Previous studies have shown that piezoelectric materials may be used to prepare bioactive electrically charged surfaces. In the current study, polyurethane/polyvinylidene fluoride (PU/PVDF) scaffolds were prepared by electrospinning. The mechanical property and piezoelectric property of the scaffolds were evaluated. The crystalline phase of PVDF in the scaffolds was characterised by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC). In vitro cell culture was performed to investigate cytocompatibility of the scaffolds. Wound-healing assay, cell-adhesion assay, quantitative RT-PCR and Western blot analyses were performed to investigate piezoelectric effect of the scaffolds on fibroblast activities. Further, the scaffolds were subcutaneously implanted in Sprague-Dawley (SD) rats to investigate their biocompatibility and the piezoelectric effect on fibrosis in vivo. The results indicated that the electrospinning process had changed PVDF crystalline phase from the nonpiezoelectric α phase to the piezoelectric β phase. The fibroblasts cultured on the scaffolds showed normal morphology and proliferation. The fibroblasts cultured on the piezoelectric-excited scaffolds showed enhanced migration, adhesion and secretion. The scaffolds that were subcutaneously implanted in SD rats showed higher fibrosis level due to the piezoelectrical stimulation, which was caused by random animal movements followed by mechanical deformation of the scaffolds. The scaffolds are potential candidates for wound healing applications.