Characterization,Stability and In - Vivo Distribution of Asialofetuin Glycopeptide Incorporating DSPC/CHOL Liposomes Prepared by Mild Cholate Incubation

In this study, a small triantennary asialoglycopeptide of fetuin (A-F₂) was used as a ligand to direct liposomes to hepatocytes. A-F₂ was cleaved from asialofetuin, purified, conjugated with fatty acids and incorporated into pre-formed sonicated DSPC/Chol (2:1) liposomes. A mild cholate incubation method for incorporating the A-F₂ ligand on pre-formed vesicles was used. In preliminary in vivo experiments ¹¹In³⁺ encapsulated in A-F₂/palmityl liposomes was seen to accumulate in the liver of mice significantly faster than when encapsulated in non-ligand bearing liposomes of the same lipid composition (studied before), justifying further investigation of this system. The presence of the A-F₂/fatty acid conjugate in a functional form on the vesicle surface was confirmed by their reversible agglutination in the presence of Ricinus communis agglutinin (RCA₁₂₀). Effects of ligand incorporation on the vesicle size distribution, z-potential, membrane integrity and stability were monitored. The results demonstrate that highest ligand incorporation was achieved when liposomes and ligand were co-incubated in the presence of 1mM sodium cholate. Incorporation increased with the length of the fatty acid used for A-F₂ conjugation. Ligand-bearing liposomes were demonstrated to be smaller in diameter (about 30%) with a more positive z-potential in comparison to control vesicles while ligand incorporation did not influence the liposome membrane integrity. The size of the ligand-incorporating vesicles was maintained after 24 hours of incubation in isotonic buffer, proving that the vesicles do not aggregate. Although the preliminary biodistribution results may suggest that ligand bearing liposomes are accumulating in the liver, further cell culture, in vivo distribution and especially liver fractionation studies are required in order to clarify the intrahepatic localization of these liposomes and the ability to target liver hepatocytes in vivo.     

Effects of temporins on molecular dynamics and membrane permeabilization in lipid vesicles

Abstract: Temporins are a novel family of small (10–13 residues) cationic antimicrobial peptides recently isolated from the skin of the European red frog Rana temporaria. Although recently acquired evidence shows that temporins have the potential to kill bacteria by permeabilizing the cytoplasmic membrane, the molecular mechanisms of membrane selectivity and permeabilization are largely unknown. In this study, it was found that temporins cause the release of fluorescent markers entrapped in phosphatidylcholine liposomes in a manner that depends significantly on the size of the solute. Temporins were also shown to lack a detergent‐like effect on lipid vesicles, indicating that marker leakage caused by these peptides is not due to total membrane disruption but to perturbation of bilayer organization on a local scale. Binding of temporins to liposomes did lead to a small increase in lipid hydrocarbon chain mobility, as revealed by EPR spectroscopy of nitroxide‐labeled fatty acids incorporated in the bilayer. Reference experiments were conducted using the bee venom peptide melittin, whose properties and behavior in natural and model membrane systems are well known. Our findings for temporins are discussed in relation to the models proposed to date to account for the action of antimicrobial peptides on membranes.     

Distribution Within the Organs of a Reticuloendothelial System of Liposomes Containing Lipid A

Abstract

Lipid A was incorporated into egg phosphatidylcholine (PC)/cholesterol liposomes which were then tested in mice in order to investigate the effect of lipid A on in vivo biodistribution of liposomes. Addition of lipid A up to 3.2 mol % decreased hepatic uptake and slightly increased splenic uptake of liposomes. When more than 3.2 mol % lipid A incorporated into liposomes, hepatic uptake increased with lipid A concentration. Liposomes containing 25 mol % lipid A were rapidly cleared from circulation and taken up mainly by the liver. Ganglioside G_M1 or N-monomethoxypoly(ethyleneglycol)-phosphatidylethanolamine (PEG-PE), which is known to prolong the half life of circulating liposomes, was included in liposomes along with lipid A. Lipid A antagonized the effect of G_M1 more effectively than that of PEG-PE. This may be due to the different mechanisms of action exerted by G_M1 and PEG-PE in liposome circulation. Hepatic uptake of liposomes containing lipid A increased with vesicle size. However, the unique splenic accumulation of large PEG-PE liposomes was only slightly affected by inclusion of a small amount of lipid A (< 7 mol %). These liposomes might be useful for intravenous delivery of antigen to the spleen for increased immune response.     

Permeability changes in liposomes modeling the stratum corneum lipid composition due to C12-alkyl betaine/sodium dodecyl sulfate mixtures

The interaction of dodecyl betaine (C₁₂-Bet)/sodium dodecyl sulfate (SDS) mixtures at different mole fractions of the zwitterionic component (X_zwitter) with liposomes modeling the stratum corneum (SC) lipid composition were investigated. Liposomes were formed by a lipid mixture of 40% ceramides, 25% cholesterol, 25% palmitic acid and 10% cholesteryl sulfate. The surfactant/lipid molar ratios (R_e) and the bilayer/aqueous phase partition coefficients (K) were determined at two sublytic levels (50 and 100% CF release) by monitoring the increase in the fluorescence intensity of liposomes due to the 5(6)-carboxyfluorescein (CF) released from the interior of vesicles. The fact that the free surfactant concentrations were always lower than their critical micelle concentrations suggests that the liposomes–surfactant sublytic interactions were mainly ruled by the action of surfactant monomers. At the two interaction levels studied the surfactant mixture for X_zwitter=0.6 showed the highest ability to alter the release of the CF entrapped in the interior of vesicles (lowest R_e values), whereas that for X_zwitter=0.4 showed the highest degree of partitioning into liposomes or affinity with these structures (highest K values). Different trends in the interaction of these mixtures with SC lipids and phosphatidylcholine (PC) liposomes were observed when comparing the present R_e and K parameters with those reported for PC ones. Thus, whereas SC lipid liposomes were more resistant to the action of C₁₂-Bet/SDS mixtures, the partitioning of these mixtures into SC lipid bilayers was always greater than that reported for PC ones.     

Study of tensioactive properties of casein signal peptides and their interactions with phospholipids

The high degree of sequence conservation in casein signal peptides reflects their unique functional properties. A series of casein signal peptides and derivatives was synthesized in order to study their insertion in phospholipidic mono- and bilayer structures. Most of these amphiphilic peptides were found to be highly tensio active. Their conformations differ and are solvent dependent. Fluorescence anisotropy measurements showed that all the peptides of the series could interact with dimyristoylphosphatidyl -choline and -glycerol when mixed with the lipids prior to hydration of the liposomes. The most soluble peptide, P6, was selected for insertion experiments in multilamellar vesicles. Its interaction with liposomes is efficient and rapid, being temperature dependent. On the one hand, the physico-chemical measures of interactions of signal peptides of casein β and α_s2 confirm their mutual genetic relationship, and on the other hand they show the divergence of casein β and α_s2 from casein K signal peptide.     

In vitro optimization of liposomal nanocarriers prepared from breast tumor cell specific phage fusion protein

Fusion proteins created by phage display peptides with tumor cell specificity and the pVIII major coat protein of filamentous phages have been explored recently as a simple and cost-effective means for preparing tumor-targeted liposomes that improve the cytotoxicity of anticancer drugs in vitro. The next step in the development of this approach is the optimization of the liposome composition for the maximum targeting activity and subsequent testing in vivo. This study aimed to investigate the impact of preparation protocols, lipid composition and phage protein content on the targeting efficiency of phage protein-modified liposomes. Analysis of size, zeta potential and morphology was used to investigate the effect of preparation protocols on the stability and homogeneity of the phage liposomes. A previously developed coculture targeting assay and a factorial design approach were used to determine the role of lipid composition of the liposomal membrane on the target cell specificity of the phage liposomes. Western blot combined with proteinase K treatment detected the orientation of targeted phage protein in liposomal membrane. Phage protein, DPPG and PEG_2k-PE showed positive effects on target specificity of phage liposomes. The results served to identify optimal formulation that offer an improved liposomal affinity for target tumor cells over the non-optimized formulation.     

Creating Functional Vesicle Assemblies from Vesicles and Nanoparticles

Vesicles (liposomes) have been shown to be excellent vehicles for drug delivery, yet assemblies of vesicles (vesicle aggregates) have been used infrequently in this context. However vesicle assemblies have useful properties not available to individual vesicles; their size can cause localisation in specific tissues and they can incorporate more functionality than is possible with individual vesicles. This article reviews progress on controlling the properties of vesicle assemblies in vitro, applications of vesicle assemblies in vivo, and our recent creation of magnetic nanoparticle–vesicle assemblies. The latter assemblies contain vesicles crosslinked by coated Fe₃O₄ nanoparticles and this inclusion of magnetic functionality makes them magnetically responsive, potentially allowing magnetically-induced contents release. This article describes further studies on the in vitro formation of these magnetic nanoparticle–vesicle assemblies, including the effect of changing magnetic nanoparticle concentration, pH, adhesive lipid structure and bilayer composition. These investigations have led to the development of thermally-sensitive magnetic nanoparticle–vesicle assemblies that release encapsulated methotrexate on warming.     

Conformation and interaction of the cyclic cationic antimicrobial peptides in lipid bilayers

Abstract: To investigate the role of peptide–membrane interactions in the biological activity of cyclic cationic peptides, the conformations and interactions of four membrane‐active antimicrobial peptides [based on Gramicidin S (GS)] were examined in neutral and negatively charged micelles and phospholipid vesicles, using CD and fluorescence spectroscopy and ultracentrifugation techniques. Moreover, the effects of these peptides on the release of entrapped fluorescent dye from unilamellar vesicles of phosphatidylcholine (PC) and phosphatidylethanolamine/phosphatidylglycerol (PE/PG) were studied. The cyclic peptides include GS10 [Cyclo(VKLdY P)₂], GS12 [Cyclo(VKLKdY PKVKLdY P)], GS14 [Cyclo(VKLKVdY PLKVKLdY P)] and [d ‐Lys]⁴GS14 [Cyclo(VKLdK VdY PLKVKLdY P)] (underlined residues are d ‐amino acids), were different in their ring size, structure and amphipathicity, and covered a broad spectrum of hemolytic and antimicrobial activities. Interaction of the peptides with the zwitterionic PC and negatively charged PE/PG vesicles were distinct from each other. The hydrophobic interaction seems to be the dominant factor in the hemolytic activity of the peptides, as well as their interaction with the PC vesicles. A combination of electrostatic and hydrophobic interactions of the peptides induces aggregation and fusion in PE/PG vesicles with different propensities in the order: [d ‐Lys]⁴GS14 > GS14 > GS12 > GS10. GS10 and GS14 are apparently located in the deeper levels of the membrane interfaces and closer to the hydrophobic core of the bilayers, whereas GS12 and [d ‐Lys]⁴GS14 reside closer to the outer boundary of the interface. Because of differing modes of interaction of the cyclic cationic peptides with lipid bilayers, the mechanism of their biological activity (and its relation to peptide–lipid interaction) proved to be versatile and complex, and dependent on the biophysical properties of both the peptides and membranes.     

Liposomes as delivery systems for nasal vaccination: strategies and outcomes

Importance of the field: Among the particulate systems that have been envisaged in vaccine delivery, liposomes are very attractive. These phospholipid vesicles can indeed deliver a wide range of molecules. They have been shown to enhance considerably the immunogenicity of weak protein antigens or synthetic peptides. Also, they offer a wide range of pharmaceutical options for the design of vaccines. In the past decade, the nasal mucosa has emerged as an effective route for vaccine delivery, together with the opportunity to develop non-invasive approaches in vaccination.

Areas covered in this review: This review focuses on the recent strategies and outcomes that have been developed around the use of liposomes in nasal vaccination.

What the reader will gain: The various formulation parameters, including lipid composition, size, charge and mucoadhesiveness, that have been investigated in the design of liposomal vaccine candidates dedicated to nasal vaccination are outlined. Also, an overview of the immunological and protective responses obtained with the developed formulations is presented.

Take home message: This review illustrates the high potential of liposomes as nasal vaccine delivery systems.     

Synthesis and Evaluation of High Functionality and Quality Cell-penetrating Peptide Conjugated Lipid for Octaarginine Modified PEGylated Liposomes In U251 and U87 Glioma Cells

The use of peptide ligand modified PEGylated liposomes has been widely investigated for tumor targeting. Peptides are mainly inserted in the liposomal lipid bilayer using PEG_2K-lipid spacer (Peptide-PEG_2K-DSPE). However, a lower cellular uptake from longer nonlinear PEG_2K spacer was reported, we here synthesized a high functionality and quality (HFQ) lipid with a short, linear serine-glycine repeated peptide [(SG)₅] spacer. The objective of the current study is to develop novel octaarginine (R₈) peptide-HFQ lipid grafted PEGylated liposomes for glioma cells targeting. In vitro liposomes characterization showed that the mean particle size of all liposomal formulations was in the nano-scale range < 120 nm, with a small PDI value (i.e. ～0.2) and had a spherical shape under Transmission Electron Microscope, indicating a homogenous particle size distribution. The flow cytometry in vitro cellular association data with U251 MG and U87 cells revealed that 1.5% R₈-(SG)₅-lipid-PEGylated liposomes exhibited significantly higher cellular association of ～15.87 and 7.59-fold than the conventional R₈-PEG_2K-lipid-PEGylated liposomes (10.4 and 6.19-fold), respectively, relative to the unmodified PEGylated liposomes. Moreover, intracellular distribution studies using confocal laser scanning microscopy (CLSM) corroborated the results of the in vitro cell association. The use of ligand-HFQ-lipid liposomes could be a potential alternative to ligand-PEG_2K-lipid-modified liposomes as a drug delivery system for tumor targeting.     

Application of 1H-N.M.R. to the design of liposomes for oral use. Synergistic activity of bile salts and pancreatic phospholipase A2 in the induced permeability of small unilamellar phospholipid vesicles

Abstract

¹H-n.m.r. spectroscopy of small unilamellar phospholipid vesicles in the presence of the lanthanide probe ion Dy³⁺ has been used to study the permeability of these liposomes induced by the bile salts (glycocholate and glycodeoxycholate) and pancreatic phospholipase A₂. A marked synergism is demonstrated in the combined effects of these digestive agents in producing permeability of the vesicles to Dy^{3 +}. Changes in the ¹H-n.m.r. spectrum of the vesicular phospholipid head-groups before permeability is induced, indicate that the products of the enzymic hydrolysis (lyso lipids and fatty acids) and transmembrane lipid exchange are involved in the permeability mechanism. The results are discussed in terms of the advantages of the use of n.m.r. techniques in the future design of liposomes for oral use.     

Essential oil from Zanthoxylum tingoassuiba loaded into multilamellar liposomes useful as antimicrobial agents

Zanthoxylum tingoassuiba essential oil (EO) is a complex mixture of organic compounds among which methyl-N-methylanthranilate and sesquiterpene alcohol α-bisabolol represent the main compounds. The essential oil antimicrobial activity was studied against bacteria and fungi cells by diffusion disk method and significant activity was observed against S. aureus, S. aureus isolated multi-resistant and the dermathophyte fungi. Essential oil from Zanthoxylum tingoassuiba loaded into DPPC multilamellar liposomes (MLV) was successfully produced through a thin film hydration method with mean diameter of 9.37 ± 4.69 µm. The EO-loaded liposomes showed adequate sphericity and narrower size distribution than empty liposomes. Results also showed that Zanthoxylum tingoassuiba essential oil can be incorporated in appreciable amounts (43.7 ± 6.0%) in the prepared vesicular dispersions. A strong interaction between essential oil and lipid bilayer was indicated by a significant decrease in T_m of the EO-loaded liposomes related to empty vesicles. Essential oil showed incomplete release profile from liposomes, suggesting that EO-loaded liposomes will be useful in pharmaceutical applications to enhance essential oil targeting to cells.     

Skin permeation behavior of elastic liposomes: role of formulation ingredients

Introduction: With the incorporation of edge activators into the lipid bilayer structure, elasticity properties are given to liposomes. Regardless of the debate over the precise permeation mechanism of elastic liposomes, these vesicles have been proven to enhance drug permeation into or through skin in most cases.

Areas covered: This article provides an overview of the formulation ingredients of elastic liposomes and their relationship with skin permeation behavior. The ingredients are divided into two categories of basic and optional ingredients. The effect of stability on permeation behavior of the vesicles is highlighted.

Expert opinion: More attention should be paid to the stability of elastic liposomes. The different stability properties of the elastic liposomes following administration can induce different skin permeation behaviors of the vesicles. It is necessary to select the optimum composition of the elastic liposomes in order to control the stability and permeation behavior of the vesicles into or through the skin. Moreover, for the development of elastic liposomes, particular attention should also be paid to the drug leakage from the vesicles during long-term storage. The application of optional ingredients to improve the stability and/or elasticity of the elastic liposomes is becoming a new trend.     

Liposome-mediated DNA Immunisation via the Subcutaneous Route

Compared to naked DNA immunisation, entrapment of plasmid-based DNA vaccines into liposomes by the dehydration–rehydration method has shown to enhance both humoural and cell-mediated immune responses to encoded antigens administered by a variety of routes. In this paper, we have investigated the application of liposome-entrapped DNA and their cationic lipid composition on such potency after subcutaneous immunisation. Plasmid pI.18Sfi/NP containing the nucleoprotein (NP) gene of A/Sichuan/2/87 (H3N2) influenza virus in the pI.18 expression vector was incorporated by the dehydration–rehydration method into liposomes composed of 16 μmol egg phosphatidylcholine (PC), 8 μmoles dioleoyl phosphatidylethanolamine (DOPE) or cholesterol (Chol) and either the cationic lipid 1,2-diodeoyl-3-(trimethylammonium) propane (DOTAP) or cholesteryl 3?-N-(dimethyl amino ethyl) carbamate (DC-Chol). This method, entailing mixing of small unilamellar vesicles (SUV) with DNA, followed by dehydration and rehydration, yielded incorporation values of 90–94% of the DNA used. Mixing or rehydration of preformed cationic liposomes with 100 μg plasmid DNA also led to similarly high complexation values (92–94%). In an attempt to establish differences in the nature of DNA association with these various liposome preparations their physico-chemical characteristics were investigated. Studies on vesicle size, zeta potential and gel electrophoresis in the presence of the anion sodium dodecyl sulphate (SDS) indicate that, under the conditions employed, formulation of liposomal DNA by the dehydration–rehydration generated submicron size liposomes incorporating most of the DNA in a manner that prevents DNA displacement through anion competition. The bilayer composition of these dehydration–rehydration vesicles (DRV(DNA)) can also further influence these physico-chemical characteristics with the presence of DOPE within the liposome bilayer resulting in a reduced vesicle zeta potential. Subcutaneous liposome-mediated DNA immunisation employing two DRV(DNA) formulations as well as naked DNA revealed that humoural responses (immunoglobulin total IgG, and subclasses IgG₁ and 1gG_2a) engendered by the plasmid encoded NP were substantially higher after dosing twice, 28 days apart with 10 μg liposome-entrapped DNA compared to naked DNA. At all time points measured, mice immunised with naked DNA showed no greater immune response compared to the control, non-immunised group. In contrast, as early as day 49, responses were significantly higher in mice injected with DNA entrapped in DRV liposomes containing DOTAP compared to the control group and mice immunised with naked DNA. By day 56, all total IgG responses from mice immunised with both DRV formulations were significantly higher. Comparison between the DRV formulations revealed no significant difference in immune responses elicited except at day 114, where the humoural responses of the group injected with liposomal formulation containing DC-Chol dropped to significantly lower levels that those measured in mice which received the DOTAP formulation. Similar results were found when the IgG₁ and IgG_2a subclass responses were determined. These results suggest that, not only can DNA be effectively entrapped within liposomes using the DRV method but that such DRV liposomes containing DNA may be a useful system for subcutaneous delivery of DNA vaccines.     

Evaluation of a Liposome System for the Delivery of Desferrioxamine to Lungs in Rats

Abstract— Liposomes with various lipid composition and sizes, prepared by two different techniques were evaluated for their potential to deliver desferrioxamine to lungs as a treatment against oxidative lung damage. Multilamellar vesicles (MLV) and reverse evaporation vesicles were prepared out of a lipid mixture containing dipalmitoyl phosphatidylcholine, stearyl amine, cholesterol and vitamin E. The administration of desferrioxamine-encapsulated liposomes to rats by the intravenous route at a dose of 100 mg kg^?1, significantly prolonged the presence of desferrioxamine in all the tested organs when compared with the administration of free desferrioxamine. The injection of reverse evaporation vesicles extruded through a 2 μm polycarbonate membrane exhibited a longer residence time of the desferrioxamine and of liposomal vitamin E in lungs compared with the other types of liposomes tested. The examination of liposome components in the bronchoalveolar lavage fluid (BALF) and the alveolar macrophages recovered from BALF revealed that about 7 × 10^?3% of the administered desferrioxamine dose was recovered by this technique at 3 and 17 h after liposome administration. This high residual concentration in the alveolar space confirms the hypothesis that liposomes can be delivered to the lung tissue when encapsulated in alveolar macrophages.     

Preparation of novel double liposomes using the glass-filter method

The glass-filter method, a newly developed preparative method for liposomes, was applied for preparation of novel double liposomes. Double liposomes were prepared by filtering a suspension of liposomes prepared using a G4 filter (pore size: 10-16 microm) into a G3 filter (pore size: 40-100 microm) coated with a similar lipid layer. The morphological structure of the double liposomes was confirmed using scanning electron microscopy by the freeze-fracture method to be multivesicular vesicles consisting of small liposomes enveloped in larger liposomes. The diameter of liposomes prepared using the G4 filter was 0.8-2 microm and that of liposomes prepared using the G3 filter or double liposomes was 5-10 microm. These results suggested that the particle size of liposomes is dependent on the pore size of the glass-filter. The encapsulation efficiencies of double liposomes for brilliant blue FCF (BB) and erythrosine (ER) were higher than those of liposomes prepared by the standard Bangham method. Double liposomes showed suppressed release of BB or ER compared with normal liposomes. In particular, no release of BB was observed from the double liposomes prepared with stearylamine. These findings implied that the outer lipid layer protects the inner liposomes. The glass-filter method is the only method that we can get the double liposomes in a short period, and double liposomes prepared by this novel method had adequate size and good stability in vitro.     

Characteristics and anti-proliferative activity of azelaic acid and its derivatives entrapped in bilayer vesicles in cancer cell lines

The hydrophilicity and lipophilicity of azelaic acid (AA) were modified to diethyl azelate (DA) which was synthesized by Fisher esterification reaction and identified by IR, MS and ¹H NMR and to azelaic acid-β-cyclodextrin complex (AACD) which was prepared by inclusion complexation and identified by IR, DSC and XRD respectively. AA, DA and AACD were entrapped in liposomes and niosomes comprising of l-α-dipalmitoyl phosphatidylcholine (DPPC)/cholesterol at 7:3 molar ratio and Tween61/cholesterol at 1:1 molar ratio, respectively, using a thin-film hydration method with sonication. The size and morphology of these bilayer vesicles were determined by optical and transmission electron microscopy. The particle size was found to be in the range of 90–190 nm. The entrapment efficiency of AA, DA and AACD in all vesicular formulations was more than 80%, as analyzed by HPLC for AA and AACD, and GC for DA. Anti-proliferative activity of AA and its derivatives (DA and AACD) both entrapped and not entrapped in bilayer vesicles, using MTT assay in three cancer cell lines (HeLa, KB and B₁₆F₁₀) comparing with vincristine, were investigated. AACD showed the highest potency comparing to AA in HeLa, KB and B₁₆F₁₀ of 1.48, 1.6 and 1.5 times, respectively. AA entrapped in liposomes was about 90 times more potent than the free AA, and about 1.5 times less potent than vincristine. When entrapped in bilayer vesicles, DA and AACD were more effective than AA in killing cancer cells. AACD entrapped in liposomes gave the highest anti-proliferation activity in HeLa cell lines with the IC₅₀ of 2.3 and 327 times more potent than vincristine and AA, respectively. DA in liposomes demonstrated the IC₅₀ of 0.03 times less potent than vincristine in KB cell lines, while in B₁₆F₁₀ AACD in niosomes showed the IC₅₀ of 0.05 times less potent than vincristine. This study has suggested that the modification of AA by derivatization and complexation as well as the entrapment in bilayer vesicles can enhance its therapeutic efficacy.     

99m-Technetium as a marker of liposomal deposition and clearance in the human lung

The clearance of pulmonary delivered ^99mTc-labelled dipalmitoyl phosphatidylcholine (DPPC) liposomes has been studied in 4 healthy volunteers using the technique of gamma-scintigraphy. The labelling process was dependent on the method of separation of free and liposomally bound activity and on the concentration of stannous chloride (SnCl₂) used. Gel chromatography resulted in lower labelling efficiencies on increased SnCl₂ concentration due to the progressive failure of liposomes to elute from the column. Higher labelling efficiencies were achieved following treatment with an anionic exchange resin as a result of efficient recovery of the liposomes. Transmission electron microscopy and X-ray microanalysis showed that liposomes (1 ml containing 40 mg phospholipid) labelled with 20–25 MBq ^99mTc in the presence of 0.3 mg SnCl₂ appeared uncontaminated with technetium-tin colloid, and were used in the in vivo studies. Pulmonary deposition of multi-lamellar and small unilamellar vesicles, delivered from an air jet nebulizer, was dependent on droplet size of the aerosol product. Short term clearance of both liposome populations was typical of muco-ciliary transport resulting in statistically equivalent retentions at 6 h. Subsequent retention data suggested that faster process(es) than those described for insoluble particulates were contributing to the clearance of alveolar deposited liposomes.     

Selection of high efficient transdermal lipid vesicle for curcumin skin delivery

Curcumin shows effective anti-inflammatory activities but is seldom used in clinic because of its poor solubility in water and vulnerablity to sunshine ultraviolet effect. Novel lipid vesicles have been developed as carriers for skin delivery. In this paper, lipid vesicles—propylene glycol liposomes (PGL), Ethosomes and traditional liposomes, were prepared as curcumin carriers respectively. Their morphology, particle size and encapsulation efficiency and drug release behavior in vitro were evaluated. Transdermal efficiency and deposition quantity in abdominal skin were also measured with Franz diffusion device. Carrageenan-induced paw edema was established to evaluate the anti-inflammatory effect. From the result, the particle size order of lipid vesicles was: PGL (182.4 ± 89.2 nm) < Ethosomes (289 ± 132.1 nm) < traditional liposomes (632.9 ± 184.1 nm). The order of particle dispersion coefficient was as the same as that of particle size. The sequence of encapsulation efficiency was: PGL > Ethosomes > traditional liposomes. PGL had the best encapsulation efficiency of 92.74 ± 3.44%. From anti-inflammatory experiment, PGL showed the highest and longest inhibition on the development of paw edema, followed by Ethosomes and Traditional liposomes. With the elevated entrapment efficiency, good transdermic ability and sustained-release behavior, PGL may represent an efficient transdermal lipid vesicle for skin delivery.     

The Shape/Morphology Balance: A Study of Stealth Liposomes via Fractal Analysis and Drug Encapsulation

Purpose

Fractal analysis was used as a tool in order to study the morphological characteristics of PEGylated liposomes. We report on the morphological characteristics of stealth liposomes composed of DPPC and DPPE-PEG 3000 in two dispersion media using fractal analysis.

Methods

Light scattering techniques were used in order to elucidate the size, the morphology and the surface charge of PEGylated liposomes as a function of PEGylated lipid concentration and temperature. Fluorescence spectroscopy studies revealed a microenvironment of low polarity inside the liposomal membranes.

Results

All formulations were found to retain their physicochemical characteristics for at least 3 weeks. The hydrodynamic radii (R_h) of stealth liposomes were stable in the process of heating up to 50°C; while the fractal dimension values (d_f) which correspond to their morphology, have been changed during heating. Hence, these results are a first indication of the presence of a heterogeneous microdomain structure of the stealth liposomal system. The amphiphilic drug indomethacin (IND) was successfully encapsulated within the liposomes and led to an increased size of stealth liposomes, while the morphology of liposomal vectors changed significantly at the highest molar ratio of PEGylated lipid.

Conclusions

We can state that this approach can promote a new analytical concept based on the morphological characteristics and quantify the shape of drug carriers complementary to that of the conventional analytical techniques.