Objectives: It is known that bioactive materials interact with the dentin to undergo biomineralization. The exact role of moisture in this interaction is unknown. Here, we investigate the effects of dentin moisture conditions on the dislocation resistance of two bioactive root canal sealers (MTA Fillapex [Angelus Solucoes Odontologicas] and GuttaFlow BioSeal [Colténe/Whaledent AG]) at 3 weeks and 3 months after obturation. Materials and Methods: Mandibular premolars (n = 120) were prepared and randomly divided into 3 groups based on the dentin condition: group 1, dry dentin; group 2, moist dentin; group 3, wet dentin. Each group was divided into 2 subgroups for root canal filling: MTA Fillapex and GuttaFlow BioSeal. Dislocation resistance was evaluated by measuring the push-out bond strength at 3 weeks and 3 months. Failure modes were examined under a stereomicroscope. Data were statistically analyzed by Kruskal-Wallis test with a significance level of 5%. Results: Moist dentin resulted in higher bond strength values for both materials at both time points. This was significantly higher than wet and dry dentin for both the sealers at the 3 months (p < 0.05), while at 3 weeks it was significant only for GuttaFlow Bioseal. The different moisture conditions demonstrated similar trends in their effects on the dislocation resistance of the 2 root canal sealers. Conclusions The dentin moisture conditions had a significant impact on its interaction with the bioactive materials tested. Maintaining moist dentin, but not dry or wet dentin, may be advantageous before the filling root canals with bioactive sealers.