Synergistic Bactericidal Effect of Zn2+ Ions and Reactive Oxygen Species Generated in Response to Either UV or X-ray Irradiation of Zn-Doped Plasma Electrolytic Oxidation TiO2 Coatings.

Zn-containing TiO2-based coatings with Na, Ca, Si, and K additives were obtained by plasma electrolytic oxidation (PEO) of Ti in order to achieve an effective and broad bactericidal protection without compromising biocompatibility. A protocol has been developed for cleaning the coating surface from electrolyte residues, ensuring the preservation of the microstructure and composition of the surface layer. Using high-resolution transmission electron microscopy, three characteristic microstructural zones in the PEO-Zn coating are well documented: zone 1 with a TiO2-based nanocrystalline structure, zone 2 with an amorphous structure, and zone 3 around pores with an amorphous-nanocrystalline structure. The excellent cytocompatibility of PEO-Zn samples was confirmed by three different methods: monitoring the proliferation of MC3T3-E1 cells, assessing the viability of sheep osteoblast cells using calcein-AM staining and fluorescence microscopy, and incubation with spheroids based on primary osteoblast cells and mouse embryonic fibroblast NIH3T3 cells. The PEO-Zn coatings absorb >60% of the incident light over the UV and Vis-NIR spectral ranges. After 24 h, the PEO-Zn coatings completely inactivate four types of strains: Gram-positive Staphylococcus aureus CSA154 and ATCC29213 and Gram-negative Escherichia coli K261 and U20, and also prevent E. coli U20 and K261 biofilm formation. The superior antibacterial activity is associated with the synergistic effect of Zn2+ ions in safe concentration and reactive oxygen species (ROS) generated in response to either UV irradiation or soft short-term X-ray irradiation. The X-ray irradiation-induced ROS formation by a PEO coating is reported for the first time. The enhanced bactericidal activity after X-ray irradiation compared to UV illumination is attributed to the more intense ROS generation in the first few hours. The results obtained significantly expand the possibilities of using PEO coatings on the surfaces of titanium implants.

X-ray and UV Irradiation-Induced Reactive Oxygen Species Mediated Antibacterial Activity in Fe and Pt Nanoparticle-Decorated Si-Doped TiCaCON Films.

Bone implants with biocompatibility and the ability to biomineralize and suppress infection are in high demand. The occurrence of early infections after implant placement often leads to repeated surgical treatment due to the ineffectiveness of antibiotic therapy. Therefore, an extremely attractive solution to this problem would be the ability to initiate bacterial protection of the implant by an external influence. Here, we present a proof-of-concept study based on the generation of reactive oxygen species (ROS) by the implant surface in response to X-ray irradiation, including through a layer of 3 mm adipose tissue, providing bactericidal protection. The effect of UV and X-ray irradiation of the implant surface on the ROS formation and the associated bactericidal activity was compared. The focus of our study was light-sensitive Si-doped TiCaCON films decorated with Fe and Pt nanoparticles (NPs) with photoinduced antibacterial activity mediated by ROS. In the visible and infrared range of 300-1600 nm, the films absorb more than 60% of the incident light. The high light absorption capacity of TiO2/TiC and TiO2/TiN heterostructures was demonstrated by density functional theory calculations. After short-term (5-10 s) low-dose X-ray irradiation, the films generated significantly more ROS than after UV illumination for 1 h. The Fe/TiCaCON-Si films showed enhanced biomineralization capacity, superior cytocompatibility, and excellent antibacterial activity against multidrug-resistant hospital Escherichia coli U20 and K261 strains and methicillin-resistant Staphylococcus aureus MW2 strain. Our study clearly demonstrates that oxidized Fe NPs are a promising alternative to the widely used Ag NPs in antibacterial coatings, and X-rays can potentially be used in ROS-regulating therapy to suppress inflammation in case of postimplant complications.

Osteoconductive, Osteogenic, and Antipathogenic Plasma Electrolytic Oxidation Coatings on Titanium Implants with BMP-2.

We report a one-pot plasma electrolytic oxidation (PEO) strategy for forming a multi-element oxide layer on the titanium surface using complex electrolytes containing Na2HPO4, Ca(OH)2, (NH2)2CO, Na2SiO3, CuSO4, and KOH compounds. For even better bone implant ingrowth, PEO coatings were additionally loaded with bone morphogenetic protein-2 (BMP-2). The samples were tested in vivo in a mouse craniotomy model. Tests for bactericidal and fungicidal activity were carried out using clinically isolated multi-drug-resistant Escherichia coli (E. coli) K261, E. coli U20, methicillin-resistant Staphylococcus aureus (S. aureus) CSA154 bacterial strains, and Neurospora crassa (N. crassa) and Candida albicans (C. albicans) D2528/20 fungi. The PEO-Cu coating effectively inactivated both Gram-positive and Gram-negative bacteria at low concentrations of Cu2+ ions: minimal bactericidal concentration for E. coli and N. crassa (99.9999%) and minimal inhibitory concentration (99.0%) for S. aureus were 5 ppm. For all studied bacterial and fungal strains, PEO-Cu coating completely prevented the formation of bacterial and fungal biofilms. PEO and PEO-Cu coatings demonstrated bone remodeling and moderate osteoconductivity in vivo, while BMP-2 significantly enhanced osteoconduction and osteogenesis. The obtained results are encouraging and indicate that Ti-based materials with PEO coatings loaded with BMP-2 can be widely used in customized medicine as implants for orthopedics and cranio-maxillofacial surgery.