The purpose of this study was to evaluate the cytotoxicity of four root canal sealers(Tubliseal, AH26, Apatite Root Canal Sealer I, Apatite Root Canal Sealer II) in Vitro. The root canal sealers were mixed and filled in molds which were $14{ imes}1.25mm$ in diameter, in height to use for cell counting and agar overlary method, and $7{ imes}1.25mm$ for millipore filter method and set for 7 days to use for experiment. Silicone and copper plate were used for negative and positive control respectively. Using the culture of L929 fibroblast, total cell number and vital cell number were counted and the ratio of vital cell number to total cell number was calculated on 2 nd, 4 th, 6 th experimental day, and the change of cell membrane permeability was tested by agar overlay method, and the succinate dehydrogenase activity was tested by millipore filter method. The obtained results were as follows. 1. In ail experimental groups, the mitotic activity of fibroblast was reduced when compared with that of negative control group, so ail experimental groups showed cytotoxicity. Apatite Root Canal Sealer I group exhibited mild cytotoxicity, and Tubliseal, AH26, Apatite Root Cenal Sealer II groups exhibited severe cytotoxicity. 2. In the test of the change of cell membrane permeability by agar overlay method, all experimental groups showed cytotoxicity. AH26 group exhibited mild cytotoxicity, and Apatite Root Canal Sealer I group exhibited moderate cytotoxicity, and Tubliseal and Apatite Root Canal Sealer II group exhibited severe cytotoxicity. 3. In the test of SDH activity by millipore filter method, there was no cytotoxicity in Apatite Root Canal Sealer I and Apatite Root Canal Sealer II group, but Tubliseal and AH26 group showed mild cytotoxicity.