Functional polymeric coatings have rapidly become one of the most efficient strategies to endow biomaterials with enhanced bioactive properties. Among the bio-inspired polymers used for biomedical applications, mussel-derived poly(dopamine) (PDA) has increasingly attracted considerable interest because of its unique characteristics. In this work, we carried out detailed physicochemical characterization of a PDA film deposited on nanoporous titanium. In particular, we employed spectroscopic techniques (Raman and ATR-FTIR) and Digital Pulsed Force Mode Atomic Force microscopy (DPFM-AFM) to probe the chemical makeup and the nanomechanical properties of PDA-coated surfaces. In addition, we investigated protein adsorption by ATR-FTIR and quantified it with ten different serum proteins by Liquid Chromatography Mass spectroscopy (LC-MS), aiming at elucidating their potential contribution to the subsequent cell colonization. Successively, we assessed the response of MG-63 human osteoblastic cells to PDA-coated titanium both the multiple- and single-cell levels. Results for this study demonstrate that, compared to bare and nanoporous titanium, the PDA coating positively influences the adhesion and proliferation of MG-63 cells. In addition, we focus on how the three different substrates influence cell morphology (i.e. aspect ratio and form factor), the establishment of focal adhesions and the expression of RhoA, a protein involved in cell contractility. In conclusion, our work provides a deeper insight on the in vitro response of human osteoblastic cells to poly(dopamine) by closing in on specific aspects of cell-PDA interactions, ultimately reaffirming the potential of this bio-inspired polymer as a functional coating for bone tissue engineering applications.