Preferential adhesion of bacterial cells onto top- and bottom-mounted nanostructured surfaces under flow conditions.

The bactericidal effect of biomimetic nanostructured surfaces has been known for a long time, with recent data suggesting an enhanced efficiency of the nanostructured surfaces under fluid shear. While some of the influential factors on the bactericidal effect of nanostructured surfaces under fluid shear are understood, there are numerous important factors yet to be studied, which is essential for the successful implementation of this technology in industrial applications. Among those influential factors, the orientation of the nanostructured surface can play an important role in bacterial cell adhesion onto surfaces. Gravitational effects can become dominant under low flow velocities, making the diffusive transport of bacterial cells more prominent than the advective transport. However, the role of nanostructure orientation in determining its bactericidal efficiency under flow conditions is still not clear. In this study, we analysed the effect of surface orientation of nanostructured surfaces, along with bacterial cell concentration, fluid flow rate, and the duration of time which the surface is exposed to flow, on bacterial adhesion and viability on these surfaces. Two surface orientations, with one on the top and the other on the bottom of a flow channel, were studied. Under flow conditions, the bactericidal efficacy of the nanostructured surface is both orientation and bacterial species dependent. The effects of cell concentration, fluid flow rate, and exposure time on cell adhesion are independent of the nanostructured surface orientation. Fluid shear showed a species-dependent effect on bacterial adhesion, while the effects of concentration and exposure time on bacterial cell adhesion are independent of the bacterial species. Moreover, bacterial cells demonstrate preferential adhesion onto surfaces based on the surface orientation, and these effects are species dependent. These results outline the capabilities and limitations of nanostructures under flow conditions. This provides valuable insights into the applications of nanostructures in medical or industrial sectors, which are associated with overlaying fluid flow.

Bactericidal Efficacy of Nanostructured Surfaces Increases under Flow Conditions.

Bacterial colonization on solid surfaces creates enormous problems across various industries causing billions of dollars' worth of economic damages and costing human lives. Biomimicking nanostructured surfaces have demonstrated a promising future in mitigating bacterial colonization and related issues. The importance of this non-chemical method has been elevated due to bacterial evolvement into antibiotic and antiseptic-resistant strains. However, bacterial attachment and viability on nanostructured surfaces under fluid flow conditions has not been investigated thoroughly. In this study, attachment and viability of Pseudomonas aeruginosa (P. aeruginosa) and Staphylococcus aureus (S. aureus) on a model nanostructured surface were studied under fluid flow conditions. A wide range of flow rates resulting in a broad spectrum of fluid wall shear stress on a nanostructured surface representing various application conditions were experimentally investigated. The bacterial suspension was pumped through a custom-designed microfluidic device (MFD) that contains a sterile Ti-6Al-4V substrate. The surface of the titanium substrate was modified using a hydrothermal synthesis process to fabricate the nanowire structure on the surface. The results of the current study show that the fluid flow significantly reduces bacterial adhesion onto nanostructured surfaces and significantly reduces the viability of adherent cells. Interestingly, the bactericidal efficacy of the nanostructured surface was increased under the flow by ∼1.5-fold against P. aeruginosa and ∼3-fold against S. aureus under static conditions. The bactericidal efficacy had no dependency on the fluid wall shear stress level. However, trends in the dead-cell count with the fluid wall shear were slightly different between the two species. These findings will be highly useful in developing and optimizing nanostructures in the laboratory as well as translating them into successful industrial applications. These findings may be used to develop antibacterial surfaces on biomedical equipment such as catheters and vascular stents or industrial applications such as ship hulls and pipelines where bacterial colonization is a great challenge.

Fluid Flow Induces Differential Detachment of Live and Dead Bacterial Cells from Nanostructured Surfaces.

Nanotopographic surfaces are proven to be successful in killing bacterial cells upon contact. This non-chemical bactericidal property has paved an alternative way of fighting bacterial colonization and associated problems, especially the issue of bacteria evolving resistance against antibiotic and antiseptic agents. Recent advancements in nanotopographic bactericidal surfaces have made them suitable for many applications in medical and industrial sectors. The bactericidal effect of nanotopographic surfaces is classically studied under static conditions, but the actual potential applications do have fluid flow in them. In this study, we have studied how fluid flow can affect the adherence of bacterial cells on nanotopographic surfaces. Gram-positive and Gram-negative bacterial species were tested under varying fluid flow rates for their retention and viability after flow exposure. The total number of adherent cells for both species was reduced in the presence of flow, but there was no flowrate dependency. There was a significant reduction in the number of live cells remaining on nanotopographic surfaces with an increasing flowrate for both species. Conversely, we observed a flowrate-independent increase in the number of adherent dead cells. Our results indicated that the presence of flow differentially affected the adherent live and dead bacterial cells on nanotopographic surfaces. This could be because dead bacterial cells were physically pierced by the nano-features, whereas live cells adhered via physiochemical interactions with the surface. Therefore, fluid shear was insufficient to overcome adhesion forces between the surface and dead cells. Furthermore, hydrodynamic forces due to the flow can cause more planktonic and detached live cells to collide with nano-features on the surface, causing more cells to lyse. These results show that nanotopographic surfaces do not have self-cleaning ability as opposed to natural bactericidal nanotopographic surfaces, and nanotopographic surfaces tend to perform better under flow conditions. These findings are highly useful for developing and optimizing nanotopographic surfaces for medical and industrial applications.