Annie Viallat

Swinging of red blood cells under shear flow

We reveal that under moderate shear stress (etagamma[over ] approximately 0.1 Pa) red blood cells present an oscillation of their inclination (swinging) superimposed to the long-observed steady tank treading (TT) motion. A model based on a fluid ellipsoid surrounded by a viscoelastic membrane initially unstrained (shape memory) predicts all observed features of the motion: an increase of both swinging amplitude and period (1/2 the TT period) upon decreasing etagamma[over ], a etagamma[over ]-triggered transition toward a narrow etagamma[over ] range intermittent regime of successive swinging and tumbling, and a pure tumbling at low etagamma[over ] values.

Vesicles and red blood cells in shear flow

We describe the similarities and the specificities of the behaviour of individual soft particles, namely, drops, lipid vesicles and red blood cells subjected to a shear flow. We highlight that their motion depends in a non-trivial way on the particle mechanical properties. We detail the effect of the presence of a wall with or without wall-particle attractive interaction from a biological perspective.

Giant lipid vesicles filled with a gel: Shape instability induced by osmotic shrinkage

We report the properties of giant lipid vesicles enclosing an agarose gel. In this system, the lipid bilayer retains some basic properties of biological membranes and the internal fluid exhibits viscoelastic properties, thus permitting us to address the question of the deformation of a cell membrane in relation to the mechanical properties of its cytoskeleton. The agarose gel (concentration c0gel = 0.07%, 0.18%, 0.36%, and 1% w/w), likely not anchored to the membrane, confers to the internal volume elastic moduli in the range of 10-10(4) Pa. Shapes and kinetics of de-swelling of gel-filled and aqueous solution-filled vesicles are compared upon either a progressive or a fast osmotic shrinkage. Both systems exhibit similar kinetics. Shapes of solution-filled vesicles are well described using the area difference elasticity model, whereas gel-filled vesicles present original patterns: facets, bumps, spikes (c0gel < 0.36%), or wrinkles (c0gel > or = 0.36%). These shapes partially vanish upon re-swelling, and some of them are reminiscent of echinocytic shapes of erythrocytes. Their characteristic size (microns) decreases upon increasing c0gel. A possible origin of these patterns, relying on the formation of a dense impermeable gel layer at the vesicle surface and associated with a transition toward a collapsed gel phase, is advanced.

Responsive viscoelastic giant lipid vesicles filled with a poly(N-isopropylacrylamide) artificial cytoskeleton

Responsive giant lipid vesicles filled with aqueous PolyNipam sol (SFV) or gel (GFV) were prepared by ultra-violet polymerisation performed in situ. Upon crossing the lower critical transition temperature of PolyNipam, SFVs and GFVs undergo a significant change of their structural and mechanical properties or a drastic volume transition, respectively. Rheometric and micropipette experiments show that both internal viscosity of SFVs and internal shear modulus of GFVs are tunable over several orders of magnitude and lie in the range observed for living cells. Moreover, the vesicle membrane is strongly bound to the internal polymer medium, making these systems interesting for mimicking the basic mechanical behaviour of passive living cells.

Adhesion induced non-planar and asynchronous flow of a giant vesicle membrane in an external shear flow

We show the existence of a flow at the surface of strongly adhering giant lipid vesicles submitted to an external shear flow. The surface flow is divided into two symmetric quadrants and presents two stagnation points (SP) on each side of the vesicle meridian plane. The position of these stagnation points depends strongly on the adhesion strength, characterized by the ratio of the contact zone diameter to the vesicle diameter. Contrary to the case of non-adhesive vesicles, streamlines do not lie in the shear plane. By avoiding the motionless contact zone, streamlines result in three-dimensional paths, strongly asymmetric away from the SP. Additional shearing dissipation may occur on the membrane surface as we observed that the mean rotational velocity of the membrane increases towards the vesicle SP, and is mainly determined by the adhesion induced vesicle shape.

Red blood cell: from its mechanics to its motion in shear flow

There is a number of publications on red blood cell deformability, that is, on the remarkable cell ability to change its shape in response to an external force and to pass through the narrowest blood capillaries and splenic sinuses. Cell deformability is postulated to be a major determinant of impaired perfusion, increase of blood viscosity, and occlusion in microvessels. Current deformability tests like ektacytometry measure global parameters, related to shape changes at the whole cell scale. Despite strong advances in our understanding of the molecular organization of red blood cells, the relationships between the rheology of each element of the cell composite structure, the global deformability tests, and the cell behavior in microflows are still not elucidated. This review describes recent advances in the description of the dynamics of red blood cells in shear flow and in the mechanistic understanding of this dynamics at the scale of the constitutive rheological and structural elements of the cell. These developments could open up new horizons for the determination of red blood cell mechanical parameters by analyzing their motion under low shear flows.

Responsive Giant Vesicles Filled with Poly(N-isopropylacrylamide) Sols or Gels

We prepared giant unilamellar vesicles (GUVs) enclosing solutions or covalent gels of Poly(Nisopropylacrylamide) (PolyNipam). Concentrated suspensions of GUVs were prepared by applying an alternative field on a lipid film hydrated by a monomer solution containing N-isopropylacrylamide, crosslinker (N,N′-methylene-bis-acrylamide), initiator and sucrose. Vesicle inner medium was polymerised and cross-linked by UV irradiation of the suspension, yielding viscous vesicles enclosing a solution of linear PolyNipam chains (when no bisacrylamide was used) or elastic vesicles filled with a covalent PolyNipam gel. We show that gel-filled vesicles are responsive systems triggered by the temperature: they shrink, reducing by a factor eight their volume below the critical temperature (32 °C in water, lower in glucose solution) and re-swell in a reversible and reproducible way upon decreasing temperature. In both cases, we show that the vesicle lipid membrane interacts with the internal polymer, resulting in an strong resistance of the vesicles to external mechanical stresses (enhanced tension of lysis).

Mimicking cell/extracellular matrix adhesion with lipid membranes and solid substrates: requirements, pitfalls and proposals

The interest in physical approaches to the study of cell adhesion has generated numerous recent works on the development of substrates mimicking the extracellular matrix and the use of giant synthetic liposomes, commonly considered as basic models of living cells. The use of well-characterized bioactive substrates and artificial cells should allow us to gain new insight into the cell–extracellular matrix interactions, provided that their biomimetic relevance has been really proved. The aim of this paper is to define some minimal requirements for effective biomimetic features and to propose simple adhesion assays. We show, for instance, that immobilization of specific ligands is sometimes not sufficient t oe nsure specific adhesion of cells expressing the corresponding receptors. B yi nvestigating comparatively the adhesive behaviour of decorated erythrocytes and vesicles, we also discuss the potentialities and limitations of synthetic vesicles as test cells.

Sensitive detection of ultra-weak adhesion states of vesicles by interferometric microscopy

We have used an original analysis of reflection interference contrast microscopy (RICM) to detect an ultra-weak specific interaction between a glycolipid vesicle and a lectin-coated substrate. The membrane height fluctuations in the contact zone are observed with a high illumination aperture; the membrane profile and the membrane-substrate distance are quantitatively determined using the new analysis, which accounts for multiple interfaces and multiple incidence rays. We showed that this refined version of RICM theory is necessary, specifically in the case of intermediate membrane-substrate distance (∼30 nm) and helped to discriminate between the ultra-weak interaction and pure gravitational sedimentation

CHAPTER 10:On the Importance of the Deformability of Red Blood Cells in Blood Flow

The evolution of macroscopic living beings on Earth required the establishment of vascular systems to transport nutrients and eliminate waste. For example, oxygen transport from the respiratory organs to tissues occurs via a high volume fraction of red blood cells (RBCs) that circulate through the vascular system. If blood was analogous to a concentrated suspension of solid particles or a suspension of droplets of similar dimensions, it would display a viscosity several orders of magnitude larger than its actual value, which would compromise the transport pathway. The amazing fluidity of blood originates from the deformability of RBCs and the microstructures they form in flow. Consequently, blood is shear-thinning. The deformability of RBCs is postulated to be a major determinant of impaired perfusion, increased blood viscosity and occlusion in microvessels. Despite advances in understanding the molecular organization of RBCs, the relationships between the rheology of each element of the cell’s composite structure, the global deformability of the cells and the behavior of the cells in microflows are not understood. In this chapter, we describe recent advances in the description of the flow of RBCs. We focus on flows for which experimental, analytical and numerical advances have been made and discuss the physics underlying hemorheological phenomena where cell deformability is important.