In this study, antibacterial self-healable polyurethanes (EBPUs series) were prepared using eugenol-based bio-polyol (EBP), which was synthesized by thiol–ene reaction between eugenol and 1,2-ethanedithiol. The effects of EBP content on the mechanical, self-healing, and antibacterial properties of polyurethanes were investigated. When EBP was added into the EBPUs series as a polyol, the tensile strength, elongation at break, and self-healing efficiency values increased to 21.78 MPa, 1462.27%, and 77.85 %, respectively. EBPU−10 shows self-healing efficiency, where its first and second self-healing efficiency reached 76.6 % and 66.1 %, respectively. For antibacterial properties, the EBPUs series show promising antibacterial activity. EBPU−10 inhibited Staphylococcus aureus growth by 99.3 %, and Bacillus cereus growth by 95.7 %, and it inhibited Escherichia coli growth by 82.3 % at 6 h, compared to EBPU−01. Biofilm mass on foams was reduced by EBPU-10, and the morphology of biofilm bacteria was altered, such as their size being reduced, distorted, shrunken, and ruptured. In conclusion, it can be used for various applications, such as packaging, antimicrobial wound dressing, catheter, tissue adhesive, and surgical implant.