The aim of the present in vitro study was to evaluate the intratubular penetration of three bioceramic sealers in root canal retreatment. Here, 30 single-rooted human teeth were instrumented with the Protaper Universal system and filled with gutta-percha and the epoxy-resin-based sealer AH Plus mixed with rhodamine B. After two weeks in a humid environment, they were re-instrumented with Reciproc Blue and divided into three groups according to the endodontic sealer to be used in the re-filling (n = 10): G1: CeraSeal, G2: TotalFill BC Sealer, G3: TotalFill BC Sealer HiFlow. For the filling, a single cone technique was used, and the respective sealers were mixed with fluorescein. The roots were then sectioned at 2, 5, and 8 mm (apical, medial, and coronal measurement points, respectively) from the apex, and the dentinal tubule penetration depth and percentage of penetration around the canal perimeter were evaluated by means of confocal laser scanning microscopy (CLSM). Penetration between groups was compared using the Kruskal−Wallis test, and within each group using the Wilcoxon test. Statistical significance was established at p < 0.05. A non-significant reduction was found in the penetration depths and in a percentage of penetration around the canal perimeter between AH Plus and the tested calcium-silicate-based sealers (p > 0.05). Consequently, this reduction may not affect the three-dimensional seal of the root canal system in a negative manner. The penetration depth and percentage of penetration around the canal perimeter at both the root canal treatment and retreatment were significantly reduced from the coronal to apical points in all groups (p < 0.05).
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