Objective: The objectives of this study were to evaluate the microtensile bond strength (mu TBS) of one-step self-etch adhesive systems to Er:YAG laser-irradiated and bur-cut dentin after water storage and thermocycling. Background data: The Er:YAG laser is a promising alternative method for cavity preparation; however, no study has compared the effect of laser irradiation and aging procedures on the adhesion of one-step self-etch adhesives to dentin. Methods: Seventy-two third molars were selected and randomly divided according to cavity preparation method (Er:YAG laser and bur-cut). One-step self-etch adhesive systems (Clearfil S-3 Bond, AdheSE One and Adper Easy One) were used to bond the composite to dentin. Following the adhesive procedure, the specimens were subdivided according to aging conditions (24 h in water control [C], 6 months of water storage [WS] and 10.000 thermocycles [TC]). The mu TBS was determined in a universal testing machine. Three-way ANOVA, independent samples t test, and post-hoc comparisons test (alpha = 0.05) were performed on all data. Results: There was no statistical difference in mu TBS between Er:YAG laser-irradiated and bur-cut dentin (p > 0.05). Similarly, no significant difference was found in mu TBS between C, WS, and TC specimens (p > 0.05). Moreover, Clearfil S-3 Bond presented the highest mu TBS to dentin in both laser-irradiated and bur-cut cavity preparation methods. Conclusions: Neither bur-cut nor Er:YAG laser-irradiated dentin was affected by the aging methods used to simulate degradation of the adhesive interface. Er: YAG laser treatment may be used as an alternative cavity preparation method.