Polypyrrole (PPy) films obtained by oxidative polymerization have positive charges in its main chains and therefore exhibit antibacterial properties to some extent. However, antibacterial activities of PPy films for biomedical applications are still insufficient. Here, to endow PPy films with excellent antibacterial properties and biocompatibility, oxygen plasma immersion ion implantation (O‐PIII) is performed to regulate their physicochemical properties, including but not limited to wettability and surface charge. Escherichia coli, Staphylococcus aureus, and MC3T3‐E1 cells are utilized to investigate the antibacterial properties and cytocompatibility, respectively. Results show that antibacterial property of PPy films against E. coli is enhanced after O‐PIII treatment, and it becomes more outstanding with the increase of implantation time, which may be ascribed to the improved hydrophilicity and change of sample surfaces from negatively charged to positively charged. The similar results cannot be found from S. aureus due to different cell membrane structure. Meanwhile, O‐PIII can significantly improve the biocompatibility of PPy films.

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