Tatsiana Lobovkina

Nanoparticle PEGylation for imaging and therapy

Nanoparticles are an essential component in the emerging field of nanomedical imaging and therapy. When deployed in vivo, these materials are typically protected from the immune system by polyethylene glycol (PEG). A wide variety of strategies to coat and characterize nanoparticles with PEG has established important trends on PEG size, shape, density, loading level, molecular weight, charge and purification. Strategies to incorporate targeting ligands are also prevalent. This article presents a background to investigators new to stealth nanoparticles, and suggests some key considerations needed prior to designing a nanoparticle PEGylation protocol and characterizing the performance features of the product.

In Vivo Sustained Release of siRNA from Solid Lipid Nanoparticles

Small interfering RNA (siRNA) is a highly potent drug in gene-based therapy with a challenge of being delivered in a sustained manner. Nanoparticle drug delivery systems allow for incorporating and controlled release of therapeutic payloads. We demonstrate that solid lipid nanoparticles can incorporate and provide sustained release of siRNA. Tristearin solid lipid nanoparticles, made by nanoprecipitation, were loaded with siRNA (4.4-5.5 wt % loading ratio) using a hydrophobic ion pairing approach that employs the cationic lipid DOTAP. Intradermal injection of these nanocarriers in mouse footpads resulted in prolonged siRNA release over a period of 10-13 days. In vitro cell studies showed that the released siRNA retained its activity. Nanoparticles developed in this study offer an alternative approach to polymeric nanoparticles for encapsulation and sustained delivery of siRNA with the advantage of being prepared from physiologically well-tolerated materials.

Shape optimization in lipid nanotube networks

Abstract.Starting from a high surface free-energy state, lipid nanotube networks are capable to self-organize into tree-like structures with particular geometrical features. In this work we analyze the process of self-organization in such networks, and report a strong similarity to the Euclidian Steiner Tree Problem (ESTP). ESTP is a well-known NP-hard optimization problem of finding a network connecting a given set of terminal points on a plane, allowing addition of auxiliary points, with the overall objective to minimize the total network length. The present study shows that aggregate lipid structures self-organize into geometries that correspond to locally optimal solutions to such problems.

Protrusive growth and periodic contractile motion in surface-adhered vesicles induced by Ca2+-gradients

Local signaling, cell polarization, and protrusive growth are key steps in directed migration of biological cells guided by chemical gradients. Here we present a minimal system which captures several key features of cellular migration from signaling-to-motion. The model system consists of flat, negatively charged phospholipid vesicles, a negatively charged surface, and a local, and controllable point-source supply of calcium ions. In the presence of a Ca2+ gradient, the surface-adhered vesicles form protrusions in the direction of the gradient. We also observe membrane shape oscillations between expanded (flattened), and spherical states as a function of the Ca2+-concentration. The observed phenomena can be of importance in explaining motile action in prebiotic, primitive, and biomimetic systems, as well as in development of novel soft-matter nano- and microscale mechanical devices.

Fractal avalanche ruptures in biological membranes

Bilayer membranes envelope cells as well as organelles, and constitute the most ubiquitous biological material found in all branches of the phylogenetic tree. Cell membrane rupture is an important biological process, and substantial rupture rates are found in skeletal and cardiac muscle cells under a mechanical load. Rupture can also be induced by processes such as cell death, and active cell membrane repair mechanisms are essential to preserve cell integrity. Pore formation in cell membranes is also at the heart of many biomedical applications such as in drug, gene and short interfering RNA delivery. Membrane rupture dynamics has been studied in bilayer vesicles under tensile stress, which consistently produce circular pores. We observed very different rupture mechanics in bilayer membranes spreading on solid supports: in one instance fingering instabilities were seen resulting in floral-like pores and in another, the rupture proceeded in a series of rapid avalanches causing fractal membrane fragmentation. The intermittent character of rupture evolution and the broad distribution in avalanche sizes is consistent with crackling-noise dynamics. Such noisy dynamics appear in fracture of solid disordered materials, in dislocation avalanches in plastic deformations and domain wall magnetization avalanches. We also observed similar fractal rupture mechanics in spreading cell membranes.

Sustained release of nucleic acids from polymeric nanoparticles using microemulsion precipitation in supercritical carbon dioxide.

A general approach for producing biodegradable nanoparticles for sustained nucleic acid release is presented. The nanoparticles are produced by precipitating a water-in-oil microemulsion in supercritical CO(2). The microemulsion consists of a transfer RNA aqueous solution (water phase), dichloromethane containing poly(l-lactic acid)-poly(ethylene glycol) (oil phase), the surfactant n-octyl β-D-glucopyranoside, and the cosurfactant n-butanol.

Membrane Tubulation in Lipid Vesicles Triggered by the Local Application of Calcium Ions

Experimental and theoretical studies on ion-lipid interactions predict that binding of calcium ions to cell membranes leads to macroscopic mechanical effects and membrane remodeling. Herein, we provide experimental evidence that a point source of Ca2+ acting upon a negatively charged membrane generates spontaneous curvature and triggers the formation of tubular protrusions that point away from the ion source. This behavior is rationalized by strong binding of the divalent cations to the surface of the charged bilayer, which effectively neutralizes the surface charge density of outer leaflet of the bilayer. The mismatch in the surface charge density of the two leaflets leads to nonzero spontaneous curvature. We probe this mismatch through the use of molecular dynamics simulations and validate that calcium ion binding to a lipid membrane is sufficient to generate inward spontaneous curvature, bending the membrane. Additionally, we demonstrate that the formed tubular protrusions can be translated along the vesicle surface in a controlled manner by repositioning the site of localized Ca2+ exposure. The findings demonstrate lipid membrane remodeling in response to local chemical gradients and offer potential insights into the cell membrane behavior under conditions of varying calcium ion concentrations.

Soft-Matter Nanotubes

This chapter provides an overview over the extended area of surfactant nanotubes research, covering theoretical as well as experimental approaches. Fabrication strategies for nanotube assemblies include surfactant or amphiphile self-assembly, forced shape transformations and, in case of appropriately functionalized building blocks, polymerization. The main body of this review is dedicated to the dynamic properties of lipid nanotubes, their role in vesicle-nanotube networks and derivatives consisting of branched, knotted, or circular nanotubes. Transport modes and enzymatic reactions in nanotube-interconnected vesicles are discussed, in particular their application as unique research tools to mimic sub-cellular conditions, and to explore concepts of unconventional, miniaturized chemical reactors in a biocompatible environment. Lipid nanotubes can be expected to further gain in importance as model systems for nanotube-based (transport) processes in biological cells. They facilitate the investigation of chemical reaction kinetics in complex structured geometries as well as studies of transport phenomena involving ultra small volumes and single molecules, and provide new insights into fundamental aspects of the biophysics of membranes. This review is particularly committed to highlight the rich opportunities to engineer and utilize surfactant nanotube assemblies, leading the way to future biological and technological applications.

Formation and release of circular lipid nanotubes

A method for formation of circular lipid nanotubes based on manipulation of nanotube-vesicle networks is presented.

Contactless Stimulation and Control of Biomimetic Nanotubes by Calcium Ion Gradients

Membrane tubular structures are important communication pathways between cells and cellular compartments. Studying these structures in their native environment is challenging, due to the complexity of membranes and varying chemical conditions within and outside of the cells. This work demonstrates that a calcium ion gradient, applied to a synthetic lipid nanotube, triggers lipid flow directed toward the application site, resulting in the formation of a bulge aggregate. This bulge can be translated in a contactless manner by moving a calcium ion source along the lipid nanotube. Furthermore, entrapment of polystyrene nanobeads within the bulge does not tamper the bulge movement and allows transporting of the nanoparticle cargo along the lipid nanotube. In addition to the synthetic lipid nanotubes, the response of cell plasma membrane tethers to local calcium ion stimulation is investigated. The directed membrane transport in these tethers is observed, but with slower kinetics in comparison to the synthetic lipid nanotubes. The findings of this work demonstrate a novel and contactless mode of transport in lipid nanotubes, guided by local exposure to calcium ions. The observed lipid nanotube behavior can advance the current understanding of the cell membrane tubular structures, which are constantly reshaped during dynamic cellular processes.