This work reports on the influence of the polarization of electroactive poly(vinylidene fluoride), PVDF, on the biological response of cells cultivated under static and dynamic conditions. Non-poled and “poled +” β-PVDF with and without a titanium layer were thus prepared. A thin titanium layer was deposited on PVDF films in order to obtain a more homogeneous surface charge. The MC3T3-E1 osteoblast cell culture exhibited different responses in the presence of PVDF films. The positively charged β-PVDF films promote higher osteoblast adhesion and proliferation, which is higher under dynamic conditions on poled samples, showing that the surface charge under mechanical stimulation improves the osteoblast growth. Therefore, electroactive membranes and scaffolds can provide the necessary electrical stimuli for the growth and proliferation of specific cells.