An HII liquid crystal-based delivery system for cyclosporin A: physical characterization

In the present study we demonstrate that large quantities of cyclosporin A and three dermal penetration enhancers (phosphatidylcholine, ethanol, or Labrasol) can be solubilized into reverse hexagonal (HII) liquid crystalline structures composed of monoolein, tricaprylin, and water. The microstructural characteristics of these complex multi-component systems were elucidated by rheological, SAXS, and DSC measurements. Addition of up to 20 wt% phosphatidylcholine improved significantly the elastic properties of the systems (lower values of tandelta) and increased the thermal stability of the mesophases enabling us to solubilize up to 6 wt% cyclosporin A and two other enhancers (Labrasol and ethanol) to obtain stable mesophases at physiological temperature. Rheological measurements revealed that solubilization of cyclosporin A alone has a destabilizing effect on the reverse hexagonal phases: it caused a deterioration in the elastic properties of the systems, leading to more liquid-like behavior and resulting in very short relaxation times (0.04-0.1 s). Labrasol, solubilized at high concentrations (up to 12 wt%) into the liquid crystals, also demonstrated a destabilizing effect on the HII structure: the decreasing elasticity of the system was attributed to Labrasol's presumed locus at the interface and its ability to bind water, as shown by DSC measurements. Ethanol had a destabilizing effect similar to that of Labrasol, yet the effect appeared to be more pronounced, probably due to its higher water-binding capability.     

Solubilization of nutraceuticals into reverse hexagonal mesophases

The solubilization of four bioactive molecules with different polarities, in three reverse hexagonal (HII) systems has been investigated. The three HII systems were a typical reverse hexagonal composed of glycerol monooleate (GMO)/tricaprylin/water and two fluid hexagonal systems containing either 2.75 wt % Transcutol or ethanol as a fourth component. The phase behavior of the liquid crystalline phases in the presence of ascorbic acid, ascorbyl palmitate, D-alpha-tocopherol and D-alpha-tocopherol acetate were determined by small-angle X-ray scattering (SAXS) and optical microscopy. Differential scanning calorimetry (DSC) and Fourier-transform infrared (FT-IR) techniques were utilized to follow modifications in the thermal behavior and in the vibrations of different functional groups upon solubilizing the bioactive molecules. The nature of each guest molecule (in both geometry and polarity) together with the different HII structures (typical and fluids) determined the corresponding phase behavior, swelling or structural transformations and its location in the HII structures. Ascorbic acid was found to act as a chaotropic guest molecule, localized in the water-rich core and at the interface. The AP was also a chaotropic guest molecule with its head located in the vicinity of the GMO headgroup while its tail embedded close to the surfactant tail. D-alpha-tocopherol and D-alpha-tocopherol acetate were incorporated between the GMO tails; however, the D-alpha-tocopherol was located closer to the interface. Once Transcutol or ethanol was present and upon guest molecule incorporation, partial migration was detected.     

Towards a mesoscopic model of water-like fluids with hydrodynamic interactions

We present a mesoscopic lattice model for non-ideal fluid flows with directional interactions, mimicking the effects of hydrogen bonds in water. The model supports a rich and complex structural dynamics of the orientational order parameter, and exhibits the formation of disordered domains whose size and shape depend on the relative strength of directional order and thermal diffusivity. By letting the directional forces carry an inverse density dependence, the model is able to display a correlation between ordered domains and low density regions, reflecting the idea of water as a denser liquid in the disordered state than in the ordered one.     

Aqueous phase behavior of a 1-O-phytanyl-beta-D-xyloside/water system. Glycolipid-based bicontinuous cubic phases of crystallographic space groups Pn3m and Ia3d

Temperature- and concentration-dependent aqueous phase diagram of a novel alkylglycoside, 1-O-phytanyl-beta-D-xyloside (beta-Xyl(Phyt)), was studied using small-angle X-ray scattering, polarizing optical microscopy, and differential scanning calorimetry. The phases found in this system include an Lc phase, an Lalpha phase, an HII phase, two inverted cubic phases of crystallographic space groups Pn3m and Ia3d, and a fluid isotropic phase, FI. The phase diagram of the beta-Xyl(Phyt)/water system is similar to that for the 1-monooleylglycerol (MO)/water system, suggesting that the phase behavior is largely determined by the overall molecular shape rather than the details of surfactant molecular structure. Moreover, the structural parameters of the beta-Xyl(Phyt) liquid crystals are also similar to those of the MO/water, due primarily to the similar molecular dimensions of two molecules. As compared to the MO/water system, however, the beta-Xyl(Phyt)/water system displays a lower value of TK ( approximately 8.(5) degrees C) and a wider temperature window for the mesophases (8.(5)-120 degrees C). Moreover, beta-Xyl(Phyt) is chemically more robust than MO, as the ether linkage is more stable against hydrolysis than the ester linkage and the phytanyl chain is fully saturated.     

Phase structures of a hydrated anionic phospholipid composition containing cationic dendrimers and pegylated lipids

The effect of 4th generation poly(amidoamine) dendrimer (4G PAMAM) present in an anionic phospholipid composition, consisting of hydrogenated soyphosphatidylcholine (HSPC), cholesterol (CH), dicetyl phosphate (DCP), and poly(ethylene glycol) (Mw approximately 2000) derivatized phosphatidylethanolamine (PEG2000-PE), on the hydration and liquid crystalline structure formation was investigated. The optical and polarized light microscopies of the liposomal dispersion obtained from the hydrated lipid composition show two types of birefringent structures (mesophases): plastic, wormlike microstructures and conventional, over-elongated lamellae. Differential scanning calorimetry (DSC) shows an increase in the liquid crystalline phase transition (Tg) of the lipid composition from 60 to 94 degrees C with increasing 4G PAMAM concentrations from 0 to 0.011 mM, respectively. The Tg values of the two microstructures were 68 and 84 degrees C, respectively, indicating that the plastic microstructures were 4G PAMAM/DCP-complexes-rich (alpha mesophases) and the conventional and elongated lamellae were dendrimer-doped HSPC/CH-rich microstructures (beta mesophases). Optical microscopy shows that the alpha mesophases convert into various other types of vesicular structures such as giant unilamellar vesicles and biliquid foams, upon heating above the phase transition temperature of the lipid composition (approximately 60-65 degrees C). The microstructure transformation is a result of an osmotic influx of water and the detergent action of PEG2000-PE present in the lipid composition. The transmission electron microscopy (TEM) images of the liposomal dispersion show particles embedding circular transparent domains that exactly correlate to the theoretical 4G PAMAM/DCP complex sizes, thus, providing evidence of 4G PAMAM interspersed within the two mesophases. Small-angle X-ray scattering (SAXS) measurements indicate that the alpha mesophases are a dendrimer-interlinked, symmetrically undulated lamellar phase and the beta mesophases are dendrimer-doped, occasionally kinked lamellae. An increase in dendrimer concentration in the lipid composition was found to decrease interlamellar spacing. On the basis of optical microscopy, DSC, TEM, and SAXS data, a model of dendrimer-doped mesophase structure and lamellae fusion is proposed. This investigation provides new self-assembled materials for drug/gene delivery and supplements the understanding of mechanisms involved in various biological processes such as membrane fusion, transmembrane permeation, and endocytosis.     

Low viscosity reversed hexagonal mesophases induced by hydrophilic additives

This study reports on the formation of a low viscosity H(II) mesophase at room temperature upon addition of Transcutol (diethylene glycol mono ethyl ether) or ethanol to the ternary mixture of GMO (glycerol monooleate)/TAG (tricaprylin)/water. The microstructure and bulk properties were characterized in comparison with those of the low viscosity HII mesophase formed in the ternary GMO/TAG/water mixture at elevated temperatures (35-40 degrees C). We characterized the role of Transcutol or ethanol as inducers of disorder and surfactant mobility. The techniques used were rheology, differential scanning calorimetry (DSC), wide- and small-angle X-ray scattering (WAXS and SAXS, respectively), NMR (self-diffusion and (2)H NMR), and Fourier transform infrared (FTIR) spectroscopies. The incorporation of either Transcutol or ethanol induced the formation of less ordered HII mesophases with smaller domain sizes and lattice parameters at room temperature (up to 30 degrees C), similar to those found for the GMO/TAG/water mixture at more elevated temperatures (35-40 degrees C). On the basis of our measurements, we suggest that Transcutol or ethanol causes dehydration of the GMO headgroups and enhances the mobility of the GMO chains. As a result, these two small molecules, which compete for water with the GMO polar headgroups, may increase the curvature of the cylindrical micelles and also perhaps reduce their length. This results in the formation of fluid H(II) structures at room temperature (up to 30 degrees C). It is possible that these phases are a prelude to the H(II)-L(2) transformation, which takes place above 35 degrees C.     

Brewster angle microscopy: A preferential method for mesoscopic characterization of monolayers at the air/water interface

Knowledge of the mesoscopic morphology of condensed phase domains formed after the main phase transition in the two-phase coexistence region of Langmuir monolayers progressed rapidly with the development of the highly-sensitive imaging techniques, particularly by Brewster angle microscopy (BAM). Latest developments of commercial BAM instruments have been developed to a high technical level and allow upgrading to imaging ellipsometers which combine optical microscopy and ellipsometry and make the assessment of small layered structures or patterned thin films possible. A large variety of condensed phase domains different in mesoscopic sizes and shapes as well as their textural features has been observed which depend sensitively on the chemical structure of the amphiphilic monolayer and the system conditions, such as surface pressure and temperature. This unsuspected morphological variety of condensed phase domains has been proven not only in Langmuir monolayers but also in adsorbed monolayers (Gibbs monolayers), in Langmuir monolayers penetrated by dissolved surfactants or in adequate molecular recognition systems. The inner textures of domains can be explained on the basis of their geometry and the two-dimensional lattice in dependence of the tilt angle of the alkyl chains and gave rise to the development of a geometric concept on the basis of the molecular packing. New knowledge has been gained about non-equilibrium structures and their transition kinetics into the equilibrium state. Combined results obtained recently by BAM have enhanced the understanding of molecular organization in phase diagrams and binary mixtures. Recent advances in model studies about chiral discrimination effects and of the highly specific structural changes of host-monolayers by recognition of non-surface active guest-components have made progress. Semi-empirical quantum chemical methods have been used to gain insight into the role of different types of interactions involved in the main characteristics of mesoscopic length scale aggregates of mimetic systems.     

Interactions of biomacromolecules with reverse hexagonal liquid crystals: drug delivery and crystallization applications

Recently, self-assembled lyotropic liquid crystals (LLCs) of lipids and water have attracted the attention of both scientific and applied research communities, due to their remarkable structural complexity and practical potential in diverse applications. The phase behavior of mixtures of glycerol monooleate (monoolein, GMO) was particularly well studied due to the potential utilization of these systems in drug delivery systems, food products, and encapsulation and crystallization of proteins. Among the studied lyotropic mesophases, reverse hexagonal LLC (H(II)) of monoolein/water were not widely subjected to practical applications since these were stable only at elevated temperatures. Lately, we obtained stable H(II) mesophases at room temperature by incorporating triacylglycerol (TAG) molecules into the GMO/water mixtures and explored the physical properties of these structures. The present feature article summarizes recent systematic efforts in our laboratory to utilize the H(II) mesophases for solubilization, and potential release and crystallization of biomacromolecules. Such a concept was demonstrated in the case of two therapeutic peptides-cyclosporin A (CSA) and desmopressin, as well as RALA peptide, which is a model skin penetration enhancer, and eventually a larger macromolecule-lysozyme (LSZ). In the course of the study we tried to elucidate relationships between the different levels of organization of LLCs (from the microstructural level, through mesoscale, to macroscopic level) and find feasible correlations between them. Since the structural properties of the mesophase systems are a key factor in drug release applications, we investigated the effects of these guest molecules on their conformations and the way these molecules partition within the domains of the mesophases. The examined H(II) mesophases exhibited great potential as transdermal delivery vehicles for bioactive peptides, enabling tuning the release properties according to their chemical composition and physical properties. Furthermore, we showed a promising opportunity for crystallization of CSA and LSZ in single crystal form as model biomacromolecules for crystallographic structure determination. The main outcomes of our research demonstrated that control of the physical properties of hexagonal LLC on different length scales is key for rational design of these systems as delivery vehicles and crystallization medium for biomacromolecules.     

Shear-induced lamellar phase of an ionic liquid crystal at room temperature

The phase behaviour of a number of N-alkylimidazolium salts was studied using polarizing optical microscopy, differential scanning calorimetry and X-ray diffraction. Two of these compounds exhibit lamellar mesophases at temperatures above 50°C. In these systems, the liquid crystalline behaviour may be induced at room temperature by shear. Sheared films of these materials, observed between crossed polarisers, have a morphology that is typical of (wet) liquid foams: they partition into dark domains separated by brighter (birefringent) walls, which are approximately arcs of circle and meet at “Plateau borders” with three or more sides. Where walls meet three at a time, they do so at approximately 120° angles. These patterns coarsen with time and both T1 and T2 processes have been observed, as in foams. The time evolution of domains is also consistent with von Neumann's law. We conjecture that the bright walls are regions of high concentration of defects produced by shear, and that the system is dominated by the interfacial tension between these walls and the uniform domains. The control of self-organised monodomains, as observed in these systems, is expected to play an important role in potential applications.     

A single parameter determines mesophase transitions in Swollen Liquid Crystals

We report how the control of a single parameter, the co-surfactant, determines the phase transitions of oil-in-water swollen liquid crystals (SLCs) prepared with cetyltrimethylammonium bromide (CTAB), from cubic to hexagonal, lamellar, and finally sponge-like structures. SLCs are complex mixtures (surfactant + co-surfactant + water + salt + oil) usually prepared to form hexagonal mesophases, with cell parameters tunable between 3 and 30 nm. These hexagonal mesophases were successfully used as nanoreactors to prepare a broad range of nanostructured materials. Because the potential of these mesophases as adaptive nanoreactors has not been extended to other liquid crystal geometries than the hexagonal, we studied in a first step the structure evolution of SLCs made with CTAB, cyclohexane, pentanol-1, water and different stabilising salts. We used small-angle X-ray scattering (SAXS), polarised light microscopy and Freeze-Fracture TEM to provide a partial phase diagram and list the different mesophases obtained as a function of composition. We report that the adjustment of a single parameter, the co-surfactant (pentanol-1), determines the phase transition between cubic, hexagonal, lamellar, and sponge-like structures, all other parameters such as the nature and concentration of salt, or amount of oil being constant.     

Thermally stable azo-substituted bent-core nematogens: observation of chiral domains in the nematic mesophases composed of smectic nano clusters

In an effort to synthesise thermally stable bent-core nematogens with fairly low transition temperatures and wide nematic mesophase range, we have synthesised five new series of azo-substituted bent-core compounds without Schiff’s base unit. Here, we studied the effect of different lateral substituents (–CH₃ and –Cl), at two different positions in one of the arms, on the mesogenic properties. We found that such variation in the molecular architecture has a clear-cut impact on the transition temperatures as well as the liquid crystalline properties. The mesogenic properties of these compounds were studied using polarising optical microscopy and differential scanning calorimetry. The exact nature of different mesophases is investigated using X-ray diffraction studies. The results obtained indicated the presence of smectic nano clusters in the nematic mesophases of these compounds. We observed domains of opposite handedness in these nematic phases. The unusual properties of these mesophases were investigated by electro-optical studies. The electro-convection pattern study showed that these nematic mesophases are of negative dielectric anisotropic in nature. The compounds synthesised here exhibit photo-switching both in solution and in their nematic mesophases, a property that can be exploited for practical applications.     

Topography and surface energy dependent calcium phosphate formation on Sol-Gel derived TiO2 coatings

Heterogeneous nucleation and growth of calcium phosphate (CaP) on sol-gel derived TiO(2) coatings was investigated in terms of surface topography and surface energy. The topography of the coatings was derived from AFM measurements, while the surface energy was determined with contact angle measurements. The degree of precipitation was examined with scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS). The precipitation of CaP was found to be dependent on both topography and surface energy. A high roughness value when combining the RMS roughness parameter S(q) with the number of local maxima per unit area parameter S(ds) enhances CaP formation. The hydrophilicity of the coating was also found to be of importance for CaP formation. We suggest that the water contact angle, which is a direct measure of the hydrophilicity of the surface, may be used to evaluate the surface energy dependent precipitation kinetics rather than using the often applied Lewis base parameter.     

Effects of oxygen plasma on cellulose surface

These studies aimed to investigate in detail changes on cellulose surfaces treated with low pressure oxygen plasma at various exposure times. Modifications of cellulose films were studied in respect to topography effects by means of atomic force microscopy and scanning electron microscopy. Chemical effects of plasma treatment were studied using X-ray photoelectron spectroscopy and X-ray diffractometry. Results show that the topographical evolution of the surfaces to rougher ones is not at all gradual. Local maxima of fractionation and the surface size regularity were investigated using surface fractal analysis and Wenzel roughness factors, respectively. It was shown, that plasma treatments decompose the cellulose material by formation of highly functionalized molecules. Such plasma-initiated and supported reactions taking place on the sample surface. The bulk phase and in particular, the crystalline domains are not influenced by plasma treatments. The studies provide useful information to understand the plasma reaction on amorphous and crystalline regions of cellulose surfaces and allow to predict effects of the plasma treatment on physical and chemical properties of much more complex cellulose systems such as cotton fibres and fabrics.     

Shear‐induced lamellar phase of an ionic liquid crystal at room temperature

The phase behaviour of a number of N‐alkylimidazolium salts was studied using polarizing optical microscopy, differential scanning calorimetry and X‐ray diffraction. Two of these compounds exhibit lamellar mesophases at temperatures above 50°C. In these systems, the liquid crystalline behaviour may be induced at room temperature by shear. Sheared films of these materials, observed between crossed polarisers, have a morphology that is typical of (wet) liquid foams: they partition into dark domains separated by brighter (birefringent) walls, which are approximately arcs of circle and meet at “Plateau borders” with three or more sides. Where walls meet three at a time, they do so at approximately 120° angles. These patterns coarsen with time and both T1 and T2 processes have been observed, as in foams. The time evolution of domains is also consistent with von Neumann's law. We conjecture that the bright walls are regions of high concentration of defects produced by shear, and that the system is dominated by the interfacial tension between these walls and the uniform domains. The control of self‐organised monodomains, as observed in these systems, is expected to play an important role in potential applications.     

Phases and phase transitions in a nematic lyotropic system with a biaxial phase

NMR spectroscopy and optical microscopy have been used to study phase transitions and structure in nematic lyotropic mesophases formed by potassium laurate, decylammonium hydrochloride and water. The different mesophases obtained in a well-defined composition range have been characterized, by deuterium NMR, following the evolution of the D₂O spectra as a function of temperature and of the orientation of the samples with respect to the magnetic field. Wide ranges of biaxiality have been found and the asymmetry parameter of the averaged electrical field gradient tensor on the deuterium of the D₂O molecule has been determined. The presence of the different mesophases has always been confirmed by observing oriented samples under the polarizing optical microscope.     

Chromonic lyomesophases formed by the self-assembly of the cyclic dinucleotide d(cGpGp)

The cyclic dinucleotide d(cGpGp) undergoes a self-association process in water to give, first, columnar aggregates similar to the four-stranded helix of poly(G). Successively, at higher concentration, these aggregates self-organize to give a cholesteric and a hexagonal mesophase, the former of which appears only in biphasic systems. The self-assembly process in isotropic solution has been studied by CD spectroscopy and the structure of the mesophases was investigated by optical microscopy and X-ray diffraction.     

Interactions of surfactants in nonionic/anionic reverse hexagonal mesophases and solubilization of α-chymotrypsinogen A

In an attempt to form H_II mesophases at room temperature we prepared lyotropic liquid crystals with two surfactants of the same lipophilic tails (glycerol monooleate, GMO, and oleyl lactate, OL) but differing in the size and charge of the headgroups.Increasing OL concentration significantly affected the hydration of the headgroups and subsequently the lipids packing. At low OL content the cubic mesophase was formed, while at higher OL contents the formation of hexagonal mesophase was favored. It was assumed that OL competed on the water binding, tuning the headgroups’ curvature and the packing parameter inducing the formation of reverse hexagonal mesophase. It was detected that cubic mesophase transformed upon heating to hexagonal structures. The hexagonal mesophases, which were formed both immediately after preparation and after aging, remained stable at elevated temperatures.α-Chymotrypsinogen was solubilized into the obtained LLCs at relatively high concentration (up to 1 wt%). The lattice parameter of the host LLCs exhibited a decrease as a function of the protein content. This process was assigned to partial dehydration of the GMO polar moieties in favor to CTA hydration.Generally speaking, the present study indicated that adding anionic to nonionic lipid is highly beneficial to gain additional compositional and structural characteristics of LLCs.     

Use of the ternary phase diagram of a mixed cationic/glucopyranoside surfactant system to predict mesostructured silica synthesis

Mixed surfactant systems have the potential to impart controlled combinations of functionality and pore structure to mesoporous metal oxides. Here, we combine a functional glucopyranoside surfactant with a cationic surfactant that readily forms liquid crystalline mesophases. The phase diagram for the ternary system CTAB/H(2)O/n-octyl-beta-D-glucopyranoside (C(8)G(1)) at 50 degrees C is measured using polarized optical microscopy. At this temperature, the binary C(8)G(1)/H(2)O system forms disordered micellar solutions up to 72 wt% C(8)G(1), and there is no hexagonal phase. With the addition of CTAB, we identify a large area of hexagonal phase, as well as cubic, lamellar and solid surfactant phases. The ternary phase diagram is used to predict the synthesis of thick mesoporous silica films via a direct liquid crystal templating technique. By changing the relative concentration of mixed surfactants as well as inorganic precursor species, surfactant/silica mesostructured thick films can be synthesized with variable glucopyranoside content, and with 2D hexagonal, cubic and lamellar structures. The domains over which different mesophases are prepared correspond well with those of the ternary phase diagram if the hydrophilic inorganic species is assumed to act as an equivalent volume of water.     

Fatty-acid monolayers at the nematic/water interface: phases and liquid-crystal alignment

The two-dimensional (2D) phases of fatty-acid monolayers (hexadecanoic, octadecanoic, eicosanoic, and docosanoic acids) have been studied at the interface of a nematic liquid crystal (LC) and water. When observed between crossed polarizers, the LC responds to monolayer structure owing to mesoscopic alignment of the LC by the adsorbed molecules. Similar to Langmuir monolayers at the air/water interface, the adsorbed monolayer at the nematic/water interface displays distinct thermodynamic phases. Observed are a 2D gas, isotropic liquid, and two condensed mesophases, each with a characteristic anchoring of the LC zenithal tilt and azimuth. By varying the monolayer temperature and surface concentration we observe reversible first-order phase transitions from vapor to liquid and from liquid to condensed. A temperature-dependent transition between two condensed phases appears to be a reversible swiveling transition in the tilt azimuth of the monolayer. Similar to monolayers at the air/water interface, the temperature of the gas/liquid/condensed triple-point temperature increased by about 10 degrees C for a two methylene group increase in chain length. However, the absolute value of the triple-point temperatures are depressed by about 40 degrees C compared to those of analogous monolayers at the air/water interface. We also observe a direct influence by the LC layer on the mesoscopic and macroscopic structure of the monolayer by analyzing the shapes and internal textures of gas domains in coexistence with a 2D liquid. An effective anisotropic line tension arises from elastic forces owing to deformation of the nematic director across phase boundaries. This results in the deformation of the domain from circular to elongated, with a distinct singularity. The LC elastic energy also gives rise to transition zones displaying mesoscopic realignment of the director tilt or azimuth between adjacent regions with a sudden change in anchoring.     

Lyotropic liquid-crystalline phase composed of aliphatic amino stearates and water

Polarization microscopy, differential calorimetry D.S.C. and X-ray diffraction have been used to examine the lyotropic mesophases which occur in solutions of amino salts in water. The following systems were investigated: methylamino, n-propylamino, n-butylamino, isobutyl-amino and n-hexylamino stearates as one component of the system and water as the other. The types of lyotropic phase and the range in which they occur were determined as functions of concentration and temperature.