Indiana University-Purdue University Indianapolis (IUPUI) / Background: Numerous studies have reported the effect of long-term use of calcium hydroxide Ca(OH)2 to dentin. Nevertheless, there is little information available about the effect of Ca(OH)2 on wettability to the dentin. Objective: To investigate the effect of Ca(OH)2 application on dentin for two and four weeks on the wettability of two root canal sealers. Methods: Polished caries-free human dentin discs (n = 156) were allocated into 12 groups; G1 and G3 had two weeks’ treatment, G4 and G6, four weeks treatment. G1 and 101 G4 were treated with sterile water. G2, G3, G5 and G6 were treated with Ca(OH)2. G1, G3, G4, and G6 were irrigated with 6.0-percent NaOCl and 17-percent EDTA while G2, and G5 were irrigated with sterile water. Then, contact angles between Tubli-Seal and the treated dentin surfaces were measured. G7 and G12 were treated in the same fashion but were treated with BC sealer. Surface morphology evaluation of G1 and G6 was carried out by scanning electron microscopy (SEM) and energy-dispersive x-ray spectroscopy (EDX). Statistics were performed using three-way ANOVA and pair-wise comparisons between groups (α = 0.05). Results: Tubli-Seal (G1 through G6) had significantly smaller values for contact angles than BC sealer (G7 through G12) (p < 0.05). For the Tubli-Seal groups (G1 through G6), G4 had the highest mean of contact angles at 104.9 ± 1.9°, whereas G5 presented the lowest mean of contact angles at 85.4 ± 15.1. For the BC sealer groups (G7 through G12), G10 had the highest mean of contact angles at 145.4 ± 1.3°, while G11 demonstrated the lowest mean of contact angles at 130.2 ± 2.6°. Groups with Ca(OH)2 treatment with water irrigation (G2, 5, 11) had significantly lower contact angle than groups with Ca(OH)2 with chemical irrigation (G3, 6, 12) (p < 0.05), except G8, 9. According to SEM and EDX, water irrigation solution showed higher remaining Ca(OH)2 than irrigation with the chemical solution while Ca(OH)2 with chemical irrigation 102 demonstrated no Ca(OH)2 remaining after irrigation, similar to the surface of the control group. Conclusion: Within the limitations of this study, Tubli-seal has better wettability on dentin than BC sealer. Remaining calcium hydroxide demonstrated a trend toward decreased contact angle between dentin and root canal sealers. Moreover, two-minute irrigation with 6-percent NaOCl and 17-percent EDTA can remove calcium hydroxide from polished dentin surfaces.