Photoswitchable vesicles

Vesicles are self-assembled nanosized structures that result from the auto-organization of amphiphilic compounds into closed bilayers. These systems are highly attractive subjects of fundamental research owing to their role as biomimetic models of cell membranes and compartmentalization systems. Vesicles also display very interesting features and properties for practical applications, namely as superior nanocontainers for encapsulation of functional molecules and materials. Exerting control over the formation, dissociation and permeability of these containers is therefore of paramount importance for many of their potentials applications. Being light the stimulus of choice for remote spatiotemporal control, we have compiled and revised selected published works on photoresponsive vesicles with the aim of providing an overview of the main strategies, advantages and limitations of such systems.     

Shrink-wrap vesicles

We describe a simple approach to the controlled removal of molecules from the membrane of large unilamellar vesicles made of fatty acids. Such vesicles shrink dramatically upon mixing with micelles composed of a mixture of fatty acid and a phospholipid (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC)), as fatty acid molecules leave the vesicle membrane and accumulate within the mixed micelles. Vesicle shrinkage was confirmed by dynamic light scattering, fluorescence recovery after photobleaching of labeled vesicles, and fluorescence resonance energy transfer between lipid dyes incorporated into the vesicle membrane. Most of the encapsulated impermeable solute is retained during shrinkage, becoming concentrated by a factor of at least 50-fold in the final small vesicles. This unprecedented combination of vesicle shrinkage with retention of contents allows for the preparation of small vesicles containing high solute concentrations, and may find applications in liposomal drug delivery.     

Formation of tubules and giant vesicles from large multilamellar vesicles

This study reports an observation of submicrometer multilamellar vesicles (MLVs) prepared by simply freeze-thawing a phospholipid dispersion at full hydration that transformed into giant vesicles (GVs) and tubules (TUs) when confined between microscope glass slides. Cover slide cleaning and surface treatment did not hamper the formation of GVs or TUs. However, when small unilamellar vesicles (SUV) were prepared or when MLVs were not confined but rather freely moved between the glass slides or when the phospholipid was in its gel phase, neither GVs nor TUs were observed. Altogether, our results suggested that MLVs would play a role as a lipid reservoir and that the liquid flow between the glass slides induces the peeling of the external bilayers, yielding the formation of tubules and giant unilamellar vesicles.     

Fatty acid vesicles

Results obtained from recent studies on the preparation and application of fatty acid vesicles are reviewed, focusing on some of the particular properties of fatty acid vesicles in comparison with conventional phospholipid vesicles (liposomes): (i) pH dependency which allows reversible transformations from non-vesicular to vesicular aggregates, and (ii) dynamic features that place fatty acid vesicles in between conventional vesicles formed from double-chain amphiphiles and micelles formed from single-chain surfactants. There are two main research areas in which fatty acid vesicles have been studied actively during the last years: (i) basic physico-chemical properties, and (ii) applications as protocell models. Applications of fatty acid vesicles in the fields of food additives and drug delivery are largely unexplored, which is at least partially due to concerns regarding the colloidal stability of fatty acid vesicles (pH- and divalent cation-sensitivity). Recently, fatty acid vesicles were prepared from highly unsaturated fatty acids (docosahexaenoic acid) and the pH range of vesicle formation could be extended to high or low pH values by preparing mixed vesicles through addition of a second type of single-chain amphiphile that stabilizes the vesicle bilayer but itself is not a fatty acid.     

Nanoparticle-loaded magnetophoretic vesicles

Magnetic nanoparticles have been assembled into the bilayer membrane of block copolymer vesicles. The nanoparticles decorate the hydrophobic/hydrophilic interface, which leads to bridging of adjacent bilayers and the formation of oligo-lamellar vesicles. The nanoparticle uptake of the vesicles is sufficiently high to become magnetophoretic in external magnetic fields as shown by video microscopy.     

Micropatterning of DNA-tagged vesicles

We present a novel concept for the creation of lipid vesicle microarrays based on a patterning approach termed Molecular Assembly Patterning by Lift-off (MAPL). A homogeneous MAPL-based single-stranded DNA microarray was converted into a vesicle array by the use of vesicles tagged with complementary DNAs, permitting sequence-specific coupling of vesicles to predefined surface regions through complementary DNA hybridization. In the multistep process utilized to fulfill this achievement, active spots consisting of PLL-g-PEGbiotin with a resistant PLL-g-PEG background, as provided by the MAPL process, was converted into a DNA array by addition of complexes of biotin-terminated DNA and NeutrAvidin. This was then followed by addition of POPC vesicles tagged with complementary cholesterol-terminated DNA, thus providing specific coupling of vesicles to the surface through complementary DNA hybridization. Quartz crystal microbalance with dissipation (QCM-D) and optical waveguide lightmode spectroscopy monitoring were used to optimize the multistep surface modification process. It was found that the amount of adsorbed biotinDNA-NeutrAvidin complexes decreases with increasing molar ratio of biotinDNA to NeutrAvidin and decreasing ionic strength of the buffer solution. Modeling of the QCM-D data showed that the shape of the immobilized vesicles depends on the amount of available anchoring groups between the vesicles and the surface. Fluorescent microscopy images confirmed the possibility to create well-defined patterns of DNA-tagged, fluorescently labeled vesicles in the micrometer range.     

Cationic beta-cyclodextrin bilayer vesicles

Cationic amphiphilic beta-cyclodextrins, substituted with hydrophobic n-alkylthio chains at the primary hydroxyl side and hydrophilic omega-amino-oligo(ethylene glycol) units at the secondary side, form bilayer vesicles with a diameter of 30-35 nm (when alkyl = hexadecyl) or nanoparticles with a diameter of ca. 120 nm (when alkyl = hexyl) in water.     

Supra-assembly of siliceous vesicles

We report a new approach to produce macroporous ( approximately 110 nm in diameter) ordered siliceous foams (MOSF) by using block copolymers as templates in the absence of any organic cosolvent. The fine three-dimensional honeycomb structure of MOSF was determined by electron tomography. A formation mechanism of MOSF that spans from the atomic to macroscopic scale is proposed, which involves the cooperative self-assembly of unilamellar vesicles followed by the supra-assembly of vesicles. The fusion of soft vesicles finally leads to MOSF with well-ordered and defined honeycomb structures.     

Self-adhesion among phospholipid vesicles

A compound was synthesized that binds to a phospholipid bilayer via a hydrophobic steroid thereby projecting a strong multi-hydrogen bonding unit into the surrounding water. As shown by light scattering, light microscopy, and cryo-HRSEM, this latter unit self-adheres and induces membrane-membrane attachments, as found in many biological systems.     

Spontaneous formation of vesicles

his review highlights the relevant issues of spontaneous formation of vesicles. Both the common characteristics and the differences between liposomes and vesicles are given. The basic concept of the molecular packing parameter as a precondition of vesicles formation is discussed in terms of geometrical factors, including the volume and critical length of the amphiphile hydrocarbon chain. According to theoretical considerations, the formation of vesicles occurs in the systems with packing parameters between 1/2 and 1. Using common as well as new methods of vesicle preparation, a variety of structures is described, and their nomenclature is given. With respect to sizes, shapes and inner structures, vesicles structures can be formed as a result of self-organisation of curved bilayers into unilamellar and multilamellar closed soft particles. Small, large and giant uni-, oligo-, or multilamellar vesicles can be distinguished. Techniques for determination of the structure and properties of vesicles are described as visual observations by optical and electron microscopy as well as the scattering techniques, notably dynamic light scattering, small angle X-ray and neutron scattering. Some theoretical aspects are described in short, viz., the scattering and the inverse scattering problem, angular and time dependence of the scattering intensity, the principles of indirect Fourier transformation, and the determination of electron density of the system by deconvolution of p(r) function. Spontaneous formation of vesicles was mainly investigated in catanionic mixtures. A number of references are given in the review.     

Properties of giant vesicles

We have discussed the specific properties of giant vesicles and their use as model systems for fluid interfaces and biomembranes. Recent advances in giant vesicle research include systematic measurements of visco-elastic parameters as a function of membrane composition, experiments with water-soluble amphiphiles and active membranes, as well as the investigation of hydrodynamic interactions. Notably, it has finally been possible to measure spontaneous curvatures of membranes for a variety of different systems. Experimentally, spontaneous curvature has been a somewhat obscure quantity so far. Furthermore, vesicles have been used to construct bioelectronic devices and new classes of vesicles made of polymers were introduced.     

Fluid vesicles in flow

We review the dynamical behavior of giant fluid vesicles in various types of external hydrodynamic flow. The interplay between stresses arising from membrane elasticity, hydrodynamic flows, and the ever present thermal fluctuations leads to a rich phenomenology. In linear flows with both rotational and elongational components, the properties of the tank-treading and tumbling motions are now well described by theoretical and numerical models. At the transition between these two regimes, strong shape deformations and amplification of thermal fluctuations generate a new regime called trembling. In this regime, the vesicle orientation oscillates quasi-periodically around the flow direction while asymmetric deformations occur. For strong enough flows, small-wavelength deformations like wrinkles are observed, similar to what happens in a suddenly reversed elongational flow. In steady elongational flow, vesicles with large excess areas deform into dumbbells at large flow rates and pearling occurs for even stronger flows. In capillary flows with parabolic flow profile, single vesicles migrate towards the center of the channel, where they adopt symmetric shapes, for two reasons. First, walls exert a hydrodynamic lift force which pushes them away. Second, shear stresses are minimal at the tip of the flow. However, symmetry is broken for vesicles with large excess areas, which flow off-center and deform asymmetrically. In suspensions, hydrodynamic interactions between vesicles add up to these two effects, making it challenging to deduce rheological properties from the dynamics of individual vesicles. Further investigations of vesicles and similar objects and their suspensions in steady or time-dependent flow will shed light on phenomena such as blood flow.     

Hydrogels from phospholipid vesicles

It is shown that phospholipid dispersions with a few percent of diacylphosphocholine PC in water can be swollen to single-phase lyotropic liquid crystalline L_α-phases by the addition of co-solvents like glycerol, 1,3-butyleneglycol BG or 1,2-propyleneglycol PG. The birefringent L_α-phases contain small unilamellar and multilamellar vesicles if the temperature of the samples is above the Krafft-Temperature T_m of the phospholipid. When such transparent birefringent viscous samples are cooled down below T_m the samples are transformed into birefringent gels. Cryo-TEM and FF-TEM measurements show that the bilayers of the vesicles are transformed from the liquid to the crystalline state during the transformation while the vesicle structure remains. The bilayers of the crystalline vesicles form adhesive contacts in the gel. Pulsed-field gradient NMR measurements show that two different kinds of water or co-solvent can be distinguished in the gels. One type of solvent molecules can diffuse like normal solvent in a continuous bulk phase. A second type of water diffuses much more slowly. This type of solvent is obviously trapped in the vesicles. The permeability of the crystalline vesicles for water and solvent molecules is much lower in the crystalline state than in the fluid state.     

Thiol-functionalized mesostructured silica vesicles

The direct supramolecular assembly of organofunctional mesostructures with a vesicular hierarchical morphology is reported for the first time for (SiO2)1-x(LSiO1.5)x compositions, where L is a mercaptopropyl group and x = 0.10-0.30.     

Synthesis of magnetic DDAB vesicles

The synthesis of magnetic vesicles is described. The vesicles are constituted by didodecyldimethylammonium bromide and have a diameter of about 1 m. An aqueous magnetic fluid, constituted by charged magnetic nanoparticles dispersed in water without surfactant, is encapsulated in the vesicle with a volume fraction in particles that may range up to 10%. The first step of the encapsulation is the synthesis of a multiple emulsion the intermediate oily phase being evaporated to obtain the DDAB bilayer. The magnetic vesicles thus synthesized align and change shape when a magnetic field is applied.     

Lysozyme binding onto cat-anionic vesicles

Mixing aqueous sodium dodecylsulfate with cetyltrimethylammonium bromide solutions in mole ratios close to (1.7/1.0) allows the formation of cat-anionic vesicles with an excess of negative charges on the outer surface. The vesicular dispersions are mixed with lysozyme, and interact electrostatically with the positive charges on the protein, forming lipo-plexes. Dielectric relaxation, zeta-potential, and light scattering indicate the occurrence of interactions between vesicles and the protein. According to CD, the vesicle-adsorbed protein retains its native conformation. Binding and surface saturation, inferred by dielectric relaxation and zeta-potential, fulfil a charge neutralisation stoichiometry. Adsorbed lysozyme promotes the vesicle clustering and is concomitant with the lipo-plexes flocculation. Above the charge neutralisation threshold, lysozyme in excess remains dispersed in molecular form. Attempts were made to determine in what conditions protein release from the vesicles occurs. Accordingly, the full neutralisation of sodium dodecylsulfate in excess by cetyltrimethylammonium bromide ensures the lipo-plexes break-up, the precipitation of the mixed surfactants and the protein release in native form.     

Giant vesicles with membranous microcompartments

Incubation of a cell-sized lipid membrane vesicle (giant vesicle, GV) in a diluted aqueous solution of neutral phosphate buffer salts or glucose transformed the GV to an oligovesicular vesicle (OVV) that encapsulates one or more smaller GVs. During the incubation, the membrane of flaccid vesicle invaginated and closed to form the inner vesicle of an OVV engulfing a part of the bulk aqueous phase. Using the GV-to-OVV transformation, an OVV that has different aqueous contents in each membranous microcompartment was constructed.