Imbalances in the eye lens proteome are linked to cataract formation.

The prevalent model for cataract formation in the eye lens posits that damaged crystallin proteins form light-scattering aggregates. The α-crystallins are thought to counteract this process as chaperones by sequestering misfolded crystallin proteins. In this scenario, chaperone pool depletion would result in lens opacification. Here we analyze lenses from different mouse strains that develop early-onset cataract due to point mutations in α-, ß-, or γ-crystallin proteins. We find that these mutant crystallins are unstable in vitro; in the lens, their levels are substantially reduced, and they do not accumulate in the water-insoluble fraction. Instead, all the other crystallin proteins, including the α-crystallins, are found to precipitate. The changes in protein composition and spatial organization of the crystallins observed in the mutant lenses suggest that the imbalance in the lenticular proteome and altered crystallin interactions are the bases for cataract formation, rather than the aggregation propensity of the mutant crystallins.

Structural Study of (Hydroxypropyl)Methyl Cellulose Microemulsion-Based Gels Used for Biocompatible Encapsulations.

(Hydroxypropyl)methyl cellulose (HPMC) can be used to form gels integrating a w/o microemulsion. The formulation in which a microemulsion is mixed with a hydrated HPMC matrix has been successfully used as a carrier of biocompatible ingredients. However, little is known about the structure of these systems. To elucidate this, scanning electron microscopy was used to examine the morphology and the bulk of the microemulsion-based gels (MBGs) and small-angle X-ray scattering to clarify the structure and detect any residual reverse micelles after microemulsion incorporation in the gel. Electron paramagnetic resonance spectroscopy was applied using spin probes to investigate the polar and non-polar areas of the gel. Furthermore, the enzyme-labelling technique was followed to investigate the location of an enzyme in the matrix. A structural model for HPMC matrix is proposed according to which, although a w/o microemulsion is essential to form the final gel, no microemulsion droplets can be detected after incorporation in the gel. Channels are formed by the organic solvent (oil), which are coated by surfactant molecules and a water layer in which the enzyme can be hosted.

Preparation and Characterization of Stabilizer-Free Phytantriol-Based Water-in-Oil Internally Liquid Crystalline Emulsions.

Despite the widespread use of surfactants, there are known issues such as allergic reactions and formulation complications in their use as emulsion stabilizers. In this study, stabilizer-free water-in-oil (W/O) emulsions containing water, phytantriol, and almond oil were prepared by an ultra-turrax homogenizer, a standard laboratory equipment, and a high specialized high-shear device. Parameters such as mixing time, stirring rate, composition, order of addition of phases, and temperature were investigated to systematically optimize the preparation of the formulations through evaluating their accelerated physical stability by a centrifugal sedimentation technique. The liquid crystalline structure of the continuous phase was studied by small-angle X-ray scattering indicating a reverse hexagonal phase (H2). Microscopy images showed the emulsions prepared via high-shear method had smaller water droplets with more uniform shape and better dispersion as confirmed by Fourier-transform infrared-attenuated total reflection spectroscopy. Rheology studies showed a larger yield stress value for emulsions with higher content of phytantriol. Our results indicated that emulsions prepared by the high-shear device with higher amount of phytantriol were the most stable formulations. Applying the correct variables in the preparation of the stabilizer-free emulsions using ultra-turrax homogenizer, one could obtain similarly stable emulsions lacking the uniformity of the droplets.

Reverse Hexosome Dispersions in Alkanes-The Challenge of Inverting Structures.

Monoglycerides form lipophilic liquid-crystalline (LC) phases when mixed with water. The corresponding LC nanostructures coexist with excess water, which is a necessary condition for the formation of internally nanostructured dispersed particles. These nanostructures comprise bicontinuous cubic phases, inverted hexagonal phases, and inverted micellar cubic phases. The dispersed particles are therefore named cubosomes, hexosomes, or micellar cubosomes. Such dispersions are usually stabilized by hydrophilic high-molecular-weight triblock (TB) copolymers. Another way to stabilize such dispersions is by forming the so-called Pickering or Ramsden emulsions using nanoparticles as stabilizers. In this contribution, we explore the possibility of forming and stabilizing inverted or reverse systems, that is, dispersions of hydrophilic LC phases in an excess oil phase like tetradecane. Our aim was to change from oil-in-water emulsions to water-in-oil emulsions, where the water phase is a LC phase in equilibrium with excess oil and where the oil is nonpolar, for example, an alkane. This work consists of three parts: (1) to find a hexagonal hydrophilic LC phase that can not only incorporate a certain amount of tetradecane but can also coexist with excess tetradecane in the case of higher oil concentration, (2) to find a suitable stabilizer-either polymeric or nanoparticle type-that can stabilize the emulsion without destroying the hexagonal LC phase, and finally (3) to check the stability of this reverse hexosome emulsion. We discovered that it is possible to create a hexagonal hydrophilic LC phase with short-chain nonionic surfactants such as polyethylene glycol alkyl ethers or with high-molecular-weight TB copolymers of type A-B-A. Furthermore, it is possible to successfully stabilize the reverse hexosomes with low hydrophilic-lipophilic balance TB copolymers-either synthesized in our laboratory or commercially available ones-as well as with hydrophobized, commercially available silica nanoparticles.

Lipid transfer between submicrometer sized Pickering ISAsome emulsions and the influence of added hydrogel.

Transfer of lipids between droplets in Pickering emulsions has been studied by time-resolved small-angle X-ray scattering (SAXS). The special features of self-assembled liquid-crystalline phases have been applied to examine the kinetics of internal phase reorganization imposed by lipid release and uptake by the droplets. The findings reveal faster transfer kinetics in Pickering emulsions than in emulsions stabilized with Pluronic F127. It is shown that the transfer kinetics can be accelerated by adding free surfactant to the dispersions and that this acceleration becomes more dominant when micelles are formed. The effect of immobilization of the droplets has been studied by incorporating them into the appropriate hydrogel network. The droplets are arrested, and the transfer slows down significantly at high enough concentrations of the hydrogel where nonergodic systems are obtained.

Lipid transfer in oil-in-water isasome emulsions: influence of arrested dynamics of the emulsion droplets entrapped in a hydrogel.

The transfer kinetics of lipids between internally self-assembled droplets of O/W emulsions is studied. The droplets (isasomes) consist of various liquid-crystalline phases or W/O microemulsions stabilized by a polymeric stabilizer F127. The various internal phases were identified by the relative peak positions in the small-angle X-ray scattering (SAXS) curves. An arrested system composed of isasomes embedded in a gel matrix actually provides an additional possibility to control these systems in terms of the release of various host molecules. These experiments have been applied to examine the kinetics of the internal phase reorganization imposed by the lipids' release and uptake by the droplets embedded in a κ-carrageenan (KC) hydrogel network. Increasing the concentration of the gelling agent slows down the transfer from one droplet to the other through the aqueous phase. We examined the region where the free diffusion is stopped. i.e., the point where the system changes from the ergodic to the nonergodic state and the kinetics is essentially slowed down. This effect can be balanced by the addition of small amounts of free polymeric stabilizer, which speeds up the kinetics. This is even possible in the case of highly arrested dynamics of the emulsion droplets, as found for the highest KC hydrogel concentrations forming nonergodic systems.

Submicrometer-sized Pickering emulsions stabilized by silica nanoparticles with adsorbed oleic acid.

Oil-water Pickering emulsions of about 200 nm were stabilized by nanosized hydrophilic silica after a simple surface treatment method. We have modified the aqueous silica nanoparticle dispersions by simple adsorption of oleic acid to their surfaces, improving the hydrophobicity of the particles while maintaining their charge and stability. The adsorption was monitored by small-angle X-ray scattering and electrophoretic measurements to estimate the interparticle interactions and surface charges. The effect of various parameters, such as nanoparticle concentration, amount of oleic acid, ionic strength, and pH, on the droplets' size and stability was investigated by dynamic light scattering. Furthermore, the ability of these modified silica nanoparticles to stabilize long-chain alkanes, liquid paraffin, and liquid-crystalline phases was examined.