A.O. Vila

Viscoelastic properties of concentrated dispersions in water of soy lecithin

Abstract Viscoelastic properties of soy lecithin dispersions at 60, 120, 180 and 240 g l−1 in water have been studied as a function of ionic strength (NaCl concentrations in the range 10−5–10−2 mol l−1) and of preparation method (sonication, freezing–unfreezing). Viscosity measurements of sonicated dispersions in a rotatory viscometer show Newtonian and plastic (Bingham) behavior, whereas the frozen–unfrozen dispersions show a spectacular change in their viscoelastic properties, that is, a non-Newtonian behavior with pseudoplastic characteristics and rheopexy. These properties have been related to electrokinetic characteristics of the segregated vesicles in each case. A decrease in ζ-potential (in absolute value) of vesicles results on an increase in the hysteresis loop area of the parents dispersions.

Transcutol containing vesicles for topical delivery of minoxidil

The aim of this work was to evaluate the ability of Transcutol (Trc) to produce elastic vesicles with soy lecithin (SL) and study the influence of the obtained vesicles on in vitro (trans)dermal delivery of minoxidil. To this purpose, so-called penetration enhancer-containing vesicles (PEVs) were prepared using Trc aqueous solutions (5–10–20–30% v/v) as hydrophilic phase. SL liposomes, without Trc, were used as control. Prepared formulations were characterized in terms of size distribution, morphology, zeta potential, deformability, and rheological behavior. The influence of the obtained PEVs on (trans)dermal delivery of minoxidil was studied by in vitro diffusion experiments through pig skin. Results showed that all prepared PEVs were able to give good entrapment efficiency (E%≈67) similar to that of conventional liposomes. Trc-containing PEVs showed to be more deformable than liposomes only when minoxidil was loaded in 5 and 10% Trc-containing vesicles. Rheological studies showed that PEVs have higher fluidity than conventional liposomes. All PEVs showed a higher stability than liposomes as shown by studying zeta potential and size distribution during three months. Results of in vitro diffusion experiments showed that Trc-containing PEVs are able to deliver minoxidil to deep skin layers without any transdermal permeation.

Chitosomes loaded with cranberry proanthocyanidins attenuate the bacterial lipopolysaccharide-induced expression of iNOS and COX-2 in raw 264.7 macrophages

Chitosan binds to negatively charged soy lecithin liposomes by an electrostatic interaction driven by its positively charged amino group. This interaction allows stable covered vesicles (chitosomes) to be developed as a suitable targeted carrier and controlled release system. This study investigated the effect of chitosomes on the activation of cranberry proanthocyanidins (PAC) in Raw 264.7 macrophages. Chitosomes were characterized according to size, zeta potential, PAC-loading, and release properties. Results showed an increase in the net positive charge and size of the liposomes as the concentration of chitosan was increased, suggesting an effective covering of the vesicles by means of electrostatic interactions, as shown by transmission electron microscopy and fluorescence microscopy. About 85% of the PAC that was loaded remained in the chitosomes after release studies for 4 hours in phosphate-buffered saline. Cyclo-oxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) are associated with inflammation. Activated RAW 264.7 macrophages increase the expression of COX-2 and iNOS in response to bacterial infection and inflammation; we, therefore, tested the ability of the PAC-loaded chitosomes to attenuate COX-2 and iNOS expression in LPS (lipopolysaccharide)-stimulated macrophages. Increasing the amount of PAC loaded into the chitosomes caused a dose-dependent attenuation of iNOS and COX-2 expression in LPS-stimulated macrophages. A 2% v/v PAC-loaded chitosomes formulation almost completely attenuated the LPS-induced expression of iNOS and COX-2. PAC-loaded chitosomes were more active than PAC alone, suggesting that the macrophage response to LPS occurs after endocytosis of the PAC-loaded chitosomes.

Geometric and thermodynamic considerations about the lipid vesicles formation in water (I. Unilamellar vesicles)

Abstract A parametrization based on the differential geometry and on the thermodynamics of dispersed systems has been developed in order to explain the processes leading to the formation of lipid vesicles in water. Several factors of known importance such as aqueous interlayer, chain-length, lipid chain-packing and membrane asymmetry, as well as the existence of two different surfaces in the vesicular bilayer have been considered. The barrier which amphiphile molecules need to surpass in order to the vesicle be formed has three contributions related to the surface energy, the mechanical energy due to overpressure and the excess of chemical potential. The theoretical results define the conditions at which unilamellar vesicles are formed and are applied to the phospholipids: dilauryl-, dimyristoyl-, dipalmytoyl-, and distearoyl-phosphatidylcholine.

Effect of the osmotic conditions on the value of ζ potential of DMPC multilamellar liposomes

Abstract The variation of the ζ potential of liposomes as a function of the ionic strength obtained under different osmotic conditions is studied. Dimyristoyl phosphatidylcholine (DMPC) multilamellar vesicles have been selected and the electrolyte used was NaCl. Three different methods were followed for the preparation of the vesicles. 1. (a) In aqueous medium at different ionic strengths. In this case, the liposomes are under iso-osmotic conditions. 2. (b) Lipidic vesicles under hypo-osmotic conditions are obtained by diluting with water a suspension of liposomes prepared in iso-osmotic conditions as in (a). 3. (c) Lipidic vesicles under hyperosmotic conditions are obtained by the addition of NaCl to liposome samples prepared as in (a). Under iso-osmotic conditions, the results allow us to obtain the ionic strength value corresponding to the maximum of the ζ potential through a parabolic fit. The maximum stability and the minimum of aggregation occur at that ionic strength value; this is a similar behaviour to that expected from DLVO theory. Under non-iso-osmotic conditions the results do not follow the above behaviour. Under hyperosmotic conditions, the results follow an opposite behaviour to the above which could be explained by a reduction of liposome size and content. Under hypo-osmotic conditions, a strong decrease of the ζ potential values takes place; therefore the tendency towards coalescence and aggregation increases.

Theoretical and experimental evaluation of decypolyglucoside vesicles as potential drug delivery systems

The aim of this work was to ascertain the capability of a commercial mixture of decylpolyglucoside (Orami.x NS10) to form vesicle structures in water, either alone or in association with cholesterol. To this purpose, we compared results obtained from a theoretical model with those acquired from the experimental characterization of different Orami.x NS10/cliolesterol mixtures. The germen vesicular formation theory was used to predict the formation process of vesicular structures. To this purpose, the energy balance involved in the vesicle formation was calculated using critical concentration for vesicle formation (ccf), surface tension and molecular area of decylpolyglucoside. The ccf was measured by turbidity measurements, and surface tension was determined by a tensiometer study. In order to con firm the validity of the theoretical model, we prepared and characterized vesicles made with decylpolyglucoside and cholesterol in several molar ratios and using different preparation methods. Vesicles were characterized using optical and light polarized microscopy, transmission electron microscopy and dynamic light scattering. The capability of vesicles to encapsulate a hydropliilic probe was also studied. Results obtained showed thai the decylpolyglucoside surfactant is able to form vesicles only in the presence of an appropriate cholesterol content (i.e. Orami.x NS10/cliolesterol molar ratio > 110.4) and that the mathematical model proposed is a useful and reproducible tool capable of predicting the type of aggregation structures given by the alkylpolyglucoside in water.

INTERACTION OF IONIC COMPOUNDS WITH MULTILAMELLAR LIPOSOMES. AN ELECTROKINETIC MODEL

Abstract We have proposed a theoretical model of interaction of ionic compounds (ionic adsorbate or ionic drugs), with multilamellar-liposomes, by means of their electrokinetic property variation with the ionic compound concentration. In this work, we show the complete development of the model proposed. Its theoretical results have been analysed to study the influence on the zeta-potential value of the following: number of membranes, size, shear-plane situation, critical concentration of formation (ccf), and the ionic compound concentration which annuls the zeta-potential value of the multilamellar-liposomes formed.

In vitro uptake of lysozyme-loaded liposomes coated with chitosan biopolymer as model immunoadjuvants

Chitosan binds to negatively charged soy lecithin liposomes by an electrostatic interaction driven by its cationic amino group. This interaction allows developing stable coated vesicles suitable as a targeted carrier and controlled release system for drugs and vaccines. In this work, we studied the effect of chitosan-coated liposomes on the uptake and antigen presentation of hen egg-white lysozyme (HEL) in Peyer’s patches peritoneal macrophages isolated from mice. Chitosan-coated liposomes were characterized according to size, zeta potential, and antigen-loading and release properties. Results showed an increase in the positive net charge and size of the liposomes as the concentration of chitosan was increased, suggesting an electrostatic interaction and an effective coating, followed by fluorescence microscopy. About 85% of the antigen loaded remained in the chitosan-coated liposomes after release studies for 4 hours in phosphate-buffered saline. After 4 hours of preincubation with a T-cell hybridoma line cocultured with murine peritoneal macrophages, only trace amounts of interleukin-2 (IL-2) were detected in the cocultures treated with HEL alone, whereas cocultures treated with HEL-liposomes had an important production of IL-2, and the HEL chitosan-coated liposomes had already reached maximum IL-2 expression. Confocal microscopy studies showed that chitosan-coated liposomes had a higher uptake rate of the fluorescently labeled HEL than uncoated liposomal vesicles after 30 minutes of incubation with the peritoneal macrophages. Since uptake by macrophage cells is the first step in vaccination, our results suggest that the chitosan-coated liposomal system is a potential candidate as an immunoadjuvant for vaccine delivery systems.

Polymer-liposome nanoparticles obtained by the electrostatic bio-adsorption of natural polymers onto soybean lecithin liposomes

This work is focused on the formulation of polymer-liposome nanoparticles based on the electrostatic bio-adsorption of natural polymers onto soybean lecithin liposomes, with potential as novel delivery system for macromolecules, such as proteins. The building up of the polymer-liposome nanoparticles was achieved through the alternating bio-adsorption of natural cationic (chitosan) and neutral (dextran) or anionic (dextran sulphate or alginate) polymer layers on a core composed by anionic nanosized soybean lecithin liposomes. The electrostatic bio-adsorption of natural polymers succeeded in building nanosized, spherical, monodisperse and stable polymer-liposome nanoparticles with cumulative sizes between 357.3 nm ± 25.3 nm and 498.2 nm ± 69.6 nm and surface charges (ζ-potential) between –30.66 mV ± 1.55 mV and –26.74 mV ± 1.04 mV for the liposomal systems composed by alternating layers of chitosan and dextran sulphate or alginate, respectively. Natural-polymer-liposome nanoparticles offer good properties for encapsulation on its liposomal aqueous core and sustained release of a model protein, BSA, in vitro.

Osmotic conditions in rheologic properties of erytrocites

Abstract The decreasing viability and in vivo survival of red blood cells, RBCs, observed in blood bank-storage are due to alterations in their rheologic properties and metabolic status. However, there is a lack of electrokinetic experiments with erythrocytes by the screening produced by the high concentration of buffer required to keep the isoosmotic conditions. Electrokinetic measurements in a wide range of sodium chloride and tetrasodium pyrophosphate are undertaken on liposomes as cell models, and in order to get an aqueous media suitable for maintaining osmotic conditions and allowing electrophoretic mobility measurements as well, advantage has been taken of dextrose properties. Measured electrokinetic values are practically constant in a wide concentration range of dextrose. With this in mind, the optimal dextrose concentration for RBCs was determined through the Drabkins method and its value was inside the above suitable dextrose range. Electrokinetic measurements of RBCs were then undertaken in this medium.