Lecithin Vesicles Research Articles

Preparation of PVP hydrogel nanoparticles using lecithin vesicles

Hydrogels micro, sub-micro and nanoparticles are of great interest for drug encapsulation and delivery or as embolotherapic agents. In this work it is described the preparation of nano and sub-microparticles of pre-formed, high molecular weight and monomer free poly(N-vinyl-2-pyrrolidone) encapsulated inside the core of lecithin vesicles. The hydrogel particles are formed with a very narrow diameter distribution, of about 800 nm, and a moderate swelling ratio, of approximately 10.

Asymmetrical stress generated by the erythrocyte lipid flippase triggers multiple bud formation on the surface of spherical giant liposomes

Proteo-giant liposomes were electroformed from a mixture of lecithin vesicles and inside-out vesicles from erythrocytes. After addition of Mg-ATP in the vicinity of the proteo-giant liposomes, small buds appeared on the liposome surfaces, which--via an increase in lipids in the outer monolayer--demonstrated the active transport of lipids from the inner to the outer monolayer, indicating flippase activity.

Application research of a novel designed peptide as a potential carrier

While a great deal of research has focused on the application of full-sequence ionic complementary peptide, detection of the capability of half-sequence ionic complementary peptide such as drug carriers, is rarely reported. This paper presents that the half-sequence ionic complementary peptide P9 (AC-Pro- Ser-Phe-Asn-Phe-Lys-Phe-Glu-Pro-NH2) can successfully stabilize a model hydrophobic drug pyrene in the aqueous solution. Soybean lecithin vesicles were used to mimic plasma membranes. Fluorescence data show that the pyrene is presented in the crystalline form when stabilized by P9 solution, and molecularly migrated from its peptide encapsulations into the membrane bilayers when the suspension is mixed with lipidosome vesicles. Slower release was observed when thicker coating was applied onto pyrene, which could be to control the wall thickness coating the cargo, and consequently the release rate. The result indicated that P9, with half-sequence ionic complement, may serve as a hydrophobic compounds carrier.

LIPOSOMAL GEL OF DICLOFENAC SODIUM AS TOPICAL DELIVERY SYSTEM

This study the preparation of diclofenac sodium in lecithin vesicles and loading in pluronic F-127 gel, the effect of sodium cholate on the diffusion of the drug through rat skin and the antiinflammatory activity of the liposomal gel formulations. Lecithin vesicles were prepared in the presence or absence of sodium cholate by the dry film method and sonication. The size of liposomal vesicles ranged from 100-700 nm and the encapsulation efficiency of the diclofenac sodium was between 60-80%. The lecithin vesicles were loaded in pluronic F-127 gel. The highest cumulative of drug diffusion through rat skin was 19.31± 1.50 (µg/cm2) for lecithin vesicles in the presence of sodium cholate (F4). Also the highest cumulative of drug diffusion through rat skin was 12.20± 0.50 (µg/cm2) for liposomal gel (F8). The antiinflammatory activity of the liposomal gel formulations was studied on carrageenan induced paw edema in rats. The results show that, F8 and F6 show superior anti-inflammatory activity in comparison with the other gel formulations. From the results, lecithin vesicles and liposomal gel of diclofenac sodium appear to be advantagesous for topical delivery of the drug.

An approach to rheological and electrokinetic behaviour of lipidic vesicles covered with chitosan biopolymer

The rheological and electrokinetic properties of soybean lecithin vesicles prepared from concentrated soy lecithin dispersions (250 g/L) obtained by slow swelling under shear conditions and mixed with chitosan biopolymer solutions were studied. The rheological behaviour of lecithin–chitosan vesicles was determined by shear stress against shear rate measurements, as well as by the variation on the hysteresis loop area. The results were compared with the rheopectic behaviour of soy lecithin dispersions without chitosan. An important change on the rheological properties of the complex dispersion was observed, depicting in a thixotropic behaviour with a plastic character in the presence of chitosan. This observation indicates that chitosan promotes the transition of planar sheets into closed structures, such as vesicles. The influence on the rheological and the electrokinetic behaviours of several electrolytes, such as NaCl, CaCl 2 and AlCl 3 with concentrations ranging between 10 −5 and 10 −2 mol/L were also studied. In all of the cases, an estimation of the diameters of the closed structures was obtained.

Binding and relaxation behavior of Coumarin-153 in lecithin–taurocholate mixed micelles: A time resolved fluorescence spectroscopic study

The microenvironment of the bile salt–lecithin mixed aggregates has been investigated using steady state and picosecond time resolved fluorescence spectroscopy. The steady state spectra show that the polarity of the bile salt is higher compared to lecithin vesicles or the mixed aggregates. We have observed slow solvent relaxation in bile salt micelles and lecithin vesicles. The solvation time is gradually slowed down due to gradual addition of the bile salt in lecithin vesicles. Addition of bile salt leads to the tighter head group packing in lecithin. Thus, mobility of the water molecules becomes slower and consequently the solvation time is also retarded. We have observed bimodal slow rotational relaxation time in all these systems.

Reduction of cytochrome b5 by NADPH–cytochrome P450 reductase

The reduction of mammalian cytochrome b 5 ( b 5) by NADPH–cytochrome P450 (P450) reductase is involved in a number of biological reactions. The kinetics of the process have received limited consideration previously, and a combination of pre-steady-state (stopped-flow) and steady-state approaches was used to investigate the mechanism of b 5 reduction. In the absence of detergent or lipid, a reductase– b 5 complex is formed and rearranges slowly to an active form. Electron transfer to b 5 is rapid within this complex (>30 s −1 at 23 °C), as fast as to cytochrome c. With excess b 5 present, a burst of reduction is observed, consistent with rapid electron transfer to one or two b 5 molecules per reductase, followed by a subsequent rate-limiting event. In detergent vesicles, the reductase and b 5 interact rapidly but electron transfer is slower (∼3 s −1 at 23 °C). Experiments with dimyristyl lecithin vesicles yielded results intermediate between the non-vesicle and detergent systems. These steady-state and pre-steady-state kinetics provide views of the different natures of the reduction of b 5 by the reductase in the absence and presence of vesicles. Without vesicles, the encounter of the reductase and b 5 is rapid, followed by a slow reorganization of the initial complex (∼0.07 s −1), very fast reduction, and dissociation. In vesicles, encounter is rapid and the slow step (∼3 s −1) is reduction within a complex less favorable for reduction than in the non-vesicle systems.

Study of Energy Transfer from 7-Amino Coumarin Donors to the Rhodamine 6G Acceptor in Lecithin Vesicles and Sodium Taurocholate−Lecithin Mixed Aggregates

The energy transfer using 7-amino coumarin dyes as the donor and rhodamine 590 (Rh6G) as the acceptor was investigated in lecithin vesicles and sodium taurocholate (NaTC)-lecithin mixed aggregates using steady-state and time-resolved fluorescence spectroscopy. All energy transfer parameters were calculated. The coumarin 153-Rh6G pair is the most efficient donor-acceptor pair as reflected by the value of k(ET). With addition of NaTC in lecithin, in the case of the coumarin 153-Rh6G pair, the energy transfer rate or efficiency does not change very much, whereas in the case of the coumarin 151-Rh6G pair, the energy transfer rate decreases 2-fold upon going from lecithin vesicles to NaTC-lecithin mixed aggregates where the molar ratio is 2.5. It is mainly due to the deeper location of coumarin 153 in the lipid bilayer or in mixed aggregates. Rotational relaxation data also support this idea.

Combined Interaction of Phospholipase C and Apolipoprotein A-I with Small Unilamellar Lecithin-Cholesterol Vesicles: Influence of Apolipoprotein A-I Concentration and Vesicle Composition

We report the combined effects of phospholipase C (PLC), a pronucleating factor, and apolipoprotein A-I (apo A-I), an antinucleating factor, in solutions of model bile. Results indicate that apo A-I inhibits cholesterol nucleation from unilamellar lecithin vesicles by two mechanisms. Initially, inhibition is achieved by apo A-I shielding of hydrophobic diacylglycerol (DAG) moieties so as to prevent vesicle aggregation. Protection via shielding is temporary. It is lost when the DAG/apo A-I molar ratio exceeds a critical value. Subsequently, apo A-I forms small ( approximately 5-15 nm) complexes with lecithin and cholesterol that coexist with lipid-stabilized (400-800 nm) DAG oil droplets. This microstructural transition from vesicles to complexes avoids nucleation of cholesterol crystals and is a newly discovered mechanism by which apo A-I serves as an antinucleating agent in bile. The critical value at which a microstructural transition occurs depends on binding of apo A-I and so varies with the cholesterol mole fraction of vesicles. Aggregation of small, unilamellar, egg lecithin vesicles (SUVs) with varying cholesterol composition (0-60 mol %) was monitored for a range of apo A-I concentrations (2 to 89 microg/mL). Suppression of aggregation persists so long as the DAG-to-bound-apo A-I molar ratio is less than 100. A fluorescence assay involving dansylated lecithin shows that the suppression is an indirect effect of apo A-I rather than a direct inhibition of PLC enzyme activity. The DAG-to-total apo A-I molar ratio at which suppression is lost increases with cholesterol because of differences in apo A-I binding. Above this value, a microstructural transition to DAG droplets and lecithin/cholesterol A-I complexes occurs, as evidenced by sudden increases in turbidity and size and enhancement of Forster resonance energy transfer; structures are confirmed by cryo TEM.

Effect of tartrates with various counterions on the precipitation of calcium oxalate in vesicle solutions

The effect of tartrates with various counterions on the crystallization of CaOxa in vesicles was investigated in the concentration range from 0.3 to 66.7 mM in lecithin vesicle solutions using X-ray diffraction (XRD) and Fourier transform infrared (FT-IR) spectroscopy. Various tartrates with hydrogen, sodium, potassium, and a mixture of sodium and potassium cations were considered. For K 2tart and NaKtart, only calcium oxalate monohydrate (COM) and calcium oxalate trihydrate (COT) were obtained depending on the concentrations. However, for H 2tart and Na 2tart, calcium oxalate dihydrate (COD) was first induced in the debt of COM, then COT was induced in the debt of COM and COD. The maximum percentages of COD were about 55% and 35% for H 2tart and Na 2tart in the concentration ranges from 6.7 to 13.3 and 1.67 to 10 mM, respectively. Linear relationships were obtained between the decrease of the mole percent (mol%) of COM crystals, or the increase of mol% of COD or COT crystals and the increase of the logarithm of the molar concentration of the tartrates. The different results are discussed in terms of the reduction in the degree of freedom associated with the adsorption process at the membrane–water interface, the ionic radii of the counterions, the solubility and the molecular structures of the hydrates, and the different adsorption of the counterions at the crystals of the calcium oxalate hydrates.

Sorbitan Monopalmitate-Based Proniosomes for Transdermal Delivery of Chlorpheniramine Maleate

A proniosomal gel for transdermal drug delivery of chlorpheniramine maleate (CPM) was developed based on Span 40 and extensively characterized in vitro. The system was evaluated for the effect of composition of formulation, type of surfactants and alcohols on the drug loading, rate of hydration, vesicle size, polydispersity, entrapment efficiency, and drug release across cellulose nitrate dialysis membrane. The stability studies were performed at 4°C and at room temperature. The results showed that lecithin produced more stable and larger vesicles with higher loading efficiency but lower dissolution efficiency than cholestrol (chol) and dicethyl phosphate (DCP). The type of alcohol had no significant effect on the stability of vesicles, but ethanol produced larger vesicles (≈44 μm) and entrapped a greater amount of drug. Drug release from vesicles of lecithin followed a first-order kinetics whereas those with DCP or without lecithin fit better with a Higuchi model. The proniosomes that contained Span 40/lecithin/chol prepared by ethanol showed optimum stability, loading efficiency, and particle size and release kinetic suitable for transdermal delivery of CPM.

2,3-Diarylimidazo[1,2-a]pyridines as potential inhibitors of UV-induced keratinocytes apoptosis: synthesis, pharmacological properties and interactions with model membranes and oligonucleotides by NMR

Four 2,3-diarylimidazo[1,2-a]pyridines (I, 1a-c) were synthesized as inhibitors of UV-induced apoptosis and showed quite different properties. First, only the pyridinyl derivative I showed protection in molt cells. From the supposed intracellular target, phospholipid membrane models were studied by (1)H, (2)H and (31)P NMR spectroscopy. All these molecules can incorporate the membrane bilayer of small unilamellar vesicles of lecithin (SUV). However, I is clearly closed to the external polar head of the lipids, and is relatively mobile in the layer. Conversely, the other molecules are strongly immobilized in the deep part of the external layer. (31)P solid-state NMR spectra recorded on phospholipid dispersions (multilayers vesicles (MLV)) completely excluded any detergent effect or any modification of temperature transition. The only structural or dynamic effect observed was a homogeneous, but limited, reduction in the chemical shift anisotropy in the presence of I, in agreement with its superficial location. (2)H NMR experiment performed on the same model using perdeuterated phospholipids showed no significant fluidity reduction at the level of terminal CD(3) groups in the presence of 1a-c, according to their deep location. Finally, their interactions with synthetic oligonucleotide, d(CGATCG)(2) was studied showing non specific interactions of 1a on the external GC pair, while no interaction was observed with the other derivatives.

Membrane transport of a polyacid-tied doxorubicin.

A model, based on fluorescence data, is developed for the poly(acrylic acid)-assisted transport of doxorubicin, a cationic antitumor drug, through a bilayer membrane. Accordingly, the doxorubicin binds to the poly(acrylic acid) via electrostatic polymer-drug interactions plus drug-drug stacking within the complex. This complex associates with neutral egg lecithin vesicles by means of hydrophobic attraction between the doxorubicin and the vesicle bilayers. In the process, the doxorubicin "destacks", providing a fluorescence change that can be monitored. Finally, the doxorubicin enters the vesicle interior which has been imparted with an acidic pH to protonate the doxorubicin and thus, in a second stage, yield an additional fluorescence change that can also be detected. A portion of the poly(acrylic acid), now devoid of doxorubicin, then leaves the outer vesicle surface and enters to external solution.

Iontophoresis of lecithin vesicles of cyclosporin A

Site-specific immunosuppression with topical cyclosporin A (CsA) has broad clinical implications in the treatment of skin disorders like psoriasis, pyoderma gangrenosum, lichen planus, cutaneous graft-versus-host disease and contact hypersensitivity and the temporary treatment of skin allografts on burn wounds. However, like any other peptide drug, its skin delivery is hindered by the barrier property of stratum corneum and the physicochemical properties of CsA. We have attempted to deliver CsA across human cadaver epidermis in vitro using colloidal systems like microemulsion and lecithin vesicles and iontophoresis. Although, passive diffusion did not result in permeation of quantifiable amounts of CsA, anodal iontophoresis of the negatively charged colloidal systems facilitated the permeation. Electroosmosis and compromised epidermis might have contributed to the higher skin flux. Lecithin vesicles were better than microemulsion for the iontophoretic delivery of CsA and appear to have potential in site-specific immunosuppression.

Interaction of apolipoprotein A-I with lecithin-cholesterol vesicles in the presence of phospholipase C

Here we study the anti-nucleating mechanism of apolipoprotein A-I (apo A-I) on model biliary vesicles in the presence of phospholipase C (PLC) utilizing dynamic light scattering (DLS), steady-state fluorescence spectroscopy, cryogenic transmission electron microscopy (cryo-TEM), and UV/Vis spectroscopy. PLC induces aggregation of cholesterol-free lecithin vesicles from an initial, average size of 100 nm to a maximal size of 600 nm. The presence of apo A-I likely inhibits vesicle aggregation by shielding the PLC-generated hydrophobic moieties, which results in vesicles of an average size of 200 nm. A similar phenomenon is observed in cholesterol-enriched lecithin vesicles. Whereas PLC alone produces aggregates of 300 nm, no aggregation is observed when apo A-I is present along with PLC. However, the ability of apo A-I to inhibit aggregation is temporary, and after 8 h, a broad particle size distribution with sizes as high as 800 nm is observed. Apo A-I possibly induces the formation of small apo A-I/lecithin/cholesterol complexes of about 5–20 nm similar to the discoidal pre-HDL complexes found in blood when it can no longer effectively shield all the DAG molecules. Concomitant with formation of complexes, DAG molecules coalesce into large oil droplets, which account for the large particles observed by light scattering. Thus, apo A-I acts as an anti-nucleating agent by two mechanisms, anti-aggregation and microstructural transition. The mode of protection is dependent on the cholesterol content and the relative amounts of DAG and apo A-I present. This study supports the possibility of apo A-I solubilizing lipids in bile in a similar fashion as it does in blood and also delineates the mechanism of formation of the complexes.

Lecithin vesicles for topical delivery of diclofenac

Skin penetration of topically applied diclofenac is important for the treatment of rheumatic diseases and actinic keratoses. We have studied the permeation of diclofenac across human cadaver epidermis in-vitro from four lecithin vesicle formulations and a few marketed semi-solid preparations. The lecithin vesicle formulations were prepared by dissolving the lipid contents (lecithin and sodium cholate) in a 1:1 mixture of methanol-chloroform, evaporating the solvents under vacuum, and hydrating the lipid layer with the drug solution in water or 10% ethanol. The vesicles were sonicated for 5 min to reduce the vesicle size and their size and Zeta potential were characterized. The cumulative amount and maximum flux of diclofenac was 69.7±40.3 μg and 4.77±3.16 μg/h cm 2 from lecithin vesicles containing sodium cholate and 10% ethanol, and is the highest of all formulations studied. The cumulative amount and mean maximum flux obtained from other formulations were in the range of 2.46±1.98–29.9±10.1 μg and 0.53±0.46–3.61±0.86 μg/h cm 2. Based on the results, lecithin vesicles of diclofenac appear to be advantageous for the topical delivery of diclofenac.

The absorption behavior of cyclosporin A lecithin vesicles in rat intestinal tissue

The purpose of the study was to investigate the absorption behavior of lecithin vesicles of cyclosporin A (CsA-VES), prepared by the rotary evaporation method and treated further with sonication. The everted gut sac technique and in situ circulation method were used to examine: (1) relationship between the CsA-VES absorption velocity and the CsA-VES content; (2) the influence of the intestinal mucus, blank vesicles, concentration of Na +, energy inhibitor and P-gp inhibitor on the absorption of CsA-VES; and (3) the respective accumulated content of CsA in the incubating medium and the sacs after incubation with Sandimmum Neoral ®(CsA-NEO) and CsA-VES. Our results showed there was a saturated absorption of CsA. Most CsA-VES accumulated in the mucus before it reached the intestinal tissue. There was no significant difference in the accumulated absorption content in the incubating medium and the sacs of CsA-NEO and CsA-VES. The addition of blank vesicles and concentration of Na + had no significant influence on the accumulated absorption of CsA ( P>0.05).The energy inhibitor and P-gp inhibitor influenced the accumulated absorption of CsA significantly ( P<0.05). CsA-VES may be transported by phagocytosis. Mucus was a barrier blocking the diffusion of CsA-VES. CsA-NEO and CsA-VES showed equal absorption levels in the intestine.

Interaction of Bile Salts with Egg-yolk Lecithin Vesicles

Interaction of Bile Salts with Egg-yolk Lecithin Vesicles

Structure of artificial cytoskeleton containing liposomes in aqueous solution studied by static and dynamic light scattering.

The structure of three types of liposomes (egg yolk phosphatidylcholine (EPC) without modification and EPC vesicles containing cross-linked N-isopropylacrylamide (NIPAM) networks of low and a high concentration inside the vesicles) were analyzed by static and dynamic light scattering. Upon polymerization the network was assumed to become attached to the membrane by reactive anchoring monomers. For the sample of high poly(NIPAM) content the polymer network was assumed to fill the whole space in the vesicles. The issue of the present study was to examine hard and hollow sphere behavior of the liposomes with networks of high and low poly(NIPAM) content. The theoretical scattering curves differ markedly for uniform hard and uniform hollow spheres by the presence of specific peaks. However, polydispersity washed out the peaks and led to smoothed asymptotes with fractal dimensions of df = 2 for hollow and df = 4 for hard spheres. The experimental data could efficiently be fitted with weakly polydisperse hollow spheres. No clear conclusion could be drawn from the angular dependence alone for the liposome of high poly(NIPAM) content. The two wavelengths from the HeNe and Ar lasers proved to be too long for the studied liposomes of about 100 nm in radius. However, evidence for hollow sphere behavior was found for fractionated liposomes from the ratio rho = Rg/Rh = 1.04 +/- 0.02 (theory rho = 1.00 for hollow spheres). Finally, from the molar mass and the sphere radius, an apparent density was determined. The analysis gave the expected density for the pure EPC lecithin vesicles and a poly(NIPAM) network density of 0.244 g/mL. For the liposome of low poly(NIPAM) content the network appeared to be attached to the inner surface of the lecithin shell to form a layer of about 18 nm thickness.

Phospholipid matrix as a target for sulfur mustard (HD): NMR study in model membrane systems.

Although the interactions of sulfur mustard (HD) with nucleic acids and proteins have been well studied, the toxic interactions with the membrane matrix and specially the phospholipid bilayer have so far been poorly investigated. We have used several NMR techniques to study these interactions: 1H NMR to observe the localization of HD in membranes of small unilamellar vesicles (SUV) of lecithin; 31P NMR to verify the hypothesis of pore formation in membranes of large unilamellar vesicles (LUV); and pseudo solid state 31P and 2H NMR to analyze the dynamic consequences of the presence of HD in multilayer dispersions of dimyristoylphosphatidylcholine (DMPC). Immediate and late modifications of the DMPC-HD complexes have been observed at the macroscopic and microscopic levels. After intoxication, HD is spontaneously incorporated into the membrane and locates at the level of the chain methylene groups. This incorporation occurs without formation of pores in the membrane. The presence of HD in the phospholipid dispersion differentially increases the membrane fluidity depending upon the level involved. Weak at the superficial level (phosphate group), this increase is dose-dependent on progression into the membrane. This increase is related to a lowering of transition temperature when measured at the chain level. Macroscopically, HD induces dose- and time-dependent modifications of the DMPC-HD complexes, leading to the formation of an optically transparent gel. This gel formation is confirmed at a microscopic level, where all structures disappear after intoxication.