The objective of this study was to address the following question: 'Which properties are modified in partially demineralized surfaces, compared with non-demineralized dentin surfaces, following orthophosphoric acid-etching as performed in clinical procedures?'. For this purpose, the complementary techniques atomic force microscopy/spectroscopy, scanning electron microscopy, energy-dispersive X-ray spectroscopy, and contact angle measurements were used to provide a multiscale characterization of the dentin substrate undergoing the acidic preconditioning designed to enhance wetting. This study underlines that after the acid etching pretreatment the contribution of the nanomechanical, morphological, and physicochemical modifications has a strong influence on the dentin adhesion properties and thus plays a significant role in the coupling of the adhesive-resin composite build-up material at the dentin surface.