Liposome destruction by hydrodynamic cavitation in comparison to chemical,physical and mechanical treatments

Liposomes are widely applied in research, diagnostics, medicine and in industry. In this study we show for the first time the effect of hydrodynamic cavitation on liposome stability and compare it to the effect of well described chemical, physical and mechanical treatments. Fluorescein loaded giant 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) lipid vesicles were treated with hydrodynamic cavitation as promising method in inactivation of biological samples. Hydrodynamic treatment was compared to various chemical, physical and mechanical stressors such as ionic strength and osmolarity agents (glucose, Na⁺, Ca²⁺, and Fe³⁺), free radicals, shear stresses (pipetting, vortex mixing, rotational shear stress), high pressure, electroporation, centrifugation, surface active agents (Triton X-100, ethanol), microwave irradiation, heating, freezing-thawing, ultrasound (ultrasonic bath, sonotrode). The fluorescence intensity of individual fluorescein loaded lipid vesicles was measured with confocal laser microscopy. The distribution of lipid vesicle size, vesicle fluorescence intensity, and the number of fluorescein loaded vesicles was determined before and after treatment with different stressors. The different environmental stressors were ranked in order of their relative effect on liposome fluorescein release. Of all tested chemical, physical and mechanical treatments for stability of lipid vesicles, the most detrimental effect on vesicles stability had hydrodynamic cavitation, vortex mixing with glass beads and ultrasound. Here we showed, for the first time that hydrodynamic cavitation was among the most effective physico-chemical treatments in destroying lipid vesicles. This work provides a benchmark for lipid vesicle robustness to a variety of different physico-chemical and mechanical parameters important in lipid vesicle preparation and application.     

Vesicles and red blood cells in flow: From individual dynamics to rheology

The rheology of suspensions of soft particles, such as red blood cells, is a long-standing problem in science and engineering due to the complex interplay between deformable microstructure and the macroscale flow. The major challenge stems from the free-boundary nature of the particle interface. Lipid bilayer membranes that envelop cells and vesicles are particularly complex interfaces because of their unusual mechanics: the molecularly thin membrane is a highly-flexible incompressible fluid sheet. As a result, particles made of closed lipid bilayers (red cells and vesicles) can exhibit richer dynamics than would capsules and drops. We overview the key experimental observations and recent advances in the theoretical modeling of the vesicles and red blood cells in flow. To cite this article: P.M. Vlahovska et al., C. R. Physique 10 (2009).     

Nanohole 3D-size tailoring through polystyrene bead combustion during thin film deposition

A novel approach is presented for nanohole 3D-size tailoring. The process starts with a monolayer of polystyrene (PS) beads spun coat on silicon wafer as a template. The holes can be directly prepared through combustion of PS beads by oxygen plasma during metal or oxide thin film deposition. The incoming particles are prevented from adhering on PS beads by H₂O and CO₂ generated from the combustion of the PS beads. The hole depth generally depends on the film thickness. The hole diameter can be tailored by the PS bead size, film deposition rate, and also the combustion speed of the PS beads. In this work, a series of holes with depth of 4-24 nm and diameter of 10-36 nm has been successfully prepared. The hole wall materials can be selected from metals such as Au or Pt and oxides such as SiO₂ or Al₂O₃. These templates could be suitable for the preparation and characterization of novel nanodevices based on single quantum dots or single molecules, and could be extended to the studies of a wide range of coating materials and substrates with controlled hole depth and diameters.     

Coarse graining of nonbonded degrees of freedom

We investigate the physical meaning of coarse-grained beads generated by coarse graining of nonbonded particles such as solvent molecules in a solution. Starting from the partition function, we analytically coarse grain an N-particle fluid to a system containing N-2 of the original particles plus a bead representing the two remaining particles. As a direct consequence of the lack of bonding interactions, the resulting effective potential becomes independent of the bead coordinates, i.e., ideal-gas-like, in the thermodynamic limit. Thus, there are no conservative forces between coarse-grained beads representing assemblies of nonbonded molecules nor between these beads and any other species in the system.     

Interparticle potential and drag coefficient in nematic colloids

Magneto-optic tweezers were used for measurements of liquid-crystal-mediated forces between spherical beads with tangential anchoring in thin nematic samples. Repulsive force, which results from the quadrupolar symmetry of defects around the immersed beads, decreases proportionally to 1/x6, with x being the bead separation. The velocity with which the particles are pushed apart also follows the same separation dependence. We thus find the effective drag coefficient gamma(eff) independent of x for surface-to-surface distances as small as 10% of the bead diameter.     

Internal magnetic gradient fields in glass bead packs from numerical simulations and constant time diffusion spin echo measurements

Internal magnetic field gradients in water saturated glass bead packs were studied by numerical simulations and a constant time spin echo (CTSE) experiment. The CTSE is comprised of two spin echo refocusing periods where each of the two evolution periods, tau1 and tau2, is varied so that the total evolution, 2(tau1 + tau2), is held constant. The experiment is similar to that introduced by Norwood and Quilter and allows the effects of dephasing due to diffusion in a magnetic field gradient to be separated from other relaxation mechanisms. In our experiments, the magnetic susceptibility difference between the pore fluid and glass beads creates the internal field gradient. CTSE measurements were performed at 7 T (300 MHz 1H) for water saturated in 50 microm diameter glass bead pack. We find that the internal gradients in the center of the pore bodies, where free diffusion applies, is in the range of 10 to 100 G/cm. This fluid volume accounts for < or =10% of the total pore volume. From direct numerical simulations of the internal magnetic field based on a first principles calculation, we find that the major fraction, >90%, of the pore volume has internal gradients of order 500 to 5,000 G/cm. Signals from water in these large gradients is not observed in our CTSE measurements.     

Theory of light scattering from a system of interacting Brownian particles

Starting from a N-particle diffusion equation for a system of N interacting spherical Brownian particles, a non-linear transport equation for concentration fluctuations δc(r, t) of the particles is derived. This dynamic equation is transformed into a hierarchy of equations for retarded propagators of increasing numbers of concentration fluctuations. A cluster expansion to lowest order in the average concentration results in a set of two coupled equations. The spectrum of light scattered by the interacting particles is in general not a Lorentzian, due to the non-linear term in the transport equation. For small scattering wave vectors k the width is D(ω)k², where ω is the transferred frequency. It is shown that D(0) = D_e, the effective diffusion coefficient. For a hardcore interaction potential the spectrum is Lorentzian and it is found that D_e = D₀(1 + φ), where D₀ is the diffusion constant for independent particles and φ the volume concentration of Brownian particles.     

Sustained Release of Protein Particle Encapsulated in Bead-on-String Electrospun Nanofibers

The feasibility of alleviating burst release of electrospun bead-on-string nanofiber scaffolds loaded with protein particles was evaluated, including an investigation of the influence of the beads number on the release profile. Bovine serum albumin–loaded dextran particles were used as the model drug and poly(lactic-co-glycolic acid) as the polymer to fabricate the bead-on-string nanofiber scaffolds by electrospinning. Both the bead structure and the distribution of the particles in the beads were examined by scanning electron, transmission electron, and fluorescence microscopy. The results of fluorescence microscopy suggested that the particles were well encapsulated by the beads of the fibers. In vitro release tests showed that a more sustainable release profile with less initial burst release could be obtained from the bead-on-string fibers than from smooth fibers with uniform diameter. In addition, when the number of the forming beads was not numerous enough to encapsulate all the particles in the suspensions, the release performance worsened because the surplus particles were not properly encapsulated.     

Electron Spin Resonance Study on the Photoinduced Electron Transfer in Chlorophyll a in Reconstituted Lipid Bilayer Vesicles

Abstract  

The photoinduced electron transfer from chlorophyll a through the interface of positively charged dioctadecyltrimethylammonium chloride (DODAC), neutral dipalmitoylphosphatidylcholine (DPPC) and negatively charged dihexadecylphosphate (DHP) headgroup of the lipid bilayers was studied. The photoinduced radicals were identified by electron spin resonance (ESR) and radical yields of chlorophyll a were determined by double integration of the ESR spectra. The formation of vesicles was identified indirectly by measuring change of the λ _max value of optical absorption spectrophotometer from diethyl ether solution to vesicle solutions, and observed directly with scanning and transmission electron microscopic images. The interaction distance between chlorophyll a and interface water (D₂O) determined by deuterium modulation depth with electron spin echo modulation (ESEM) showed a decreasing order DODAC > DPPC > DHP. The interface charge of each vesicle was determined with zeta potential measurement. The interface charge of the lipid bilayers affected the radical yields of chlorophyll a more critically than the interaction distance between chlorophyll a and interface water.     

A method for quantitative interpretation of fluorescence detection of poly(ethylene glycol)-mediated 1-palmitoyl-2-[[[2-[4-(phenyl-trans-1,3,5-hexatrienyl) phenyl]ethyl]oxyl]carbonyl]3-sn-phosphatidylcholine (DPHpPC) transfer and fusion between phospholipid vesicles in the dehydrated state

A method has been developed for calculating the expected fluorescence lifetime of the DPH _p PC probe distributed between different membrane environments. We show how this method can be used to distinguish between lipid transfer and fusion between large unilamellar vesicles occurring in the presence of poly(ethylene glycol) (PEG). This application of the calculation took into consideration the heterogeneity of microenvironments experienced by the probe in a sample containing vesicle aggregates of different sizes. Assuming that the aggregate size distribution was a delta function of the aggregate size, comparison of the calculated and observed lifetimes yielded an estimate of the vesicle aggregate size. For vesicles of varying compositions in the presence of dehydrating concentrations of PEG, this method suggested that only small aggreggates formed. For vesicles that could be demonstrated by other means not to have fused, the data were consistent with lipid transfer occurring only between the outer leaflets of two to four vesicles, even at high PEG concentrations. For vesicles that could be demonstrated to fuse by contents mixing and size changes, the fluorescence lifetime data were consistent with lipid transfer between both the inner and the outer leaflets of two to four fused vesicles. At very high PEG concentrations, where extensive rupture and large, multilamellar products were previously observed, the lifetime data were consistent with much more extensive lipid transfer within larger aggregates. The agreement of predictions made on the basis of lifetime measurements with other observations attests to the validity of the fluorescence lifetime method. In addition, the model and data presented here provide evidence that fusion occurs between small numbers of PEG-aggregated vesicles before the removal of PEG.     

Diffusion of a single particle in a 3D random packing of spheres

We report an experimental study of the dispersion properties of individual spherical particles of size d, moving under gravity in a dry random packing of large spheres of size D. The diameter ratio d/D is below the critical value 0.1547 above which beads get pinned inside the packing . They move in this regime at a constant mean velocity decreasing with the ratio d/D. We analyse dispersion parallel and transverse to the mean velocity by studying the bead distribution in the x-y plane at the exit of the packing (radial dispersion) and the transit time distribution (longitudinal dispersion) while varying the height H of the bed. Diffusion in both directions is found to be governed essentially by the diameter D of packed spheres and not by the size d of the small beads. A dispersivity length characterising the spreading amplitude is determined. Comparisons between transverse and longitudinal dispersion demonstrate that both processes have similar properties. A key parameter is the diameter D which controls the path length of the particles. Received 5 November 1999 and Received in final form 30 March 2000     

Dynamics of multicomponent vesicles in a viscous fluid

We develop and investigate numerically a thermodynamically consistent model of two-dimensional multicomponent vesicles in an incompressible viscous fluid. The model is derived using an energy variation approach that accounts for different lipid surface phases, the excess energy (line energy) associated with surface phase domain boundaries, bending energy, spontaneous curvature, local inextensibility and fluid flow via the Stokes equations. The equations are high-order (fourth order) nonlinear and nonlocal due to incompressibility of the fluid and the local inextensibility of the vesicle membrane. To solve the equations numerically, we develop a nonstiff, pseudo-spectral boundary integral method that relies on an analysis of the equations at small scales. The algorithm is closely related to that developed very recently by Veerapaneni et al. [81] for homogeneous vesicles although we use a different and more efficient time stepping algorithm and a reformulation of the inextensibility equation. We present simulations of multicomponent vesicles in an initially quiescent fluid and investigate the effect of varying the average surface concentration of an initially unstable mixture of lipid phases. The phases then redistribute and alter the morphology of the vesicle and its dynamics. When an applied shear is introduced, an initially elliptical vesicle tank-treads and attains a steady shape and surface phase distribution. A sufficiently elongated vesicle tumbles and the presence of different surface phases with different bending stiffnesses and spontaneous curvatures yields a complex evolution of the vesicle morphology as the vesicle bends in regions where the bending stiffness and spontaneous curvature are small.     

Label-acquired magnetorotation for biosensing: An asynchronous rotation assay

This paper presents a novel application of magnetic particles for biosensing, called label-acquired magnetorotation (LAM). This method is based on a combination of the traditional sandwich assay format with the asynchronous magnetic bead rotation (AMBR) method. In label-acquired magnetorotation, an analyte facilitates the binding of a magnetic label bead to a nonmagnetic solid phase sphere, forming a sandwich complex. The sandwich complex is then placed in a rotating magnetic field, where the rotational frequency of the sandwich complex is a function of the amount of analyte attached to the surface of the sphere. Here, we use streptavidin-coated beads and biotin-coated particles as analyte mimics, to be replaced by proteins and other biological targets in future work. We show this sensing method to have a dynamic range of two orders of magnitude.     

Phase separation transition in a driven diffusive system with anti-ferromagnetic interaction

One-dimensional non-equilibrium systems with short-range interaction can undergo phase transitions from homogeneous states to phase separated states as interaction (?) among particles is increased. One of the model systems where such a transition has been observed is the extended Katz-Lebowitz-Spohn (KLS) model with ferromagnetically interacting particles at ?=4/5. Here, the system remains homogeneous for small interaction strength (?<4/5), and for anti-ferromagnetic interactions (?<0). We show that the phase separation transitions can also occur in anti-ferromagnetic systems if interaction among particles depends explicitly on the size of the block (n) they belong to. We study this transition in detail for a specific case ?=δ/n, where phase separation occurs for δ<−1.     

Vesicle migration and spatial organization driven by flow line curvature

Cross-streamline migration of deformable entities is essential in many problems such as industrial particulate flows, DNA sorting, and blood rheology. Using two-dimensional numerical experiments, we have discovered that vesicles suspended in a flow with curved flow lines migrate towards regions of high flowline curvature, which are regions of high shear rates. The migration velocity of a vesicle is found to be a universal function of the normal stress difference and the flow curvature. This finding quantitatively demonstrates a direct coupling between a microscopic quantity (migration) and a macroscopic one (normal stress difference). Furthermore, simulations with multiple vesicles revealed a self-organization, which corresponds to segregation, in a rim closer to the inner cylinder, resulting from a subtle interaction among vesicles. Such segregation effects could have a significant impact on the rheology of vesicle flows.     

About the influence of friction and polydispersity on the jamming behavior of bead assemblies

We study the jamming of bead assemblies placed in a cylindrical container whose bottom is pierced with a circular hole. Their jamming behavior is quantified here by the median jamming diameter, that is the diameter of the hole for which the jamming probability is 0.5. Median jamming diameters of monodisperse assemblies are obtained numerically using the Distinct Element Method and experimentally with steel beads. We obtain good agreement between numerical and experimental results. The influence of friction is then investigated. In particular, the formation of concentric bead rings is observed for low frictions. We identify this phenomenon as a boundary effect and study its influence on jamming. Relying on measures obtained from simulation runs, the median jamming diameter of bidisperse bead assemblies is finally found to depend only on the volume-average diameter of their constituting beads. We formulate this as a tentative law and validate it using bidisperse assemblies of steel beads.     

On the Interplay of Magnetic and Molecular Forces in Curie–Weiss Ferrofluid Models

We consider a mean-field continuum model of classical particles in R ^d with Ising or Heisenberg spins. The interaction has two ingredients, a ferromagnetic spin coupling and a spin-independent molecular force. We show that a feedback between these forces gives rise to a first-order phase transition with simultaneous jumps of particle density and magnetization per particle, either at the threshold of ferromagnetic order or within the ferromagnetic region. If the direct particle interaction alone already implies a phase transition, then the additional spin coupling leads to an even richer phase diagram containing triple (or higher order) points.     

Dynamic strength of fluid membranes

Rupturing fluid membrane vesicles with a steady ramp of micropipette suction yields a tension distribution that images the kinetic process of membrane failure. When plotted on a log scale of tension loading rate, the distribution peaks (membrane strengths) define a dynamic tension spectrum with distinct regimes that reflect passage of prominent energy barriers along the pathway to rupture. Demonstrated here by tests on giant PC lipid vesicles over loading rates from 0.06–60 mN/m/s, the stochastic process of rupture can be modelled as a causal sequence of two thermally-activated transitions where each transition governs membrane strength on separate scales of loading rate. Under fast ramps of tension, a steep linear regime appears in each spectrum at high strengths which implies that failure requires nucleation of a rare nanoscale defect. The slope and projected intercept yield defect size and spontaneous production rate respectively. However, under slow ramps of loading, the spectrum crosses over to a shallow-curved regime at lower strength, which is consistent with the kinetic impedance to opening an unstable hole in a fluid film. The dependence of rupture tension on rate reveals hole edge energy and frequency scale for thermal fluctuations in size. To cite this article: E. Evans, V. Heinrich, C. R. Physique 4 (2003).