Multiparametric Sensing of Outer Membrane Vesicle-Derived Supported Lipid Bilayers Demonstrates the Specificity of Bacteriophage Interactions.

The use of bacteriophages, viruses that specifically infect bacteria, as antibiotics has become an area of great interest in recent years as the effectiveness of conventional antibiotics recedes. The detection of phage interactions with specific bacteria in a rapid and quantitative way is key for identifying phages of interest for novel antimicrobials. Outer membrane vesicles (OMVs) derived from Gram-negative bacteria can be used to make supported lipid bilayers (SLBs) and therefore in vitro membrane models that contain naturally occurring components of the bacterial outer membrane. In this study, we employed Escherichia coli OMV derived SLBs and use both fluorescent imaging and mechanical sensing techniques to show their interactions with T4 phage. We also integrate these bilayers with microelectrode arrays (MEAs) functionalized with the conducting polymer PEDOT:PSS and show that the pore forming interactions of the phages with the SLBs can be monitored using electrical impedance spectroscopy. To highlight our ability to detect specific phage interactions, we also generate SLBs using OMVs derived from Citrobacter rodentium, which is resistant to T4 phage infection, and identify their lack of interaction with the phage. The work presented here shows how interactions occurring between the phages and these complex SLB systems can be monitored using a range of experimental techniques. We believe this approach can be used to identify phages that work against bacterial strains of interest, as well as more generally to monitor any pore forming structure (such as defensins) interacting with bacterial outer membranes, and thus aid in the development of next generation antimicrobials.

Survival of polymeric microstructures subjected to interrogatory touch.

Polymeric arrays of microrelief structures have a range of potential applications. For example, to influence wettability, to act as biologically inspired adhesives, to resist biofouling, and to play a role in the "feel" of an object during tactile interaction. Here, we investigate the damage to micropillar arrays comprising pillars of different modulus, spacing, diameter, and aspect ratio due to the sliding of a silicone cast of a human finger. The goal is to determine the effect of these parameters on the types of damage observed, including adhesive failure and ploughing of material from the finger onto the array. Our experiments point to four principal conclusions [1]. Aspect ratio is the dominant parameter in determining survivability through its effect on the bending stiffness of micropillars [2]. All else equal, micropillars with larger diameter are less susceptible to breakage and collapse [3]. The spacing of pillars in the array largely determines which type of adhesive failure occurs in non-surviving arrays [4]. Elastic modulus plays an important role in survivability. Clear evidence of elastic recovery was seen in the more flexible polymer and this recovery led to more instances of pristine survivability where the stiffer polymer tended to ablate PDMS. We developed a simple model to describe the observed bending of micropillars, based on the quasi-static mechanics of beam-columns, that indicated they experience forces ranging from 10-4-10-7 N to deflect into adhesive contact. Taken together, results obtained using our framework should inform design considerations for microstructures intended to be handled by human users.