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Abstract Introduction Convencional resin cements can be used in combination with a total-etch system in a conventional mode or as self-adhesive resin cements. The latter are less technique sensitive and able to bond to dental tissues without previous treatment or adhesive layer and requires only a single step to be applied to dental structures. Objective To compare qualitatively the adhesive interfaces of two self-adhesive resin cements and one conventional resin cement after different tooth surface treatments under scanning electron microscopy. Material and method 42 crowns of bovine incisors were sectioned and flattened exposing enamel (E) or dentine (D) substrate. Subgroups were defined according to conditioning type and time: E1—no treatment, E2—37% phosphoric acid for 15 seconds, E3—37% phosphoric acid for 30 seconds; D1—no treatment, D2—37% phosphoric acid for 5 seconds; D3—11.5% polyacrylic acid for 15 seconds. A resin block was bonded to each substrate using the self-adhesive resin cements RelyX U100 (3M ESPE) and RelyX U200 (3M ESPE). As a reference hybrid layer, six resin blocks were luted with RelyX ARC and Scotchbond Multi-Purpose adhesive system (3M ESPE) (enamel—EA; dentine—DA). After aging for 7 days in a moist environment at 37±1°C, samples were prepared for microscopy analysis. Result and Discussion In the ARC specimens, there was hybrid layer formation in both EA and DA. U100 E1 showed gaps at the adhesive interface, while E2 and E3 showed interaction for both self-adhesive cements. There was superficial interaction with bothU100 and U200 in D1, while in D2 and D3, resin tags were only observed in the case of U100. Conclusion It was concluded that substrate conditioning may enhance the interaction between self-adhesive resin cements and dental tissues, although this is not the case for RelyX U200 and dentine.