Dimyristoylphosphatidylcholine Liposomes Research Articles

A cochleate formulation optimized by D-optimal mixture design enhances oral bioavailability of Revaprazan

A cochleate formulation was developed to enhance the oral bioavailability of revaprazan (RVP). Dimyristoyl phosphatidylcholine (DMPC) liposome containing dicetyl phosphate (DCP) successfully formed a cochleate after treatment with CaCl2, whereas that containing sodium deoxycholate did not. Cochleate was optimised using a D-optimal mixture design with three independent variables—DMPC (X1, 70.58 mol%), cholesterol (X2, 22.54 mol%), and DCP (X3, 6.88 mol%)—and three response variables: encapsulation efficiency (Y1, 76.92%), released amount of free fatty acid at 2 h (Y2, 39.82%), and released amount of RVP at 6 h (Y3, 73.72%). The desirability function was 0.616, showing an excellent agreement between the predicted and experimental values. The cylindrical morphology of the optimised cochleate was visualised, and laurdan spectroscopy confirmed the dehydrated membrane interface, showing an increased generalised polarisation value (approximately 0.5) over small unilamellar vesicle of RVP (RVP-SUV; approximately 0.1). The optimised cochleate showed greater resistance to pancreatic enzyme than RVP-SUV. RVP was released in a controlled manner, achieving approximately 94% release in 12 h. Following oral administration in rats, the optimised cochleate improved the relative bioavailability of RVP by approximately 274%, 255%, and 172% compared to RVP suspension, a physical mixture of RVP and the cochleate, and RVP-SUV, respectively. Thus, the optimised cochleate formulation might be a good candidate for the practical development of RVP.

A Biophysical Research on the Determination of Morphological and Structural Properties of Coumarin-Loaded Liposomes

The major goal of this research was to examine how coumarin affects lipid model membranes. For this reason, liposome membranes were formed using dimyristoyl phosphatidylcholine (DMPC) as zwitterionic lipid. The influence of coumarin on the morphology, packing order, fluidity, and hydration state of lipid membranes was specifically investigated by means of microscopic (field emission scanning electron microscopy (FE-SEM)) and spectroscopic (attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy) techniques. Taken into account the results obtained with FE-SEM images and analysis, liposomes without and with coumarin have uniform structures and spherical shapes in appearance. However, coumarin-loaded liposomes are observed with an increase in size when compared to a mean diameter of unloaded-liposomes. Considering ATR-FTIR analysis, the investigation of the vibrational bands which belong to the hydrophobic and hydrophilic parts of DMPC lipid reveals that coumarin alters the physical features of the DMPC liposomes by decreasing the order and increasing the fluidity of the system and making hydrogen bonding with the interfacial and headgroup regions of zwitterionic lipid DMPC. Finally, performing more biophysical studies on the interactions of biologically active compounds with model membranes plays an important role in determining the molecular action mechanisms of these compounds in drug discovery and formulations.

Application of Fixed-Length Ultrasonic Interferometry to Determine the Kinetics of Light-/Heat-Induced Damage to Biological Membranes and Protein Complexes

This manuscript describes the application of a fixed-length ultrasonic spectrometer to determine the kinetics of heat- and photo-induced damage to biological membranes and protein complexes and provides examples of the test measurements. We implemented a measurement scheme using the digital analysis of harmonic signals. To conduct the research, the fixed-length ultrasonic spectrometer was modernized: the speed was increased; lighting was supplied to the sample cells; the possibility of changing the gas atmosphere and mixing the sample was given. Using solutions containing natural concentrations of deuterium oxide, a high sensitivity of the spectrometer was shown. The spectrometer performed well in the measurement of phase state of dimyristoylphosphatidylcholine liposomes, both in the absence and in the presence of additions, which are capable of changing the lipid properties (sodium dodecyl sulfate, palmitic acid, and calcium ions). The heat- and photo-induced changes in the state of photosystem II core complexes were demonstrated using a fixed-length ultrasonic spectrometer. Transitions at 35.5 °C, 43.5 °C, 56.5 °C, and 66.7 °C were revealed. It is proposed that the transitions reflect the disassembly of the complexes and protein denaturation. Thus, the present study demonstrates that a fixed-length ultrasonic spectrometer can be applied to determine the kinetics of heat- and photo-induced damage to biological membranes and protein complexes.

THE INFLUENCE OF CHLORPROMAZINE HYDROCHLORIDE ON THE THERMOTROPIC BEHAVIOR OF DIMYRISTOYL PHOSPHATIDYLCHOLINE LIPOSOMES AS REVEALED BY DIFFERENTIAL SCANNING CALORIMETRY

Objective: The aim of this study is to investigate the influence of the model cationic, amphiphilic, drug chlorpromazine hydrochloride (CPZ-HCl) on the thermotropic behavior of dimyristoyl phosphatidylcholine (DMPC) liposomes, using differential scanning calorimetry (DSC). The effect of sonication, charged lipids and CPZ-HCl at concentrations known to cause anesthesia on the enthalpy (ΔHt), entropy (ΔSt), phase transition (Tc), pre-transition (pre-TC) and half-height width (HHW) of DSC thermograms were examined. Methods: The experiments conducted, using the Perkin Elmer (DSC-2C), include the effect of a wide range of CPZ-HCl concentrations on ΔHt, ΔSt, Tc, pre-Tc and HHW of DSC thermograms of DMPC liposomes. The effect of sonication on ΔHt, ΔSt, Tc and HHW of DSC thermograms of DMPC/CPZ-HCl liposomes as a function of sonication time. The effect of both positively charged stearyl amine (ST) and negatively charged diacetyl phosphate (DCP) lipids on ΔHt, ΔSt and Tc of DMPC/CPZ-HCl liposomes. In addition, the effect of CPZ-HCl at concentrations known to cause anesthesia on ΔHt, ΔSt. and Tc of DMPC liposomes in the presence and absence of ST and DCP in phosphate buffer (pH 7.4), was also carried out. Results: Using DSC, CPZ-HCl concentrations as low as 1×10-7M were observed to alter the gel-liquid crystalline phase transition and thus to possess a membrane destabilizing effect. CPZ-HCl reduces ΔHt, ΔSt, TC, the pre-TC and increases HHW of DMPC liposomes. ΔHt and ΔSt of DMPC liposomes were observed to decrease with increasing CPZ-HCl concentrations, exhibiting an inflection point at 5×10-5M. ΔHt of DMPC liposomes was observed to decrease linearly in the absence and presence of and CPZ-HCl as a function of sonication time. Both ΔHt and ΔSt of DMPC liposomes were observed to increase in the presence of cationic lipid (ST) and to decrease in the presence of anionic lipid (DCP). ΔSt and Tc of DMPC, DMPC/ST, DMPC/DCP liposomes, were found to decrease as a function of CPZ-HCl concentrations known to cause anesthesia. Conclusion: Using DSC, CPZ-HCl concentrations, as low as 1×10-7 M were observed to influence the enthalpy, entropy, phase transition, pre-transition and half-height width of DSC thermograms of DMPC liposomes, altering the gel-liquid crystalline phase transition and thus possessing a membrane destabilizing effect. It can also be inferred that CPZ-HCl interacts with both the polar head group and the hydrophobic interior of the phospholipid bilayer. These results could support the hypothesis that the addition of local anesthetics might trigger a change in the lipid surrounding the sodium channel from the gel to the liquid crystalline state, allowing the sodium channel to close with the resulting anesthesia.

Interaction of triblock copolymers (Pluronic®) with DMPC vesicles: a photophysical and computational study

Pluronic/lipid mix promises stealth liposomes with long circulation time and long-term stability for pharmaceutical applications. However, the influence of Pluronics on several aspects of lipid membranes has not been fully elucidated. Herein it was described the effect of Pluronics on the structured water, alkyl chain conformation, and kinetic stability of dimyristoylphosphatidylcholine (DMPC) liposomes using interfacial and deeper fluorescent probes along with computational molecular modeling data. Interfacial water changed as a function of Pluronics’ hydrophobicity with polypropylene oxide (PPO) anchoring the copolymers in the lipid bilayer. Pluronics with more than 30–40 PO units had facilitated penetration at the bilayer while shorter PPO favored a more interfacial interaction. Low Pluronic concentrations provided long-term stability of vesicles by steric effects of polyethylene oxide (PEO), but high amounts destabilized the vesicles as a sum of water-bridge cleavage at the polar head group and the reduced alkyl-alkyl interactions among the lipids. The high kinetic stability of Pluronic/DMPC vesicles is a proof-of-concept of its advantages and applicability in nanotechnology over conventional liposome-based pharmaceutical products for future biomedical applications.

Comparison of functionality and structural stability of bacteriorhodopsin reconstituted in partially fluorinated dimyristoylphosphatidylcholine liposomes with different perfluoroalkyl chain lengths

Amphiphilic molecules with one or more perfluoroalkyl groups (Rf, CnF2n+1), which show peculiar interfacial properties, are attracting much attention in membrane protein science. We recently have developed a partially fluorinated dimyristoylphosphatidylcholine (DMPC) with a perfluorobutyl group in the hydrophobic chain terminal (F4-DMPC) and demonstrated that F4-DMPC is a promising material for incorporating membrane proteins. Moreover, we have found out that membrane properties of a series of partially fluorinated DMPCs with different Rf chain lengths (Fn-DMPCs) vary in a significant Rf chain length-dependent manner. In the present study, structural and functional properties of a membrane protein bacteriorhodopsin (bR) in the Fn-DMPC (n = 4, 6, and 8) membranes (bR/Fn-DMPC) are investigated using several physicochemical techniques. Regardless of the Rf chain lengths, bR/Fn-DMPCs retain native-like structural and functional properties at 30 °C, unlike bR molecules in DMPC vesicles. In particular, bR/F6-DMPC, which is in the fluid phase at 30 °C, shows flash-induced transient absorption changes very similar to the native purple membrane (PM) and very high thermal stability of bR trimers comparable to the PM. Structural and functional properties of bR/Fn-DMPCs are discussed compared to the PM and bR/DMPC.

Spherical and tubular dimyristoylphosphatidylcholine liposomes

A physicochemical study of the interaction of pinocembrin with dimyristoylphosphatidylcholine (DMPC) liposomes was carried out. Unilamellar vesicles of about 100 nm were prepared. Pinocembrin was incorporated into the lipid membrane by interacting strongly with the acyl groups of the phospholipid chains, favoring the fluid phase. The incorporation efficiency was 89%. At concentrations higher than 0.436 mg mL−1 of pinocembrin, a change of structure from spherical to tubular liposomes was observed, and the enthalpy of this transition was determined by differential scanning calorimetry. After 0.744 mg mL−1, there was no change in the transition enthalpy from gel phase to liquid crystalline, so it is possible to assume that the lipid membrane reached saturation. With the findings presented in this research, it is possible to increase the solubility and bioavailability of the pinocembrin by using DMPC as a molecular carrier, enhancing its use as drug in the pharmaceutical industry to treat cancer, ischemic stroke, intracerebral hemorrhage, Alzheimer’s disease, cardiovascular diseases and atherosclerosis as well as other diseases.

The THE INFLUENCE OF CHLORPROMAZINE HYDROCHLORIDE ON THE SIZE OF SMALL UNILAMELLAR DIMYROSTYLPHOSPHATIDYLCHOLINE LIPOSOMES AS REVEALED BY LIGHT SCATTERING PHOTOMETRY

Objectives: This study aims to investigate the possible influence of the model, cationic, surface-active solute chlorpromazine hydrochloride (CPZ-HCl) on the size of small unilamellar dimyristoyl phosphatidylcholine (DMPC) liposomes as a function of temperature and CPZ-HCl concentration, below and above the critical micelle concentration (CMC).
 Methods: Small unilamellar DMPC liposomes were prepared by dissolving DMPC in chloroform and the solvent was rota-evaporated in a water bath adjusted at 40 °C. The lipid film was then dispersed in 0.1 M KCl solution adjusted at pH 6.2 to form large multilamellar liposomes which are then sonicated and fractionated via Sepharose 2B-Cl gel. The elution profile was followed spectrophotometrically at λ 260 nm. Combined fractions from the trailing edge of the included peak which is due to small unilamellar liposomes, were used as a source throughout this study. The SOFICA light scattering photometer (Model 42000) was used to determine the weight average liposomes weight (Lw) of small unilamellar DMPC liposomes. The Lw was determined in the absence and presence of CPZ-HCl both above and below the CMC over the temperature range of 25 °C to 40 °C.
 Results: The Lw was observed to increase linearly in the absence and presence of CPZ-HCl.
 The Lw was observed to increase linearly in the absence of CPZ-HCl, from 1.88×106+0.02 g/mol at 25 °C to 3.25×106+0.03 g/mol at 40 °C. Similarly, the Lw was observed to increase linearly in the present of CPZ-HCl, for example at 18 mmol drug concentration, the Lw increases from 11×106+0.04 g/mol at 25 °C to 13.75×106+0.03 g/mol at 40 °C. When the data are presented as a function of CPZ-HCl concentration, a gradual increase in Lw was observed below the CMC. Little increase in Lw however, was observed at post-micellar concentrations of 14 mmol and 18 mmol.

Membrane structural change of dimyristoylphosphatidylcholine liposome on the interaction with polyethyleneimine.

Polyethyleneimine (PEI) has long been considered as "golden standard" for polymeric gene delivery carriers. To get a better understanding on the molecular basis of PEI cytotoxicity, dynamic light scattering, zeta-potential measurement, fluorescence emission, Fröster resonance energy transfer and anisotropy measurement were conducted to reveal the interaction between PEI and dimyristoylphosphatidylcholine (DMPC) liposome and the influence on the structural properties of the membrane. PEI was found to bind onto the surface of the liposome, inducing an aggregation of the vesicle and an increase in surface potential at low PEI concentration up to 0.05 mg mL-1. A further increase in PEI concentration made little change on the surface potential, however reduced the aggregation of the vesicle due to the repulsion between the adsorbed PEI chains. PEI binding slightly increased the fluidity of lipid in interface region and decreased its packing density, and thus resulted in an enhanced leakage of calcein through the membrane. The polymer size played an important role in PEI-DMPC liposome interaction. PEI of higher molecular weight was more favorable to interact with DMPC and more efficient to perturb the structural properties of the membrane.

Partial molecular volumes of cholesterol and phosphatidylcholine in mixed bilayers

Abstract Dispersion of multilamellar liposomes of dimyristoylphosphatidylcholine (DMPC) and cholesterol (CHOL) were studied by vibrational densitometer for the CHOL mole fractions X = 0−0.54 in the temperature range 18−50 °C, both below and above the main phase transition. DMPC-CHOL bilayers served as a simple model for lipidic part of biological membrane. Volumetric parameters are essential not only to evaluate the data obtained by scattering and diffraction methods on model membranes but can provide valuable information about molecular packing in bilayers and the phase behaviour of lipid-CHOL mixtures. In this paper, preliminary results regarding the changes in the specific volume of lipid bilayer with increasing temperature and CHOL content are presented. Different values of apparent molecular volume of CHOL for different CHOL mole fraction pointed out the non-ideal mixing of DMPC and CHOL.

Physicochemical interactions among α-eleostearic acid-loaded liposomes applied to the development of drug delivery systems

In this study, α-eleostearic acid-loaded (α-ESA-loaded) dimyristoylphosphatidylcholine (DMPC) liposomes had their physicochemical properties characterized by horizontal attenuated total reflectance Fourier transform infrared (HATR-FTIR) spectroscopy, nuclear magnetic resonance (NMR) and differential scanning calorimetry (DSC). In vitro thiobarbituric acid reactive substance (TBARS) assays were performed to obtain preliminary information on the oxidative potential of the system. An α-ESA-promoted ordering effect in the lipid phosphate region was observed. It was associated with a rotation restriction due to an increase in the amount of lipid group hydrogen bonds. The fatty acid was responsible for the reduction in the degree of hydration of carbonyl groups located in the interfacial region of lipids. α-ESA disordered the DMPC methylene acyl chains by trans-gauche isomerization and increased its rotation rate. TBARS results showed pro-oxidant behavior on liposomes, induced by α-ESA. The discussion about the responses considered the degree of saturation of phosphatidylcholines and suggested that the α-ESA oxidative effects may be modulated by the liposome lipid composition. The versatility of liposomal carriers may be promising for the development of efficacious α-ESA-based drug delivery systems. Results described in this study contribute to the selection of adequate material to produce them.

Electron paramagnetic resonance and transmission electron microscopy study of the interactions between asbestiform zeolite fibers and model membranes

ABSTRACTDifferent asbestiform zeolite fibers of the erionite (termed GF1 and MD8, demonstrated carcinogenic) and offretite (termed BV12, suspected carcinogenic) families were investigated by analyzing the electron paramagnetic resonance (EPR) spectra of selected surfactant spin probes and transmission electron microscopy (TEM) images in the presence of model membranes—cetyltrimethylammonium (CTAB) micelles, egg-lecithin liposomes, and dimyristoylphosphatidylcholine (DMPC) liposomes. This was undertaken to obtain information on interactions occurring at a molecular level between fibers and membranes which correlate with entrance of fibers into the membrane model or location of the fibers at the external or internal membrane interfaces. For CTAB micelles, all fibers were able to enter the micelles, but the hair-like structure and chemical surface characteristics of GF1 modified the micelle structure toward a bilayer-like organization, while MD8 and BV12, being shorter fibers and with a high density of surface interacting groups, partially destroyed the micelles. For liposomes, GF1 fibers partially penetrated the core solution, but DMPC liposomes showed increasing rigidity and organization of the bilayer. Conversely, for MD8 and BV12, the fibers did not cross the membrane demonstrating a smaller membrane structure perturbation. Scolecite fibers (termed SC1), used for comparison, presented poor interactions with the model membranes. The carcinogenicity of the zeolites, as postulated in the series SC1<BV12<MD8<GF1, may be related to the structural modifications of the model membranes when interacting with these zeolite fibers.

DSC and Raman study of DMPC liposomes in presence of Ibuprofen at different pH

Ibuprofen (IbuH), 2-(4-isobutylphenyl) propionic acid, is a well-known nonsteroidal anti-inflammatory drug and is a promising antiatherosclerotic agent. Since IbuH has demonstrated to affect cell membranes structure, the study of the interaction between the membrane components, like phospholipids, and the drug, is of paramount interest. Moreover, liposomes can be used as drug carriers. In this paper, the effect of increasing amounts of Ibuprofen at neutral and acidic pH on the behaviour of dimyristoylphosphatidylcholine (DMPC) liposomes was investigated by means of Raman and differential scanning calorimetry (DSC) techniques. The results showed that pH influenced noticeably the liposome structure. A simple ‘solution-like’ model can explain the system when small or high IbuH or IbuNa (IbuH sodium salt forming at neutral and basic pH) amounts are present (IbuH/DMPC molar ratio ≤1/8.3 or ≥1/2.2 or IbuNa/DMPC molar ratio ≤1/17.5 or ≥1/4), whereas at intermediate amounts, two DSC peaks appeared: these systems could be described as a mixture of ‘phase II’ domains inserted within a structure of smaller and further ramified ‘phase I’ domains. Both DSC and Raman data suggested IbuH can penetrate within the apolar bilayer at pH 3, while at pH 7, the setting up of polar interactions of different strength between the carboxylic groups of IbuNa and the choline head of the DMPC, hamper a further penetration of the drug. Similar results were obtained both inserting directly IbuH molecules into liposomes as well as acidifying IbuNa water solutions, suggesting a possible use of liposomes as drug carriers when polarity is pH dependent.

Extraction of Phospholipids from Human Erythrocyte Membranes by Hemoglobin Oxidation Products.

This investigation examines oxidation conditions under which hemoglobin extracts membrane phospholipid from erythrocytes and model membranes. In erythrocytes made echinocytic with exogenous phospholipid, addition of hemoglobin oxidized with hydrogen peroxide (H2O2) or Vitamin C (conditions that result in the formation of significant quantities of choleglobin), but not ferricyanide (which produces predominantly methemoglobin), induced dose-dependent shape reversion to less echinocytic forms, consistent with extraction of phospholipids from the exofacial side of the membrane. Erythrocytes preloaded with radiolabeled phosphatidylcholine or NBD-labeled phosphatidylcholine, phosphatidylglycerol or phosphatidic acid, exhibited greatest extraction of radiolabel or fluorescence signal with exogenous hemoglobin oxidized via H2O2 or Vitamin C, but not ferricyanide. However, with NBD-phosphatidylserine (a preferential inner monolayer intercalator), significantly less extraction of labeled lipid occurred with oxidized hemoglobin prepared under all three oxidizing conditions. In dimyristoylphosphatidylcholine liposomes containing radiolabeled phosphatidylcholine, phosphatidylserine or phosphatidylethanolamine, subsequent addition of hemoglobin oxidized with H2O2 or Vitamin C extracted radiolabeled lipid with significantly greater efficiency than ferricyanide-treated hemoglobin, with enhanced extraction detectable at levels approaching physiological plasma oxidant concentrations. Radiolabeled lipid extraction was comparable for phospholipids containing saturated acyl chains between 12 and 18 carbons but diminished significantly for oleoyl-containing phospholipids. Hemoglobin dimerization occurred at very low levels with H2O2 treatment, and even lower levels with Vitamin C treatment, and thus did not correlate to the high efficiency and consistent levels of lipid extraction observed with these treatments. These findings indicate that choleglobin extracts lipids from cell membranes regardless of headgroup or acyl chain length, through a process of direct hydrophobic interaction with the membrane surface.

Small-Angle Neutron Scattering investigation of cholesterol-doped DMPC liposomes interacting with β-cyclodextrin

The Small-Angle Neutron Scattering technique has been applied to characterize the influence of a randomly methylated β-cyclodextrin, called RAMEB, on dimyristoylphosphatidylcholine (DMPC) liposomes doped with cholesterol. From the modelling of the experimental neutron scattering cross sections, the detailed response of the vesicle structure upon addition of increasing amounts of RAMEB up to 30 mM has been assessed. This study has been performed below and above the DMPC bilayer phase transition temperature and shows that cholesterol extraction by RAMEB is linked to a decrease of the average radius and of the aggregation number of the vesicles. This extraction takes place in a dose-dependent way until a more monodisperse population of cholesterol-free liposomes is obtained. In addition, the bilayer thickness evolution was inferred, as well as the liposome coverage by RAMEB.

Investigation of the interaction between a β-cyclodextrin and DMPC liposomes: a small angle neutron scattering study

The small angle neutron scattering technique has been applied to investigate the interaction between a cyclodextrin (CD) and liposomes. From the modelling of the experimental neutron scattering cross sections, the detailed structure of dimyristoylphosphatidylcholine (DMPC) liposomes is assessed upon addition of increasing amounts of randomly methylated β-CD (RAMEB). This study has been performed at two temperatures bracketing the phase transition of the DMPC bilayers. The fraction of DMPC molecules incorporated into the vesicles is inferred. The dose-dependent phospholipidic extraction by RAMEB is quantified as well as the concomitant evolution of the liposome radius and of the thickness of the hydrophobic and hydrophilic parts of the membrane. The possible formation of CD-DMPC inclusion complexes is also assessed. The data suggest the dose-dependent coverage by RAMEB of the outer liposome interface. Our analysis highlights the important role of temperature on the mechanism of action of RAMEB. These results are discussed in the framework of the area-difference-elasticity model.

Location, Partitioning Behavior, and Interaction of Capsaicin with Lipid Bilayer Membrane: Study Using Its Intrinsic Fluorescence.

Capsaicin is an ingredient of a wide variety of red peppers, and it has various pharmacological and biological applications. The present study explores the interaction of capsaicin with dimyristoylphosphatidylcholine (DMPC) lipid bilayer membrane by monitoring various photophysical parameters using its intrinsic fluorescence. In order to have a clearer understanding of the photophysical responses of capsaicin, studies involving (i) its solvation behavior in different solvents, (ii) the partition coefficient of capsaicin in different thermotropic phase states of lipid bilayer membrane, and (iii) its location inside lipid bilayer membrane have been carried out. Capsaicin has a reasonably high partition coefficient for DMPC liposome membrane, in both solid gel (2.8 ± 0.1 × 10(5)) and liquid crystalline (2.6 ± 0.1 × 10(5)) phases. Fluorescence quenching study using cetylpyridinium chloride (CPC) as quencher suggests that the phenolic group of capsaicin molecule is generally present near the headgroup region and hydrophobic tail present inside hydrophobic core region of the lipid bilayer membrane. The intrinsic fluorescence intensity and lifetime of capsaicin sensitively respond to the temperature dependent phase changes of liposome membrane. Above 15 mol %, capsaicin in the aqueous liposome suspension medium lowers the thermotropic phase transition temperature by about 3 °C, and above 30 mol %, the integrity of the membrane is significantly lost.

Interaction study between maltose-modified PPI dendrimers and lipidic model membranes

The influence of maltose-modified poly(propylene imine) (PPI) dendrimers on dimyristoylphosphatidylcholine (DMPC) or dimyristoylphosphatidylcholine/dimyristoylphosphatidylglycerol (DMPC/DMPG) (3%) liposomes was studied. Fourth generation (G4) PPI dendrimers with primary amino surface groups were partially (open shell glycodendrimers — OS) or completely (dense shell glycodendrimers — DS) modified with maltose residues. As a model membrane, two types of 100nm diameter liposomes were used to observe differences in the interactions between neutral DMPC and negatively charged DMPC/DMPG bilayers. Interactions were studied using fluorescence spectroscopy to evaluate the membrane fluidity of both the hydrophobic and hydrophilic parts of the lipid bilayer and using differential scanning calorimetry to investigate thermodynamic parameter changes. Pulsed-filed gradient NMR experiments were carried out to evaluate common diffusion coefficient of DMPG and DS PPI in D2O when using below critical micelle concentration of DMPG. Both OS and DS PPI G4 dendrimers show interactions with liposomes. Neutral DS dendrimers exhibit stronger changes in membrane fluidity compared to OS dendrimers. The bilayer structure seems more rigid in the case of anionic DMPC/DMPG liposomes in comparison to pure and neutral DMPC liposomes. Generally, interactions of dendrimers with anionic DMPC/DMPG and neutral DMPC liposomes were at the same level. Higher concentrations of positively charged OS dendrimers induced the aggregation process with negatively charged liposomes. For all types of experiments, the presence of NaCl decreased the strength of the interactions between glycodendrimers and liposomes. Based on NMR diffusion experiments we suggest that apart from electrostatic interactions for OS PPI hydrogen bonds play a major role in maltose-modified PPI dendrimer interactions with anionic and neutral model membranes where a contact surface is needed for undergoing multiple H-bond interactions between maltose shell of glycodendrimers and surface membrane of liposome.

N-homocysteinylation of apolipoprotein A-I impairs the protein’s antioxidant ability but not its cholesterol efflux capacity

A high homocysteine (Hcy) level is a risk factor for atherosclerosis. Hcy can be added to proteins through a process known as N-homocysteinylation. This is thought to be a potential cause of atherosclerosis induction. We previously reported that N-homocysteinylated apolipoprotein A-I (N-Hcy-apoA-I) was identified in normal human plasma. In this study, the effect of N-homocysteinylation on the functions of apoA-I was examined. A kinetic study using dimyristoyl phosphatidylcholine (DMPC) liposomes indicated that N-Hcy-apoA-I showed increased lipid-binding activity compared to wild-type apoA-I. Two reconstituted high-density lipoprotein (rHDL) particles of different sizes (approximately 8.2 nm and 7.6 nm in diameter) were produced by mixing apoA-I and 1-palmitoyl-2-oleoyl phosphatidylcholine (POPC). However, an increased ratio of large to small particles was found in rHDL prepared with N-Hcy-apoA-I. The normal apoA-I antioxidant ability, estimated by the suppression of conjugated diene formation in low-density lipoprotein (LDL) induced by copper sulfate oxidation, was considerably impaired when using N-Hcy-apoA-I. Although N-Hcy-apoA-I functioned as an oxidant, no significant difference was observed in the cholesterol efflux capacity from THP-1 macrophages between wild-type apoA-I and N-Hcy-apoA-I. These results suggest that N-Hcy-apoA-I might be proatherogenic due to its oxidative behavior but not an attenuation of cholesterol efflux capacity.

The effect of furazolidone on the physico-chemical properties of dimyristoylphosphatidylcholine bilayers: Relevance to anti-leishmanial therapy

In this study, the influence of furazolidone, an anti-leishmanial drug, on dimyristoylphosphatidylcholine (DMPC) liposome hydration degree, mobility and thermodynamics was investigated by Fourier transform infrared spectroscopy (FTIR), nuclear magnetic resonance (NMR) and differential scanning calorimetry (DSC). FTIR results showed that furazolidone was responsible for an increase in the hydrogen bound number and mobility of the lipid phosphate group. Furazolidone also affected the lipid choline group by increasing its motional freedom, as shown by FTIR and 1H NMR spin–lattice relaxation time measurements. At the DMPC interfacial region, FTIR results showed a drug-induced reduction of the carbonyl hydration and order degrees. Very weak interaction among furazolidone and the hydrophobic lipid chains was also observed. However, no furazolidone-induced changes on thermodynamical parameters, such as phase transition temperature (Tm) and enthalpy variation (ΔH), were detected by the DSC technique. Thus, furazolidone seems to interact preferentially with lipid polar and interfacial regions, enhancing the freedom for gauche-trans isomerization of the first methylene groups of DMPC acyl chains. Responses described in this paper may explain the improved activity of furazolidone-encapsulated liposomes by comparison with the effect of the free drug, described in literature. The findings can also improve further strategies for the potential therapeutic application of liposomal furazolidone as a drug delivery system and minimize the risk of drug resistance and collateral effects related to high toxicity.