Apicoectomy failure is generally related to inappropriate marginal sealing of the retrocavity, which allows percolation of microorganisms and their products from root canal system to periapex. This study evaluated tubular penetration of canal sealers and marginal permeability after retrocavity irradiation with Er;Cr:YSGG laser and retrofilling with MTA or cyanoacrylate. Twenty-two singlerooted teeth were decoronated and endodontically treated, their apical 3 mm were resected and the root ends were retroprepared with a low-speed bur. Twenty roots were randomly assigned to 4 groups (n=5): GI and GII -retrofilling with MTA and cyanoacrylate, respectively; GIII and GIV -retrocavity irradiation with Er;Cr:YSGG laser (2.78 μm, 4 W, 20 Hz, 70.8 J/cm 2 ) and retrofilling with MTA and cyanoacrylate, respectively. The remaining 2 roots served as positive and negative controls. The analysis of rhodamine B dye infiltration (p=0.05) demonstrated that laser irradiation and MTA retrofilling presented significantly higher permeability rates (p<0.05). Retrofilling with cyanoacrylate showed significantly lower permeability, either when laser was used or not on retrocavity. SEM analysis depicted more cyanoacrylate penetration through dentinal tubules when compared to MTA, suggesting a more efficient marginal sealing. Based on these results, it may be concluded that cyanoacrylate provided a less permeable retrofilling regardless of the retropreparation method, suggesting a more favorable condition to the establishment of the periapical healing.