Hierarchical patterning of hydrogels by replica molding of impregnated breath figures leads to superoleophobicity.

The surface chemistry and topography govern the spreading of liquids on a solid. When an oil drop makes a contact angle, θ > 90° on a solid surface, the solid is termed as oleophobic. Adding roughness to an inherently oleophobic surface enhances its oil dewetting and can lead to superoleophobicity when θ > 150°. In this study, we introduce the concept of a two-tier hierarchical roughness on the surface of soft materials such as hydrogels by forming the patterned inverse replica of breath figure polymer films impregnated with nanoparticles. The directed deposition of nanoparticles in the breath figure pores is accomplished by an aerosol assisted technique that exclusively leads to deposition within the pores and filling of the pores. The inverse replica of such impregnated films exhibits a close packed hexagonally structured second tier of surface roughness which directly leads to a superoleophobic surface. Since these structures have well defined geometries, it is possible to estimate the contact angle by assuming a partial wetting of the oil drop in a 'fakir' state on the rough surface. The estimation is in good agreement with the experimental contact angle value. While the work demonstrates a facile method to impart superoleophobicity to a hydrogel surface, it also demonstrates new methods to imbue breath figure pores with functional materials that can be easily transferred to the pores of the inverse replica.

Spatially directed vesicle capture in the ordered pores of breath-figure polymer films.

This work describes a new method to selectively capture liposomes and other vesicle entities in the patterned pores of breath-figure polymer films. The process involves the deposition of a hydrophobe containing biopolymer in the pores of the breath figure, and the tethering of vesicles to the biopolymer through hydrophobic interactions. The process is versatile, can be scaled up and extended to the deposition of other functional materials in the pores of breath figures.

Fenofibrate subcellular distribution as a rationale for the intracranial delivery through biodegradable carrier.

Fenofibrate, a well-known normolipidemic drug, has been shown to exert strong anticancer effects against tumors of neuroectodermal origin including glioblastoma. Although some pharmacokinetic studies were performed in the past, data are still needed about the detailed subcellular and tissue distribution of fenofibrate (FF) and its active metabolite, fenofibric acid (FA), especially in respect to the treatment of intracranial tumors. We used high performance liquid chromatography (HPLC) to elucidate the intracellular, tissue and body fluid distribution of FF and FA after oral administration of the drug to mice bearing intracranial glioblastoma. Following the treatment, FF was quickly cleaved to FA by blood esterases and FA was detected in the blood, urine, liver, kidney, spleen and lungs. We have also detected small amounts of FA in the brains of two out of six mice, but not in the brain tumor tissue. The lack of FF and FA in the intracranial tumors prompted us to develop a new method for intracranial delivery of FF. We have prepared and tested in vitro biodegradable poly-lactic-co-glycolic acid (PLGA) polymer wafers containing FF, which could ultimately be inserted into the brain cavity following resection of the brain tumor. HPLC-based analysis demonstrated a slow and constant diffusion of FF from the wafer, and the released FF abolished clonogenic growth of glioblastoma cells. On the intracellular level, FF and FA were both present in the cytosolic fraction. Surprisingly, we also detected FF, but not FA in the cell membrane fraction. Electron paramagnetic resonance spectroscopy applied to spin-labeled phospholipid model-membranes revealed broadening of lipid phase transitions and decrease of membrane polarity induced by fenofibrate. Our results indicate that the membrane-bound FF could contribute to its exceptional anticancer potential in comparison to other lipid-lowering drugs, and advocate for intracranial delivery of FF in the combined pharmacotherapy against glioblastoma.

Liposomes tethered to a biopolymer film through the hydrophobic effect create a highly effective lubricating surface.

Liposomal coatings are formed on films of a biopolymer, hydrophobically modified chitosan (hm-chitosan), containing dodecyl groups as hydrophobes along the polymer backbone. The alkyl groups insert themselves into the liposome bilayer through hydrophobic interactions and thus tether liposomes, leading to a densely packed liposome layer on the film surface. Such liposomal surfaces exhibit effective lubrication properties due to their high degree of hydration, and reduce the coefficient of friction to the biologically-relevant range. The compliancy and robustness of these tethered liposomes allow retention on the film surface upon repeated applications of shear. Such liposome coated films have potential applications in biolubrication.

Microstructure determination of AOT + phenol organogels utilizing small-angle X-ray scattering and atomic force microscopy.

Dry reverse micelles of the anionic twin-tailed surfactant bis(2-ethylhexyl) sulfosuccinate (AOT) dissolved in nonpolar solvents spontaneously form an organogel when p-chlorophenol is added in a 1:1 AOT:phenol molar ratio. The solvents used were benzene, toluene, m-xylene, 2,2,4-trimethylpentane (isooctane), decane, dodecane, tetradecane, hexadecane, and 2,6,10,14-tetramethylpentadecane (TMPD). The proposed microstructure of the gel is based on strands of stacked phenols linked to AOT through hydrogen bonding. Small-angle X-ray scattering (SAXS) spectra of the organogels suggest a characteristic length scale for these phenol-AOT strands that is independent of concentration but dependent on the chemical nature of the nonpolar solvent used. Correlation lengths determined from the SAXS spectra indicate that the strands self-assemble into fibers. Direct visualization of the gel in its native state is accomplished by using tapping mode atomic force microscopy (AFM). It is shown that these organogels consist of fiber bundle assemblies. The SAXS and AFM data reinforce the theory of a molecular architecture consisting of three length scales-AOT/phenolic strands (ca. 2 nm in diameter) that self-assemble into fibers (ca. 10 nm in diameter), which then aggregate into fiber bundles (ca. 20-100 nm in diameter) and form the organogel.

EPR characterizations of alpha-chymotrypsin active site dynamics in reversed micelles at enhanced gas pressures and after subjection to clathrate formation conditions.

Electron paramagnetic resonance spectroscopy is used to characterize the active site dynamics of alpha-chymotrypsin solubilized in reversed micelles. Of particular interest is the behavior of the enzyme when the micellar system is subjected to enhanced gas pressures and low temperatures. At specific thermodynamic conditions, clathrate hydrates from from the intramicellar water, reducing the micelle size and water content. Also, beyond a critical pressure, micellar instbility results. The EPR spectra under these conditions indicate that the rotational correlation times increase appreciably only when the water-to-surfactant molar ratio, W(0), is reduced to values lower than 10. The EPR characterization also reveals a remarkable resilience of the enzyme when subjected to pressure-induced changes; when returned to ambient conditions, activity and active site dynamics are fully restored. (c) 1994 John Wiley & Sons, Inc.

Catalytic and interfacial aspects of enzymatic polymer synthesis in reversed micellar systems.

The enzyme horseradish peroxidase, when encapsulated in reversed micelles, is capable of catalyzing the synthesis of phenolic and aromatic amine polymers. The synthesis of polyethylphenol is specifically considered in this article and is found to be extremely feasible in the micellar system. Polymer chain growth can be controlled to some degree by manipulating the ability of the solvent to sustain chain solubility; this is effectively done by adjusting the surfactant concentration. This results in a degree of control of polymer molecular weight. The synthesized polymer drops out of solution and can be easily recovered.

The use of pressure to modify enzyme activity in reversed micelles.

Pressurization of enzyme-containing AOT-water-isooctane reversed micelles with low molecular weight gases leads to markedly different responses in activity characteristics. Microbial lipases exhibit a total cutoff in activity with as low a pressure as 2 MPa and a remarkable activity regain with depressurization. The observation also holds for reaction in monophasic organic solvents. The protease, alpha-chymotrypsin, is unaffected by pressurization until a critical pressure wherein micellar instability occurs. The use of pressure as a switch for lipase reaction in nonaqueous media is discussed.

Protein recovery from reversed micellar solutions through contact with a pressurized gas phase.

We describe a new process for the recovery of encapsulated protein from reversed micellar solution in concentrated form. The method involves desolubilization of the protein by decreasing solvent density through gas dissolution. Under appropriate thermodynamic conditions, the micellar water pool can be converted to clathrate hydrates. Protein recovery is facilitated by clathrate hydrate formation, which causes the desolubilized protein to exist in a solid phase, distinct from the micellar supernatant. The process is carried out without any ionic strength or pH modification.

Modification of enzyme activity in reversed micelles through clathrate hydrate formation.

The physical phenomenon of clathrate hydrate formation in protein-containing reversed micelles is described. Hydrate formation in reversed micelles is a method of adjusting the water to surfactant molar ratio, wo, which influences micellar size. Lipase and alpha-chymotrypsin encapsulated in large reversed micelles of high wo show significant enhancements in activity when the micelle size is reduced through hydrate formation. Alternate methods of micelle size adjustments also show enhancements in activity. The implications for improving the activity of such encapsulated enzymes recovered from fermentation media through phase transfer into reversed micelles are discussed.

Some conditions affecting the intracellular arrangement and concentration of tobacco mosaic virus particles in local lesions.

Tobacco mosaic virus particles were found in small packets and in small numbers, with the electron microscope, in necrotic leaf cells of Nicotiana glutinosa when the samples were fixed in glutaraldehyde and postfixed in OsO(4), and the sections were stained with heavy metals. The numbers and size of the virus packets were increased greatly when the leaves were detached from the plant after inoculation Assay of concentration showed that detachment resulted in a 30-fold increase of virus. A similar increase in the number of virus particles detected by electron microscopy was produced by keeping inoculated plants at an air temperature of 26 degrees C. A still greater increase in concentration was effected by incubating detached inoculated leaves at 26 degrees C. Moreover the arrangement of virus particles in these cells resembled that of a systemic virus infection. Cells in local lesions of Chenopodium amaranticolor contained large numbers of virus particles both as packets and in the loose arrangement characteristic of systemic infection. Neither the number of particles nor their arrangement was affected in this host by detaching the leaf or by changing the air temperature. It is suggested that there may be two types of localized virus infections, one of which produces virus in low concentration and is amenable to changes in virus concentration and arrangement as a result of environmental manipulation.