Active particles induce large shape deformations in giant lipid vesicles.

Biological cells generate intricate structures by sculpting their membrane from within to actively sense and respond to external stimuli or to explore their environment1-4. Several pathogenic bacteria also provide examples of how localized forces strongly deform cell membranes from inside, leading to the invasion of neighbouring healthy mammalian cells5. Giant unilamellar vesicles have been successfully used as a minimal model system with which to mimic biological cells6-11, but the realization of a minimal system with localized active internal forces that can strongly deform lipid membranes from within and lead to dramatic shape changes remains challenging. Here we present a combined experimental and simulation study that demonstrates how self-propelled particles enclosed in giant unilamellar vesicles can induce a plethora of non-equilibrium shapes and active membrane fluctuations. Using confocal microscopy, in the experiments we explore the membrane response to local forces exerted by self-phoretic Janus microswimmers. To quantify dynamic membrane changes, we perform Langevin dynamics simulations of active Brownian particles enclosed in thin membrane shells modelled by dynamically triangulated surfaces. The most pronounced shape changes are observed at low and moderate particle loadings, with the formation of tether-like protrusions and highly branched, dendritic structures, whereas at high volume fractions globally deformed vesicle shapes are observed. The resulting state diagram predicts the conditions under which local internal forces generate various membrane shapes. A controlled realization of such distorted vesicle morphologies could improve the design of artificial systems such as small-scale soft robots and synthetic cells.

Collective behavior of squirmers in thin films.

Bacteria in biofilms form complex structures and can collectively migrate within mobile aggregates, which is referred to as swarming. This behavior is influenced by a combination of various factors, including morphological characteristics and propulsive forces of swimmers, their volume fraction within a confined environment, and hydrodynamic and steric interactions between them. In our study, we employ the squirmer model for microswimmers and the dissipative particle dynamics method for fluid modeling to investigate the collective motion of swimmers in thin films. The film thickness permits a free orientation of non-spherical squirmers, but constraints them to form a two-layered structure at maximum. Structural and dynamic properties of squirmer suspensions confined within the slit are analyzed for different volume fractions of swimmers, motility types (e.g., pusher, neutral squirmer, puller), and the presence of a rotlet dipolar flow field, which mimics the counter-rotating flow generated by flagellated bacteria. Different states are characterized, including a gas-like phase, swarming, and motility-induced phase separation, as a function of increasing volume fraction. Our study highlights the importance of an anisotropic swimmer shape, hydrodynamic interactions between squirmers, and their interaction with the walls for the emergence of different collective behaviors. Interestingly, the formation of collective structures may not be symmetric with respect to the two walls. Furthermore, the presence of a rotlet dipole significantly mitigates differences in the collective behavior between various swimmer types. These results contribute to a better understanding of the formation of bacterial biofilms and the emergence of collective states in confined active matter.

Motion of microswimmers in cylindrical microchannels.

Biological and artificial microswimmers often have to propel through a variety of environments, ranging from heterogeneous suspending media to strong geometrical confinement. Under confinement, local flow fields generated by microswimmers, and steric and hydrodynamic interactions with their environment determine the locomotion. We propose a squirmer-like model to describe the motion of microswimmers in cylindrical microchannels, where propulsion is generated by a fixed surface slip velocity. The model is studied using an approximate analytical solution for cylindrical swimmer shapes, and by numerical hydrodynamics simulations for spherical and spheroidal shapes. For the numerical simulations, we employ the dissipative particle dynamics method for modelling fluid flow. Both the analytical model and simulations show that the propulsion force increases with increasing confinement. However, the swimming velocity under confinement remains lower than the swimmer speed without confinement for all investigated conditions. In simulations, different swimming modes (i.e. pusher, neutral, puller) are investigated, and found to play a significant role in the generation of propulsion force when a swimmer approaches a dead end of a capillary tube. Propulsion generation in confined systems is local, such that the generated flow field generally vanishes beyond the characteristic size of the swimmer. These results contribute to a better understanding of microswimmer force generation and propulsion under strong confinement, including the motion in porous media and in narrow channels.

Competition between deformation and free volume quantified by 3D image analysis of red blood cell.

Cells in living organisms are subjected to mechanical strains caused by external forces like overcrowding, resulting in strong deformations that affect cell function. We study the interplay between deformation and crowding of red blood cells (RBCs) in dispersions of nonabsorbing rod-like viruses. We identify a sequence of configurational transitions of RBC doublets, including configurations that can only be induced by long-ranged attraction: highly fluctuating T-shaped and face-to-face configurations at low, and doublets approaching a complete spherical configuration at high, rod concentrations. Complementary simulations are used to explore different energy contributions to deformation as well as the stability of RBC doublet configurations. Our advanced analysis of 3D reconstructed confocal images of RBC doublets quantifies the depletion interaction and the resulting deformation energy. Thus, we introduce a noninvasive, high-throughput platform that is generally applicable to investigate the mechanical response of biological cells to external forces and characterize their mechanical properties.

Dynamic shapes of floppy vesicles enclosing active Brownian particles with membrane adhesion.

Recent advances in micro- and nano-technologies allow the construction of complex active systems from biological and synthetic materials. An interesting example is active vesicles, which consist of a membrane enclosing self-propelled particles, and exhibit several features resembling biological cells. We investigate numerically the behavior of active vesicles, where the enclosed self-propelled particles can adhere to the membrane. A vesicle is represented by a dynamically triangulated membrane, while the adhesive active particles are modelled as active Brownian particles (ABPs) that interact with the membrane via the Lennard-Jones potential. Phase diagrams of dynamic vesicle shapes as a function of ABP activity and particle volume fraction inside the vesicle are constructed for different strengths of adhesive interactions. At low ABP activity, adhesive interactions dominate over the propulsion forces, such that the vesicle attains near static configurations, with protrusions of membrane-wrapped ABPs having ring-like and sheet-like structures. At moderate particle densities and strong enough activities, active vesicles show dynamic highly-branched tethers filled with string-like arrangements of ABPs, which do not occur in the absence of particle adhesion to the membrane. At large volume fractions of ABPs, vesicles fluctuate for moderate particle activities, and elongate and finally split into two vesicles for large ABP propulsion strengths. We also analyze membrane tension, active fluctuations, and ABP characteristics (e.g., mobility, clustering), and compare them to the case of active vesicles with non-adhesive ABPs. The adhesion of ABPs to the membrane significantly alters the behavior of active vesicles, and provides an additional parameter for controlling their behavior.

Erythrocyte Sedimentation: Collapse of a High-Volume-Fraction Soft-Particle Gel.

The erythrocyte sedimentation rate is one of the oldest medical diagnostic methods whose physical mechanisms remain debatable today. Using both light microscopy and mesoscale cell-level simulations, we show that erythrocytes form a soft-particle gel. Furthermore, the high volume fraction of erythrocytes, their deformability, and weak attraction lead to unusual properties of this gel. A theoretical model for the gravitational collapse is developed, whose predictions are in agreement with detailed macroscopic measurements of the interface velocity.

Correction: Non-equilibrium shapes and dynamics of active vesicles.

Correction for 'Non-equilibrium shapes and dynamics of active vesicles' by Priyanka Iyer et al., Soft Matter, 2022, 18, 6868-6881, https://doi.org/10.1039/D2SM00622G.

Non-equilibrium shapes and dynamics of active vesicles.

Active vesicles, constructed through the confinement of self-propelled particles (SPPs) inside a lipid membrane shell, exhibit a large variety of non-equilibrium shapes, ranging from the formation of local tethers and dendritic conformations, to prolate and bola-like structures. To better understand the behavior of active vesicles, we perform simulations of membranes modelled as dynamically triangulated surfaces enclosing active Brownian particles. A systematic analysis of membrane deformations and SPP clustering, as a function of SPP activity and volume fraction inside the vesicle is carried out. Distributions of membrane local curvature, and the clustering and mobility of SPPs obtained from simulations of active vesicles are analysed. There exists a feedback mechanism between the enhancement of membrane curvature, the formation of clusters of active particles, and local or global changes in vesicle shape. The emergence of active tension due to the activity of SPPs can well be captured by the Young-Laplace equation. Furthermore, a simple numerical method for tether detection is presented and used to determine correlations between the number of tethers, their length, and local curvature. We also provide several geometrical arguments to explain different tether characteristics for various conditions. These results contribute to the future development of steerable active vesicles or soft micro-robots whose behaviour can be controlled and used for potential applications.

Effect of cytosol viscosity on the flow behavior of red blood cell suspensions in microvessels.

OBJECTIVE: The flow behavior of blood is strongly affected by red blood cell (RBC) properties, such as the viscosity ratio C between cytosol and suspending medium, which can significantly be altered in several pathologies (e.g. sickle-cell disease, malaria). The main objective of this study is to understand the effect of C on macroscopic blood flow properties such as flow resistance in microvessels, and to link it to the deformation and dynamics of single RBCs. METHODS: We employ mesoscopic hydrodynamic simulations to investigate flow properties of RBC suspensions with different cytosol viscosities for various flow conditions in cylindrical microchannels. RESULTS: Starting from a dispersed cell configuration which approximates RBC dispersion at vessel bifurcations in the microvasculature, we find that the flow convergence and development of RBC-free layer (RBC-FL) depend only weakly on C, and require a convergence length in the range of 25D-50D, where D is channel diameter. In vessels with D≤20µm , the final resistance of developed flow is nearly the same for C = 5 and C = 1, while for D=40µm , the flow resistance for C = 5 is about 10% larger than for C = 1. The similarities and differences in flow resistance can be explained by viscosity-dependent RBC-FL thicknesses, which are associated with the viscosity-dependent dynamics of single RBCs. CONCLUSIONS: The weak effect on the flow resistance and RBC-FL explains why RBCs can contain a high concentration of hemoglobin for efficient oxygen delivery, without a pronounced increase in the flow resistance. Furthermore, our results suggest that significant alterations in microvascular flow in various pathologies are likely not due to mere changes in cytosolic viscosity.

Dissipative particle dynamics with energy conservation: Isoenergetic integration and transport properties.

Simulations of nano- to micro-meter scale fluidic systems under thermal gradients require consistent mesoscopic methods accounting for both hydrodynamic interactions and proper transport of energy. One such method is dissipative particle dynamics with energy conservation (DPDE), which has been used for various fluid systems with non-uniform temperature distributions. We propose an easily parallelizable modification of the velocity-Verlet algorithm based on local energy redistribution for each DPDE particle such that the total energy in a simulated system is conserved up to machine precision. Furthermore, transport properties of a DPDE fluid are analyzed in detail. In particular, an analytical approximation for the thermal conductivity coefficient is derived, which allows its a priori estimation for a given parameter set. Finally, we provide approximate expressions for the dimensionless Prandtl and Schmidt numbers, which characterize fluid transport properties and can be adjusted independently by a proper selection of model parameters. In conclusion, our results strengthen the DPDE method as a very robust approach for the investigation of mesoscopic systems with temperature inhomogeneities.

Importance of Erythrocyte Deformability for the Alignment of Malaria Parasite upon Invasion.

Invasion of erythrocytes by merozoites is an essential step for the survival and progression of malaria parasites. To invade red blood cells (RBCs), apicomplexan parasites have to adhere with their apex to the RBC membrane. This necessary apex-membrane contact (or alignment) is not immediately established because the orientation of a free merozoite with respect to the RBC membrane is random when an adhesion contact first occurs. Therefore, it has been suggested that after the initial adhesion, merozoites facilitate their proper alignment by inducing considerable membrane deformations, frequently observed before the invasion process. This proposition is based on a positive correlation between RBC membrane deformation and successful invasion; however, the role of RBC mechanics and its deformation in the alignment process remains elusive. Using a mechanically realistic model of a deformable RBC, we investigate numerically the importance of RBC deformability for merozoite alignment. Adhesion between the parasite and RBC membrane is modeled by an attractive potential that might be inhomogeneous, mimicking possible adhesion gradients at the surface of a parasite. Our results show that RBC membrane deformations are crucial for successful merozoite alignment and require interaction strengths comparable to adhesion forces measured experimentally. Adhesion gradients along the parasite body further improve its alignment. Finally, an increased membrane rigidity is found to result in poor merozoite alignment, which can be a possible reason for a reduction in the invasion susceptibility of RBCs in several blood diseases associated with membrane stiffening.

High-Throughput Microfluidic Characterization of Erythrocyte Shapes and Mechanical Variability.

The motion of red blood cells (RBCs) in microchannels is important for microvascular blood flow and biomedical applications such as blood analysis in microfluidics. The current understanding of the complexity of RBC shapes and dynamics in microchannels is mainly based on several simulation studies, but there are a few systematic experimental investigations. Here, we present a combined study that systematically characterizes RBC behavior for a wide range of flow rates and channel sizes. Even though simulations and experiments generally show good agreement, experimental observations demonstrate that there is no single well-defined RBC state for fixed flow conditions but rather a broad distribution of states. This result can be attributed to the inherent variability in RBC mechanical properties, which is confirmed by a model that takes the variation in RBC shear elasticity into account. This represents a significant step toward a quantitative connection between RBC behavior in microfluidic devices and their mechanical properties, which is essential for a high-throughput characterization of diseased cells.

State diagram for wall adhesion of red blood cells in shear flow: from crawling to flipping.

Red blood cells in shear flow show a variety of different shapes due to the complex interplay between hydrodynamics and membrane elasticity. Malaria-infected red blood cells become generally adhesive and less deformable. Adhesion to a substrate leads to a reduction in shape variability and to a flipping motion of the non-spherical shapes during the mid-stage of infection. Here, we present a complete state diagram for wall adhesion of red blood cells in shear flow obtained by simulations, using a particle-based mesoscale hydrodynamics approach, multiparticle collision dynamics. We find that cell flipping at a substrate is replaced by crawling beyond a critical shear rate, which increases with both membrane stiffness and viscosity contrast between the cytosol and suspending medium. This change in cell dynamics resembles the transition between tumbling and tank-treading for red blood cells in free shear flow. In the context of malaria infections, the flipping-crawling transition would strongly increase the adhesive interactions with the vascular endothelium, but might be suppressed by the combined effect of increased elasticity and viscosity contrast.

Red cells' dynamic morphologies govern blood shear thinning under microcirculatory flow conditions.

Blood viscosity decreases with shear stress, a property essential for an efficient perfusion of the vascular tree. Shear thinning is intimately related to the dynamics and mutual interactions of RBCs, the major component of blood. Because of the lack of knowledge about the behavior of RBCs under physiological conditions, the link between RBC dynamics and blood rheology remains unsettled. We performed experiments and simulations in microcirculatory flow conditions of viscosity, shear rates, and volume fractions, and our study reveals rich RBC dynamics that govern shear thinning. In contrast to the current paradigm, which assumes that RBCs align steadily around the flow direction while their membranes and cytoplasm circulate, we show that RBCs successively tumble, roll, deform into rolling stomatocytes, and, finally, adopt highly deformed polylobed shapes for increasing shear stresses, even for semidilute volume fractions of the microcirculation. Our results suggest that any pathological change in plasma composition, RBC cytosol viscosity, or membrane mechanical properties will affect the onset of these morphological transitions and should play a central role in pathological blood rheology and flow behavior.

Flow-Induced Transitions of Red Blood Cell Shapes under Shear.

A recent study of red blood cells (RBCs) in shear flow [Lanotte et al., Proc. Natl. Acad. Sci. U.S.A. 113, 13289 (2016)PNASA60027-842410.1073/pnas.1608074113] has demonstrated that RBCs first tumble, then roll, transit to a rolling and tumbling stomatocyte, and finally attain polylobed shapes with increasing shear rate, when the viscosity contrast between cytosol and blood plasma is large enough. Using two different simulation techniques, we construct a state diagram of RBC shapes and dynamics in shear flow as a function of shear rate and viscosity contrast, which is also supported by microfluidic experiments. Furthermore, we illustrate the importance of RBC shear elasticity for its dynamics in flow and show that two different kinds of membrane buckling trigger the transition between subsequent RBC states.

Effect of spectrin network elasticity on the shapes of erythrocyte doublets.

Red blood cell (RBC) aggregates play an important role in determining blood rheology. RBCs in plasma or polymer solution interact attractively to form various shapes of RBC doublets, where the attractive interactions can be varied by changing the solution conditions. A systematic numerical study on RBC doublet formation is performed, which takes into account the shear elasticity of the RBC membrane due to the spectrin cytoskeleton, in addition to the membrane bending rigidity. RBC membranes are modeled by two-dimensional triangular networks of linked vertices, which represent three-dimensional cell shapes. The phase space of RBC doublet shapes in a wide range of adhesion strengths, reduced volumes, and shear elasticities is obtained. The shear elasticity of the RBC membrane changes the doublet phases significantly. Experimental images of RBC doublets in different solutions show similar configurations. Furthermore, we show that rouleau formation is affected by the doublet structure.

Microvascular blood flow resistance: Role of red blood cell migration and dispersion.

Microvascular blood flow resistance has a strong impact on cardiovascular function and tissue perfusion. The flow resistance in microcirculation is governed by flow behavior of blood through a complex network of vessels, where the distribution of red blood cells across vessel cross-sections may be significantly distorted at vessel bifurcations and junctions. In this paper, the development of blood flow and its resistance starting from a dispersed configuration of red blood cells is investigated in simulations for different hematocrit levels, flow rates, vessel diameters, and aggregation interactions between red blood cells. Initially dispersed red blood cells migrate toward the vessel center leading to the formation of a cell-free layer near the wall and to a decrease of the flow resistance. The development of cell-free layer appears to be nearly universal when scaled with a characteristic shear rate of the flow. The universality allows an estimation of the length of a vessel required for full flow development, lc ≲ 25D, for vessel diameters in the range 10 µm < D < 100 µm. Thus, the potential effect of red blood cell dispersion at vessel bifurcations and junctions on the flow resistance may be significant in vessels which are shorter or comparable to the length lc. Aggregation interactions between red blood cells generally lead to a reduction of blood flow resistance. The simulations are performed using the same viscosity for both external and internal fluids and the RBC membrane viscosity is not considered; however, we discuss how the viscosity contrast may affect the results. Finally, we develop a simple theoretical model which is able to describe the converged cell-free-layer thickness at steady-state flow with respect to flow rate. The model is based on the balance between a lift force on red blood cells due to cell-wall hydrodynamic interactions and shear-induced effective pressure due to cell-cell interactions in flow. We expect that these results can also be used to better understand the flow behavior of other suspensions of deformable particles such as vesicles, capsules, and cells.

Effect of fluid-colloid interactions on the mobility of a thermophoretic microswimmer in non-ideal fluids.

Janus colloids propelled by light, e.g., thermophoretic particles, offer promising prospects as artificial microswimmers. However, their swimming behavior and its dependence on fluid properties and fluid-colloid interactions remain poorly understood. Here, we investigate the behavior of a thermophoretic Janus colloid in its own temperature gradient using numerical simulations. The dissipative particle dynamics method with energy conservation is used to investigate the behavior in non-ideal and ideal-gas like fluids for different fluid-colloid interactions, boundary conditions, and temperature-controlling strategies. The fluid-colloid interactions appear to have a strong effect on the colloid behavior, since they directly affect heat exchange between the colloid surface and the fluid. The simulation results show that a reduction of the heat exchange at the fluid-colloid interface leads to an enhancement of colloid's thermophoretic mobility. The colloid behavior is found to be different in non-ideal and ideal fluids, suggesting that fluid compressibility plays a significant role. The flow field around the colloid surface is found to be dominated by a source-dipole, in agreement with the recent theoretical and simulation predictions. Finally, different temperature-control strategies do not appear to have a strong effect on the colloid's swimming velocity.