Gutta-percha (GP) has been the most widely used root canal filling material because of its well-known low toxicity. The inertness of GP, however, was challenged recently. The purpose of this study was to evaluate the toxicity of marketed endodontic GP using the radiochromium release test. Fourteen commercially available and three experimental GP brands were tested. Raw GP, zinc oxide, and barium sulfate, which were considered major components of GP points, and zinc ions were also evaluated. The material was spread to cover the bottom of testing wells after being dissolved in chloroform or warmed. A labeled suspension of L929 cells was added to the wells. After incubation at 37 degrees C for 4 and 24 h, extracellular radiochromium in the culture medium was measured and calculated in percentage of the total intracellular label. Spontaneous release of radiochromium was used as control and the results were considered to be within normal limits either at 4 or 24 h. All chloroform-dissolved GP showed low toxicity at 4 h, whereas warmed GP showed statistically significant differences at 4 h. Both dissolved and warmed GP were toxic at 24 h. The raw materials and barium sulfate were not toxic, whereas zinc oxide and zinc ions showed marked toxicity. All GP points tested were toxic at longer observation periods, and the toxicity was attributed to leakage of zinc ions into the fluids.