Jafar Hasan

Antibacterial surfaces: the quest for a new generation of biomaterials

In this review we attempt to clarify the notion of what is meant by the term antibacterial surfaces and categorise the approaches that are commonly used in the design of antibacterial surfaces. Application of surface coatings and the modification of the surface chemistry of substrata are generally considered to be a chemical approach to surface modification (as are surface polymerisation, functionalisation, and derivatisation), whereas, modification of the surface architecture of a substrate can be considered a physical approach. Here, the antifouling and bactericidal effects of antibacterial surfaces are briefly discussed. Finally, several recent efforts to design a new generation of antibacterial surfaces, which are based on mimicking the surface nanotopography of natural surfaces, are considered.

Bactericidal activity of black silicon

Black silicon is a synthetic nanomaterial that contains high aspect ratio nanoprotrusions on its surface, produced through a simple reactive-ion etching technique for use in photovoltaic applications. Surfaces with high aspect-ratio nanofeatures are also common in the natural world, for example, the wings of the dragonfly Diplacodes bipunctata. Here we show that the nanoprotrusions on the surfaces of both black silicon and D. bipunctata wings form hierarchical structures through the formation of clusters of adjacent nanoprotrusions. These structures generate a mechanical bactericidal effect, independent of chemical composition. Both surfaces are highly bactericidal against all tested Gram-negative and Gram-positive bacteria, and endospores, and exhibit estimated average killing rates of up to ~450,000 cells min−1 cm−2. This represents the first reported physical bactericidal activity of black silicon or indeed for any hydrophilic surface. This biomimetic analogue represents an excellent prospect for the development of a new generation of mechano-responsive, antibacterial nanomaterials.

Natural Bactericidal Surfaces: Mechanical Rupture of Pseudomonas aeruginosa Cells by Cicada Wings

Natural superhydrophobic surfaces are often thought to have antibiofouling potential due to their self-cleaning properties. However, when incubated on cicada wings, Pseudomonas aeruginosa cells are not repelled; instead they are penetrated by the nanopillar arrays present on the wing surface, resulting in bacterial cell death. Cicada wings are effective antibacterial, as opposed to antibiofouling, surfaces.

Biophysical model of bacterial cell interactions with nanopatterned cicada wing surfaces

The nanopattern on the surface of Clanger cicada (Psaltoda claripennis) wings represents the first example of a new class of biomaterials that can kill bacteria on contact based solely on their physical surface structure. The wings provide a model for the development of novel functional surfaces that possess an increased resistance to bacterial contamination and infection. We propose a biophysical model of the interactions between bacterial cells and cicada wing surface structures, and show that mechanical properties, in particular cell rigidity, are key factors in determining bacterial resistance/sensitivity to the bactericidal nature of the wing surface. We confirmed this experimentally by decreasing the rigidity of surface-resistant strains through microwave irradiation of the cells, which renders them susceptible to the wing effects. Our findings demonstrate the potential benefits of incorporating cicada wing nanopatterns into the design of antibacterial nanomaterials.

Surface topographical factors influencing bacterial attachment

Substratum surface roughness is known to be one of the key factors in determining the extent of bacterial colonization. Understanding the way by which the substratum topography, especially at the nanoscale, mediates bacterial attachment remains ambiguous at best, despite the volume of work available on the topic. This is because the vast majority of bacterial attachment studies do not perform comprehensive topographical characterization analyses, and typically consider roughness parameters that describe only one aspect of the surface topography. The most commonly reported surface roughness parameters are average and root mean square (RMS) roughness (R(a) and R(q) respectively), which are both measures of the typical height variation of the surface. They offer no insights into the spatial distribution or shape of the surface features. Here, a brief overview of the current state of research on topography-mediated bacterial adhesion is presented, as well as an outline of the suite of roughness characterization parameters that are available for the comprehensive description of the surface architecture of a substratum. Finally, a set of topographical parameters is proposed as a new standard for surface roughness characterization in bacterial adhesion studies to improve the likelihood of identifying direct relationships between substratum topography and the extent of bacterial adhesion.

Selective bactericidal activity of nanopatterned superhydrophobic cicada Psaltoda claripennis wing surfaces

The nanopattern on the surface of Clanger cicada (Psaltoda claripennis) wings represents the first example of a new class of biomaterials that can kill bacteria on contact based solely on its physical surface structure. As such, they provide a model for the development of novel functional surfaces that possess an increased resistance to bacterial contamination and infection. Their effectiveness against a wide spectrum of bacteria, however, is yet to be established. Here, the bactericidal properties of the wings were tested against several bacterial species, possessing a range of combinations of morphology and cell wall type. The tested species were primarily pathogens, and included Bacillus subtilis, Branhamella catarrhalis, Escherichia coli, Planococcus maritimus, Pseudomonas aeruginosa, Pseudomonas fluorescens, and Staphylococcus aureus. The wings were found to consistently kill Gram-negative cells (i.e., B. catarrhalis, E. coli, P. aeruginosa, and P. fluorescens), while Gram-positive cells (B. subtilis, P. maritimus, and S. aureus) remained resistant. The morphology of the cells did not appear to play any role in determining cell susceptibility. The bactericidal activity of the wing was also found to be quite efficient; 6.1 ± 1.5 × 106P. aeruginosa cells in suspension were inactivated per square centimeter of wing surface after 30-min incubation. These findings demonstrate the potential for the development of selective bactericidal surfaces incorporating cicada wing nanopatterns into the design.

Molecular Organization of the Nanoscale Surface Structures of the Dragonfly Hemianax papuensis Wing Epicuticle

The molecular organization of the epicuticle (the outermost layer) of insect wings is vital in the formation of the nanoscale surface patterns that are responsible for bestowing remarkable functional properties. Using a combination of spectroscopic and chromatographic techniques, including Synchrotron-sourced Fourier-transform infrared microspectroscopy (FTIR), x-ray photoelectron spectroscopy (XPS) depth profiling and gas chromatography-mass spectrometry (GCMS), we have identified the chemical components that constitute the nanoscale structures on the surface of the wings of the dragonfly, Hemianax papuensis. The major components were identified to be fatty acids, predominantly hexadecanoic acid and octadecanoic acid, and n-alkanes with even numbered carbon chains ranging from C14 to C30. The data obtained from XPS depth profiling, in conjunction with that obtained from GCMS analyses, enabled the location of particular classes of compounds to different regions within the epicuticle. Hexadecanoic acid was found to be a major component of the outer region of the epicuticle, which forms the surface nanostructures, and was also detected in deeper layers along with octadecanoic acid. Aliphatic compounds were detected throughout the epicuticle, and these appeared to form a third discrete layer that was separate from both the inner and outer epicuticles, which has never previously been reported.

Physico-mechanical characterisation of cells using atomic force microscopy — Current research and methodologies

Atomic force microscopy (AFM) is a technique that has long been employed in materials science, but is now increasingly being used in the biological sciences. AFM provides excellent topographical information on prokaryotic and eukaryotic cell surfaces, and the extracellular material produced by the cells. It helps to generate important data on the mechanical properties of cells, such as hardness and elasticity. AFM can also be used to measure the strength of adhesion, attraction, and repulsion forces between cells and surfaces or even between individual molecules. Additionally, by combining AFM with other complementary techniques such as fluorescence microscopy or Raman spectroscopy, the chemistry of given surface structures can be identified. This review aims to provide an update on the AFM techniques currently used in cell biology studies, along with a description of the range of recently developed research methodologies in which AFM plays a key role.

Dual role of outer epicuticular lipids in determining the wettability of dragonfly wings

Numerous natural surfaces possess superhydrophobicity and self-cleaning properties that would be extremely beneficial when applied in industry. Dragonfly wings are one example of such surfaces, and while their general surface structure is known, their precise chemical composition is not. Here, the epicuticular lipids of dragonfly wing membranes were characterized to investigate their significance in contributing to self-cleaning and superhydrophobic properties. After just 10s of lipid extraction using chloroform, the water contact angles exhibited by the wings decreased below the accepted threshold for superhydrophobicity (150°). Infrared spectra collected at the Australian Synchrotron contained characteristic absorption bands of amide, ester and aliphatic hydrocarbons moieties on the wing surfaces, the latter of which was decreased post-extraction with chloroform. GC-MS data analysis revealed that the epicuticular wax components were dominated by n-alkanes with even-numbered carbons, especially n-hexacosane, and palmitic acid. SEM and AFM data analysis conducted on the untreated and chloroform-extracted wing surfaces demonstrated that surface topography changed after extraction; the surface nanostructure was progressively lost with extended extraction times. The data presented here indicate that epicuticular lipids contribute not only to self-cleaning and superhydrophobic properties through their inherent hydrophobic nature, but also by forming the physical structure of the wing surface. This knowledge will be extremely valuable for reconstruction of dragonfly wing structures as a biomimetic template.

Spatial variations and temporal metastability of the self-cleaning and superhydrophobic properties of damselfly wings

Self-cleaning surfaces found in nature show great potential for application in many fields, ranging from industry to medicine. The ability for a surface to self-clean is intimately related to the wetting properties of the surface; for a surface to possess self-cleaning ability it must exhibit extremely high water contact angles and low water adhesion. While investigating the self-cleaning properties of damselfly wings, significant spatial variations in surface wettability were observed. Within an area of 100 μm × 100 μm of the wing surface the water contact angle was found to vary up to 17.8°, while remaining consistently superhydrophobic. The contributions of both surface chemistry and topography to the hydrophobicity of the wings were assessed in an effort to explain these variations. Synchrotron-sourced Fourier-transform infrared microspectroscopy revealed that some of the major components of the wing were aliphatic hydrocarbons and esters, which are attributable to epicuticular lipids. The wing topography, as determined by optical profilometry and atomic force microscopy (AFM), also showed only minor levels of heterogeneity arising from irregular ordering of surface nanostructures. The measured contact angle of a single droplet of water was also found to decrease over time as it evaporated, reaching a minimum of 107°. This is well below the threshold value for superhydrophobicity (i.e., 150°), demonstrating that when the surface is in contact with water for a prolonged period, the damselfly wings lose their superhydrophobicity and subsequently their ability to self-clean. This decrease in hydrophobicity over time can be attributed to the surface undergoing a transition from the Cassie-Baxter wettability state toward the Wenzel wettability state.