Jurriaan J. J. Gillissen

Effect of Glucose on the Mobility of Membrane-Adhering Liposomes

Enclosed lipid bilayer structures, referred to as liposomes or lipid vesicles, have a wide range of biological functions, such as cellular signaling and membrane trafficking. The efficiency of cellular uptake of liposomes, a key step in many of these functions, is strongly dependent on the contact area between a liposome and a cell membrane, which is governed by the adhesion force w, the membrane bending energy κ, and the osmotic pressure Δp. Herein, we investigate the relationship between these forces and the physicochemical properties of the solvent, namely, the presence of glucose (a nonionic osmolyte). Using fluorescence microscopy, we measure the diffusivity D of small (∼50 nm radius), fluorescently labeled liposomes adhering to a supported lipid bilayer or to the freestanding membrane of a giant (∼10 μm radius) liposome. It is observed that glucose in solution reduces D on the supported membrane, while having negligible effect on D on the freestanding membrane. Using well-known hydrodynamic theory for the diffusivity of membrane inclusions, these observations suggest that glucose enhances the contact area between the small liposomes and the underlying membrane, while not affecting the viscosity of the underlying membrane. In addition, quartz crystal microbalance experiments showed no significant change in the hydrodynamic height of the adsorbed liposomes, upon adding glucose. This observation suggests that instead of osmotic deflation, glucose enhances the contact area via adhesion forces, presumably due to the depletion of the glucose molecules from the intermembrane hydration layer.

A Numerical Study on the Effect of Particle Surface Coverage on the Quartz Crystal Microbalance Response

The quartz crystal microbalance (QCM) is a surface-sensitive measurement technique to characterize adsorption processes at solid-fluid interfaces. While QCM measurements are routinely applied to study homogeneous thin films, characterizing heterogeneous films of adsorbed particles remains challenging because QCM is sensitive to not only the mass of adsorbed particles but also to that of hydrodynamically coupled fluid. To extract information about adsorbed particles, it is necessary to model these hydrodynamic effects, however, current QCM models are restricted to the limit of either a very low surface coverage or to the extrapolated limit of saturation coverage. Herein, we investigated QCM measurement responses in the intermediate surface coverage regime, by conducting lattice Boltzmann simulations of monodisperse, spherical particles that are attached to an oscillating surface. From the simulations, we relate the overtone-dependent QCM frequency and bandwidth shifts to particle size, interparticle distance, and the relevant hydrodynamic length scale. The corresponding results are in qualitative agreement with experimental QCM data for sub-100 nm, gel-phase liposomes. Furthermore, the data provide a theoretical basis for extracting particle sizes from QCM data in the high surface coverage limit.

Quartz Crystal Microbalance Model for Quantitatively Probing the Deformation of Adsorbed Particles at Low Surface Coverage

Characterizing the deformation of nanoscale, soft-matter particulates at solid-liquid interfaces is a demanding task, and there are limited experimental options to perform quantitative measurements in a nonperturbative manner. Previous attempts, based on the quartz crystal microbalance (QCM) technique, focused on the high surface coverage regime and modeled the adsorbed particles as a homogeneous film, while not considering the coupling between particles and surrounding fluid and hence resulting in an underestimation of the known particle height. In this work, we develop a model for the hydrodynamic coupling between adsorbed particles and surrounding fluid in the limit of a low surface coverage, which can be used to extract shape information from QCM measurement data. We tackle this problem by using hydrodynamic simulations of an ellipsoidal particle on an oscillating surface. From the simulation results, we derived a phenomenological relation between the aspect ratio r of the absorbed particles and the slope and intercept of the line that fits instantaneous, overtone-dependent QCM data on (δ/a, -Δf/n) coordinates where δ is the viscous penetration depth, a is the particle radius, Δf is the QCM frequency shift, and n is the overtone number. The model was applied to QCM measurement data pertaining to the adsorption of 34 nm radius, fluid-phase and gel-phase liposomes onto a titanium oxide-coated surface. The osmotic pressure across the liposomal bilayer was varied to induce shape deformation. By combining these results with a membrane bending model, we determined the membrane bending energy for the gel-phase liposomes, and the results are consistent with literature values. In summary, a phenomenological model is presented and validated in order to show for the first time that QCM experiments can quantitatively measure the deformation of adsorbed particles at low surface coverage.

Probing Membrane Viscosity and Interleaflet Friction of Supported Lipid Bilayers by Tracking Electrostatically Adsorbed, Nano‐Sized Vesicles

Particle tracking is used to measure the diffusional motion of nanosized (≈100 nm), lipid vesicles that are electrostatically adsorbed onto a solid supported lipid bilayer. It is found that the motion of membrane-adhering vesicles is Brownian and depends inversely on the vesicle size, but is insensitive to the vesicle surface charge. The measured diffusivity agrees well with the Evans-Sackmann model for the diffusion of inclusions in supported, fluidic membranes. The agreement implies that the vesicle motion is coupled to that of a nanoscopic lipid cluster in the upper leaflet, which slides over the lower leaflet. The diffusivity of membrane-adhering vesicles is therefore predominantly governed by the interleaflet friction coefficient, while the diffusivity of single lipids is mainly governed by the membrane viscosity. Combined with fluorescence recovery after photobleaching analysis, the interleaflet friction coefficient and the membrane viscosity are determined by applying the Evans-Sackmann model to the measured diffusivity of membrane adhering vesicles and that of supported membrane lipids. This approach provides an alternative to existing methods for measuring the interleaflet friction coefficient and the membrane viscosity.

Hydrodynamic Propulsion of Liposomes Electrostatically Attracted to a Lipid Membrane Reveals Size-Dependent Conformational Changes.

The efficiency of lipid nanoparticle uptake across cellular membranes is strongly dependent on the very first interaction step. Detailed understanding of this step is in part hampered by the large heterogeneity in the physicochemical properties of lipid nanoparticles, such as liposomes, making conventional ensemble-averaging methods too blunt to address details of this complex process. Here, we contribute a means to explore whether individual liposomes become deformed upon binding to fluid cell-membrane mimics. This was accomplished by using hydrodynamic forces to control the propulsion of nanoscale liposomes electrostatically attracted to a supported lipid bilayer. In this way, the size of individual liposomes could be determined by simultaneously measuring both their individual drift velocity and diffusivity, revealing that for a radius of ∼45 nm, a close agreement with dynamic light scattering data was observed, while larger liposomes (radius ∼75 nm) displayed a significant deformation unless composed of a gel-phase lipid. The relevance of being able to extract this type of information is discussed in the context of membrane fusion and cellular uptake.

Brownian Dynamics of Electrostatically Adhering Small Vesicles to a Membrane Surface Induces Domains and Probes Viscosity

Using single-particle tracking, we investigate the interaction of small unilamellar vesicles (SUVs) that are electrostatically tethered to the freestanding membrane of a giant unilamellar vesicle (GUV). We find that the surface mobility of the GUV-riding SUVs is Brownian, insensitive to the bulk viscosity, vesicle size, and vesicle fluidity but strongly altered by the viscosity of the underlying membrane. Analyzing the diffusional behavior of SUVs within the Saffman-Delbrück model for the dynamics of membrane inclusions supports the notion that the mobility of the small vesicles is coupled to that of dynamically induced lipid clusters within the target GUV membrane. The reversible binding also offers a nonperturbative means for measuring the viscosity of biomembranes, which is an important parameter in cell physiology and function.