eluates of Traitment SPAD, Tubli-Seal, Endomethasone N, CRCS, and Ketac-Endo were not mutagenic in the Ames test. Both eluates of silver-free AH26 set for 24 h were weakly mutagenic in Salmonella typhimurium TA100. Weak mutagenicity of saline eluates of the material was also observed in TA97a and TA102. These results point to the possibility that mixed silver-free AH26 might contain small amounts of two mutagenic substances: bisphenol A diglycidyl ether and formaldehyde.