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This study evaluated setting time (ST), radiopacity, pH, flow, solubility, and volumetric change (VC) of a silicone, gutta-percha, and bioactive glass-based sealer, GuttaFlow Bioseal (GFB), and a calcium silicate-based sealer, TotalFill BC Sealer (TFBC), in comparison with AH Plus.ST and flow were evaluated in accordance with the ISO 6876 Standard. pH was evaluated after different time intervals (1, 3, 7, 14, 21, and 28 days). Radiopacity was evaluated by radiographic analysis in millimeters of aluminum. Solubility was evaluated by means of mass loss (%) after 7 and 30 days of immersion in distilled water. VC was evaluated by micro-computed tomography, by using cavities 3 mm deep and 1 mm in diameter in acrylic resin, filled with the materials. The materials were evaluated after setting and after 7 and 30 days of immersion in distilled water. The data were submitted to analysis of variance and Tukey statistical tests (P < .05).TFBC demonstrated the highest pH and solubility. GFB had the shortest ST, and lowest radiopacity and flow values. VC was similar for the sealers in both time intervals.TFBC presented the highest pH and solubility, but showed similar VC to GFB and AH Plus. GFB showed proper physicochemical properties. Micro-computed tomography complements the physicochemical analysis of endodontic sealers.A silicone, gutta-percha, and bioactive glass-based sealer, GuttaFlow Bioseal, was evaluated.Endodontic sealer volumetric change was evaluated by using micro-computed tomography (micro-CT).GuttaFlow Bioseal showed volumetric change similar to TotalFill BC Sealer and AH Plus.Micro-CT complements the physicochemical analysis of endodontic sealers.