Conductive polymers (CPs), such as polypyrrole (PPy), have shown promising properties for use as electro-responsive bioactive scaffolds for tissue regeneration. PPy can be synthesized by chemical electrosynthesis and doped with biomolecules such as hyaluronic acid (HA). Taking advantage of the electrochemical synthesis versatility, nanofibers for surface-modified indium tin oxide (ITO) electrodes can be used as templates to produce tridimensional HA-doped PPy scaffolds. In this study, polyvinyl alcohol/chitosan (PVA/CTS) electrospun nanofibers deposited on ITO electrodes were used as a 3D template for the in situ electrosynthesis of HA-doped PPy to produce a bioactive scaffold for tissue engineering. The final material gathers the advantages of each biopolymer, the porous morphology of the nanofiber, and the conductivity of the electrosynthetized polymer. Furthermore, the biological activity of the NF-PVA/CTS@PPy:HA composite was evaluated in NIH-3T3 fibroblasts by MTT, resulting in a cell viability of 146 ± 40% and wound-healing capacity of 97 ± 1.9% at 24 h of culture.