1-2-oleoylphosphatidylcholine and 1-2-dipalmitoylphosphatidylglycerol

Sickle cell disease (SCD) is a mortal erythrocyte-based disease which is hard to treat effectively. Development of a treatment method that can prevent deoxygenation of erythrocytes or reduce the oxidative stress of sickle erythrocytes is one of the important issues towards SCD. Among a wide variety of potential drug carriers, liposomes are advantageous and preferable with their easy preparation and biocompatibility. In this study, L-Glutamine (Gln) loaded liposomes were prepared with 1,2-Dioleoyl-sn-glycero-3-phosphocholine (DOPC) and 1,2-Dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt (DPPG). Liposomes were characterized via zeta potential, size measurements, differential scanning calorimetry, Fourier Transform Infra-red Spectroscopy and they were visualized via transmission electron microscopy and scanning electron microscopy. Effect of the encapsulated amount of Gln was investigated by encapsulating Gln at three different concentrations (i.e0.20 mM, 40 mM and 60 mM). Drug encapsulation and release studies were implemented with high pressure liquid chromatography (HPLC). The encapsulation efficiency of Gln was determined to be the higher than the ones reported in the literature: 83.6%, 87.1% and 84.9% for 20 mM, 40 mM and 60 mM Gln, respectively. It was found that after 6 hours, liposomes loaded with 60 mM of Gln had released 45.7% of Gln. Optical microscopy images of the erythrocytes after 3 hours of incubation and haemolysis measurements proved that presence of liposomes did not cause any structural changes on the erythrocyte shape. Overall, it was concluded that L-Gln loaded PC/PG liposomes provide promising results in terms of developing a new drug delivery platform for SCD.

Topics: Erythrocytes; Glutamine; Liposomes; Phosphatidylcholines; Phosphatidylglycerols

Non-fluidic bio-layer interferometry (BLI) has rapidly become a standard tool for monitoring almost all biomolecular interactions in a label-free, real-time and high-throughput manner. High-efficiency screening methods which measure the kinetics of liposomes with a variety of compounds require the immobilization of liposomes. In this work, a method is described for immobilizing liposomes for interaction studies, based on the biophysical principles of this biosensor platform. The immobilization approach includes the loading of DSPE-PEG

Topics: 1,2-Dipalmitoylphosphatidylcholine; Adsorption; Biosensing Techniques; Biotin; Cardiolipins; Cytochromes c; Drosophila Proteins; Fluoresceins; Fluorescent Dyes; High-Throughput Screening Assays; Hydrophobic and Hydrophilic Interactions; Interferometry; Kinetics; Liposomes; Micelles; Microscopy, Fluorescence; Phosphatidylcholines; Phosphatidylethanolamines; Phosphatidylglycerols; Polyethylene Glycols; Protein Phosphatase 1; Reproducibility of Results

Topics: 1,2-Dipalmitoylphosphatidylcholine; Biodegradation, Environmental; Cadmium; Cell Membrane; Environmental Pollutants; Indoleacetic Acids; Models, Biological; Naphthaleneacetic Acids; Phosphatidylcholines; Phosphatidylglycerols; Plant Leaves; Plants; Surface Tension; Thermodynamics; Unilamellar Liposomes; Water

Dimethyl sulfoxide (DMSO) has been broadly used in biology as a cosolvent, a cryoprotectant, and an enhancer of membrane permeability, leading to the general assumption that DMSO-induced structural changes in cell membranes and their hydration water play important functional roles. Although the effects of DMSO on the membrane structure and the headgroup dehydration have been extensively studied, the mechanism by which DMSO invokes its effect on lipid membranes and the direct role of water in this process are unresolved. By directly probing the translational water diffusivity near unconfined lipid vesicle surfaces, the lipid headgroup mobility, and the repeat distances in multilamellar vesicles, we found that DMSO exclusively weakens the surface water network near the lipid membrane at a bulk DMSO mole fraction (XDMSO) of <0.1, regardless of the lipid composition and the lipid phase. Specifically, DMSO was found to effectively destabilize the hydration water structure at the lipid membrane surface at XDMSO <0.1, lower the energetic barrier to dehydrate this surface water, whose displacement otherwise requires a higher activation energy, consequently yielding compressed interbilayer distances in multilamellar vesicles at equilibrium with unaltered bilayer thicknesses. At XDMSO >0.1, DMSO enters the lipid interface and restricts the lipid headgroup motion. We postulate that DMSO acts as an efficient cryoprotectant even at low concentrations by exclusively disrupting the water network near the lipid membrane surface, weakening the cohesion between water and adhesion of water to the lipid headgroups, and so mitigating the stress induced by the volume change of water during freeze-thaw.

Topics: 1,2-Dipalmitoylphosphatidylcholine; Diffusion; Dimethyl Sulfoxide; Fatty Acids, Monounsaturated; Magnetic Resonance Spectroscopy; Membranes, Artificial; Phosphatidylcholines; Phosphatidylglycerols; Quaternary Ammonium Compounds; Scattering, Small Angle; Water; X-Ray Diffraction

Phase separation in lipid bilayers that include negatively charged lipids is examined experimentally. We observed phase-separated structures and determined the membrane miscibility temperatures in several binary and ternary lipid mixtures of unsaturated neutral lipid, dioleoylphosphatidylcholine (DOPC), saturated neutral lipid, dipalmitoylphosphatidylcholine (DPPC), unsaturated charged lipid, dioleoylphosphatidylglycerol (DOPG((-))), saturated charged lipid, dipalmitoylphosphatidylglycerol (DPPG((-))), and cholesterol. In binary mixtures of saturated and unsaturated charged lipids, the combination of the charged head with the saturation of the hydrocarbon tail is a dominant factor in the stability of membrane phase separation. DPPG((-)) enhances phase separation, while DOPG((-)) suppresses it. Furthermore, the addition of DPPG((-)) to a binary mixture of DPPC/cholesterol induces phase separation between DPPG((-))-rich and cholesterol-rich phases. This indicates that cholesterol localization depends strongly on the electric charge on the hydrophilic head group rather than on the ordering of the hydrocarbon tails. Finally, when DPPG((-)) was added to a neutral ternary system of DOPC/DPPC/cholesterol (a conventional model of membrane rafts), a three-phase coexistence was produced. We conclude by discussing some qualitative features of the phase behaviour in charged membranes using a free energy approach.

Topics: Cholesterol; Lipid Bilayers; Models, Chemical; Phosphatidylcholines; Phosphatidylglycerols

By using Fourier transform infrared (FT-IR) spectroscopy in combination with differential scanning calorimetry (DSC) coupled with pressure perturbation calorimetry (PPC), ultrasound velocimetry, Laurdan fluorescence spectroscopy, fluorescence microscopy and atomic force microscopy (AFM), the temperature and pressure dependent phase behavior of the five-component anionic model raft lipid mixture DOPC/DOPG/DPPC/DPPG/cholesterol (20:5:45:5:25 mol%) was investigated. A temperature range from 5 to 65 °C and a pressure range up to 16 kbar were covered to establish the temperature-pressure phase diagram of this heterogeneous model biomembrane system. Incorporation of 10-20 mol% PG still leads to liquid-ordered (l(o))-liquid-disordered (l(d)) phase coexistence regions over a wide range of temperatures and pressures. Compared to the corresponding neutral model raft mixture (DOPC/DPPC/Chol 25:50:25 mol%), the p,T-phase diagram is - as expected and in accordance with the Gibbs phase rule - more complex, the phase sequence as a function of temperature and pressure is largely similar, however. This anionic heterogeneous model membrane system will serve as a more realistic model biomembrane system to study protein interactions with anionic lipid bilayers displaying liquid-disordered/liquid-ordered domain coexistence over a wide range of the temperature-pressure plane, thus allowing also studies of biologically relevant systems encountered under extreme environmental conditions.

Topics: 1,2-Dipalmitoylphosphatidylcholine; Algorithms; Calorimetry; Calorimetry, Differential Scanning; Cholesterol; Lipid Bilayers; Membrane Microdomains; Microscopy, Atomic Force; Microscopy, Fluorescence; Models, Chemical; Phosphatidylcholines; Phosphatidylglycerols; Pressure; Rheology; Spectrometry, Fluorescence; Spectroscopy, Fourier Transform Infrared; Temperature

Liposomal locked-in dendrimers (LLDs), the combination of liposomes and dendrimers in one formulation, represents a relatively new term in the drug carrier technology. LLDs undergone appropriate physicochemical investigation can merge the benefits of liposomal and dendrimeric nanocarriers. In this study generation 1 and 2 hydroxy-terminated dendrimers were synthesized and were then "locked" in liposomes consisting of DOPC/DPPG. The anticancer drug doxorubicin (Dox) was loaded into pure liposomes or LLDs and the final products were subjected to lyophilization. The loading of Dox as well as its in vitro release rate from all systems was determined and the interaction of liposomes with dendrimers was assessed by thermal analysis and fluorescence spectroscopy. The results were very promising in terms of drug encapsulation and release rate, factors that can alter the therapeutic profile of a drug with low therapeutic index such as Dox. Physicochemical methods revealed a strong, generation dependent, interaction between liposomes and dendrimers that probably is the basis for the higher loading and slower drug release from the LLDs comparing to pure liposomes.

Topics: Antibiotics, Antineoplastic; Antineoplastic Agents; Dendrimers; Differential Thermal Analysis; Doxorubicin; Drug Carriers; Drug Compounding; Drug Delivery Systems; Freeze Drying; Indicators and Reagents; Liposomes; Membrane Fluidity; Neoplasms; Phosphatidylcholines; Phosphatidylglycerols; Solubility; Spectrometry, Fluorescence

We investigate the permeability of lipid membranes for fluorescence dyes and ions. We find that permeability reaches a maximum close to the chain melting transition of the membranes. Close to transitions, fluctuations in area and compressibility are high, leading to an increased likelihood of spontaneous lipid pore formation. Fluorescence correlation spectroscopy reveals the permeability for rhodamine dyes across 100-nm vesicles. Using fluorescence correlation spectroscopy, we find that the permeability of vesicle membranes for fluorescence dyes is within error proportional to the excess heat capacity. To estimate defect size we measure the conductance of solvent-free planar lipid bilayer. Microscopically, we show that permeation events appear as quantized current events very similar to those reported for channel proteins. Further, we demonstrate that anesthetics lead to a change in membrane permeability that can be predicted from their effect on heat capacity profiles. Depending on temperature, the permeability can be enhanced or reduced. We demonstrate that anesthetics decrease channel conductance and ultimately lead to blocking of the lipid pores in experiments performed at or above the chain melting transition. Our data suggest that the macroscopic increase in permeability close to transitions and microscopic lipid ion channel formation are the same physical process.

Topics: 1,2-Dipalmitoylphosphatidylcholine; Algorithms; Anesthetics; Calorimetry; Cell Membrane Permeability; Fluorescent Dyes; Ion Channel Gating; Ion Channels; Ions; Kinetics; Lipid Bilayers; Membrane Potentials; Phosphatidylcholines; Phosphatidylglycerols; Spectrum Analysis; Temperature

To optimize the formulation and preparation method of multivesicular liposome of thymopentin and to investigate its pharmacokinetics in rats, the multivesicular liposome of thymopentin was prepared by double emulsification method and the formulation was optimized by orthogonal design. The release characteristics of thymopentin from multivesicular liposome in PBS (pH 7.4) and in plasma were investigated. The multivesicular liposome of thymopentin labeled with fluorescein isothiocyanate was prepared by double emulsification method. Its pharmacokinetics was evaluated following intramuscular injection in rats. The optimal formulation of multivesicular liposome of thymopentin were formulated with 7.5% glucose in aqueous phase and 2.25 mol x L(-1) triolein, 2.68 mol x L(-1) DPPG and 16.96 mol x L(-1) DOPC in organic phase. The entrapment efficiency of the multivesicular liposome of thymopentin was above 85% and the mean particle size was about 22 microm. The in vitro release of thymopentin from multivesicular liposome in PBS (pH 7.4) and in plasma was found to be in a sustained manner. The release curves were fitted to Higuchi equation. The pharmacokinetics following intramuscular injection of the multivesicular liposome of thymopentin labeled with fluorescein isothiocyanate in rats showed that the peak concentration of thymopentin was lower and elimination of it was slower significantly than that of thymopentin labeled with fluorescein isothiocyanate solution in the same dose. The plasma concentration of thymopentin maintained above quantitative limitation at 120 h after administration of multivesicular liposome of thymopentin. The optimized formulation and preparation technology of multivesicular liposome of thymopentin with higher entrapment efficiency are feasible with good reproducibility. Multivesicular liposome of thymopentin showed significant sustained-release property following intramuscular injection in rats.

Topics: Adjuvants, Immunologic; Animals; Area Under Curve; Delayed-Action Preparations; Drug Carriers; Drug Compounding; Drug Delivery Systems; Glucose; Liposomes; Male; Particle Size; Phosphatidylcholines; Phosphatidylglycerols; Rats; Rats, Sprague-Dawley; Thymopentin; Triolein

Using the Langmuir technique, we have studied the properties at the air/water interface and the interaction of the hepatitis G virus synthetic peptide E1(53-66) and its palmitoyl derivative with membrane phospholipids. These phospholipids had different characteristics referring to the net charge and saturation of the acyl chain. The palmitoyl derivative was more stable at the air/water interface and in the kinetic at constant area measurements showed higher incorporation to the monolayer. The interaction was higher for saturated phospholipids and those with a negative net charge. When the peptides were in the subphase, they produced changes in the miscibility of mixed monolayers composed of DPPC/DPPG or DOPC/DOPG. It can be deduced from the results obtained that electrostatic interactions play a major role, but when the peptide is derivatized with the palmitoyl chain, hydrophobic interactions are added to the former ones. The interaction is also influenced by the saturation of the acyl chain.

Topics: 1,2-Dipalmitoylphosphatidylcholine; Adsorption; Capsid; GB virus C; Membranes, Artificial; Palmitates; Peptides; Phosphatidylcholines; Phosphatidylglycerols; Phospholipids; Surface Properties; Thermodynamics