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Hypothesis: Nanomaterials have been extensively investigated for a wide range of biomedical applications, including as antimicrobial agents, drug delivery vehicles, and diagnostic devices. The commonality between these biomedical applications is the necessity for the nanoparticle to interact with or pass through the cellular wall and membrane. Cell-nanomaterial interactions/uptake can occur in various ways, including adhering to the cell wall, forming aggregates on the surface, becoming absorbed within the cell wall itself, or transversing into the cell cytoplasm. These interactions are common to mammalian cells, bacteria, and yeast cells. This variety of interactions can cause changes to the integrity of the cell wall and the cell overall, but the precise mechanisms underpinning such interactions remain poorly understood. Here, we investigate the interaction between commonly investigated gold nanoparticles (AuNPs) and the cell wall/membrane of a model fungal cell to explore the general effects of interaction and uptake., Experiments: The interactions between 100 nm citrate-capped AuNPs and the cell wall of Candida albicans fungal cells were studied using a range of advanced microscopy techniques, including atomic force microscopy, confocal laser scanning microscopy, scanning electron microscopy, transmission electron microscopy, and synchrotron-FTIR micro-spectroscopy., Findings: In most cases, particles adhered on the cell surface, although instances of particles being up-taken into the cell cytoplasm and localised within the cell wall and membrane were also observed. There was a measurable increase in the stiffness of the fungal cell after AuNPs were introduced. Analysis of the synchrotron-FTIR data showed significant changes in spectral features associated with phospholipids and proteins after exposure to AuNPs., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)

Hypothesis: Titanium and its alloys are commonly used implant materials. Once inserted into the body, the interface of the biomaterials is the most likely site for the development of implant-associated infections. Imparting the titanium substrate with high-aspect-ratio nanostructures, which can be uniformly achieved using hydrothermal etching, enables a mechanical contact-killing (mechanoresponsive) mechanism of bacterial and fungal cells. Interaction between cells and the surface shows cellular inactivation via a physical mechanism meaning that careful engineering of the interface is needed to optimse the technology. This mechanism of action is only effective towards surface adsorbed microbes, thus any cells not directly in contact with the substrate will survive and limit the antimicrobial efficacy of the titanium nanostructures. Therefore, we propose that a dual-action mechanoresponsive and chemical-surface approach must be utilised to improve antimicrobial activity. The addition of antimicrobial silver nanoparticles will provide a secondary, chemical mechanism to escalate the microbial response in tandem with the physical puncture of the cells., Experiments: Hydrothermal etching is used as a facile method to impart variant nanostrucutres on the titanium substrate to increase the antimicrobial response. Increasing concentrations (0.25 M, 0.50 M, 1.0 M, 2.0 M) of sodium hydroxide etching solution were used to provide differing degrees of nanostructured morphology on the surface after 3 h of heating at 150 °C. This produced titanium nanospikes, nanoblades, and nanowires, respectively, as a function of etchant concentration. These substrates then provided an interface for the deposition of silver nanoparticles via a reduction pathway. Methicillin-resistant Staphylococcous aureus (MRSA) and Candida auris (C. auris) were used as model bacteria and fungi, respectively, to test the effectiveness of the nanostructured titanium with and without silver nanoparticles, and the bio-interactions at the interface., Findings: The presence of nanostructure increased the bactericidal response of titanium against MRSA from ∼ 10 % on commercially pure titanium to a maximum of ∼ 60 % and increased the fungicidal response from ∼ 10 % to ∼ 70 % in C. auris. Introducing silver nanoparticles increased the microbiocidal response to ∼ 99 % towards both bacteria and fungi. Importantly, this study highlights that nanostructure alone is not sufficient to develop a highly antimicrobial titanium substrate. A dual-action, physical and chemical antimicrobial approach is better suited to produce highly effective antibacterial and antifungal surface technologies., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Inc. All rights reserved.)