Influence of sterilization on the performance of anodized nanoporous titanium implants

Highlights: • Fabrication of aligned titania nanopores on micro-rough titanium via anodization • Investigating the influence of various sterilization techniques on the performance of nano-engineered titanium • In-depth topographical, chemical, mechanical and bioactivity characterization of sterilized nanopores • Autoclaving (both wet and dry) compromises nanopores • UV irradiation is the most optimized sterilization technique for nano-engineered implants. Towards clinical translation of bioactive nano-engineered titanium implants, achieving appropriate sterilization and understanding its influence on the modified implant characteristics is essential. With limited studies exploring the influence of sterilization techniques on electrochemically anodized titanium with TiO₂ nanostructures, we aimed to advance this domain by performing an in-depth evaluation of the influence of common sterilization techniques (ethanol immersion, various UV irradiation times, gamma irradiation, and dry/wet autoclaving) on TiO₂ nanopores fabricated on micro-rough Ti surfaces (dual micro-nano) via single step anodization. Various sterilized surfaces were systematically compared in terms of topographical, chemical, crystalline, wettability and mechanical characteristics. Next, we investigated the protein adhesion capacity and functions of primary gingival fibroblasts (proliferation, adhesion/alignment and spreading morphology) to compare the bioactivity of the sterilized nanopores. Ethanol immersion, gamma irradiation and UV irradiation conserved the topography of the fabricated nanopores, while autoclave sterilization (both dry and wet) compromised the nanoporous structures. Various duration of UV-sterilization resulted in no significant changes in the surface topography and chemistry of the fabricated TNPs. Our findings revealed that UV irradiation is the most appropriate technique to sterilize nano-engineered titanium implants for appropriate wettability, protein adhesion capacity and enhanced metabolism and proliferation of human gingival fibroblasts (hGFs). This study systematically investigated the influence of sterilization on anodized nano-engineered titanium implants towards achieving reproducible outcomes (in terms of topography, chemistry and bioactivity), and found that UV irradiation holds great promise for application across different nano-engineered metal surfaces.