Flower-shaped 2D crystals grown in curved fluid vesicle membranes.

The morphologies of two-dimensional (2D) crystals, nucleated, grown, and integrated within 2D elastic fluids, for instance in giant vesicle membranes, are dictated by an interplay of mechanics, permeability, and thermal contraction. Mitigation of solid strain drives the formation of crystals with vanishing Gaussian curvature (i.e., developable domain shapes) and, correspondingly, enhanced Gaussian curvature in the surrounding 2D fluid. However, upon cooling to grow the crystals, large vesicles sustain greater inflation and tension because their small area-to-volume ratio slows water permeation. As a result, more elaborate shapes, for instance, flowers with bendable but inextensible petals, form on large vesicles despite their more gradual curvature, while small vesicles harbor compact planar crystals. This size dependence runs counter to the known cumulative growth of strain energy of 2D colloidal crystals on rigid spherical templates. This interplay of intra-membrane mechanics and processing points to the scalable production of flexible molecular crystals of controllable complex shape.

Contact Area and Deformation of Escherichia coli Cells Adhered on a Cationic Surface.

When bacteria adhere to surfaces, the chemical and mechanical character of the cell-substrate interface guides cell function and the development of microcolonies and biofilms. Alternately on bactericidal surfaces, intimate contact is critical to biofilm prevention. The direct study of the buried cell-substrate interfaces at the heart of these behaviors is hindered by the small bacterial cell size and inaccessibility of the contact region. Here, we present a total internal reflectance fluorescence depletion approach to measure the size of the cell-substrate contact region and quantify the gap separation and curvature near the contact zone, providing an assessment of the shapes of the near-surface undersides of adhered bacterial cells. Resolution of the gap height is about 10%, down to a few nanometers at contact. Using 1 and 2 µm silica spheres as calibration standards we report that, for flagella-free Escherichia coli (E. coli) adhering on a cationic poly-l-lysine layer, the cell-surface contact and apparent cell deformation vary with adsorbed cell configuration. Most cells adhere by their ends, achieving small contact areas of 0.15 µm2, corresponding to about 1-2% of the cell's surface. The altered Gaussian curvatures of end-adhered cells suggest the flattening of the envelope within the small contact region. When cells adhere by their sides, the contact area is larger, in the range 0.3-1.1 µm2 and comprising up to ∼12% of the cell's total surface. A region of sharper curvature, greater than that of the cells' original spherocylindrical shape, borders the flat contact region in cases of side-on or end-on cell adhesion, suggesting envelope stress. From the measured curvatures, precise stress distributions over the cell surface could be calculated in future studies that incorporate knowledge of envelope moduli. Overall the small contact areas of end-adhered cells may be a limiting factor for antimicrobial surfaces that kill on contact rather than releasing bactericide.

Depletion attractions drive bacterial capture on both non-fouling and adhesive surfaces, enhancing cell orientation.

Depletion attractions, occurring between surfaces immersed in a polymer solution, drive bacteria adhesion to a variety of surfaces. The latter include the surfaces of non-fouling coatings such as hydrated polyethylene glycol (PEG) layers but also, as demonstrated in this work, surfaces that are bacteria-adhesive, such as that of glass. Employing a flagella free E. coli strain, we demonstrate that cell adhesion on glass is enhanced by dissolved polyethylene oxide (PEO), exhibiting a faster rate and greater numbers of captured cells compared with the slower capture of the same cells on glass from a buffer solution. After removal of depletant, any cell retention appears to be governed by the substrate, with cells immediately released from non-fouling PEG surfaces but retained on glass. A distinguishing feature of cells captured by depletion on PEG surfaces is their orientation parallel to the surface and very strong alignment with flow. This suggests that, in the moments of capture, cells are able to rotate as they adhere. By contrast on glass, captured cells are substantially more upright and less aligned by flow. On glass the free polymer exerts forces that slightly tip cells towards the surface. Free polymer also holds cells still on adhesive and non-fouling surfaces alike but, upon removal of free PEO, cells retained on glass tend to be held by one end and exhibit a Brownian type rotational rocking.

Motility Increases the Numbers and Durations of Cell-Surface Engagements for Escherichia coli Flowing near Poly(ethylene glycol)-Functionalized Surfaces.

Bacteria are keenly sensitive to properties of the surfaces they contact, regulating their ability to form biofilms and initiate infections. This study examines how the presence of flagella, interactions between the cell body and the surface, or motility itself guides the dynamic contact between bacterial cells and a surface in flow, potentially enabling cells to sense physicochemical and mechanical properties of surfaces. This work focuses on a poly(ethylene glycol) biomaterial coating, which does not retain cells. In a comparison of four Escherichia coli strains with different flagellar expressions and motilities, cells with substantial run-and-tumble swimming motility exhibited increased flux to the interface (3 times the calculated transport-limited rate which adequately described the non-motile cells), greater proportions of cells engaging in dynamic nanometer-scale surface associations, extended times of contact with the surface, increased probability of return to the surface after escape and, as evidenced by slow velocities during near-surface travel, closer cellular approach. All these metrics, reported here as distributions of cell populations, point to a greater ability of motile cells, compared with nonmotile cells, to interact more closely, forcefully, and for greater periods of time with interfaces in flow. With contact durations of individual cells exceeding 10 s in the window of observation and trends suggesting further interactions beyond the field of view, the dynamic contact of individual cells may approach the minute timescales reported for mechanosensing and other cell recognition pathways. Thus, despite cell translation and the dynamic nature of contact, flow past a surface, even one rendered non-cell arresting by use of an engineered coating, may produce a subpopulation of cells already upregulating virulence factors before they arrest on a downstream surface and formally initiate biofilm formation.

Interplay of physico-chemical and mechanical bacteria-surface interactions with transport processes controls early biofilm growth: A review.

Biofilms initiate when bacteria encounter and are retained on surfaces. The surface orchestrates biofilm growth through direct physico-chemical and mechanical interactions with different structures on bacterial cells and, in turn, through its influence on cell-cell interactions. Individual cells respond directly to a surface through mechanical or chemical means, initiating "surface sensing" pathways that regulate gene expression, for instance producing extra cellular matrix or altering phenotypes. The surface can also physically direct the evolving colony morphology as cells divide and grow. In either case, the physico-chemistry of the surface influences cells and cell communities through mechanisms that involve additional factors. For instance the numbers of cells arriving on a surface from solution relative to the generation of new cells by division depends on adhesion and transport kinetics, affecting early colony density and composition. Separately, the forces experienced by adhering cells depend on hydrodynamics, gravity, and the relative stiffnesses and viscoelasticity of the cells and substrate materials, affecting mechanosensing pathways. Physical chemistry and surface functionality, along with interfacial mechanics also influence cell-surface friction and control colony morphology, in particular 2D and 3D shape. This review focuses on the current understanding of the mechanisms in which physico-chemical interactions, deriving from surface functionality, impact individual cells and cell community behavior through their coupling with other interfacial processes.

Depletion forces drive reversible capture of live bacteria on non-adhesive surfaces.

Because bacterial adhesion to surfaces is associated with infections and biofilm growth, it has been a longstanding goal to develop coatings that minimize biomolecular adsorption and eliminate bacteria adhesion. We demonstrate that, even on carefully-engineered non-bioadhesive coatings such as polyethylene glycol (PEG) layers that prevent biomolecule adsorption and cell adhesion, depletion interactions from non-adsorbing polymer in solution (such as 10 K PEG or 100 K PEO) can cause adhesion and retention of Escherichia coli cells, defeating the antifouling functionality of the coating. The cells are immobilized and remain viable on the timescale of the study, at least up to 45 minutes. When the polymer solution is replaced by buffer, cells rapidly escape from the surface, consistent with expectations for the reversibility of depletion attractions. The dissolved polymer additionally causes cells to aggregate in solution and aggregates rapidly dissociate to singlets upon tenfold dilution in buffer, also consistent with depletion. Hydrodynamic forces can substantially reduce the adhesion of aggregates on surfaces in conditions where single cells adhere via depletion. The findings reported here suggest that because bacteria thrive in polymer-rich environments both in vivo and in situ, depletion interactions may make it impossible to avoid bacterial retention on surfaces.

Surface Chemistry Guides the Orientations of Adhering E. coli Cells Captured from Flow.

Motivated by observations of cell orientation at biofilm-substrate interfaces and reports that cell orientation and adhesion play important roles in biofilm evolution and function, we investigated the influence of surface chemistry on the orientation of Escherichia coli cells captured from flow onto surfaces that were cationic, hydrophobic, or anionic. We characterized the initial orientations of nonmotile cells captured from gentle shear relative to the surface and flow directions. The broad distribution of captured cell orientations observed on cationic surfaces suggests that rapid electrostatic attractions of cells to oppositely charged surfaces preserve the instantaneous orientations of cells as they rotate in the near-surface shearing flow. By contrast, on hydrophobic and anionic surfaces, cells were oriented slightly more in the plane of the surface and in the flow direction compared with that on the cationic surface. This suggests slower development of adhesion at hydrophobic and anionic surfaces, allowing cells to tip toward the surface as they adhere. Once cells were captured, the flow was increased by 20-fold. Cells did not reorient substantially on the cationic surface, suggesting a strong cell-surface bonding. By contrast, on hydrophobic and anionic surfaces, increased shear forced cells to tip toward the surface and align in the flow direction, a process that was reversible upon reducing the shear. These findings suggest mechanisms by which surface chemistry may play a role in the evolving structure and function of microbial communities.

Escherichia coli Swimming back Toward Stiffer Polyetheylene Glycol Coatings, Increasing Contact in Flow.

Bacterial swimming in flow near surfaces is critical to the spread of infection and device colonization. Understanding how material properties affect flagella- and motility-dependent bacteria-surface interactions is a first step in designing new medical devices that mitigate the risk of infection. We report that, on biomaterial coatings such as polyethylene glycol (PEG) hydrogels and end-tethered layers that prevent adhesive bacteria accumulation, the coating mechanics and hydration control the near-surface travel and dynamic surface contact of E. coli cells in gentle shear flow (order 10 s-1). Along relatively stiff (order 1 MPa) PEG hydrogels or end-tethered layers of PEG chains of similar polymer correlation length, run-and-tumble E. coli travel nanometrically close to the coating's surface in the flow direction in distinguishable runs or "engagements" that persist for several seconds, after which cells leave the interface. The duration of these engagements was greater along stiff hydrogels and end-tethered layers compared with softer, more-hydrated hydrogels. Swimming cells that left stiff hydrogels or end-tethered layers proceeded out to distances of a few microns and then returned to engage the surface again and again, while cells engaging the soft hydrogel tended not to return after leaving. As a result of differences in the duration of engagements and tendency to return to stiff hydrogel and end-tethered layers, swimming E. coli experienced 3 times the integrated dynamic surface contact with stiff coatings compared with softer hydrogels. The striking similarity of swimming behaviors near 16-nm-thick end-tethered layers and 100-µm-thick stiff hydrogels argues that only the outermost several nanometers of a highly hydrated coating influence cell travel. The range of material stiffnesses, cell-surface distance during travel, and time scales of travel compared with run-and-tumble time scales suggests the influence of the coating derives from its interactions with flagella and its potential to alter flagellar bundling. Given that restriction of flagellar rotation is known to trigger increased virulence, bacteria influenced by surfaces in one region may become predisposed to form a biofilm downstream.

Switchable positioning of plate-like inclusions in lipid membranes: Elastically mediated interactions of planar colloids in 2D fluids.

We demonstrate how manipulating curvature in an elastic fluid lamella enables the reversible relative positioning of flat, rigid, plate-like micrometer-scale inclusions, with spacings from about a micrometer to tens of micrometers. In an experimental model comprising giant unilamellar vesicles containing solid domain pairs coexisting in a fluid membrane, we adjusted vesicle inflation to manipulate membrane curvature and mapped the interdomain separation. A two-dimensional model of the pair potential predicts the salient experimental observations and reveals both attractions and repulsions, producing a potential minimum entirely a result of the solid domain rigidity and bending energy in the fluid membrane. The impact of vesicle inflation on domain separation in vesicles containing two solid domains was qualitatively consistent with observations in vesicles containing many domains. The behavior differs qualitatively from the pure repulsions between fluid membrane domains or interactions between nanoscopic inclusions whose repulsive or attractive character is not switchable.

Rapid Electrostatic Capture of Rod-Shaped Particles on Planar Surfaces: Standing up to Shear.

We compare the electrostatically driven capture of flowing rod-shaped and spherical silica particles from dilute solutions onto a flow chamber wall that carries the opposite electrostatic charge from the particles. Particle accumulation and orientation are measured in time at a fixed region on the wall of a shear flow chamber. Rod-shaped particle aspect ratios are 2.5-3.2 and particle lengths are 1.3 and 2.67 µm for two samples, while sphere diameters were 0.72, 0.96, and 2.0 µm for three samples. At a moderate wall shear rate of 22 s-1, the particle accumulation for both rods and spheres is well described by diffusion-limited kinetics, demonstrating the limiting effect of particle diffusion in the near-wall boundary layer for electrostatically driven capture in this particle shape and size range. The significance of this finding is demonstrated in a calculation that shows that for delivery applications, nearly the same (within 10%) particle volume or mass is delivered to a surface at the diffusion-limited rate by rods and spheres. Therefore, in the absence of other motivating factors, the expense of developing rod-shaped microscale delivery packages to enhance capture from flow in the diffusion-limited simple shear regime is unwarranted. It is also interesting that the captured orientations of the larger rods, 2.6 µm in average length, were highly varied and insensitive to flow: a substantial fraction of rods were trapped in standing and slightly leaning orientations, touching the surface by their ends. Additionally, for particles that were substantially tipped over, there was only modest orientation in the flow direction. Taken together, these findings suggest that on the time scale of near-surface particle rotations, adhesion events are fast, trapping particles in orientations that do not necessarily maximize their favored adhesive contact or reduce hydrodynamic drag.

Mechanical Properties and Concentrations of Poly(ethylene glycol) in Hydrogels and Brushes Direct the Surface Transport of Staphylococcus aureus.

Surface-associated transport of flowing bacteria, including cell rolling, is a mechanism for otherwise immobile bacteria to migrate on surfaces and could be associated with biofilm formation or the spread of infection. This work demonstrates how the moduli and/or local polymer concentration play critical roles in sustaining contact, dynamic adhesion, and transport of bacterial cells along a hydrogel or hydrated brush surface. In particular, stiffer more concentrated hydrogels and brushes maintained the greatest dynamic contact, still allowing cells to travel along the surface in flow. This study addressed how the mechanical properties, molecular architectures, and thicknesses of minimally adhesive poly(ethylene glycol) (PEG)-based coatings influence the flow-driven surface motion of Staphylococcus aureus MS2 cells. Three protein-repellant PEG-dimethylacrylate hydrogel films (∼100 µm thick) and two protein-repellant PEG brushes (8-16 nm thick) were sufficiently fouling-resistant to prevent the accumulation of flowing bacteria. However, the rolling or hopping-like motions of gently flowing S. aureus cells along the surfaces were specific to the particular hydrogel or brush, distinguishing these coatings in terms of their mechanical properties (with moduli from 2 to 1300 kPa) or local PEG concentrations (in the range 10-50% PEG). On the stiffer hydrogel coatings having higher PEG concentrations, S. aureus exhibited long runs of surface rolling, 20-50 µm in length, an increased tendency of cells to repeatedly return to some surfaces after rolling and escaping, and relatively long integrated contact times. By contrast, on the softer more dilute hydrogels, bacteria tended to encounter the surface for brief periods before escaping without return. The dynamic adhesion and motion signatures of the cells on the two brushes were bracketed by those on the soft and stiff hydrogels, demonstrating that PEG coating thickness was not important in these studies where the vertically oriented surfaces minimized the impact of gravitational forces. Control studies with similarly sized poly(ethylene oxide)-coated rigid spherical microparticles, that also did not arrest on the PEG coatings, established that the bacterial skipping and rolling signatures were specific to the S. aureus cells and not simply diffusive. Dynamic adhesion of the S. aureus cells on the PEG hydrogel surfaces correlated well with quiescent 24 h adhesion studies in the literature, despite the orientation of the flow studies that eliminated the influence of gravity on bacteria-coating normal forces.

Nanoscale Functionalized Particles with Rotation-Controlled Capture in Shear Flow.

Important processes in nature and technology involve the adhesive capture of flowing particles or cells on the walls of a conduit. This paper introduces engineered spherical microparticles whose capture rates are limited by their near surface motions in flow. Specifically, these microparticles are sparsely functionalized with nanoscopic regions ("patches") of adhesive functionality, without which they would be nonadhesive. Not only is particle capture on the wall of a shear-chamber limited by surface chemistry as opposed to transport, but also the capture rates depend specifically on particle rotations that result from the vorticity of the shear flow field. These particle rotations continually expose new particle surface to the opposing chamber wall, sampling the particle surface for an adhesive region and controlling the capture rate. Control studies with the same patchy functionality on the chamber wall rather than the particles reveal a related signature of particle capture but substantially faster (still surface limited) particle capture rates. Thus, when the same functionality is placed on the wall rather than the particles, the capture is faster because it depends on the particle translation past a functionalized wall rather than on the particle rotations. The dependence of particle capture on functionalization of the particles versus the wall is consistent with the faster near-wall particle translation in shearing flow compared with the velocity of the rotating particle surface near the wall. These findings, in addition to providing a new class of nanoscopically patchy engineered particles, provide insight into the capture and detection of cells presenting sparse distinguishing surface features and the design of delivery packages for highly targeted pharmaceutical delivery.

Tunable fluorescence quenching near the graphene-aqueous interface.

With increased interest in graphene-based sensors for biomolecules and other targets, we investigated the impact of ionic strength on the steady-state emissions from fluorescein labels on proteins adsorbing on pristine CVD (chemical vapor deposited)-graphene on a silica support. Using the model system of fluorescein-tagged fibrinogen we demonstrated that, for fluorescein tags on adsorbed fibrinogen, emission intensity was very sensitive to the salt concentration. This behavior was not seen for fluorescein-tagged fibrinogen in solution. We demonstrated that fluorescein "quenching" in this system was a result of fluorescein's pH sensitivity: with changes in salt concentration, near-surface fluorescein experiences either the neutral bulk pH or, with a negatively charged surface, an acidic environment. The findings carry the important implication that the aqueous environment near silica-supported graphene is substantially acidic as a result of near-surface negative charge. This further implies, because of the purity of the graphene in this study and its lack of oxidation, that negative charge arises from ion adsorption and/or from the underlying silica support, which may be hydrated and present dissociated surface silanols. That is, the electrostatic potential from silica beneath the graphene may pass through the graphene, much as van der Waals interactions have been proven to do. Results were semi-quantitatively consistent with calculations that employed a Guoy Chapman model of the interface and the established pKa of the fluorescein. While these findings were obtained with adsorbed proteins, similar fluorescence quenching would be expected for any fluorescein-tagged species in the vicinity of silica-supported graphene. Thus, because of the negative charge at the aqueous graphene interface, ionic strength can be exploited as means of creating a molecular ruler of fluorescein emissions, and the emissions can be assessed within different distances, corresponding to the Debye length, from the graphene interface.

Evidence for negative charge near large area supported graphene in water: A study of silica microsphere interactions.

This study addresses the electrostatic and van der Waals interactions at the aqueous interface of large area CVD graphene, 1-3 layers thick on a silica support and assessed by Raman spectroscopy to have exclusive sp2 character. Ionic strength was found to substantially alter the interactions of silica microspheres with silica-supported graphene. Particles were nonadhesive at large Debye lengths but became irreversibly adherent at reduced Debye lengths about 2nm or less. This was demonstrated to be qualitatively parallel to the influence of ionic strength on silica-silica interactions. The observed ionic strength effects are best explained by negative charges in the vicinity, within a few nanometers, of the supported graphene. DLVO-based modeling of the silica-water-supported graphene interaction suggests that van der Waals interactions drive particle capture and that the surface potential at the supported graphene surface is at least -10 to -15mV (corresponding to a charge density of 0.02-0.06/nm2). This charge could result from ion adsorption or from charges on silica beneath the graphene. The conclusions are not substantially affected by inclusion of nanometer-scale interfacial roughness in the modeling.

Surfaces for competitive selective bacterial capture from protein solutions.

Active surfaces that form the basis for bacterial sensors for threat detection, food safety, or certain diagnostic applications rely on bacterial adhesion. However, bacteria capture from complex fluids on the active surfaces can be reduced by the competing adsorption of proteins and other large molecules. Such adsorption can also interfere with device performance. As a result, multiple upstream processing steps are frequently employed to separate macromolecules from any cells, which remain in the buffer. Here, we present an economical approach to capture bacteria, without competitive adsorption by proteins, on engineered surfaces that do not employ biomolecular recognition, antibodies, or other molecules with engineered sequences. The surfaces are based on polyethylene glycol (PEG) brushes that, on their own, repel both proteins and bacteria. These PEG brushes backfill the surface around sparsely adsorbed cationic polymer coils (here, poly-L-lysine (PLL)). The PLL coils are effectively embedded within the brush and produce locally cationic nanoscale regions that attract negatively charged regions of proteins or cells against the steric background repulsion from the PEG brush. By carefully designing the surfaces to include just enough PLL to capture bacteria, but not enough to capture proteins, we achieve sharp selectivity where S. aureus is captured from albumin- or fibrinogen-containing solutions, but free albumin or fibrinogen molecules are rejected from the surface. Bacterial adhesion on these surfaces is not reduced by competitive protein adsorption, in contrast to performance of more uniformly cationic surfaces. Also, protein adsorption to the bacteria does not interfere with capture, at least for the case of S. aureus, to which fibrinogen binds through a specific receptor.

Statistically-based DLVO approach to the dynamic interaction of colloidal microparticles with topographically and chemically heterogeneous collectors.

Electrostatic surface heterogeneity on the order of a few nanometers is common in colloidal and bacterial systems, dominating adhesion and aggregation and inducing deviations from classical DLVO theory based on a uniform distribution of surface charge. Topographical heterogeneity and roughness also strongly influence adhesion. In this work, a model is introduced to quantify the spatial fluctuations in the interaction of microparticles in a flowing suspension with a wall aligned parallel to the flow. The wall contains nanoscale chemical and topographical heterogeneities ("patches") that are randomly distributed and produce localized attraction and repulsion. These attractive and repulsive regions induce fluctuations in the trajectories of the flowing particles that are critical to particle capture by the wall. The statistical distribution of patches is combined with mean-field DLVO calculations between a particle and two homogeneous surfaces: one with the surface potential of the patches and one with the potential of the underlying wall. These surface potentials could be obtained in experiments from zeta potential measurements for the bare wall and for one saturated with patches. This simple model reproduces the mean DLVO interaction force or energy vs. particle-wall separation distance, its variance, and particle adhesion thresholds from direct simulations of particle trajectories over patchy surfaces. The predictions of the model are consistent with experimental findings of significant microparticle deposition onto patchy, net-repulsive surfaces whose apparent zeta potential has the same sign as that of the particles. Deposition is significantly enhanced if the patches protrude even slightly from the surface. The model predictions are also in agreement with the observed variation of the adhesion threshold with the shear rate in published studies of dynamic microparticle adhesion on patchy surfaces.

Engineering nanoscale surface features to sustain microparticle rolling in flow.

Nanoscopic features of channel walls are often engineered to facilitate microfluidic transport, for instance when surface charge enables electro-osmosis or when grooves drive mixing. The dynamic or rolling adhesion of flowing microparticles on a channel wall holds potential to accomplish particle sorting or to selectively transfer reactive species or signals between the wall and flowing particles. Inspired by cell rolling under the direction of adhesion molecules called selectins, we present an engineered platform in which the rolling of flowing microparticles is sustained through the incorporation of entirely synthetic, discrete, nanoscale, attractive features into the nonadhesive (electrostatically repulsive) surface of a flow channel. Focusing on one example or type of nanoscale feature and probing the impact of broad systematic variations in surface feature loading and processing parameters, this study demonstrates how relatively flat, weakly adhesive nanoscale features, positioned with average spacings on the order of tens of nanometers, can produce sustained microparticle rolling. We further demonstrate how the rolling velocity and travel distance depend on flow and surface design. We identify classes of related surfaces that fail to support rolling and present a state space that identifies combinations of surface and processing variables corresponding to transitions between rolling, free particle motion, and arrest. Finally we identify combinations of parameters (surface length scales, particle size, flow rates) where particles can be manipulated with size-selectivity.

Hybrid copolymer-phospholipid vesicles: phase separation resembling mixed phospholipid lamellae, but with mechanical stability and control.

Vesicles whose bilayer membranes contain phospholipids mixed with co-polymers or surfactants comprise new hybrid materials having potential applications in drug delivery, sensors, and biomaterials. Here we describe a model polymer-phospholipid hybrid membrane system exhibiting strong similarities to binary phospholipid mixtures, but with more robust membrane mechanics. A lamella-forming graft copolymer, PDMS-co-PEO (polydimethylsiloxane-co-polyethylene oxide) was blended with a high melting temperature phospholipid, DPPC (1,2-dipalmitoyl-sn-glycero-3-phosphocholine), over a broad compositional range. The resulting giant hybrid unilamellar vesicles were compared qualitatively and quantitatively to analogous mixed phospholipid membranes in which a low melting temperature phospholipid, DOPC (1,2-dioleoyl-sn-glycero-3-phosphocholine), was blended with DPPC. The mechanical properties of the hybrid vesicles, even when phase separated, were robust with high lysis stresses and strains approaching those of the pure copolymer vesicles. The temperature-composition phase diagram of the hybrid vesicles closely resembled that of the mixed phospholipids; with only slightly greater nonidealities in the hybrid compared with DOPC/DPPC mixed membranes. In both systems, it was demonstrated that tension could be used to manipulate DPPC solidification into domains of patchy or striped morphologies that exhibited different tracer incorporation. The patch and stripe-shaped domains are thought to be different solid DPPC polymorphys: ripple and tilt (or gel). This work demonstrates that in mixed-phospholipid bilayers where a high-melting phospholipid solidifies on cooling, the lower-melting phospholipid may be substituted by an appropriate copolymer to improve mechanical properties while retaining the underlying membrane physics.

Antimicrobial surfaces containing cationic nanoparticles: how immobilized, clustered, and protruding cationic charge presentation affects killing activity and kinetics.

This work examines how the antimicrobial (killing) activity of net-negative surfaces depends on the presentation of antimicrobial cationic functionality: distributed versus clustered, and flat clusters versus raised clusters. Specifically, the ability to kill Staphylococcus aureus by sparsely distributed 10 nm cationic nanoparticles, immobilized on a negative surface and backfilled with a PEG (polyethylene glycol) brush, was compared with that for a dense layer of the same immobilized nanoparticles. Additionally, sparsely distributed 10 nm poly-L-lysine (PLL) coils, adsorbed to a surface to produce flat cationic "patches" and backfilled with a PEG brush were compared to a saturated adsorbed layer of PLL. The latter resembled classical uniformly cationic antimicrobial surfaces. The protrusion of the cationic clusters substantially influenced killing but the surface concentration of the clusters had minor impact, as long as bacteria adhered. When surfaces were functionalized at the minimum nanoparticle and patch densities needed for bacterial adhesion, killing activity was substantial within 30 min and nearly complete within 2 h. Essentially identical killing was observed on more densely functionalized surfaces. Surfaces containing protruding (by about 8 nm) nanoparticles accomplished rapid killing (at 30 min) compared with surfaces containing similarly cationic but flat features (PLL patches). Importantly, the overall surface density of cationic functionality within the clusters was lower than reported thresholds for antimicrobial action. Also surprising, the nanoparticles were far more deadly when surface-immobilized compared with free in solution. These findings support a killing mechanism involving interfacial stress.