The aim of this study was to analyse the effect of preliminary phosphoric acid etching of enamel and dentine before the application of two, one-step self-etch adhesive systems. The systems were applied onto acid-etched or smear-layer-covered enamel and dentine. The treatment groups were as follows: group 1, Adper Easy Bond (3M ESPE) on etched substrate; group 2, Adper Easy Bond (control); group 3, iBond Self-Etch (Heraeus Kulzer) on etched substrate; and group 4, iBond Self-Etch (control). Enamel and dentine bond strengths were calculated using microshear and microtensile bond-strength tests. Additional specimens were prepared to evaluate nanoleakage at the dentine-adhesive interface and were investigated using light microscopy or transmission electron microscopy. Both adhesives demonstrated higher microshear bond strengths when enamel was pre-acid-etched with phosphoric acid (Adper Easy Bond 28.7 ± 4.8 MPa; iBond Self-Etch 19.7 ± 3.6 MPa) compared with controls (Adper Easy Bond 19.2 ± 3.3 MPa; iBond Self-Etch 17.5 ± 2.7 MPa) and increased microtensile bond strength when applied on acid-etched (Adper Easy Bond 35.8 ± 5.7 MPa; iBond Self-Etch 24.3 ± 7.9 MPa) vs. smear-layer-covered dentine (Adper Easy Bond 26.9 ± 6.2 MPa; iBond Self-Etch 17.6 ± 4.3 MPa). Adper Easy Bond showed lower nanoleakage than iBond Self-Etch, irrespective of preliminary etching. The results of this study support the use of phosphoric acid etching before the application of one-step self-etch adhesive systems.