Developing antibacterial and biocompatible coatings remains a challenge. A promising new thought is addressed to control the ion release behaviors through the design of the phases, structures, and morphologies of the products. In this study, TiO2-CuxO/Ag coatings were prepared via co-sputtering combined with annealing, the changes brought by different annealing temperatures were characterized, the physical, electrochemical, and antibacterial properties were concerned, and the relationship between biological performance and ions-releasing behavior was investigated. Results showed that the TiO2-CuxO/Ag coatings performed the morphologies of larger Ag particles seeded like islands on the dense CuO, such features improved the hydrophilicity and corrosion resistance of the Ti substrate. Rising the annealing temperature further enhanced the surface roughness, hydrophilicity, Young's modulus, hardness, and dynamic coefficient of friction of the coatings. Within the initial 3 h of soaking, both metal ions contributed to playing the antibacterial roles and produced a synergistic enhancement effect to satisfy the reduction of Staphylococcus aureus to 99.9%. In the next 3–120 h soaking, Ag releasing was suppressed and Cu releasing maintained its slow and stable trend, which made sure that the MC3T3-E1 cells could proliferate and adhere on the negatively charged coating surface, and copper oxide nanoparticles were phagocytized by cells. Among all TiO2-CuxO/Ag coatings, the M500 sample displayed superior performances on the surface roughness, contact angle, mechanical Young's modulus, and surface hardness, also realized the aim of killing pathogenic bacteria first and then activating cell survival, which enabled it to be potentially applied in clinical treatment.
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