Aim: This in vitro study evaluated and compared sealing ability of Diadent BioAggregate (DBA) as a new root-end filling material (REFM) versus amalgam, intermediate restorative material (IRM) and white mineral trioxide aggregate (WMTA). Materials and Methods: Crowns of sixty extracted human maxillary incisors were sectioned at the cemento-enamel junction (CEJ). All the roots were instrumented and obturated with gutta-percha and resin sealer. Obturated roots were divided randomly into 2 control groups and 4 experimental groups of 10 samples each. In the negative control group (group I), roots were kept without any further preparation. In the positive and experimental groups roots, were apically resected and root-end cavities were prepared and filled with: (a) gutta-percha (group 2-positive control group); (b) amalgam (group 3); (c) IRM (group 4); (d) WMTA (group 5); (e) DBA (group 6). Apical leakage was assessed using dye penetration technique. Data were submitted to statistical analysis by the one-way analysis of variance (ANOVA) and Tukey′s test. Results: Significant difference of sealing ability was found among 4 tested groups. DBA followed by MTA showed the highest sealing ability. Conclusions: DBA with its high sealing ability can be considered a possible alternative to MTA.