The aim of the present in vitro study was to evaluate specific mechanical and physicochemical properties of two calcium silicate based sealers, (AH Plus Bioceramic “AHPB”; Well-Root ST “WRST”), and a conventional resin sealer (AH Plus “AHP”). These aims were accomplished by assessing the porosity, pH, compression strength, roughness, wettability and cell attachment of the tested materials. The results were compared statistically using the one-way ANOVA test. Higher pH values were obtained in both AHPB and WRST compared to AHP at 3, 24 and 72 h (p < 0.05). A greater level of porosity and wettability was detected for both AHPB and WRST compared to the resin sealer AHP (p < 0.05). Evident cell growth characterized by elongated morphology was observed on the surface of AHPB and WRST, while only a thin layer of cells was seen on the surface of AHP. A significant lower compression strength and modulus were obtained in the specimens created using AHPB compared to those made with AHP and WRST (p < 0.05). The removal of calcium silicates may be quite tricky during endodontic retreatment. In conclusion, considering the limitations of the present in vitro study, both calcium silicate sealers demonstrated good physicochemical properties. However, the lower compression strength and modulus of AHPB may facilitate its removal and make the retreatment procedures considerably easier.