The purpose of this study was to evaluate the ability of two dentin adhesive systems to induce remineralization in the bonded dentin interface after in vitro thermo-cycling. Dentin surfaces were treated with two different adhesive approaches: (1) 37% phosphoric acid (PA) plus an "etch-and-rinse" dentin adhesive (single bond, SB) (PA+SB) or (2) application of a "self-etch" dentin adhesive (Clearfil SE bond, SEB). Three groups were established: (i) 24 h or (ii) 3 m storage, and (iii) specimens submitted to thermal cycling (100,000 cy/5 and 55ºC). Atomic force microscopy imaging/nanoindentation, Raman spectroscopy/cluster analysis with dye-assisted confocal laser scanning microscopy (CLSM) evaluation and Masson's trichrome staining assessments were implemented for characterization. Thermo-cycling increased nanohardness in PA+SB at the hybrid layer (HL) and in SEB at the bottom of the HL (BHL). Young's modulus increased at both the HL and BHL in SEB and at the HL in PA+SB, after thermal stress. Cluster analysis demonstrated an augmentation of the mineral-matrix ratio in thermo-cycled specimens. CLSM showed a decrease of both micropermeability and nanoleakage after thermo-cycling in PA+SB, and were completely absent in SEB. Trichrome staining reflected a scarce demineralized front in PA+SB after thermo-cycling and total remineralization in SEB.