Dima Libster

In vitro permeation of diclofenac salts from lyotropic liquid crystalline systems.

In this paper we examined feasible correlations between the structure of different lyotropic mesophases and transdermal administration of three diclofenac derivatives with varying degrees of kosmotropic or chaotropic properties, solubilized within the mesophases. It was found that the most chaotropic derivative of diclofenac diethyl amine (DEA-DFC) interacted with the polar heads of glycerol monooleate (GMO), thus expanding the water-lipid interface of the lamellar and cubic mesophases. This effect was detected by an increase in the lattice parameter of both mesophases, enhanced elastic properties, and increased solid-like response of the systems in the presence of DEA. Potassium diclofenac (K-DFC), a less chaotropic salt, had less pronounced effect on the structural features of the mesophases. Kosmotropic Na+ salt (Na-DFC) had only minor influence on both lamellar and cubic structures. The locus of solubilization of the molecules with the host mesophases was correlated with their delivery. It was suggested that transdermal delivery of kosmotropic Na-DFC was accelerated by the aqueous phase and less constrained by the interaction with monoglyceride. On the other hand, the chaotropic cations (K+ and DEA+), presumably entrapped in the water-lipid interface, interacted with monoglyceride headgroups, which is likely to be the key cause for their sustained administration.

Lysozyme entrapped within reverse hexagonal mesophases: physical properties and structural behavior.

A model protein (lysozyme) was incorporated into monoolein-based reverse hexagonal (H(II)) mesophase and its structure effects were characterized by small angle X-ray scattering, ATR-FTIR spectroscopy, and rheological measurements. Modifications in molecular organization of the H(II) mesophases as well as the conformational stability of lysozyme (LSZ) as a function of pH and denaturating agent (urea) were clarified. Up to 3 wt.% LSZ can be solubilized into the H(II). The vibration FTIR analysis revealed that LSZ interacted with OH groups of glycerol monooleate (GMO) in the outer interface region, resulting in strong hydrogen bonding between the surfactant and its environment. Simultaneously, the decrease in the hydrogen-bonded carbonyl population of GMO was monitored, indicating dehydration of the monoolein carbonyls. These molecular interactions yielded a minor decrease in the lattice parameter of the systems, as detected by small angle X-ray scattering. Furthermore, LSZ was crystallized within the medium of the hexagonal structures in a single crystal form. The alpha-helix conformation of lysozyme was stabilized at high pH conditions, demonstrating greater helical structure content, compared to D(2)O solution. Moreover, the hexagonal phase decreased the unfavorable alpha-->beta transition in lysozyme, thereby increasing the stability of the protein under chemical denaturation. The rheological behavior of the hexagonal structures varied with the incorporation of LSZ, reflected in stronger elastic properties and pronounced solid-like response of the systems. The hydrogen bonding enhancement in the interface region of the structures was most likely responsible for these phenomena. The results of this study provided valuable information on the use of hexagonal systems as a carrier for incorporation and stabilization of proteins for various applications.

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.

Lipid polymorphism in lyotropic liquid crystals for triggered release of bioactives

In this review we present recent progress on lyotropic liquid crystals (LLC) as delivery vehicles for cosmetoceuticals, nutraceuticals, and drugs. LLC have been known for decades and their potential as delivery vehicles is well recognized. Yet, the two major mesophases, reverse hexagonal (H(II)) and bicontinuous cubic (primitive, gyroid, and diamond), are relatively hard gels with very slow release kinetics of the bioactives. In recent years a discontinuous cubic micellar mesophase (Q(L)) was characterized and studied, showing significant potential as a delivery vehicle. In addition, the H(II) mesophase formed could be much more fluid and produced at room temperature. Recent studies concentrated on establishing methods to evaluate solubilization capacity and relationship between the diameter and length of the cylinders and the nature of the solubilizates. Special attention was given to finding methods to target the vehicles to the lumen and to trigger the release of the bioactives. This review summarizes the efforts of our group along with work by numerous other scientists in this area. All these efforts suggest that the lyotropic mesophases and the corresponding dispersed soft particles (cubosomes, hexosomes, micellosomes) are now more than ever ready to become drug delivery vehicles for transport across the skin and the gut.

Penetratin-induced transdermal delivery from HII mesophases of sodium diclofenac

Penetratin, a cell penetrating peptide is embedded within a reversed hexagonal (H(II)) mesophase for improved transdermal delivery of sodium diclofenac (Na-DFC). The H(II) mesophase serves as the solubilization reservoir and gel matrix whereas penetratin is the transdermal penetration enhancer for the drug. The systems were characterized and the interactions between the components were determined by SAXS, ATR-FTIR and SD-NMR. High affinity of Na-DFC to glycerol monooleate (GMO) was revealed, associated with increasing the order within the water channels. This affinity is enhanced upon heating and seems to be associated with GMO dehydration. Penetratin (PEN) is entrapped at the hydrophilic region of the H(II) mesophase, between the GMO headgroups, reducing the order of the system and decreasing the size of the hexagonal domains. The transdermal delivery rate of Na-DFC through porcine skin, from the H(II) mesophases, was enhanced by PEN and so also the cumulative transport crossing the skin. PEN induced accelerated drug diffusion through the stratum corneum, towards the different skin layers. The transdermal delivery enhancement is explained from the results of the ATR-FTIR analysis. It seems that PEN accelerates the structural transition of skin lipids from hexagonal to liquid. The disordering results in enhanced diffusion of Na-DFC through the stratum corneum, followed by enhanced overall penetration of the drug.

Concentration- and temperature-induced effects of incorporated desmopressin on the properties of reverse hexagonal mesophase.

In this paper we report on the solubilization of desmopressin, as a model for peptide drugs, into reverse hexagonal (H(II)) liquid crystals. Concentration- and temperature-induced interactions of desmopressin, as well as the conformation of the peptide, were studied using small-angle X-ray scattering, ATR-FTIR spectroscopy, SD-NMR, and rheological measurements. A considerable increase (up to 6 A) in the lattice parameter of the mesophases was obtained upon incorporation of the peptide. According to the ATR-FTIR analysis, the chaotropic effect of peptide embedment was assigned to its interactions with hydroxyls of monoglyceride in the outer interface region. These interactions had only a minor influence on the conformation of the peptide; weakening or opening the gamma-turns resulted in partial binding of the peptides free carbonyls to monoolein. Temperature-dependent SAXS measurements displayed a chaotropic destabilizing effect of desmopressin on the structure, shifting toward the lower temperature H(II)-L(2) structural transition. Temperature increase resulted in an increase of the domain size in the presence of the peptide, in contrast to the trend observed in the empty mesophase. SD-NMR analysis enabled distinguishing between two factors impeding the diffusion of the peptide: the restriction of motion due to the geometrical constrain of diffusion within the water tubes, and the interactions of the guest molecule with monoglyceride. The onset of the critical behavior at 45 degrees C was found to be significant, indicating considerable weakening of the monoglyceride and desmopressin interactions and the destabilizing effect of the peptide on the mesophase above this temperature. Similar temperature-dependent behavior was revealed by rheological measurements displaying the same onset of the critical behavior. It was demonstrated by Franz diffusion cell measurements that hexagonal mesophases can potentially be used as delivery vehicles for sustained delivery of desmopressin.

Soft matter dispersions with ordered inner structures, stabilized by ethoxylated phytosterols

This paper describes the formation and characterization of liquid crystalline dispersions based on the hexagonal phase of GMO/tricaprylin/water. As a stabilizer of the soft particles dispersed in the aqueous phase, a non-ionic, non-polymeric surfactant--ethoxylated phytosterol with 30 oxyethylene units (PhEO) was utilized. In contrast to Pluronic copolymers, normally utilized in the stabilization of liquid crystalline dispersions with ordered inner structure, use of such non-polymeric surfactant is not a common practice in this field. We revealed how properties of these particles, such as internal structure, size, and stability, can be rationally modified by the concentration of the stabilizing agent and processing conditions. The physical stability of the hexosomes was further examined by the LUMiFuge technique. Structural effect of PhEO solubilization on the properties of the bulk H(II) mesophase system showed that phase behavior was greatly influenced following phase transitions: H(II)-->H(II)+cubic-->cubic+L(alpha)-->L(alpha). The decrease of hydrogen bonding of the hydroxyl and carbonyl groups of monoolein with water and simultaneous hydration of EO groups of PhEO appeared to be important for the observed behavior. The use of PhEO as a dispersant resulted in a soft matter multi-phase water dispersion with bimodal distribution of the particle population. Effective stabilization of hexosomes was obtained in an extremely narrow concentration range of PhEO (0.1-0.2 wt%), coexisting with small vesicles and disordered particles. At higher PhEO content, particles had disordered inner structure, and unilamellar and multilamellar vesicles, at the expense of hexosomes in consequence of incorporation of the dispersant into the hexosome structure. PhEO was found to induce lamellar phase formation, introducing disorder into the hexagonal LLC and reducing their domain size. Finally, hexosomes were evaluated as delivery vehicles for the therapeutic peptide desmopressin. Sustained release of this drug was observed during the first 10 h; however, permeation drastically increased in the 10-24 h range.

Sodium diclofenac and cell-penetrating peptides embedded in H(II) mesophases: physical characterization and delivery.

Glycerol monooleate (GMO)-based mesophases offer extensive prospects for incorporation of various bioactive molecules. This work deals with the solubilization of selected cell-penetrating peptides (CPPs) together with sodium diclofenac (Na-DFC) within the H(II) mesophase for transdermal applications. The effect of CPPs such as RALA (an amphipatic CPP), penetratin (PEN), and oligoarginine (NONA) on Na-DFC skin permeation kinetics to provide controlled release and tune the drug transdermal diffusion was studied. The location of the drug and the CPPs within the mesophase was probed by DSC and FTIR. Na-DFC was found to be located at the interfacial region between the surfactant chains, leading to denser H(II) mesophase. The hydrophilic NONA was intercalated into the aqueous cylinders and caused their swelling. It induced a significant decrease in the hydrogen binding between the GMO carbonyls and their surrounding. The amphiphilic PEN was entrapped within two different regions, depending on its concentration. PEN and NONA improved Na-DFC permeation by 100%, whereas RALA enhanced permeation by 50%. When estimating Na-DFC migration rate out of the mesophase toward surrounding aqueous media, it appeared to be slower with the CPPs. The peptides were not involved at this diffusion-controlled step. It seems that their effect on skin permeation is based on their specific interaction with the skin.

From the microscopic to the mesoscopic properties of lyotropic reverse hexagonal liquid crystals.

In the present study we aimed to explore a correlation between the microstructural properties of the lyotropic reverse hexagonal phase (HII) of the GMO/tricaprylin/phosphatidylcholine/water system and its mesoscopic structure. The mesoscopic organization of discontinuous and anisotropic domains was examined, in the native state, using environmental scanning electron microscopy. The topography of the HII mesophases was imaged directly in their hydrated state, as a function of aqueous-phase concentration and composition, when a proline amino acid was solubilized into the systems as a kosmotropic (water-structure maker) guest molecule. The domain structures of several dozen micrometers in size, visualized in the environmental scanning electron microscopy, were found to possess fractal characteristics, indicating a discontinuous and disordered alignment of the corresponding internal water rods on the mesoscale. On the microstructural level, SAXS measurements revealed that as water content (Cw) increases the characteristic lattice parameter of the mesophases increases as well. Using the water concentration as the mass measure of the mixtures, a scaling relationship between the lattice parameter and the concentration was found to obey a power law whereby the derived fractal dimension was the relevant exponent, confirming the causal link between the microscopic and mesoscopic organizations. The topography of the HII mesophase was found to be affected by the microstructural parameters and the composition of the samples. Thermal analysis experiments involving these systems further confirmed that the behavior of water underpins both microscopical and mesoscopic features of the systems. It was shown that both the swelling of the lattice parameter and the mesoscopic domains is correlated to the bulk water concentration in the water rods.

Solubilization of Gabapentin into HII Mesophases

In the present work, we report on the solubilization of gabapentin (GBP) into lyotropic hexagonal mesophases composed of monoolein, tricaprylin, and water. It was demonstrated that the hexagonal structure remained intact up to 2 wt % gabapentin, whereas the lamellar phase coexisted with the hexagonal one in the concentration range of 3-4 wt % of the drug. At gabapentin content of 5-6 wt %, only lamellar phases containing defects such as dislocations and multilamellar vesicles were detected. Incorporation of GBP decreased the lattice parameter of the H(II) mesophase from 56.6 to 50.6 Å, while the structural dimensions of the lamellar phase were not affected. ATR-FTIR analysis suggested enhanced hydrogen bonding between the protonated amine of GBP and the O-H groups of the GMO and the water surrounding in the inner hydrophilic interface region. This led to intercalation of the drug into the water-lipid interface. At higher GBP loads of 4-6 wt %, thermal analysis revealed disordering within the lipid packing, apparently induced by the spatially altered interface area. Rheological measurements correlated the macroscopic features of the systems with alterations on the molecular level and allowed distinguishing between closely related mesophases due to their different rheological characteristics. In vitro transdermal delivery studies showed that the examined mesophases enabled a sustained release of GBP compared to its aqueous solution. Sustained release was more pronounced in the case of the hexagonal mesophase, compared to the lamellar one.