ObjectiveTo evaluate the impact of sterilization methods on the structural integrity and antimicrobial properties of graphene nanocoating on titanium (GN). MethodsGN was transferred to titanium using wet (WT) or dry transfer (DT) techniques and sterilized using an autoclave (AC), glutaraldehyde (GA), or ethylene oxide (EtO). The GN structure was characterized using Raman spectroscopy before and after sterilization. Additional specimens were characterized by Raman after AC and water jetting. Biofilm formation was assessed before and after AC using colony-forming units (CFU), biofilm biomass, and SEM (uncoated titanium was the control). Three independent samples were used for structural characterization and biofilm quantification. Statistical analyses were conducted using one-way analysis of variance (ANOVA) and Tukey's test (α = 0.05). ResultsWT and DT demonstrated high structural stability after sterilization and water jetting, with negligible coating quality or coverage loss. GN exhibited lower biofilm formation even after AC sterilization, as shown by the reduction in CFU counts, biofilm biomass, and SEM images compared to the control. SignificanceGN demonstrated high resistance to the stresses imposed by all sterilization methods tested, maintaining its structural integrity, resistance to water-jet cleaning, and antibiofilm potential. The findings suggest that standard industrial practices can effectively sterilize highly resilient GN on titanium implants and possibly other biomaterials.
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