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Using a Stokesian dynamics simulation, the microstructure of particle aggregates at an oil/water interface with an applied Couette flow is studied. The results of the aggregation are consistent with previously published experimental work demonstrating multiple regimes of behavior based on the relative strength of shear and capillary forces. In previous work, densification of aggregates at low shear rates was theorized to occur due to short time scale fragmentation/reaggregation of aggregates with rigid particle bonds. In simulations, densification is observed at low shear rates but occurs due to local reorganization of particles due to capillary torques over long time scales. Moderate shear rates create mobile bonds between particles at shorter time scales, allowing aggregates to fragment without reaggregation into smaller isolated clusters, consistent with prior experimental work. At the highest shear rates, aggregation is inhibited completely.
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A new technique was used to measure the viscoelasticity of in vivo Pseudomonas aeruginosa biofilms. This was done through ex vivo microrheology measurements of in vivo biofilms excised from mouse wound beds. To our knowledge, this is the first time that the mechanics of in vivo biofilms have been measured. In vivo results are then compared to typical in vitro measurements. Biofilms grown in vivo are more relatively elastic than those grown in a wound-like medium in vitro but exhibited similar compliance. Using various genetically mutated P. aeruginosa strains, it is observed that the contributions of the exopolysaccharides Pel, Psl, and alginate to biofilm viscoelasticity were different for the biofilms grown in vitro and in vivo. In vitro experiments with collagen containing medium suggest this likely arises from the incorporation of host material, most notably collagen, into the matrix of the biofilm when it is grown in vivo. Taken together with earlier studies that examined the in vitro effects of collagen on mechanical properties, we conclude that collagen may, in some cases, be the dominant contributor to biofilm viscoelasticity in vivo.
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Biopelículas , Pseudomonas aeruginosa , Animales , Colágeno/metabolismo , Colágeno/farmacología , Ratones , Polisacáridos Bacterianos/metabolismo , Pseudomonas aeruginosa/fisiología , Sustancias Viscoelásticas , Heridas y Lesiones/microbiologíaRESUMEN
Pseudomonas aeruginosa is an opportunistic pathogen that causes thousands of deaths every year in part due to its ability to form biofilms composed of bacteria embedded in a matrix of self-secreted extracellular polysaccharides (EPS), e-DNA, and proteins. In chronic wounds, biofilms are exposed to the host extracellular matrix, of which collagen is a major component. How bacterial EPS interacts with host collagen and whether this interaction affects biofilm viscoelasticity is not well understood. Since physical disruption of biofilms is often used in their removal, knowledge of collagen's effects on biofilm viscoelasticity may enable new treatment strategies that are better tuned to biofilms growing in host environments. In this work, biofilms are grown in the presence of different concentrations of collagen that mimic in vivo conditions. In order to explore collagen's interaction with EPS, nine strains of P. aeruginosa with different patterns of EPS production were used to grow biofilms. Particle tracking microrheology was used to characterize the mechanical development of biofilms over two days. Collagen is found to decrease biofilm compliance and increase relative elasticity regardless of the EPS present in the system. However, this effect is minimized when biofilms overproduce EPS. Collagen appears to become a de facto component of the EPS, through binding to bacteria or physical entanglement.
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Biopelículas , Pseudomonas aeruginosa , Colágeno , Polisacáridos Bacterianos , ViscosidadRESUMEN
Hydrophobic/hydrophilic mixtures of latex particles at an air/water interface self-assemble, creating space filling, interconnected aggregates as the relative surface fractions of the dissimilar particles approach 0.5, which is reflected both in qualitative observation and fractal dimension of the microstructure. It is hypothesized that this change in microstructure occurs due to an asymmetry in the electrostatic interaction between similar and dissimilar particles caused by polarization of hydrophilic particles by hydrophobic particles. The changes in both microstructure and interparticle interactions significantly impact the interfacial viscoelasticity. As greater shape complexity is observed, interfacial complex moduli can increase by as much as 3 orders of magnitude and interfaces become more elastic.
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HYPOTHESIS: Aggregation of particles on a liquid interface is controlled by inter-particle forces and hydrodynamic interactions. Previous experimental work has shown atypical structures despite diffusion limited cluster aggregation like behavior. It is likely that this is primarily due to the role of capillary quadrupoles in allowing particle repositioning after aggregation, which is tested here. EXPERIMENTS: Using Stokesian dynamics and inter-particle forces unique to particles at liquid interfaces, aggregation of particles adsorbed to a liquid interface is studied. Simulations' parameters are adjusted to control hydrodynamic interaction strength, initial particle position, and inter-particle forces magnitudes to compare to existing experimental results and hypothesis. FINDINGS: It is found that initial particle position plays a small role on equilibrium interfacial microstructure but has a significant impact on aggregation kinetics. Interfacial hydrodynamic interactions and inter-particle forces have a strong impact on equilibrium microstructure by altering the amount particles can reposition, which is consistent with published results. Capillary forces that allow significant repositioning after contact appear to play a key role in previously observed fractal dimensions of particle laden interfaces.
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Microbial biofilms are viscoelastic materials formed by bacteria, which occur on solid surfaces, at liquid interfaces, or in free solution. Although solid surface biofilms have been widely studied, pellicles, biofilms at liquid interfaces, have had significantly less focus. In this work, interfacial shear rheology and scanning electron microscopy imaging are used to characterize how flagella, type IV pili, biosurfactants, and extracellular polymeric substance polysaccharides affect the formation of pellicles by Pseudomonas aeruginosa at an air/water interface. Pellicles still form with the loss of a single biological attachment mechanism, which is hypothesized to be due to surface tension-aided attachment. Changes in the surface structure of the pellicles are observed when changing both the function/structure of type IV pili, removing the flagella, or stopping the expression of biosurfactants. However, these changes do not appear to affect pellicle elasticity in a consistent way. Traits that affect adsorption and growth/spreading appear to affect pellicles in a manner consistent with literature results for solid surface biofilms; small differences are seen in attachment-related mechanisms, which may occur due to surface tension.
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Proteínas Bacterianas/metabolismo , Fimbrias Bacterianas/metabolismo , Pseudomonas aeruginosa/metabolismo , Matriz Extracelular de Sustancias Poliméricas/metabolismo , Flagelos/metabolismo , Microscopía Electrónica de Rastreo , Tensoactivos/metabolismoRESUMEN
HYPOTHESIS: It is possible to control the absolute and relative magnitude of repulsive and attractive interactions and hence microstructure of interfacial particles at and air/water interface by adjusting subphase composition. It should be possible to modify interfacial viscoelasticity from elastic to viscous behavior through these changes to interfacial microstructure. EXPERIMENTS: Particle laden interfaces are made from micron sized polystyrene at an air/water interface. The inter-particle interactions are controlled by the subphase salt concentration and addition of both non-ionic and ionic surfactants. These interfaces are then characterized using an interfacial rheometer with a custom visualization system. FINDINGS: Three distinct microstructures are observed. Low repulsion and high attraction systems exhibit a soft glassy rheology with a disordered but dense microstructure. Creating high repulsion results in a dense hexagonal crystal. Finally, in systems with reduced repulsion and attraction, a hexatic phase can be observed. Each of these microstructures exhibit unique interfacial viscoelastic behavior. These results indicate that control over the properties of these interfaces, and hence Pickering emulsions, is possible through manipulation of interparticle forces.
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Valvular interstitial cells (VICs) constitute the major cell population in heart valves. Quiescent fibroblastic VICs are seen in adult healthy valves. They become activated myofibroblastic VICs during development, in diseased valves and in vitro. 2D substrate stiffness within a 5-15 kPa range along with high passage numbers promote VIC activation in vitro. In this study, we characterize VIC quiescence and activation across a 1-21 kPa range of substrate stiffness and passages. We define a cell morphology characterization system for VICs as they transform. We hypothesize that VICs show distinct morphological characteristics in different activation states and the morphology distribution varies with substrate stiffness and passage number. Four VIC morphologies - tailed, spindle, rhomboid and triangle - account for the majority of VIC in this study. Using α-smooth muscle actin (α-SMA), non-muscle myosin heavy chain B (SMemb) and transforming growth factor ß (TGF-ß) as activation markers for validation, we developed a system where we categorize morphology distribution of VIC cultures, to be potentially used as a non-destructive detection method of activation state. We also show that this system can be used to force stiffness-induced deactivation. The reversibility in VIC activation has important implications in in vitro research and tissue engineering.
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Diferenciación Celular/fisiología , Fibroblastos/citología , Fibroblastos/metabolismo , Válvulas Cardíacas/citología , Animales , Válvulas Cardíacas/metabolismo , Miofibroblastos/citología , Miofibroblastos/metabolismo , Fenotipo , PorcinosRESUMEN
A Stokesian dynamics simulation of the effect of surface Couette flow on the microstructure of particles irreversibly adsorbed to an interface is presented. Rather than modeling both bulk phases, the interface, and particles in a full 3D simulation, known interfacial interactions between adsorbed particles are used to create a 2D model from a top down perspective. This novel methodology is easy to implement and computationally inexpensive, which makes it favorable to simulate behavior of particles under applied flow at fluid-fluid interfaces. The methodology is used to examine microstructure deformation of monodisperse, rigid spherical colloids with repulsive interactions when a surface Couette flow is imposed. Simulation results compare favorably to experimental results taken from literature, showing that interparticle forces must be 1 order of magnitude greater than viscous drag for microstructure to transition from aligned particle strings to rotation of local hexagonal domains. Additionally, it is demonstrated that hydrodynamic interactions between particles play a significant role in the magnitude of these microstructure deformations.
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Hyaluronic acid solutions have been widely studied due to their relevance to the rheological behavior of synovial fluid and joint lubrication. Ambulatory joint motion is typically large oscillatory deflections; therefore, large amplitude oscillatory shear strain experiments are used to examine the relevant non-linear viscoelastic properties of these solutions. Using the sequence of physical processes method to analyze data provides time dependent viscoelastic moduli, which exhibit a clear physiologically relevant behavior to hyaluronic acids non-linear viscoelasticity. In particular, it is seen that during peak strain/acceleration, the time dependent elastic modulus peaks and the loss modulus is at a minimum. The hyaluronic acid can provide an immediate elastic response to sudden forces, acting like a shock absorber during sudden changes in direction of motion or maximum deflection. However, during peak rate, the elastic modulus is at a minimum and the loss modulus is at a maximum, which provides greater efficacy to hydrodynamic shear lubrication.
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Elasticidad , Ácido Hialurónico/metabolismo , Dinámicas no Lineales , Líquido Sinovial/metabolismo , Humanos , Rodilla/fisiología , Modelos Biológicos , Movimiento , Reología , Estrés Mecánico , Factores de Tiempo , ViscosidadRESUMEN
Recent measurements have implied a distribution of interfacially adsorbed particles' contact angles; however, it has been impossible to measure statistically significant numbers for these contact angles noninvasively in situ. Using a new microscopy method that allows nanometer-scale resolution of particle's 3D positions on an interface, we have measured the contact angles for thousands of latex particles at an oil/water interface. Furthermore, these measurements are dynamic, allowing the observation of the particle contact angle with high temporal resolution, resulting in hundreds of thousands of individual contact angle measurements. The contact angle has been found to fit a normal distribution with a standard deviation of 19.3°, which is much larger than previously recorded. Furthermore, the technique used allows the effect of measurement error, constrained interfacial diffusion, and particle property variation on the contact angle distribution to be individually evaluated. Because of the ability to measure the contact angle noninvasively, the results provide previously unobtainable, unique data on the dynamics and distribution of the adsorbed particles' contact angle.
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The role of interfacial rheology on the bulk linear viscoelastic moduli of low concentration bovine albumin solutions is probed. Previously reported soft gel properties of these systems were attributed to either protein aggregation or organization within the bulk. Instead, these behaviors are shown to be attributable to the measurement error caused by interfacial rheology due to adsorption of bovine serum albumin to the air and water interface. Even at low bulk concentrations, fast interfacial adsorption results in erroneous measurements. When these effects are removed, the solutions are viscous dominated with a dynamic viscosity slightly larger than water.
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Modelos Químicos , Pliegue de Proteína , Reología/métodos , Albúmina Sérica Bovina/química , Soluciones/química , Agua/química , Simulación por Computador , Módulo de Elasticidad , Modelos Lineales , Modelos Biológicos , Propiedades de Superficie , ViscosidadRESUMEN
The study of particle laden interfaces has increased significantly due to the increasing industrial use of particle stabilized foams and Pickering emulsions, whose bulk rheology and stability are highly dependent on particle laden interface's interfacial rheology, which is a function of interfacial microstructure. To understand the physical mechanisms that dictate interfacial rheology of particle laden interfaces requires correlating rheology to microstructure. To achieve this goal, a double wall ring interfacial rheometer has been modified to allow real time, simultaneous interfacial visualization and shear rheology measurements. The development of this tool is outlined, and its ability to provide novel and unique measurements is demonstrated on a sample system. This tool has been used to examine the role of microstructure on the steady shear rheology of densely packed, aggregated particle laden interfaces at three surface concentrations. Through examination of the rheology and analysis of interfacial microstructure response to shear, a transition from shear thinning due to aggregated cluster breakup to yielding at a slip plane within the interface has been identified. Interestingly, it is found that aggregated interfaces transition to yielding well before they reached a jammed state. Furthermore, these systems undergo significant shear induced order when densely packed. These results indicate that the mechanics of these interfaces are not simply jammed or unjammed and that the interfacial rheology relationship with microstructure can give us significant insight into understanding how to engineer particle laden interfaces in the future. By examining both rheology and microstructure, the mechanisms that dictate observed rheology are now understood and can be used to predict and control the rheology of the interface.
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Rheological methods that interrogate nanolitre scale volumes of fluids and solids have advanced considerably over the past decade, yet there remains a need for methods that probe the frequency-dependent complex rheological moduli through application of homogenous strain fields. Here we describe a Micro-Electro-Mechanical System (MEMS) based approach for the measurement of dynamic rheology of soft matter where oscillatory strain is produced in a sample sandwiched between an oscillating MEMS stage and a glass plate. The resulting stress-strain relationships are revealed by measurement and analysis of the stage motion. We present preliminary data on simple viscous fluids and on viscoelastic thin films. In this proof-of-principle device, we measure moduli in the range of 50 Pa to 10 kPa over a range of 3 rad s(-1) to 3000 rad s(-1) using less than 5 nL of sample material. The device's measurement window is limited primarily by our current ability to measure the motion of the stage. This device will provide a new way to characterize dynamic microrheology of an array of novel materials and will prove useful in a number of areas including biorheology, microfluidics and polymer thin films.
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An experimental study of droplet breakup in T-shaped microfluidic junctions is presented in which the capillary number and flow rate ratio are varied over a wide range for several different viscosity ratios and several different ratios of the inlet channel widths. The range of conditions corresponds to the region in which both the squeezing pressure that arises when the emerging interface obstructs the channel and the viscous shear stress on the emerging interface strongly influence the process. In this regime, the droplet volume depends on the capillary number, the flow rate ratio, and the ratio of inlet channel widths, which controls the degree of confinement of the droplets. The viscosity ratio influences the droplet volume only when the viscosities are similar. When there is a large viscosity contrast in which the dispersed-phase liquid is at least 50 times smaller than the continuous-phase liquid, the resulting size is independent of the viscosity ratio and no transition to a purely squeezing regime appears. In this case, both the droplet volume and the droplet production frequency obey power-law behavior with the capillary number, consistent with expectations based on mass conservation of the dispersed-phase liquid. Finally, scaling arguments are presented that result in predicted droplet volumes that depend on the capillary number, flow rate ratio, and width ratio in a qualitatively similar way to that observed in experiments.