BACKGROUND Successful root canal treatment depends on proper cleaning, disinfecting and shaping of the root canal space. Pulpless teeth have lower dentin microhardness value compared to that of vital teeth. A material which can cause change in dentin composition may affect the microhardness. Thus the aim of this study was to evaluate and compare the effect of two root canal sealers on dentin microhardness. METHODS Forty two single rooted teeth were selected and divided into 3 equal groups; Apexit, iRootSP and control groups (n=14) Each group was then divided into 2 subgroups according to the post evaluation period; 1 week and 2 months (n=7). Root canal procedure was done in the experimental groups and obturation was made using either; Apexit, iRootSP or left unprepared and unobturated in the control group. Roots were sectioned transversely into cervical, middle and apical segments. The three sections of each root were mounted in a plastic chuck with acrylic resin. The coronal dentin surfaces of the root segments werepolished. Microhardness of each section was measured at 500 µm and 1000 µm from the canal lumen. RESULTS Four way-ANOVA revealed that different tested sealer materials, canal third, measuring distance from the pulp and time as independent variables had statistically non significant effect on mean microhardness values (VHN) at p≤0.001. Among iRootSP groups there was a statistically significant difference between iRoot SP at coronal root portion (87.79±17.83) and iRoot SP at apical root portion (76.26±9.33) groups where (p=0.01). IRoot SP at coronal canal third had higher statistically significant mean microhardness value (87.79±17.83) compared to Apexit at coronal third (73.61±13.47) where (p=0.01). CONCLUSIONS Root canal sealers do not affect dentin microhardness. Key words:Root canal, dentin, sealers, microhardness, bioceramic.