Progress in liquid crystalline dispersions: Cubosomes

Dispersed particles of bicontinuous cubic liquid crystalline phase, cubosomes, are self-assembled nanostructured particles that can be formed in aqueous lipid and surfactant systems. Contributions to cubosome research have come from the fields of biology, material science, medicine, and mathematics and much is known about their formation and properties. At the center of much of the discovery and innovation is the technique of cryo-transmission electron microscopy. Most of the research into cubosomes is motivated by potential applications in drug delivery and material synthesis although no commercialized product based on cubosomes is known. Recent advances in understanding and use of cubosomes are discussed in the context of some of the more promising application areas and the opportunities for microscopy techniques to make unique contributions to these areas.     

Cryo-TEM studies of DNA and DNA–lipid structures

Studies published between 2001 and 2005 that utilise cryogenic transmission electron microscopy as a tool for investigating solutions containing DNA with or without amphiphiles present are reviewed. DNA or oligonucleotides form complexes with amphiphilic molecules such as lipids, so-called lipoplexes, and the structures and morphologies are excellent objects to study by cryogenic transmission electron microscopy. Recent studies show that this technique is an effective tool for identification of a range of structures such as unilamellar or multilamellar vesicles or dispersed liquid crystalline phases.     

Periodic minimal surface organizations of the lipid bilayer at the lung surface and in cubic cytomembrane assemblies

The existence of infinite periodic lipid bilayer structures in biological systems was first demonstrated in cell membrane assemblies. Such periodicity is only possible in symmetric bilayers, and their occurrence is discussed here in relation to the asymmetry of cell membranes in vivo. A periodic membrane conformation in the prolamellar body of plants corresponds to a dormant state without photosynthesis. A similar reversible formation of a dormant state has also been observed in the mitochondria of the amoeba Chaos. In these cases the energy production has become insufficient to maintain the membrane asymmetry. Formation of membranes that are symmetric over the bilayer is proposed to be a principal mechanism behind formation of cubic membrane systems.     

Effects of chemical structures of para-halobenzoates on micelle nanostructure,drag reduction and rheological behaviors of dilute CTAC solutions

Counterion chemical structure and counterion to cationic surfactant molar ratio, ξ, control counterion binding, micelle nanostructures, drag reduction (DR) effectiveness and rheological behavior of quaternary ammonium surfactant systems. The effects of chemical structures of four sodium para-halobenzoate (F, Cl, Br, I) counterions with different ξ values on these properties were compared for dilute solutions of cetyltrimethylammonium chloride (CTAC). Counterion binding was determined by zeta-potential and ¹H NMR measurements. Nanostructures were determined by ¹H NMR and cryo-TEM imaging. Nanostructures, drag reduction effectiveness measured over a range of temperatures and Reynolds numbers, shear viscosities and first normal stress differences N1 were related to the chemical structures of the four counterions and their molar ratios to CTAC.     

Milk membrane lipid vesicle structures studied with Cryo-TEM

Milk fat globule membrane (MFGM) lipids have been studied in the presence and absence of proteins β-lactoglobulin and β-casein. The aim of this study was to relate the self-assembly structure, e.g. vesicles, formed in aqueous dispersions of MFGM lipids to the lipid composition, electrolyte composition as well as the effect of added milk proteins, i.e. β-lactoglobulin and β-casein. For this purpose, vesicles of phospholipid mixtures, containing dioleoylphosphatidylcholine (DOPC), sphingomyelin (SM), dioleoylphosphatidylethanolamine (DOPE), phosphatidylinositol (PI) and dioleoylphosphatidylserine (DOPS) at composition corresponding to that of the MFGM, were prepared by extrusion. The morphology of the formed structures of different sample compositions was studied with cryogenic transmission electron microscopy (Cryo-TEM). Mixtures of membrane lipid with a composition (e.g. 80% DOPE, 12% DOPC and 8% SM) that at high lipid content give liquid crystalline phases at the boundary of lamellar to reversed hexagonal phase rather formed microtubular structures than vesicles at high water content. A large proportion of multilamellar vesicles is formed in buffer and divalent salts than in pure water. A small increase in the interlayer spacing of the multilamellar vesicle was observed in the presence of β-casein.     

Spontaneous vesicles of sodium dihexadecylphosphate in HEPES buffer

The formation of spontaneous vesicles of dihexadecylphosphate (DHP) in a HEPES buffered solution at pH 7.4, the size, morphology and melting temperature, obtained by cryogenic transmission electron microscopy (cryo-TEM) and differential scanning calorimetry (DSC), are reported. The vesicles were formed by simply mixing a 5.0 mM lipid-solvent mixture at a temperature (75 degrees C) safely above the higher melting temperature T(m)=70.4 degrees C of DHP. The vesicle diameter ranges from 100 to 332 nm and their geometry is spherical, faceted or oblong. T(m) increases from 66.8 to 70.4 as DHP concentration is raised from 0.6 to 5.0 mM.     

Polymeric Nanoparticles Stabilized by Surfactants Investigated by Light Scattering,Small-Angle Neutron Scattering,and Cryo-TEM Methods

We have investigated self-organization of polymers with surfactants through solvent shifting process resulting in formation of stable and uniform nanoparticles. We studied polymeric nanoparticles made of poly(methylmethacrylate) and of polystyrene dispersed in water. The dispersion was prepared by a fast mixing of a solution of the polymers with a solution of several ionic and nonionic surfactants in pure water. We observed the formation of well defined nanoparticles by light scattering, small-angle neutron scattering (SANS), and cryogenic transmission electron microscopy (Cryo-TEM) methods. The study shows how nanoparticle properties are changed by the chemical composition of surfactants, molar mass of polymers, concentrations of both components and finally, by variations in method of nanoparticles preparation. Dynamic light scattering (DLS) and static light scattering (SLS) provide the hydrodynamic radii and radii of gyration for selected types of nanoparticles. Cryo-TEM experiments prove that the nanoparticles have good spherical shape. Analysis of SANS data and Cryo-TEM micrographs suggest that the prepared particles are composed of polymer and surfactant that are evenly distributed.     

Redox-active lipid-incorporating proteins as a novel immobilisation technique

A novel chiral redox-active ferrocene compound (FcV1) with amphiphilic properties has been synthesised and used successfully to form protein + lipid ‘clusters’ with ferritin and gold-labelled BSA. These structures could be observed by cryogenic transmission electron microscopy and had diameters between 40 and 300 nm. It was shown that the novel lipid formed reasonably stable films on an edge plane graphite electrode surface and also mediated the electron transfer between flavocytochrome b₂ and this electrode. Consequently, the enzyme was incorporated into an FcV1 film and its response towards (+)lactate in solution investigated. It could be shown that the electrode indeed was sensitive to (+)lactate.     

Silicone Nano/Microstructures Obtained in Ionic Polymerization

Summary: This work presents a novel surfactant-free method of preparation of silicone hollow nano/microstructures in water. New silicone monomer was synthesized by attaching the unsaturated fatty acid ester – methyl 10-undecenoate (UDM) to the commercial silicone monomer - 1,3,5,7-tetramethylcyclotetrasiloxane (D) using hydrosilylation reaction. The reaction was catalyzed by Karstedt's catalyst [Pt(0)-divinyltetramethyldisiloxane complex]. That modified monomer, tetra(11-methoxy-11-oxoundecyl)-tetramethylcyclotetrasiloxane (D), undergoes self-organization in water. The anionic ring-opening polymerization of D resulted in formation of the silicone polymer (poly-D). Both, the silicone monomer and the polymer were dispersed in aqueous medium using sonication. The structures in the dispersions were visualized by cryo-transmission electron microscopy (Cryo-TEM) and optical microscopy. The size of the particles was also determined from dynamic light scattering (DLS) measurements.     

The influence of lignin on the self-assembly behaviour of xylan rich fractions from birch (Betula pendula)

A crude xylan isolate obtained by prehydrolysis and mild alkaline extraction from birch wood chips (Betula pendula), and a carefully delignified xylan fraction from the same source, were examined by dynamic light scattering (DLS) and cryogenic transmission electron microscopy (cryo-TEM) with regard to their propensity to self-assemble in water into insoluble aggregates. The delignification involved the extraction with chloroform of a crude xylan solution in a pyridine/acetic acid/water mixture. It resulted in a purified xylan fraction in a yield of 23% in which 75 and 90% of the lignin had been removed as indicated by Klason and UV-determination, respectively. It was found that both xylan fractions formed agglomerates by self-assembly in water. However, DLS and cryo-TEM indicated that the aggregates were larger in size (90 vs. 40 nm) and greater in mass when more lignin was present. The addition of an alkaline solution of isolated lignin (obtained by steam explosion) to increasing concentrations of a delignified xylan revealed increasing turbidity. Our conclusion is that lignin induces agglomeration of xylan in aqueous solutions, but xylan concentration plays an active role in the aggregation phenomena. An agglomeration mechanism for lignin rich xylan fractions is proposed.