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While there has been significant research conducted on bacterial colonization on implant materials, with a focus on developing surface modifications to prevent the formation of bacterial biofilms, the study of Candida albicans biofilms on implantable materials is still in its infancy, despite its growing relevance in implant-associated infections. C. albicans fungal infections represent a significant clinical concern due to their severity and associated high fatality rate. Pathogenic yeasts account for an increasing proportion of implant-associated infections, since Candida spp. readily form biofilms on medical and dental device surfaces. In addition, these biofilms are highly antifungal-resistant, making it crucial to explore alternative solutions for the prevention of Candida implant-associated infections. One promising approach is to modify the surface properties of the implant, such as the wettability and topography of these substrata, to prevent the initial Candida attachment to the surface. This review summarizes recent research on the effects of surface wettability, roughness, and architecture on Candida spp. attachment to implantable materials. The nanofabrication of material surfaces are highlighted as a potential method for the prevention of Candida spp. attachment and biofilm formation on medical implant materials. Understanding the mechanisms by which Candida spp. attach to surfaces will allow such surfaces to be designed such that the incidence and severity of Candida infections in patients can be significantly reduced. Most importantly, this approach could also substantially reduce the need to use antifungals for the prevention and treatment of these infections, thereby playing a crucial role in minimizing the possibility contributing to instances of antimicrobial resistance. STATEMENT OF SIGNIFICANCE: In this review we provide a systematic analysis of the role that surface characteristics, such as wettability, roughness, topography and architecture, play on the extent of C. albicans cells attachment that will occur on biomaterial surfaces. We show that exploiting bioinspired surfaces could significantly contribute to the prevention of antimicrobial resistance to antifungal and chemical-based preventive measures. By reducing the attachment and growth of C. albicans cells using surface structure approaches, we can decrease the need for antifungals, which are conventionally used to treat such infections., 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 © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Despite recent advances in the development of orthopedic devices, implant-related failures that occur as a result of poor osseointegration and nosocomial infection are frequent. In this study, we developed a multiscale titanium (Ti) surface topography that promotes both osteogenic and mechano-bactericidal activity using a simple two-step fabrication approach. The response of MG-63 osteoblast-like cells and antibacterial activity toward Pseudomonas aeruginosa and Staphylococcus aureus bacteria was compared for two distinct micronanoarchitectures of differing surface roughness created by acid etching, using either hydrochloric acid (HCl) or sulfuric acid (H 2 SO 4 ), followed by hydrothermal treatment, henceforth referred to as either MN-HCl or MN-H 2 SO 4 . The MN-HCl surfaces were characterized by an average surface microroughness ( S a ) of 0.8 ± 0.1 μm covered by blade-like nanosheets of 10 ± 2.1 nm thickness, whereas the MN-H 2 SO 4 surfaces exhibited a greater S a value of 5.8 ± 0.6 μm, with a network of nanosheets of 20 ± 2.6 nm thickness. Both micronanostructured surfaces promoted enhanced MG-63 attachment and differentiation; however, cell proliferation was only significantly increased on MN-HCl surfaces. In addition, the MN-HCl surface exhibited increased levels of bactericidal activity, with only 0.6% of the P. aeruginosa cells and approximately 5% S. aureus cells remaining viable after 24 h when compared to control surfaces. Thus, we propose the modulation of surface roughness and architecture on the micro- and nanoscale to achieve efficient manipulation of osteogenic cell response combined with mechanical antibacterial activity. The outcomes of this study provide significant insight into the further development of advanced multifunctional orthopedic implant surfaces.

A variant of the cold spray (CS) technique was applied for the functionalization of polymer-based materials such as polydimethylsiloxane (PDMS) to improve the extent of mammalian cell interactions with these substrates. This was demonstrated by the embedment of porous titanium (pTi) into PDMS substrates using a single-step CS technique. CS processing parameters such as gas pressure and temperature were optimized to achieve the mechanical interlocking of pTi in the compressed PDMS to fabricate a unique hierarchical morphology possessing micro-roughness. As evidenced by the preserved porous structure, the pTi particles did not undergo any significant plastic deformation upon impact with the polymer substrate. The thickness of the particle embedment layer was determined, by cross-sectional analysis, ranging from 120 μm to over 200 μm. The behavior of osteoblast-like cells MG63 coming into contact with the pTi-embedded PDMS was examined. The results showed that the pTi-embedded PDMS samples promoted 80-96% of cell adhesion and proliferation during the early stages of incubation. The low cytotoxicity of the pTi-embedded PDMS was confirmed, with cell viability of the MG63 cells being above 90%. Furthermore, the pTi-embedded PDMS facilitated the production of alkaline phosphatase and calcium deposition in the MG63 cells, as demonstrated by the higher amount of alkaline phosphatase (2.6 times) and calcium (10.6 times) on the pTi-embedded PDMS sample fabricated at 250 °C, 3 MPa. Overall, the work demonstrated that the CS process provided flexibility in the parameters used for the production of the modified PDMS substrates and is highly efficient for the fabrication of coated polymer products. The results obtained in this study suggest that a tailorable porous and rough architecture could be achieved that promoted osteoblast function, indicating that the method has promise in the design of titanium-polymer composite materials applied to biomaterials used in musculoskeletal applications.

The mechano-bactericidal action of nanostructured surfaces is well-documented; however, synthetic nanostructured surfaces have not yet been explored for their antifungal properties toward filamentous fungal species. In this study, we developed a biomimetic nanostructured surface inspired by dragonfly wings. A high-aspect-ratio nanopillar topography was created on silicon (nano-Si) surfaces using inductively coupled plasma reactive ion etching (ICP RIE). To mimic the superhydrophobic nature of insect wings, the nano-Si was further functionalised with trichloro(1H,1H,2H,2H-perfluorooctyl)silane (PFTS). The viability of Aspergillus brasiliensis spores, in contact with either hydrophobic or hydrophilic nano-Si surfaces, was determined using a combination of standard microbiological assays, confocal laser scanning microscopy (CLSM), and focused ion beam scanning electron microscopy (FIB-SEM). Results indicated the breakdown of the fungal spore membrane upon contact with the hydrophilic nano-Si surfaces. By contrast, hydrophobised nano-Si surfaces prevented the initial attachment of the fungal conidia. Hydrophilic nano-Si surfaces exhibited both antifungal and fungicidal properties toward attached A. brasisiensis spores via a 4-fold reduction of attached spores and approximately 9-fold reduction of viable conidia from initial solution after 24 h compared to their planar Si counterparts. Thus, we reveal, for the first time, the physical rupturing of attaching fungal spores by biomimetic hydrophilic nanostructured surfaces.

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.)

Nanomaterials have the potential to transform biological and biomedical research, with applications ranging from drug delivery and diagnostics to targeted interference of specific biological processes. Most existing research is aimed at developing nanomaterials for specific tasks such as enhanced biocellular internalization. However, fundamental aspects of the interactions between nanomaterials and biological systems, in particular, membranes, remain poorly understood. In this study, we provide detailed insights into the molecular mechanisms governing the interaction and evolution of one of the most common synthetic nanomaterials in contact with model phospholipid membranes. Using a combination of atomic force microscopy (AFM) and molecular dynamics (MD) simulations, we elucidate the precise mechanisms by which citrate-capped 5 nm gold nanoparticles (AuNPs) interact with supported lipid bilayers (SLBs) of pure fluid (DOPC) and pure gel-phase (DPPC) phospholipids. On fluid-phase DOPC membranes, the AuNPs adsorb and are progressively internalized as the citrate capping of the NPs is displaced by the surrounding lipids. AuNPs also interact with gel-phase DPPC membranes where they partially embed into the outer leaflet, locally disturbing the lipid organization. In both systems, the AuNPs cause holistic perturbations throughout the bilayers. AFM shows that the lateral diffusion of the particles is several orders of magnitude smaller than that of the lipid molecules, which creates some temporary scarring of the membrane surface. Our results reveal how functionalized AuNPs interact with differing biological membranes with mechanisms that could also have implications for cooperative membrane effects with other molecules.

Atomic force microscopy (AFM) was used to investigate the morphology and rigidity of the opportunistic pathogenic yeast, Candida albicans ATCC 10231, during its attachment to surfaces of three levels of nanoscale surface roughness. Non-polished titanium (npTi), polished titanium (pTi), and glass with respective average surface roughness ( S a ) values of 389 nm, 14 nm, and 2 nm, kurtosis ( S kur ) values of 4, 16, and 4, and skewness ( S skw ) values of 1, 4, and 1 were used as representative examples of each type of nanoarchitecture. Thus, npTi and glass surfaces exhibited similar S skw and S kur values but highly disparate S a . C. albicans cells that had attached to the pTi surfaces exhibited a twofold increase in rigidity of 364 kPa compared to those yeast cells attached to the surfaces of npTi (164 kPa) and glass (185 kPa). The increased rigidity of the C. albicans cells on pTi was accompanied by a distinct round morphology, condensed F-actin distribution, lack of cortical actin patches, and the negligible production of cell-associated polymeric substances; however, an elevated production of loose extracellular polymeric substances (EPS) was observed. The differences in the physical response of C. albicans cells attached to the three surfaces suggested that the surface nanoarchitecture (characterized by skewness and kurtosis), rather than average surface roughness, could directly influence the rigidity of the C. albicans cells. This work contributes to the next-generation design of antifungal surfaces by exploiting surface architecture to control the extent of biofilm formation undertaken by yeast pathogens and highlights the importance of performing a detailed surface roughness characterization in order to identify and discriminate between the surface characteristics that may influence the extent of cell attachment and the subsequent behavior of the attached cells.

Hypothesis: The ability exhibited by insect wings to resist microbial infestation is a unique feature developed over 400 million years of evolution in response to lifestyle and environmental pressures. The self-cleaning and antimicrobial properties of insect wings may be attributed to the unique combination of nanoscale structures found on the wing surface., Experiments: In this study, we characterised the wetting characteristics of superhydrophobic damselfly Calopteryx haemorrhoidalis wings. We revealed the details of air entrapment at the micro- and nano scales on damselfly wing surfaces using a combination of spectroscopic and electron microscopic techniques. Cryo-focused-ion-beam scanning electron microscopy was used to directly observe fungal spores and conidia that were unable to cross the air-liquid interface. By contrast, bacterial cells were able to cross the air-water interface to be ruptured upon attachment to the nanopillar surface. The robustness of the air entrapment, and thus the wing antifungal behaviour, was demonstrated after 1-week of water immersion. A newly developed wetting model confirmed the strict Cassie-Baxter wetting regime when damselfly wings are immersed in water., Findings: We provide evidence that the surface nanopillar topography serves to resist both fungal and bacterial attachment via a dual action: repulsion of fungal conidia while simultaneously killing bacterial cells upon direct contact. These findings will play an important role in guiding the fabrication of biomimetic, anti-fouling surfaces that exhibit both bactericidal and anti-fungal properties., 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 © 2021 Elsevier Inc. All rights reserved.)

Plasma polymer coatings fabricated from Melaleuca alternifolia essential oil and its derivatives have been previously shown to reduce the extent of microbial adhesion on titanium, polymers, and other implantable materials used in dentistry. Previous studies have shown these coatings to maintain their performance under standard operating conditions; however, when used in e.g., a dental implant, these coatings may inadvertently become subject to in situ cleaning treatments, such as those using an atmospheric pressure plasma jet, a promising tool for the effective in situ removal of biofilms from tissues and implant surfaces. Here, we investigated the effect of such an exposure on the antimicrobial performance of the Melaleuca alternifolia polymer coating. It was found that direct exposure of the polymer coating surface to the jet for periods less than 60 s was sufficient to induce changes in its surface chemistry and topography, affecting its ability to retard subsequent microbial attachment. The exact effect of the jet exposure depended on the chemistry of the polymer coating, the length of plasma treatment, cell type, and incubation conditions. The change in the antimicrobial activity for polymer coatings fabricated at powers of 20-30 W was not statistically significant due to their limited baseline bioactivity. Interestingly, the bioactivity of polymer coatings fabricated at 10 and 15 W against Staphylococcus aureus cells was temporarily improved after the treatment, which could be attributed to the generation of loosely attached bioactive fragments on the treated surface, resulting in an increase in the dose of the bioactive agents being eluted by the surface. Attachment and proliferation of Pseudomonas aeruginosa cells and mixed cultures were less affected by changes in the bioactivity profile of the surface. The sensitivity of the cells to the change imparted by the jet treatment was also found to be dependent on their origin culture, with mature biofilm-derived P. aeruginosa bacterial cells showing a greater ability to colonize the surface when compared to its planktonic broth-grown counterpart. The presence of plasma-generated reactive oxygen and nitrogen species in the culture media was also found to enhance the bioactivity of polymer coatings fabricated at power levels of 10 and 15 W, due to a synergistic effect arising from simultaneous exposure of cells to reactive oxygen and nitrogen species (RONS) and eluted bioactive fragments. These results suggest that it is important to consider the possible implications of inadvertent changes in the properties and performance of plasma polymer coatings as a result of exposure to in situ decontamination, to both prevent suboptimal performance and to exploit possible synergies that may arise for some polymer coating-surface treatment combinations.

Polymer matrix composite materials have the capacity to aid the indirect transmission of viral diseases. Published research shows that respiratory viruses, including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2 or COVID-19), can attach to polymer substrata as a result of being contacted by airborne droplets resulting from infected people sneezing or coughing in close proximity. Polymer matrix composites are used to produce a wide range of products that are "high-touch" surfaces, such as sporting goods, laptop computers and household fittings, and these surfaces can be readily contaminated by pathogens. This article reviews published research on the retention of SARS-CoV-2 and other virus types on plastics. The factors controlling the viral retention time on plastic surfaces are examined and the implications for viral retention on polymer composite materials are discussed. Potential strategies that can be used to impart antiviral properties to polymer composite surfaces are evaluated. These strategies include modification of the surface composition with biocidal agents (e.g., antiviral polymers and nanoparticles) and surface nanotexturing. The potential application of these surface modification strategies in the creation of antiviral polymer composite surfaces is discussed, which opens up an exciting new field of research for composite materials., Competing Interests: The authors declare no conflict of interest., (© 2021 The Authors. Nano Select published by Wiley‐VCH GmbH.)

Membrane model systems capable of mimicking live cell membranes were used for the first time in studying the effects arising from electromagnetic fields (EMFs) of 18 GHz where membrane permeability was observed following exposure. A present lack of understanding of the mechanisms that drive such a rapid change in membrane permeabilization as well as any structural or dynamic changes imparted on biomolecules affected by high-frequency electromagnetic irradiation limits the use of 18 GHz EMFs in biomedical applications. A phospholipid, 1,2-dioleoyl- sn -glycero-3-phosphocholine (DOPC) labelled with a fluorescent marker 1,2-dioleoyl- sn -glycero-3-phosphoethanolamine- N -(lissamine rhodamine B sulfonyl) (rhodamine-DOPE) was used in constructing the giant unilamellar vesicles (GUVs). After three cycles of exposure, enhanced membrane permeability was observed by the internalisation of hydrophilic silica nanospheres of 23.5 nm and their clusters. All-atom molecular dynamics simulations of 1-palmitoyl-2-oleoyl- sn -glycero-3-phosphocholine (POPC) membranes exposed to high frequency electric fields of different field strengths showed that within the simulation timeframe only extremely high strength fields were able to cause an increase in the interfacial water dynamics characterized by water dipole realignments. However, a lower strength, high frequency EMF induced changes of the water hydrogen bond network, which may contribute to the mechanisms that facilitate membrane permeabilization in a longer timeframe., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

Biofilms are assemblages of microbial cells, extracellular polymeric substances (EPS), and other components extracted from the environment in which they develop. Within biofilms, the spatial distribution of these components can vary. Here we present a fundamental characterization study to show differences between biofilms formed by Gram-positive methicillin-resistant Staphylococcus aureus (MRSA), Gram-negative Pseudomonas aeruginosa , and the yeast-type Candida albicans using synchrotron macro attenuated total reflectance-Fourier transform infrared (ATR-FTIR) microspectroscopy. We were able to characterise the pathogenic biofilms' heterogeneous distribution, which is challenging to do using traditional techniques. Multivariate analyses revealed that the polysaccharides area (1200-950 cm -1 ) accounted for the most significant variance between biofilm samples, and other spectral regions corresponding to amides, lipids, and polysaccharides all contributed to sample variation. In general, this study will advance our understanding of microbial biofilms and serve as a model for future research on how to use synchrotron source ATR-FTIR microspectroscopy to analyse their variations and spatial arrangements.

A major health concern of the 21 st century is the rise of multi-drug resistant pathogenic microbial species. Recent technological advancements have led to considerable opportunities for low-dimensional materials (LDMs) as potential next-generation antimicrobials. LDMs have demonstrated antimicrobial behaviour towards a variety of pathogenic bacterial and fungal cells, due to their unique physicochemical properties. This review provides a critical assessment of current LDMs that have exhibited antimicrobial behaviour and their mechanism of action. Future design considerations and constraints in deploying LDMs for antimicrobial applications are discussed. It is envisioned that this review will guide future design parameters for LDM-based antimicrobial applications.

Antimicrobial resistance has rendered many conventional therapeutic measures, such as antibiotics, ineffective. This makes the treatment of infections from pathogenic micro-organisms a major growing health, social, and economic challenge. Recently, nanomaterials, including two-dimensional (2D) materials, have attracted scientific interest as potential antimicrobial agents. Many of these studies, however, rely on the input of activation energy and lack real-world utility. In this work, we present the broad-spectrum antimicrobial activity of few-layered black phosphorus (BP) at nanogram concentrations. This property arises from the unique ability of layered BP to produce reactive oxygen species, which we harness to create this unique functionality. BP is shown to be highly antimicrobial toward susceptible and resistant bacteria and fungal species. To establish cytotoxicity with mammalian cells, we showed that both L929 mouse and BJ-5TA human fibroblasts were metabolically unaffected by the presence of BP. Finally, we demonstrate the practical utility of this approach, whereby medically relevant surfaces are imparted with antimicrobial properties via functionalization with few-layer BP. Given the self-degrading properties of BP, this study demonstrates a viable and practical pathway for the deployment of novel low-dimensional materials as antimicrobial agents without compromising the composition or nature of the coated substrate.

Antibiotic resistance is a global human health threat, causing routine treatments of bacterial infections to become increasingly difficult. The problem is exacerbated by biofilm formation by bacterial pathogens on the surfaces of indwelling medical and dental devices that facilitate high levels of tolerance to antibiotics. The development of new antibacterial nanostructured surfaces shows excellent prospects for application in medicine as next-generation biomaterials. The physico-mechanical interactions between these nanostructured surfaces and bacteria lead to bacterial killing or prevention of bacterial attachment and subsequent biofilm formation, and thus are promising in circumventing bacterial infections. This Review explores the impact of surface roughness on the nanoscale in preventing bacterial colonization of synthetic materials and categorizes the different mechanisms by which various surface nanopatterns exert the necessary physico-mechanical forces on the bacterial cell membrane that will ultimately result in cell death.

The microbial contamination of surfaces presents a significant challenge due to the adverse effects associated with biofilm formation, particularly on implantable devices. Here, the attachment and biofilm formation of the opportunistic human pathogen, Candida albicans ATCC 10231, were studied on surfaces with decreasing magnitudes of nanoscale roughness. The nanoscale surface roughness of nonpolished titanium, polished titanium, and glass was characterized according to average surface roughness, skewness, and kurtosis. Nonpolished titanium, polished titanium, and glass possessed average surface roughness ( S a ) values of 350, 20, and 2.5 nm; skewness ( S skw ) values of 1.0, 4.0, and 1.0; and ( S kur ) values of 3.5, 16, and 4, respectively. These unique characteristics of the surface nanoarchitecture were found to play a key role in limiting C. albicans attachment and modulating the functional phenotypic changes associated with biofilm formation. Our results suggest that surfaces with a specific combination of surface topographical parameters could prevent the attachment and biofilm formation of C. albicans . After 7 days, the density of attached C. albicans cells was recorded to be 230, 70, and 220 cells mm -2 on nonpolished titanium, polished titanium, and glass surfaces, respectively. Despite achieving a very low attachment density, C. albicans cells were only observed to produce hyphae associated with biofilm formation on nonpolished titanium surfaces, possessing the highest degree of surface roughness ( S a = 350 nm). This study provides a more comprehensive picture of the impact of surface architectures on C. albicans attachment, which is beneficial for the design of antifungal surfaces.

The formation and proliferation of bacterial biofilms on surfaces, particularly those on biomedical devices, is a significant issue that results in substantial economic losses, presenting severe health risks to patients. Furthermore, heterogeneous biofilms consisting of different bacterial species can induce the increase in pathogenicity, and the resistance to antimicrobial agents due to the synergistic interactions between the different species. Heterogeneous bacterial biofilms are notoriously difficult to treat due to the presence of extracellular polymeric substances (EPS) and, in conjunction with the rapid rise of multi-drug resistant pathogens, this means that new solutions for anti-biofilm treatment are required. In this study, we investigate the application of magneto-responsive gallium-based liquid metal (GLM-Fe) nanomaterials against a broad range of Gram-positive and Gram-negative bacterial mono-species and multi-species biofilms. The GLM-Fe particles exhibit a magneto-responsive characteristic, causing spherical particles to undergo a shape transformation to high-aspect-ratio nanoparticles with sharp asperities in the presence of a rotating magnetic field. These shape-transformed particles are capable of physically removing bacterial biofilms and rupturing individual cells. Following treatment, both mono-species and multi-species biofilms demonstrated significant reductions in their biomass and overall cell viability, demonstrating the broad-spectrum application of this antibacterial technology. Furthermore, the loss of integrity of the bacterial cell wall and membranes was visualized using a range of microscopy techniques, and the leakage of intracellular components (such as nucleic acids and protein) was observed. Insights gained from this study will impact the design of future liquid metal-based biofilm treatments, particularly those that rely on magneto-responsive properties.

It is commonly accepted that nanoparticles (NPs) can kill bacteria; however, the mechanism of antimicrobial action remains obscure for large NPs that cannot translocate the bacterial cell wall. It is demonstrated that the increase in membrane tension caused by the adsorption of NPs is responsible for mechanical deformation, leading to cell rupture and death. A biophysical model of the NP-membrane interactions is presented which suggests that adsorbed NPs cause membrane stretching and squeezing. This general phenomenon is demonstrated experimentally using both model membranes and Pseudomonas aeruginosa and Staphylococcus aureus, representing Gram-positive and Gram-negative bacteria. Hydrophilic and hydrophobic quasi-spherical and star-shaped gold (Au)NPs are synthesized to explore the antibacterial mechanism of non-translocating AuNPs. Direct observation of nanoparticle-induced membrane tension and squeezing is demonstrated using a custom-designed microfluidic device, which relieves contraction of the model membrane surface area and eventual lipid bilayer collapse. Quasi-spherical nanoparticles exhibit a greater bactericidal action due to a higher interactive affinity, resulting in greater membrane stretching and rupturing, corroborating the theoretical model. Electron microscopy techniques are used to characterize the NP-bacterial-membrane interactions. This combination of experimental and theoretical results confirm the proposed mechanism of membrane-tension-induced (mechanical) killing of bacterial cells by non-translocating NPs., (© 2020 Wiley-VCH GmbH.)

The recent rise of antibiotic resistance amongst Staphylococcus aureus (S. aureus) populations has made treating Staph-based infections a global medical challenge. Therapies that specifically target the peptidoglycan layer of S. aureus have emerged as new treatment avenues, towards which bacteria are less likely to develop resistance. While the majority of antibacterial polymers/oligomers have the ability to disrupt bacterial membranes, the design parameters for the enhanced disruption of peptidoglycan outer layer of Gram-positive bacteria remain unclear. Here, the design of oligomeric structures with favorable conformational characteristics for improved disruption of the peptidoglycan outer layer of Gram-positive bacteria is reported. Molecular dynamics simulations were employed to inform the structure design and composition of cationic oligomers displaying collapsed and expanded conformations. The most promising diblock and triblock cationic oligomers were synthesized by photo-induced atom transfer radical polymerization (photo ATRP). Following synthesis, the diblock and triblock oligomers displayed average antibacterial activity of ~99% and ~98% for S. aureus and methicillin-resistant S. aureus (MRSA), respectively, at the highest concentrations tested. Importantly, triblock oligomers with extended conformations showed significantly higher disruption of the peptidoglycan outer layer of S. aureus compared to diblock oligomers with more collapsed conformation, as evidenced by a number of characterization techniques including scanning electron, confocal and atomic force microscopy. This work provides new insight into the structure/property relationship of antibacterial materials and advances the design of functional materials for combating the rise of drug-resistant bacteria., 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 © 2020 Elsevier Inc. All rights reserved.)

A fungal biofilm refers to the agglomeration of fungal cells surrounded by a polymeric extracellular matrix (ECM). The ECM is composed primarily of polysaccharides that facilitate strong surface adhesion, proliferation, and cellular protection from the surrounding environment. Biofilms represent the majority of known microbial communities, are ubiquitous, and are found on a multitude of natural and synthetic surfaces. The compositions, and in-turn nanomechanical properties, of fungal biofilms remain poorly understood, because these systems are complex, composed of anisotropic cellular and extracellular material, and importantly are species and environment dependent. Therefore, genomic variation, and/or mutations, as well as environmental and growth factors can change the composition of a fungal cell's biofilm. In this work, we probe the physico-mechanical and biochemical properties of two fungal species, Candida albicans (C. albicans) and Cryptococcus neoformans (C. neoformans), as well as two antifungal resistant sub-species of C. neoformans, fluconazole-resistant C. neoformans (Fluc R C. neoformans) and amphotericin B-resistant C. neoformans (AmB R C. neoformans). A new experimental methodology of characterization is proposed, employing a combination of atomic force microscopy (AFM), instrumented nanoindentation, and Synchrotron ATR-FTIR measurements. This allowed the nano-mechanical and chemical characterisation of each fungal biofilm.

Environmental monitoring is necessary to ensure the overall health and conservation of an ecosystem. However, ecosystems (e.g. air, water, soil), are complex, involving numerous processes (both native and external), inputs, contaminants, and living organisms. As such, monitoring an environmental system is not a trivial task. The data obtained from natural systems is often multifaceted and convoluted, as a multitude of inputs can be intertwined within the matrix of the information obtained as part of a study. This means that trends and important results can be easily overlooked by conventional and single dimensional data analysis protocols. Recently, chemometric methods have emerged as a powerful method for maximizing the details contained within a chemical data set. Specifically, chemometrics refers to the use of mathematical and statistical analysis methods to evaluate chemical data, beyond univariant analysis. This type of analysis can provide a quantitative description of environmental measurements, while also having the capacity to reveal previously overlooked trends in data sets. Applying chemometrics to environmental data allows us to identify and describe the inter-relationship of environmental drivers, sources of contamination, and their potential impact upon the environment. This review aims to provide a detailed understanding of chemometric techniques, how they are currently used in environmental monitoring, and how these techniques can be used to improve current practices. An enhanced ability to monitor environmental conditions and to predict trends would be greatly beneficial to government and research agencies in their ability to develop environmental policies and analytical procedures.

For many years, an extensive array of chemometric methods have provided a platform upon which a quantitative description of environmental conditions can be obtained. Applying chemometric methods to environmental data allows us to identify and describe the interrelations between certain environmental drivers. They also provide an insight into the interrelationships between these drivers and afford us a greater understanding of the potential impact that these drivers can place upon the environment. However, an effective marriage of these two systems has not been performed. Therefore, it is the aim of this review to highlight the advantages of using chemometrics and sensors to identify hidden trends in environmental parameters, which allow the state of the environment to be effectively monitored. Despite the combination of chemometrics and sensors, to capture new developments and applications in the field of environmental sciences, these methods have not been extensively used. Importantly, although different parameters and monitoring procedures are required for different environments (e.g., air, water, soil), they are not distinct, separate entities. Contemporary developments in the use of chemometrics afford us the ability to predict changes in different aspects of the environment using instrumental methods. This review also provides an insight into the prevailing trends and the future of environmental sensing, highlighting that chemometrics can be used to enhance our ability to monitor the environment. This enhanced ability to monitor environmental conditions and to predict trends would be beneficial to government and research agencies in their ability to develop environmental policies and analysis procedures.

The mechano-bactericidal activity of nanostructured surfaces has become the focus of intensive research toward the development of a new generation of antibacterial surfaces, particularly in the current era of emerging antibiotic resistance. This work demonstrates the effects of an incremental increase of nanopillar height on nanostructure-induced bacterial cell death. We propose that the mechanical lysis of bacterial cells can be influenced by the degree of elasticity and clustering of highly ordered silicon nanopillar arrays. Herein, silicon nanopillar arrays with diameter 35 nm, periodicity 90 nm and increasing heights of 220, 360, and 420 nm were fabricated using deep UV immersion lithography. Nanoarrays of 360-nm-height pillars exhibited the highest degree of bactericidal activity toward both Gram stain-negative Pseudomonas aeruginosa and Gram stain-positive Staphylococcus aureus bacteria, inducing 95 ± 5% and 83 ± 12% cell death, respectively. At heights of 360 nm, increased nanopillar elasticity contributes to the onset of pillar deformation in response to bacterial adhesion to the surface. Theoretical analyses of pillar elasticity confirm that deflection, deformation force, and mechanical energies are more significant for the substrata possessing more flexible pillars. Increased storage and release of mechanical energy may explain the enhanced bactericidal action of these nanopillar arrays toward bacterial cells contacting the surface; however, with further increase of nanopillar height (420 nm), the forces (and tensions) can be partially compensated by irreversible interpillar adhesion that reduces their bactericidal effect. These findings can be used to inform the design of next-generation mechano-responsive surfaces with tuneable bactericidal characteristics for antimicrobial surface technologies., Competing Interests: The authors declare no competing interest.

The fabrication of antimicrobial surfaces that exhibit enhanced activity toward a large variety of microbial species is one of the major challenges of our time. In fact, the negative effects associated with both bacterial and fungal infections are enormous, especially considering that many microbial species are developing resistance to known antibiotics. In this work, we show how a combination of a specific surface morphology and surface chemistry can create a surface that exhibits nearly 100% antimicrobial activity toward both Gram-negative and Gram-positive bacteria and fungal cells. Arrays of vertically aligned, oxygen-deficient zinc oxide (ZnO) nanowires grown on a substrate exhibit enhanced antimicrobial activity compared with surfaces containing either less defective nanowires or highly oxygen-deficient flat films. This synergistic effect between physical activity (morphology) and chemical activity (surface composition) has been shown to be responsible for the outstanding antimicrobial activity of our surfaces, especially toward notoriously resilient bacterial or fungal species. These findings provide a series of design rules for tuning the activities of antibacterial and antifungal nanomaterials. These rules constitute an excellent platform for the development of next-generation antimicrobial surfaces.

Microbial contamination of polymer surfaces has become a significant challenge in domestic, industrial, and biomedical applications. Recent progress in our understanding of how topographical features of different length scales can be used to effectively and selectively control the attachment and proliferation of different cell types has provided an alternative strategy for imparting antibacterial activity to these surfaces. Among the well-recognized engineered models of antibacterial surface topographies, self-organized wrinkles have shown particular promise with respect to their antimicrobial characteristics. Here, we critically review the mechanisms by which wrinkles form on the surface of different types of polymer material and how they interact with various biomolecules and cell types. We also discuss the feasibility of using this antimicrobial strategy in real-life biomedical applications., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The development of antimicrobial drug resistance among pathogenic bacteria and fungi is one of the most significant health issues of the 21st century. Recently, advances in nanotechnology have led to the development of nanomaterials, particularly metals that exhibit antimicrobial properties. These metal nanomaterials have emerged as promising alternatives to traditional antimicrobial therapies. In this review, a broad overview of metal nanomaterials, their synthesis, properties, and interactions with pathogenic micro-organisms is first provided. Secondly, the range of nanomaterials that demonstrate passive antimicrobial properties are outlined and in-depth analysis and comparison of stimuli-responsive antimicrobial nanomaterials are provided, which represent the next generation of microbiocidal nanomaterials. The stimulus applied to activate such nanomaterials includes light (including photocatalytic and photothermal) and magnetic fields, which can induce magnetic hyperthermia and kinetically driven magnetic activation. Broadly, this review aims to summarize the currently available research and provide future scope for the development of metal nanomaterial-based antimicrobial technologies, particularly those that can be activated through externally applied stimuli., Competing Interests: The authors declare no conflict of interest., (© 2020 The Authors. Published by WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim.)

Hypothesis: Titanium and titanium alloys are often the most popular choice of material for the manufacture of medical implants; however, they remain susceptible to the risk of device-related infection caused by the presence of pathogenic bacteria. Hydrothermal etching of titanium surfaces, to produce random nanosheet topologies, has shown remarkable ability to inactivate pathogenic bacteria via a physical mechanism. We expect that systematic tuning of the nanosheet morphology by controlling fabrication parameters, such as etching time, will allow for optimisation of the surface pattern for superior antibacterial efficacy., Experiments: Using time-dependent hydrothermal processing of bulk titanium, we fabricated bactericidal nanosheets with variable nanoedge morphologies according to a function of etching time. A systematic study was performed to compare the bactericidal efficiency of nanostructured titanium surfaces produced at 0.5, 1, 2, 3, 4, 5, 6, 24 and 60 h of hydrothermal etching., Findings: Titanium surfaces hydrothermally treated for a period of 6 h were found to achieve maximal antibacterial efficiency of 99 ± 3% against Gram-negative Pseudomonas aeruginosa and 90 ± 9% against Gram-positive Staphylococcus aureus bacteria, two common human pathogens. These surfaces exhibited nanosheets with sharp edges of approximately 10 nm. The nanotopographies presented in this work exhibit the most efficient mechano-bactericidal activity against both Gram-negative and Gram-positive bacteria of any nanostructured titanium topography reported thus far., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Antibiotic resistance has made the treatment of biofilm-related infections challenging. As such, the quest for next-generation antimicrobial technologies must focus on targeted therapies to which pathogenic bacteria cannot develop resistance. Stimuli-responsive therapies represent an alternative technological focus due to their capability of delivering targeted treatment. This study provides a proof-of-concept investigation into the use of magneto-responsive gallium-based liquid metal (LM) droplets as antibacterial materials, which can physically damage, disintegrate, and kill pathogens within a mature biofilm. Once exposed to a low-intensity rotating magnetic field, the LM droplets become physically actuated and transform their shape, developing sharp edges. When placed in contact with a bacterial biofilm, the movement of the particles resulting from the magnetic field, coupled with the presence of nanosharp edges, physically ruptures the bacterial cells and the dense biofilm matrix is broken down. The antibacterial efficacy of the magnetically activated LM particles was assessed against both Gram-positive and Gram-negative bacterial biofilms. After 90 min over 99% of both bacterial species became nonviable, and the destruction of the biofilms was observed. These results will impact the design of next-generation, LM-based biofilm treatments.

The resistance of pathogenic bacteria toward traditional biocidal treatment methods is a growing concern in various settings, including that of water treatment and in the medical industry. As such, advanced antibacterial technologies are needed to prevent infections, against which current antibiotics are failing. This study introduces copper oxide nanoparticles (CuONPs) doped in graphene oxide (GO) as a potential pathogenic bacterial treatment. The aim of the study was to evaluate the antibacterial properties of the GO-CuONP hybridized material against pathogenic Escherichia coli ATCC 8739 ( E. coli ) and Salmonella typhimurium ATCC 14028 ( S. typhimurium ). GO was synthesized using a modified Hummer's method and doped with 40% w/w CuONPs using a series of thermal chemical reactions. The resulting hybrids were then characterized using scanning electron microscopic (SEM) and spectroscopic studies. These studies revealed that the hybrid material was considerably altered by the inclusion of CuONPs. The live and dead bacteria attached to the GO-CuONP material were detected using confocal laser scanning microscopy (CLSM). The antibacterial activity assay of the GO-CuONP material was conducted using a standard plate count method. Importantly, the GO-CuONP nanocomposite was determined to be an effective antibacterial nanomaterial, significantly inhibiting the growth of both E. coli and S. typhimurium bacteria compared to that observed on the pristine GO material. This study suggests that GO-CuONP composites are a promising high-efficacy antibacterial nanomaterial.

In this work, we report on the incorporation of a siloxane copolymer additive, poly((2-phenylethyl) methylsiloxane)-co(1-phenylethyl) methylsiloxane)-co-dimethylsiloxane), which is fully soluble at room temperature, in a rapid-cure thermoset polyester coating formulation. The additive undergoes polymerization-induced phase segregation (PIPS) to self-assemble on the coating surface as discrete discoid nanofeatures during the resin cure process. Moreover, the copolymer facilitates surface co-segregation of titanium dioxide pigment microparticulate present in the coating. Depending on the composition, the coatings can display persistent superhydrophobicity and self-cleaning properties and, surprisingly, the titanium dioxide pigmented coatings that include the siloxane copolymer additive display high levels of antibacterial performance against Gram-positive ( Staphylococcus aureus) and Gram-negative ( Pseudomonas aeruginosa ) bacteria. This antibacterial performance is believed to be associated with the unique surface topology of these coatings, which comprise stimuli-responsive discoid nanofeatures. This paper provides details of the surface morphology of the coatings and how these relates to the antimicrobial properties of the coating.

Nanostructured mechano-bactericidal surfaces represent a promising technology to prevent the incidence of microbial contamination on a variety of surfaces and to avoid bacterial infection, particularly with antibiotic resistant strains. In this work, a regular array of silicon nanopillars of 380 nm height and 35 nm diameter was used to study the release of bacterial cell debris off the surface, following inactivation of the cell due to nanostructure-induced rupture. It was confirmed that substantial bactericidal activity was achieved against Gram-negative Pseudomonas aeruginosa (85% non-viable cells) and only modest antibacterial activity towards Staphylococcus aureus (8% non-viable cells), as estimated by measuring the proportions of viable and non-viable cells via fluorescence imaging. In situ time-lapse AFM scans of the bacteria-nanopillar interface confirmed the removal rate of the dead P. aeruginosa cells from the surface to be approximately 19 minutes per cell, and approximately 11 minutes per cell for dead S. aureus cells. These results highlight that the killing and dead cell detachment cycle for bacteria on these substrata are dependant on the bacterial species and the surface architecture studied and will vary when these two parameters are altered. The outcomes of this work will enhance the current understanding of antibacterial nanostructures, and impact upon the development and implementation of next-generation implants and medical devices.

Additively manufactured selective laser melted titanium (SLM-Ti) opens the possibility of tailored medical implants for patients. Despite orthopedic implant advancements, significant problems remain with regard to suboptimal osseointegration at the interface between the implant and the surrounding tissue. Here, we show that applying a nanodiamond (ND) coating onto SLM-Ti scaffolds provides an improved surface for mammalian cell growth while inhibiting colonization of Staphylococcus aureus bacteria. Owing to the simplicity of our methodology, the approach is suitable for coating SLM-Ti geometries. The ND coating achieved 32 and 29% increases in cell density of human dermal fibroblasts and osteoblasts, respectively, after 3 days of incubation compared with the uncoated SLM-Ti substratum. This increase in cell density complements an 88% reduction in S. aureus detected on the ND-coated SLM-Ti substrata. This study paves a way to create facile antifouling SLM-Ti structures for biomedical implants.

The attachment of single-celled organisms, namely bacteria and fungi, to abiotic surfaces is of great interest to both the scientific and medical communities. This is because the interaction of such cells has important implications in a range of areas, including biofilm formation, biofouling, antimicrobial surface technologies, and bio-nanotechnologies, as well as infection development, control, and mitigation. While central to many biological phenomena, the factors which govern microbial surface attachment are still not fully understood. This lack of understanding is a direct consequence of the complex nature of cell-surface interactions, which can involve both specific and non-specific interactions. For applications involving micro- and nano-structured surfaces, developing an understanding of such phenomenon is further complicated by the diverse nature of surface architectures, surface chemistry, variation in cellular physiology, and the intended technological output. These factors are extremely important to understand in the emerging field of antibacterial nanostructured surfaces. The aim of this perspective is to re-frame the discussion surrounding the mechanism of nanostructured-microbial surface interactions. Broadly, the article reviews our current understanding of these phenomena, while highlighting the knowledge gaps surrounding the adhesive forces which govern bacterial-nanostructure interactions and the role of cell membrane rigidity in modulating surface activity. The roles of surface charge, cell rigidity, and cell-surface adhesion force in bacterial-surface adsorption are discussed in detail. Presently, most studies have overlooked these areas, which has left many questions unanswered. Further, this perspective article highlights the numerous experimental issues and misinterpretations which surround current studies of antibacterial nanostructured surfaces., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Biomaterials that have been newly implanted inside the body are the substratum targets for a "race for the surface", in which bacterial cells compete against eukaryotic cells for the opportunity to colonize the surface. A victory by the former often results in biomaterial-associated infections, which can be a serious threat to patient health and can undermine the function and performance of the implant. Moreover, bacteria can often have a 'head start' if implant contamination has taken place either prior to or during the surgery. Current prevention and treatment strategies often rely on systemic antibiotic therapies, which are becoming increasingly ineffective due to a growing prevalence of antibiotic-resistant bacteria. Nanostructured surfaces that kill bacteria by physically rupturing bacterial cells upon contact have recently emerged as a promising solution for the mitigation of bacterial colonization of implants. Furthermore, these nanoscale features have been shown to enhance the adhesion and proliferation of eukaryotic cells, which is a key to, for example, the successful osseointegration of load-bearing titanium implants. The bactericidal activity and biocompatibility of such nanostructured surfaces are often, however, examined separately, and it is not clear to what extent bacterial cell-surface interactions would affect the subsequent outcomes of host-cell attachment and osseointegration processes. In this study, we investigated the ability of bactericidal nanostructured titanium surfaces to support the attachment and growth of osteoblast-like MG-63 human osteosarcoma cells, despite them having been pre-infected with pathogenic bacteria. MG-63 is a commonly used osteoblastic model to study bone cell viability, adhesion, and proliferation on the surfaces of load-bearing biomaterials, such as titanium. The nanostructured titanium surfaces used here were observed to kill the pathogenic bacteria, whilst simultaneously enhancing the growth of MG-63 cells in vitro when compared to that occurring on sterile, flat titanium surfaces. These results provide further evidence in support of nanostructured bactericidal surfaces being used as a strategy to help eukaryotic cells win the "race for the surface" against bacterial cells on implant materials.

Recent technological advances have allowed the development of a new generation of nanostructured materials, such as those displaying both mechano-bactericidal activity and substrata that favor the growth of mammalian cells. Nanomaterials that come into contact with biological media such as blood first interact with proteins, hence understanding the process of adsorption of proteins onto these surfaces is highly important. The Random Sequential Adsorption (RSA) model for protein adsorption on flat surfaces was modified to account for nanostructured surfaces. Phenomena related to the nanofeature geometry have been revealed during the modelling process; e.g., convex geometries can lead to lower steric hindrance between particles, and hence higher degrees of surface coverage per unit area. These properties become more pronounced when a decrease in the size mismatch between the proteins and the surface nanostructures occurs. This model has been used to analyse the adsorption of human serum albumin (HSA) on a nano-structured black silicon (bSi) surface. This allowed the Blocking Function (the rate of adsorption) to be evaluated. The probability of the protein to adsorb as a function of the occupancy was also calculated.

The interface between water and a textured hydrophobic surface can exist in two regimes; either the Wenzel (surface-engulfed) or Cassie-Baxter (water-suspended) state. Better understanding of the influence of pattern geometry and spacing is crucial for the design of functional (super)hydrophobic surfaces, as inspired by numerous examples in nature. In this work, we have employed amplitude modulated - atomic force microscopy to visualize the air-water interface with an unprecedented degree of clarity on a superhydrophobic and a highly hydrophobic nanostructured surface. The images obtained provide the first real-time experimental visualization of the Cassie-Baxter wetting on the surface of biomimetic silicon nanopillars and a naturally superhydrophobic cicada wing. For both surfaces, the air-water interface was found to be remarkably well-defined, revealing a distinctly nanostructured air-water interface in the interstitial spacing. The degree of interfacial texture differed as a function of surface geometry. These results reveal that the air-water interface is heterogeneous in its structure and confirmed the presence of short-range interfacial ordering. Additionally, the overpressure values for each point on the interface were calculated, quantifying the difference in wetting behavior for the biomimetic and natural surface. Results suggest that highly-ordered, closely spaced nanofeatures facilitate robust Cassie-Baxter wetting states and therefore, can enhance the stability of (super)hydrophobic surfaces., (Copyright © 2018 Elsevier Inc. All rights reserved.)

The waxy epicuticle of dragonfly wings contains a unique nanostructured pattern that exhibits bactericidal properties. In light of emerging concerns of antibiotic resistance, these mechano-bactericidal surfaces represent a particularly novel solution by which bacterial colonization and the formation of biofilms on biomedical devices can be prevented. Pathogenic bacterial biofilms on medical implant surfaces cause a significant number of human deaths every year. The proposed mechanism of bactericidal activity is through mechanical cell rupture; however, this is not yet well understood and has not been well characterized. In this study, we used giant unilamellar vesicles (GUVs) as a simplified cell membrane model to investigate the nature of their interaction with the surface of the wings of two dragonfly species, Austrothemis nigrescens and Trithemis annulata, sourced from Victoria, Australia, and the Baix Ebre and Terra Alta regions of Catalonia, Spain. Confocal laser scanning microscopy and cryo-scanning electron microscopy techniques were used to visualize the interactions between the GUVs and the wing surfaces. When exposed to both natural and gold-coated wing surfaces, the GUVs were adsorbed on the surface, exhibiting significant deformation, in the process of membrane rupture. Differences between the tensile rupture limit of GUVs composed of 1,2-dioleoyl- sn-glycero-3-phosphocholine and the isotropic tension generated from the internal osmotic pressure were used to indirectly determine the membrane tensions, generated by the nanostructures present on the wing surfaces. These were estimated as being in excess of 6.8 mN m -1 , the first experimental estimate of such mechano-bactericidal surfaces. This simple model provides a convenient bottom-up approach toward understanding and characterizing the bactericidal properties of nanostructured surfaces.

High frequency (HF) electromagnetic fields (EMFs) have been widely used in many wireless communication devices, yet within the terahertz (THz) range, their effects on biological systems are poorly understood. In this study, electromagnetic radiation in the range of 0.3⁻19.5 × 10 12 Hz, generated using a synchrotron light source, was used to investigate the response of PC 12 neuron-like pheochromocytoma cells to THz irradiation. The PC 12 cells remained viable and physiologically healthy, as confirmed by a panel of biological assays; however, exposure to THz radiation for 10 min at 25.2 ± 0.4 °C was sufficient to induce a temporary increase in their cell membrane permeability. High-resolution transmission electron microscopy (TEM) confirmed cell membrane permeabilization via visualisation of the translocation of silica nanospheres ( d = 23.5 ± 0.2 nm) and their clusters ( d = 63 nm) into the PC 12 cells. Analysis of scanning electron microscopy (SEM) micrographs revealed the formation of atypically large (up to 1 µm) blebs on the surface of PC 12 cells when exposed to THz radiation. Long-term analysis showed no substantial differences in metabolic activity between the PC 12 cells exposed to THz radiation and untreated cells; however, a higher population of the THz-treated PC 12 cells responded to the nerve growth factor (NGF) by extending longer neurites (up to 0⁻20 µm) compared to the untreated PC12 cells (up to 20 µm). These findings present implications for the development of nanoparticle-mediated drug delivery and gene therapy strategies since THz irradiation can promote nanoparticle uptake by cells without causing apoptosis, necrosis or physiological damage, as well as provide a deeper fundamental insight into the biological effects of environmental exposure of cells to electromagnetic radiation of super high frequencies.

Background: Effects of man-made electromagnetic fields (EMF) on living organisms potentially include transient and permanent changes in cell behaviour, physiology and morphology. At present, these EMF-induced effects are poorly defined, yet their understanding may provide important insights into consequences of uncontrolled ( e.g. , environmental) as well as intentional ( e.g. , therapeutic or diagnostic) exposure of biota to EMFs. In this work, for the first time, we study mechanisms by which a high frequency (18 GHz) EMF radiation affects the physiology of membrane transport in pheochromocytoma PC 12, a convenient model system for neurotoxicological and membrane transport studies., Methods and Results: Suspensions of the PC 12 cells were subjected to three consecutive cycles of 30s EMF treatment with a specific absorption rate (SAR) of 1.17 kW kg -1 , with cells cooled between exposures to reduce bulk dielectric heating. The EMF exposure resulted in a transient increase in membrane permeability for 9 min in up to 90 % of the treated cells, as demonstrated by rapid internalisation of silica nanospheres (diameter d ≈ 23.5 nm) and their clusters ( d ≈ 63 nm). In contrast, the PC 12 cells that received an equivalent bulk heat treatment behaved similar to the untreated controls, showing lack to minimal nanosphere uptake of approximately 1-2 %. Morphology and growth of the EMF treated cells were not altered, indicating that the PC 12 cells were able to remain viable after the EMF exposure. The metabolic activity of EMF treated PC 12 cells was similar to that of the heat treated and control samples, with no difference in the total protein concentration and lactate dehydrogenase (LDH) release between these groups., Conclusion: These results provide new insights into the mechanisms of EMF-induced biological activity in mammalian cells, suggesting a possible use of EMFs to facilitate efficient transport of biomolecules, dyes and tracers, and genetic material across cell membrane in drug delivery and gene therapy, where permanent permeabilisation or cell death is undesirable., Competing Interests: Disclosure The authors report no conflicts of interest in this work.

The incorporation of high-aspect-ratio nanostructures across surfaces has been widely reported to impart antibacterial characteristics to a substratum. This occurs because the presence of such nanostructures can induce the mechanical rupture of attaching bacteria, causing cell death. As such, the development of high-efficacy antibacterial nano-architectures fabricated on a variety of biologically relevant materials is critical to the wider acceptance of this technology. In this study, we report the antibacterial behavior of a series of substrata containing multi-directional electrodeposited gold (Au) nanospikes, as both a function of deposition time and precursor concentration. Firstly, the bactericidal efficacy of substrata containing Au nanospikes was assessed as a function of deposition time to elucidate the nanopattern that exhibited the greatest degree of biocidal activity. Here, it was established that multi-directional nanospikes with an average height of ∼302 nm ± 57 nm (formed after a deposition time of 540 s) exhibited the greatest level of biocidal activity, with ∼88% ± 8% of the bacterial cells being inactivated. The deposition time was then kept constant, while the concentration of the HAuCl 4 and Pb(CH 3 COO) 2 precursor materials (used for the formation of the Au nanospikes) was varied, resulting in differing nanospike architectures. Altering the Pb(CH 3 COO) 2 precursor concentration produced multi-directional nanostructures with a wider distribution of heights, which increased the average antibacterial efficacy against both Gram-negative Pseudomonas aeruginosa and Gram-positive Staphylococcus aureus bacteria. Importantly, the in situ electrochemical fabrication method used in this work is robust and straightforward, and is able to produce highly reproducible antibacterial surfaces. The results of this research will assist in the wider utilization of mechano-responsive nano-architectures for antimicrobial surface technologies., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

Dragonfly wings are of great interest to researchers investigating biomimetic designs for antiwetting and antibacterial surfaces. The waxy epicuticular layer on the membrane of dragonfly wings possesses a unique surface nanoarchitecture that consists of irregular arrays of nanoscale pillars. This architecture confers superhydrophobic, self-cleaning, antiwetting, and antibiofouling behaviors. There is some evidence available that suggests that lifestyle factors may have influenced the evolution of the wing nanostructures and, therefore, the resulting properties of the wings; however, it appears that no systematic studies have been performed that have compared the wing surface features across a range of dragonfly species. Here, we provided a comparison of relevant wing surface characteristics, including chemical composition, wettability, and nanoarchitecture, of seven species of dragonfly from three families including Libellulidae , Aeshnidae , and Gomphidae . The characteristic nanopillar arrays were found to be present, and the chemical composition and the resultant wing surface superhydrophobicity were found to be well-conserved across all of the species studied. However, subtle differences were observed between the height, width, and density of nanofeatures and water droplet bouncing behavior on the wing surfaces. The results of this research will contribute to an understanding of the physical and chemical surface features that are optimal for the design of antiwetting and antibacterial surfaces., Competing Interests: The authors declare no competing financial interest.

Damselflies Calopteryx haemorrhoidalis exhibiting black wings are found in the western Mediterranean, Algeria, France, Italy, Spain and Monaco. Wing pigmentation is caused by the presence of melanin, which is involved in physiological processes including defence reactions, wound healing and sclerotization of the insect. Despite the important physiological roles of melanin, the presence and colour variation among males and females of the C. haemorrhoidalis species and the localization of the pigment within the wing membrane remain poorly understood. In this study, infrared (IR) microspectroscopy, coupled with the highly collimated synchrotron IR beam, was employed in order to identify the distribution of the pigments in the wings at a high spatial resolution. It was found that the melanin is localized in the procuticle of the C. haemorrhoidalis damselfly wings, distributed homogeneously within this layer, and not associated with the lipids of the epicuticle.

Titanium is a biocompatible material that is frequently used for making implantable medical devices. Nanoengineering of the surface is the common method for increasing material biocompatibility, and while the nanostructured materials are well-known to represent attractive substrata for eukaryotic cells, very little information has been documented about the interaction between mammalian cells and bactericidal nanostructured surfaces. In this study, we investigated the effect of bactericidal titanium nanostructures on PC12 cell attachment and differentiation—a cell line which has become a widely used in vitro model to study neuronal differentiation. The effects of the nanostructures on the cells were then compared to effects observed when the cells were placed in contact with non-structured titanium. It was found that bactericidal nanostructured surfaces enhanced the attachment of neuron-like cells. In addition, the PC12 cells were able to differentiate on nanostructured surfaces, while the cells on non-structured surfaces were not able to do so. These promising results demonstrate the potential application of bactericidal nanostructured surfaces in biomedical applications such as cochlear and neuronal implants.

Wrinkled patterns, which possess an extensive surface area over a limited planar space, can provide surface features ranging across the nano- and microscale that have become an engineering material with the flexibility to be tuneable for a number of technologies. Here, we investigate the surface parameters that influence the attachment response of two model bacteria (P. aeruginosa and S. aureus) to wrinkled gold-coated polystyrene surfaces having topologies at the nano- and microscale. Together with flat gold films as the controls, surface feature heights spanned 2 orders of magnitude (15 nm, 200 nm, and 1 micron). The surface wrinkle topology was shown through confocal laser scanning microscopic, atomic force microscopic and scanning electron microscopic image analyses to consist of air-water interfacial areas unavailable for bacterial attachment, which were also shown to be stable by time-lapsed contact angle measurements. Imposition of the nanoscale wrinkles reduced P. aeruginosa attachment to 57% and S. aureus attachment to 20% of their flat equivalent surfaces whereas wrinkles at the microscale further reduced these attachments to 7.5% and 14.5%, respectively. The density of attachments indicated an inherent species specific selectivity that changed with feature dimension, attributable to the scale of the air-water interfaces in contact with the bacterial cell. Parameters influencing static bacterial attachment were the total projected surface areas minus the air-water interface areas and the scale of these respective air-water interfaces (area distribution) with respect to the cell morphology. The range of these controlling parameters may provide new design principles for the evolving suite of physical anti-biofouling materials not reliant on biocidal agents under development.

Additive manufacturing using selective laser melted titanium (SLM-Ti) is used to create bespoke items across many diverse fields such as medicine, defense, and aerospace. Despite great progress in orthopedic implant applications, such as for "just in time" implants, significant challenges remain with regards to material osseointegration and the susceptibility to bacterial colonization on the implant. Here, we show that polycrystalline diamond coatings on these titanium samples can enhance biological scaffold interaction improving medical implant applicability. The highly conformable coating exhibited excellent bonding to the substrate. Relative to uncoated SLM-Ti, the diamond coated samples showed enhanced mammalian cell growth, enriched apatite deposition, and reduced microbial S. aureus activity. These results open new opportunities for novel coatings on SLM-Ti devices in general and especially show promise for improved biomedical implants.

One of the major challenges faced by the biomedical industry is the development of robust synthetic surfaces that can resist bacterial colonization. Much inspiration has been drawn recently from naturally occurring mechano-bactericidal surfaces such as the wings of cicada ( Psaltoda claripennis ) and dragonfly ( Diplacodes bipunctata ) species in fabricating their synthetic analogs. However, the bactericidal activity of nanostructured surfaces is observed in a particular range of parameters reflecting the geometry of nanostructures and surface wettability. Here, several of the nanometer-scale characteristics of black silicon (bSi) surfaces including the density and height of the nanopillars that have the potential to influence the bactericidal efficiency of these nanostructured surfaces have been investigated. The results provide important evidence that minor variations in the nanoarchitecture of substrata can substantially alter their performance as bactericidal surfaces.

Cellular attachment plays a vital role in the differentiation of pheochromocytoma (PC12) cells. PC12 cells are noradrenergic clonal cells isolated from the adrenal medulla of Rattus norvegicus and studied extensively as they have the ability to differentiate into sympathetic neuron-like cells. The effect of several experimental parameters including (i) the concentration of nerve growth factor (NGF); (ii) substratum coatings, such as poly-L-lysine (PLL), fibronectin (Fn), and laminin (Lam); and (iii) double coatings composed of PLL/Lam and PLL/Fn on the differentiation process of PC12 cells were studied. Cell morphology was visualised using brightfield phase contrast microscopy, cellular metabolism and proliferation were quantified using a 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay, and the neurite outgrowth and axonal generation of the PC12 cells were evaluated using wide field fluorescence microscopy. It was found that double coatings of PLL/Lam and PLL/Fn supported robust adhesion and a two-fold enhanced neurite outgrowth of PC12 cells when treated with 100 ng/mL of NGF while exhibiting stable metabolic activity, leading to the accelerated generation of axons., Competing Interests: The authors declare no conflict of interest.

Surface modification of polymers and paints is a popular and effective way to enhance the properties of these materials. This can be achieved by introducing a thin coating that preserves the bulk properties of the material, while protecting it from environmental exposure. Suitable materials for such coating technologies are inorganic oxides, such as alumina, titania and silica; however, the fate of these materials during long-term environmental exposure is an open question. In this study, polymer coatings that had been enhanced with the addition of silica nanoparticles (SiO2NPs) and subsequently subjected to environmental exposure, were characterized both before and after the exposure to determine any structural changes resulting from the exposure. High-resolution synchrotron macro ATR-FTIR microspectroscopy and surface topographic techniques, including optical profilometry and atomic force microscopy (AFM), were used to determine the long-term effect of the environment on these dual protection layers after 3 years of exposure to tropical and sub-tropical climates in Singapore and Queensland (Australia). Principal component analysis (PCA) based on the synchrotron macro ATR-FTIR spectral data revealed that, for the 9% (w/w) SiO2NP/polymer coating, a clear discrimination was observed between the control group (no environmental exposure) and those samples subjected to three years of environmental exposure in both Singapore and Queensland. The PCA loading plots indicated that, over the three year exposure period, a major change occurred in the triazine ring vibration in the melamine resins. This can be attributed to the triazine ring being very sensitive to hydrolysis under the high humidity conditions in tropical/sub-tropical environments. This work provides the first direct molecular evidence, acquired using a high-resolution mapping technique, of the climate-induced chemical evolution of a polyester coating. The observed changes in the surface topography of the coating are consistent with the changes in chemical composition.

The scientific and industrial interest in antimicrobial surfaces has significantly increased in recent times. This interest is largely in response to the persistent microbial contamination of industrial and, importantly, medical implant surfaces. Bacterial contamination of implant surfaces often leads to infection at the implant-tissue interface, and with the prevalence of increasing levels of antimicrobial resistance, the treatment of these infections is becoming far more challenging. Recently, many naturally occurring, high-aspect-ratio surface topographies have been discovered that exhibit high levels of biocidal efficacy. These include epicuticular lipid nano-architectures that are formed on the surfaces of insect wings, such as cicadae and dragonflies. The antimicrobial activity of such surfaces has been found to be a consequence of the physical interactions between the nanoscale topography of the substrate and the attaching pathogenic cells, meaning that the activity is independent of biochemical surface functionality. Importantly, these desirable surface properties can be translated to synthetic biomimetic surfaces, which, when mimicked, lead to a substantial increase in the antimicrobial properties of such surfaces. This paper reviews the recent advances in understanding the basis of these mechanical antimicrobial mechanisms, and discusses the progress being made towards the fabrication of optimised, biocompatible, synthetic analogues., (Crown Copyright © 2017. Published by Elsevier Inc. All rights reserved.)

Self-organized bacteria have been the subject of interest for a number of applications, including the construction of microbial fuel cells. In this paper, we describe the formation of a self-organized, three-dimensional network that is constructed using Gluconobacter oxydans B-1280 cells in a hydrogel consisting of poly(vinyl alcohol) (PVA) with N -vinyl pyrrolidone (VP) as a cross-linker, in which the bacterial cells are organized in a particular side-by-side alignment. We demonstrated that nonmotile G. oxydans cells are able to reorganize themselves, transforming and utilizing PVA-VP polymeric networks through the molecular interactions of bacterial extracellular polysaccharide (EPS) components such as acetan, cellulose, dextran, and levan. Molecular dynamics simulations of the G. oxydans EPS components interacting with the hydrogel polymeric network showed that the solvent-exposed loops of PVA-VP extended and engaged in bacterial self-encapsulation., Competing Interests: The authors declare no competing financial interest.