In this study, biodegradable piezoelectric poly(3-hydroxybuthyrate) (PHB) and PHB-TiO₂ as well as non-piezoelectric poly(ε-caprolactone) (PCL) fibrous scaffolds were successfully fabricated. First, TiO₂ nanoparticles (NPs) with various content (1%, 2%, and 3% wt) were loaded into the PHB matrix to improve its tensile and wettability properties as well as piezoelectric performances. The piezoelectric property of the fibrous scaffolds was examined and a significant improvement in the piezoelectric property of hybrid fibrous scaffolds compared to pure PHB was detected. For the PHB-2%TiO₂ sample, a maximum of in the range 4.5–5 V electricity production from a height of 10 cm and a mass drop of 35 g was observed after 150°C heat treatment. Then, an in vitro bactericidal analysis was carried out to test the bacteriostatic effect of the produced piezoelectric biomaterials against E-cherichia coli (E coli) under ultrasound treatment. It was observed that E. coli appeared to be the most sensitive to the PHB-%2TiO₂ sample and consequently the antibacterial activity of all the samples against E. coli was dependent on the piezoelectric properties of the samples. The results indicated that the fabricated fibrous scaffolds could be considered as a promising piezoelectric biomaterial with ultrasonically-controlled bacteriostatic activity for various tissue engineering applications.