Vi Khanh Truong

The influence of nano-scale surface roughness on bacterial adhesion to ultrafine-grained titanium.

We discuss the effect of extreme grain refinement in the bulk of commercial purity titanium (CP, Grade-2) on bacterial attachment to the mechano-chemically polished surfaces of the material. The ultrafine crystallinity of the bulk was achieved by severe plastic deformation by means of equal channel angular pressing (ECAP). The chemical composition, wettability, surface topography and roughness of titanium surfaces were characterized using X-ray photoelectron spectroscopy (XPS) and water contact angle (WCA) measurements, as well as atomic force microscopy (AFM) with 3D interactive visualization of the titanium surface morphology. It was found that physico-chemical surface characteristics of the as-received and the ECAP-modified CP titanium did not differ in any significant way, while the surface roughness at the nano-scale did. Optical profilometry performed on large scanning areas of approximately 225 mum x 300 mum showed that there was no significant difference between the roughness parameters R(a) and R(q) for surfaces in the two conditions, the overall level of roughness being lower for the ECAP-processed one. By contrast, topographic profile analysis at the nano-scale by AFM did reveal a difference in these parameters. This difference was sensitive to the size of the scanned surface area. A further two surface roughness parameters, skewness (R(skw)) and kurtosis (R(kur)), were also used to describe the morphology of titanium surfaces. It was found that the bacterial strains used in this study as adsorbates, viz. Staphylococcus aureus CIP 65.8 and Pseudomonas aeruginosa ATCC 9025, showed preference for surfaces of ECAP-processed titanium. S. aureus cells were found to have a greater propensity for attachment to surfaces of ECAP-modified titanium, while the attachment of P. aeruginosa, while also showing some preference for the ECAP-processed material, was less sensitive to the ECAP processing.

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.

Bacterial Retention on Superhydrophobic Titanium Surfaces Fabricated by Femtosecond Laser Ablation

Two-tier micro- and nanoscale quasi-periodic self-organized structures, mimicking the surface of a lotus Nelumbo nucifera leaf, were fabricated on titanium surfaces using femtosecond laser ablation. The first tier consisted of large grainlike convex features between 10 and 20 μm in size. The second tier existed on the surface of these grains, where 200 nm (or less) wide irregular undulations were present. The introduction of the biomimetic surface patterns significantly transformed the surface wettabilty of the titanium surface. The original surface possessed a water contact angle of θ(W) 73 ± 3°, whereas the laser-treated titanium surface became superhydrophobic, with a water contact angle of θ(W) 166 ± 4°. Investigations of the interaction of S. aureus and P. aeruginosa with these superhydrophobic surfaces at the surface-liquid interface revealed a highly selective retention pattern for two pathogenic bacteria. While S. aureus cells were able to successfully colonize the superhydrophobic titanium surfaces, no P. aeruginosa cells were able to attach to the surface (i.e., any attached bacterial cells were below the estimated lower detection limit).

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.

Impact of nanoscale roughness of titanium thin film surfaces on bacterial retention

Two human pathogenic bacteria, Staphylococcus aureus CIP 68.5 and Pseudomonas aeruginosa ATCC 9025, were adsorbed onto surfaces containing Ti thin films of varying thickness to determine the extent to which nanoscale surface roughness influences the extent of bacterial attachment. A magnetron sputter thin film system was used to deposit titanium films with thicknesses of 3, 12, and 150 nm on glass substrata with corresponding surface roughness parameters of R(q) 1.6, 1.2, and 0.7 nm (on a 4 microm x 4 microm scanning area). The chemical composition, wettability, and surface architecture of titanium thin films were characterized using X-ray photoelectron spectroscopy, contact angle measurements, atomic force microscopy, three-dimensional interactive visualization, and statistical approximation of the topographic profiles. Investigation of the dynamic evolution of the Ti thin film topographic parameters indicated that three commonly used parameters, R(a), R(q), and R(max), were insufficient to effectively characterize the nanoscale rough/smooth surfaces. Two additional parameters, R(skw) and R(kur), which describe the statistical distributions of roughness character, were found to be useful for evaluating the surface architecture. Analysis of bacterial retention profiles indicated that bacteria responded differently to the surfaces on a scale of less than 1 nm change in the R(a) and R(q) Ti thin film surface roughness parameters by (i) an increased number of retained cells by a factor of 2-3, and (ii) an elevated level of secretion of extracellular polymeric substances.

Graphene Induces Formation of Pores That Kill Spherical and Rod-Shaped Bacteria

Pristine graphene, its derivatives, and composites have been widely reported to possess antibacterial properties. Most of the studies simulating the interaction between bacterial cell membranes and the surface of graphene have proposed that the graphene-induced bacterial cell death is caused either by (1) the insertion of blade-like graphene-based nanosheets or (2) the destructive extraction of lipid molecules by the presence of the lipophilic graphene. These simulation studies have, however, only take into account graphene-cell membrane interactions where the graphene is in a dispersed form. In this paper, we report the antimicrobial behavior of graphene sheet surfaces in an attempt to further advance the current knowledge pertaining to graphene cytotoxicity using both experimental and computer simulation approaches. Graphene nanofilms were fabricated to exhibit different edge lengths and different angles of orientation in the graphene sheets. These substrates were placed in contact with Pseudomonas aeruginosa and Staphylococcus aureus bacteria, where it was seen that these substrates exhibited variable bactericidal efficiency toward these two pathogenic bacteria. It was demonstrated that the density of the edges of the graphene was one of the principal parameters that contributed to the antibacterial behavior of the graphene nanosheet films. The study provides both experimental and theoretical evidence that the antibacterial behavior of graphene nanosheets arises from the formation of pores in the bacterial cell wall, causing a subsequent osmotic imbalance and cell death.

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.

Accelerated stem cell attachment to ultrafine grained titanium.

Commercial purity titanium with an average grain size in the low sub-micron range was produced by equal channel angular pressing (ECAP). Attachment of human bone marrow-derived mesenchymal stem cells (hMSCs) to the surface of conventional coarse grained and ECAP-modified titanium was studied. It was demonstrated that the attachment and spreading of hMSCs in the initial stages (up to 24h) of culture was enhanced by grain refinement. Surface characterization by a range of techniques showed that the main factor responsible for the observed acceleration of hMSC attachment and spreading on titanium due to grain refinement in the bulk is the attendant changes in surface topography on the nanoscale. These results indicate that, in addition to its superior mechanical properties, ECAP-modified titanium possesses improved biocompatibility, which makes it to a potent candidate for applications in medical implants.