Preparation of liposomes containing Ceramide 3 and their membrane characteristics

Liposomes composed of Ceramide 3, [2S,3S,4R-2-stearoylamide-1,3,4-octadecanetriol], and L-alpha-dipalmitoylphosphatidylcholine (DPPC) were prepared by varying the amount of Ceramide 3, and the effects of Ceramide 3 on the liposome formation, particle size, dispersibility, microviscosity and phase transition temperature were examined by means of a microscopy, a dynamic light scattering method, a fluorescence polarization method, a differential scanning calorimetry (DSC) and so on. All the DPPC was able to contribute to the formation of liposomes up to 0.130 mol fraction of Ceramide 3. The particle size of liposomes was almost unaffected by the addition of Ceramide 3. The dispersibility of liposomes containing Ceramide 3 was maintained for at least 15 days. The microviscosity of liposomal bilayer membranes in the liquid crystalline state was increased with increasing the mole fraction of Ceramide 3, while that in the gel state was independent of the mole fraction of Ceramide 3. The phase transition temperature from gel to liquid crystalline states of DPPC bilayer membranes was shifted upwards with the addition of Ceramide 3, indicating a cooperative interaction between DPPC and Ceramide 3 molecules. However, a sharp DSC peak became broad and split at higher mole fractions of Ceramide 3, suggesting a phase separation in the mixed DPPC/Ceramide 3 liposomal bilayer membranes. These phenomena were suggested to be related to the previously observed fact for the mixed DPPC/Ceramide 3 monolayers that Ceramide 3 interacts with DPPC in the liquid-expanded phase with consequent phase separation accompanied with domain formation.     

二棕榈酰磷脂酰胆碱/豆固醇脂质体液态有序相的结构性质:同步辐射X光衍射研究

用同步辐射小角和宽角X光衍射实验技术研究了由二棕榈酰磷脂酰胆碱(DPPC)和豆固醇所形成的脂质体的液态有序相的结构性质. 结果表明液态有序相的小角X光衍射d值(d-spacing)随着固醇温度和浓度的变化仅有微小的改变. 与凝胶相及液晶相的宽角X光衍射d值相比, 液态有序相的宽角X光衍射d值有更宽的变化范围, 在30到52 °C的温度范围内, 液态有序相的宽角X光衍射d值从0.422 nm变化到0.460 nm. 电子云密度计算表明液态有序相的脂双层厚度和水层厚度都要大于与之平衡共存的液晶相的脂双层厚度和水层厚度. 电子云密度计算结果还表明液态有序相的脂双层厚度随温度升高而降低. 本研究结果对于从定量的角度认识 生物膜的相态及深入认识生物膜中的有序结构具有重要意义.     

Liposome fluidization and melting point depression by pressurized CO2 determined by fluorescence anisotropy

The influence of CO2 on the bilayer fluidity of liposomes, which are representative of model cellular membranes, was examined for the first time at the elevated pressures (up to 13.9 MPa) associated with CO2-based processing of liposomes and microbial sterilization. Fluidization and melting point depression of aqueous dipalmitoylphosphatidylcholine (DPPC) liposomes by pressurized CO2 (present as an excess phase) were studied by steady-state fluorescence anisotropy using the membrane probe 1,6-diphenyl-1,3,5-hexatriene (DPH). Isothermal experiments revealed reversible, pressure-dependent fluidization of DPPC bilayers at temperatures corresponding to near-gel (295 K) and fluid (333 K) phases at atmospheric pressure, where the gel-to-fluid phase transition (Tm) occurs at approximately 315 K. Isobaric measurements (PCO2 =1.8, 7.0, and 13.9 MPa) of DPH anisotropy demonstrate substantial melting point depression (DeltaTm = -4.8 to -18.5 K) and a large broadening of the gel-fluid phase transition region, which were interpreted using conventional theories of melting point depression. Liposome fluidity is influenced by CO2 accumulation in the hydrocarbon core and polar headgroup region, as well as the formation of carbonic acid and/or the presence of buffering species under elevated CO2 pressure.     

Freeze-fracture electron microscopic and calorimetric studies on microscopic states of surface-modified liposomes with poly(ethylene glycol) chains

The differential scanning calorimetry (DSC) and the freeze-fracture electron microscopy of dipalmitoyl phosphatidylcholine (DPPC) liposomes containing distearoyl-N-monomethoxy poly(ethylene glycol)-succinyl-phosphatidylethanolamines (PEG-DSPE) were carried out. The DSC peak of DPPC liposomes containing PEG-DSPE had a shoulder. The main phase transition temperature of DPPC bilayer membranes containing PEG-DSPE whose molecular weight of PEG is less than 3000 was slightly shifted to a higher temperature, while that containing PEG-DSPE whose molecular weight of PEG is more than 5000 was slightly shifted to a lower temperature. The electron micrographs of freeze-fracture replicas of DPPC liposomes containing PEG-DSPE quenched from 37±2°C exhibited banded and planar textures, suggesting the lateral phase separation in the bilayer membranes.     

Fluorescence studies on radiation oxidative damage to membranes with implications to cellular radiosensitivity

Radiation oxidative damage to plasma membrane and its consequences to cellular radiosensitivity have received increasing attention in the past few years. This review gives a brief account of radiation oxidative damage in model and cellular membranes with particular emphasis on results from our laboratory. Fluorescence and ESR spin probes have been employed to investigate the structural and functional alterations in membranes after y-irradiation. Changes in the lipid bilayer in irradiated unilamellar liposomes prepared from egg yolk lecithin (EYL) were measured by using diphenylhexatriene (DPH) as a probe. The observed increase in DPH polarization and decrease in fluorescence intensity after γ-irradiation of liposomes imply radiation-induced decrease in bilayer fluidity. Inclusion of cholesterol in liposome was found to protect lipids against radiation damage, possibly by modulation of bilayer organization e.g. lipid packing. Measurements on dipalmitoyl phosphatidylcholine (DPPC) liposomes loaded with 6-carboxyfluorescein (CF) showed radiation dose-dependent release of the probe indicating radiation-induced increased permeability. Changes in plasma membrane permeability of thymocytes were monitored by fluorescein diacetate (FDA) and induced intracellular reactive oxygen species (ROS) were determined by 2,7-dichlorodihydro fluorescein diacetate (DCH-FDA). Results suggest a correlation between ROS generation and membrane permeability changes induced by radiation within therapeutic doses (0-10 Gy). It is concluded that increase in membrane permeability was the result of ROS-mediated oxidative reactions, which might trigger processes leading to apoptotic cell death after radiation exposure.     

Characterization of the interactions between synthetic nematic LCs and model cell membranes

Differential scanning calorimetry (DSC) was used to characterize interactions of synthetic LCs, 4‐pentyl‐4′‐cyanobiphenyl (5CB) and TL205 (a mixture of cyclohexane‐fluorinated biphenyls and fluorinated terphenyls) with simple mimics of cell membranes. The investigation was motivated by reports that living cells can be placed into contact with TL205 without apparent toxicity, whereas contact of cells with 5CB leads to cell death. The tendency was examined for 5CB and TL205 to spontaneously partition into and influence the organization for model cell membranes composed of phospholipids. Upon contact of an aqueous dispersion of DPPC liposomes with neat LC for 4 h, 5CB partitioned into the liposomes at a weight ratio of 5:1 DPPC:5CB, whereas TL205 partitioned at a ratio of 310:1 DPPC:TL205. DSC endotherms indicated that the 5CB spontaneously partitioned into the liposomes was far more perturbing than TL205. DSC endotherms of DPPC bilayers containing the same concentration of either 5CB or TL205 also revealed 5CB to be more perturbing than TL205. The effect of up to 7.8 wt % of TL205 was small, resulting in a shift in the melting transition from 41.4°C to 40.1°C and a minor change in peak width, indicating only minor effects on the organization of the bilayer. These effects are similar to those caused by cholesterol in DPPC bilayers. In contrast, 5CB shifted the DPPC melting transition from 41.4°C to ～36°C and increased the width of the transition peak by a factor of ten, indicating a destabilization of the ordered phase in the bilayer and a disruption of the cooperative nature of the gel‐to‐LC transition of the phospholipid bilayer. Taken together, the results demonstrate that 5CB and TL205 differ significantly in their interactions with model cell membranes, which suggests one possible origin of their different toxicities toward cells.     

不同固醇与DPPC二元体系的液态有序相

应用同步辐射X射线衍射和差示扫描量热法研究了由不同结构的固醇(胆固醇、脱氢胆固醇、豆固醇、谷固醇、麦角固醇以及固醇核)和二棕榈酰磷脂酰胆碱(DPPC)二元体系形成的液态有序相. 研究表明, 胆固醇比植物固醇(豆固醇和谷固醇)和真菌固醇(麦角固醇)能更有效地与DPPC形成液态有序相(Lo); 有胆固醇或者脱氢胆固醇参与的液态有序相能够在较宽的温度范围内保持稳定, 而由植物固醇和真菌固醇参与的液态有序相对温度有较强的依赖性, 在DPPC主相变温度附近有明显的热致相变过程, 因此这一液态有序相应该进一步区分为Loβ和Loα相. 研究结果有助于阐明固醇尾链在液态有序相以及脂筏中的作用, 也有助于理解在进化过程中动物细胞膜为何选择胆固醇作为主要固醇.     

Effects of poly(ethylene glycol) (PEG) chain length of PEG-lipid on the permeability of liposomal bilayer membranes

The effects of poly(ethylene glycol) (PEG) chain length of PEG-lipid on the membrane characteristics of liposomes were investigated by differential scanning calorimetry (DSC), freeze-fracture electron microscopy (FFEM), fluorescence polarization measurement and permeability measurement using carboxyfluorescein (CF). PEG-liposomes were prepared from mixtures of dipalmitoyl phosphatidylcholine (DPPC) and distearoyl phosphatidylethanolamines with covalently attached PEG molecular weights of 1000, 2000, 3000 and 5000 (DSPE-PEG). DSC and FFEM results showed that the addition of DSPE-PEG to DPPC in the preparation of liposomes caused the lateral phase separation both in the gel and liquid-crystalline states. The fluidity in the hydrocarbon region of liposomal bilayer membranes was not significantly changed by the addition of DSPE-PEG, while that in the interfacial region was markedly increased. From these results, it was anticipated that the CF leakage from PEG-liposomes is accelerated compared with DPPC liposomes. However, CF leakage from liposomes containing DSPE-PEG with a 0.060 mol fraction was depressed compared with regular liposomes, and the leakage decreased with increasing PEG chain length. Furthermore, the CF leakage from liposomes containing DSPE-PEG with a 0.145 mol fraction was slightly increased compared with that of liposomes containing DSPE-PEG with a 0.060 mol fraction. It is suggested that the solute permeability from the PEG-liposomes was affected by not only properties of the liposomal bilayer membranes such as phase transition temperature, phase separation and membrane fluidity, but also the PEG chain of the liposomal surface.     

Characterization of the interactions between synthetic nematic LCs and model cell membranes

Differential scanning calorimetry (DSC) was used to characterize interactions of synthetic LCs, 4-pentyl-4'-cyanobiphenyl (5CB) and TL205 (a mixture of cyclohexane-fluorinated biphenyls and fluorinated terphenyls) with simple mimics of cell membranes. The investigation was motivated by reports that living cells can be placed into contact with TL205 without apparent toxicity, whereas contact of cells with 5CB leads to cell death. The tendency was examined for 5CB and TL205 to spontaneously partition into and influence the organization for model cell membranes composed of phospholipids. Upon contact of an aqueous dispersion of DPPC liposomes with neat LC for 4 h, 5CB partitioned into the liposomes at a weight ratio of 5:1 DPPC:5CB, whereas TL205 partitioned at a ratio of 310:1 DPPC:TL205. DSC endotherms indicated that the 5CB spontaneously partitioned into the liposomes was far more perturbing than TL205. DSC endotherms of DPPC bilayers containing the same concentration of either 5CB or TL205 also revealed 5CB to be more perturbing than TL205. The effect of up to 7.8 wt % of TL205 was small, resulting in a shift in the melting transition from 41.4°C to 40.1°C and a minor change in peak width, indicating only minor effects on the organization of the bilayer. These effects are similar to those caused by cholesterol in DPPC bilayers. In contrast, 5CB shifted the DPPC melting transition from 41.4°C to ∼36°C and increased the width of the transition peak by a factor of ten, indicating a destabilization of the ordered phase in the bilayer and a disruption of the cooperative nature of the gel-to-LC transition of the phospholipid bilayer. Taken together, the results demonstrate that 5CB and TL205 differ significantly in their interactions with model cell membranes, which suggests one possible origin of their different toxicities toward cells.     

Chain-melting phase transition and short-range molecular interactions in phospholipid foam bilayers

Occurrence of two-dimensional chain melting phase transition in foam bilayers was established for the first time. Microscopic horizontal foam bilayers [Newton black films (NBF)] were investigated by the microinterferometric method of Scheludko-Exerowa. The foam bilayers were formed from water-ethanol solutions of dimyristoylphosphatidylcholine (DMPC) and dipalmitoylphosphatidylcholine (DPPC) and egg phosphatidylcholine (Egg PC) and samples of amniotic fluid (AF) at different temperatures. The influence of temperature on the foam bilayer thickness h(w) and on the critical concentration Cc for formation of foam bilayer was studied. It was shown that in the range of the main phase transition the temperature dependence of h(w) and C(c) changed specifically in the case of DMPC and DPPC foam bilayers. The thickness of the foam bilayers increased with decreasing temperature in the range of the main phase transition due to the melting of hydrocarbon tails of phospholipid molecules. These changes took place at the temperatures of the bulk chain-melting phase transitions, as determined by differential scanning calorimetry (DSC) for both aqueous, and water/ethanol DMPC, DPPC, and DPPC dispersions. An effect of the 'disperse medium' on h(w) was found for foam bilayers from DPPC. The results that foam bilayers could have different thickness at different temperatures disproved the current concept that NBF acquired constant thickness at concentrations higher than C(el,cr). The data for Cc were analysed on the basis of the hole-nucleation theory of bilayer stability of Kashchiev and Exerowa. This theory considered the amphiphile bilayer as a two-dimensional ordered system with short-range molecular interactions between the first neighbour molecules (as in a crystal). The short-range molecular interactions were presented by the parameter binding energy Q of an amphiphile molecule in the bilayer. The binding energy Q of two neighbouring phospholipids was calculated for the gel (30-60 kT) and liquid crystalline state (16-18 kT) of the bilayers from DMPC, DPPC, Egg PC, AF. Concentration/temperature phase diagram of DPPC foam bilayers that defined regions of gaseous (ruptured), gel and liquid crystalline foam bilayers were drawn. The values of Q obtained for various samples were very close and vary from 5.3 x 10(-20) to 9.4 x 10(-20) (approx. 13-22 kT) which indicated that in all cases the foam bilayers were in liquid-crystalline state. This is an important result since the parameter studied-threshold concentration (threshold dilution) is crucial for a very successful assessment of the risk for respiratory distress syndrome (RDS) in newborns and could be employed in medicine for assessment of other respiratory disturbances. It is to be expected that foam bilayers from phospholipids could be used as a model for investigation of short-range forces in biological structures, of interaction between membranes, etc.     

Remodeling of ordered membrane domains by GPI-anchored intestinal alkaline phosphatase

Glycosylphosphatidyl-inositol (GPI)-anchored proteins preferentially localize in the most ordered regions of the cell plasma membrane. Acyl and alkyl chain composition of GPI anchors influence the association with the ordered domains. This suggests that, conversely, changes in the fluid and in the ordered domains lipid composition affect the interaction of GPI-anchored proteins with membrane microdomains. Validity of this hypothesis was examined by investigating the spontaneous insertion of the GPI-anchored intestinal alkaline phophatase (BIAP) into the solid (gel) phase domains of preformed supported membranes made of dioleoylphosphatidylcholine/dipalmitoylphosphatidylcholine (DOPC/DPPC), DOPC/sphingomyelin (DOPC/SM), and palmitoyloleoylphosphatidylcholine/SM (POPC/SM). Atomic force microscopy (AFM) showed that BIAP inserted in the gel phases of the three mixtures. However, changes in the lipid composition of membranes had a marked effect on the protein containing bilayer topography. Moreover, BIAP insertion was associated with a net transfer of phospholipids from the fluid to the gel (DOPC/DPPC) or from the gel to the fluid (POPC/SM) phases. For DOPC/SM bilayers, transfer of lipids was dependent on the homogeneity of the gel SM phase. The data strongly suggest that BIAP interacts with the most ordered lipid species present in the gel phases of phase-separated membranes. They also suggest that GPI-anchored proteins might contribute to the selection of their own microdomain environment.     

Methotrexate interaction with a lipid membrane model of DPPC

Dipalmitoylphosphatidylcholine (DPPC) liposomes were employed as membrane models for the investigation of the interaction occurring between methotrexate (MTX) and bilayer lipid matrix. Liposomes were obtained by hydrating a lipid film with 50 mM Tris buffer (pH 7.4). The differential scanning calorimetry (DSC) evaluation of the thermotropic parameters associated with the phase transitions of DPPC liposomes gave useful information about the kind of drug-membrane interaction. The results showed an electrostatic interaction taking place with the negatively charged molecules of MTX and the phosphorylcholine head groups, constituting the outer part of DPPC bilayers. No interaction with the hydrophobic phospholipid bilayer domains was detected, revealing a poor capability of MTX to cross through lipid membranes to reach the interior compartment of a lipid bounded structure. These findings correlate well within vitro biological experiments on MTX cell susceptibility.     

Relationship between lipid peroxidation and rigidity in L-alpha-phosphatidylcholine-DPPC vesicles

DPPC incorporation into egg-PC unilamellar vesicles reduces their oxidation rate beyond that expected from the unsaturated lipid dilution. Addition of the unsaturated lipids produces changes in the physical properties of the inner parts of the lipid bilayer, as sensed by fluorescence anisotropy of DPH, and in the hydrophilic/hydrophobic region, as sensed by the generalized polarization of laurdan. DPPC (30 mol%) incorporation into egg-PC vesicles produces a decrease in alkyl chain mobility in the inner part of the bilayer, evaluated by the increase of DPH fluorescence anisotropy, and a rise of the generalized polarization value of laurdan in the bilayer interface. It also leads to a decrease in the rate of water efflux promoted by a hypertonic shock. Oxidation of PC LUVs, promoted by AAPH, as sensed by oxygen uptake and MDA formation, leads to qualitatively similar results than DPPC addition: rigidification at the inner part and the surface of the liposomes, and a lower rate of water permeation. It is suggested that these changes could contribute to the observed decrease in oxidation rate with conversion.     

Sodium Taurocholate-Induced Lamellar-Micellar Phase Transitions of DPPC

The thermotropic transitions of 1,2-dipalmitoylphosphatidylcholine (DPPC) and the structural changes of its lamellar phases have been studied between 0 and 50°C by both DSC and synchrotron small angle X-ray diffraction/scattering as a function of temperature (XRDT) and sodium taurocholate concentration [TC] in the 0–40 mM range ([DPPC]=50 mM) at pH 7.4. The existence of multiple phase transitions (up to 5 peaks within a 5°C interval) in a narrow domain of temperature between 25 and 42°C depending on the [TC]/[lipid] ratio was observed in the DSC curves. XRDT showed that at low ratios they might correspond to transitions between lamellar phases, the structural characteristics of which are given. At higher ratios a lamellar to micellar transition was observed, and the temperature at which it was observed decreased as a function of the TC content. The relationships with DPPC vesicle bilayer permeabilization and solubilization are discussed.     

Molecular aspects of the interaction between plants sterols and DPPC bilayers: an experimental and theoretical approach

The effect of sterols composition in a lipid bilayer was investigated on membranes of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and mixtures with the plant sterols β-sitosterol and stigmasterol. Differential scanning calorimetry, 1,6-diphenyl-1,3,5-hexatriene (DPH) fluorescence polarization and infrared spectroscopy studies showed that both sterols changed the packing of the membrane and the hydration of the polar headgroup of the phospholipids, disordering the gel phase and, vice versa, ordering the membrane in the liquid crystalline phase. In all cases some differences among β-sitosterol and stigmasterol could be observed, being β-sitosterol slightly more efficient than stigmasterol in ordering a fluid membrane, bringing the membrane to a more packed liquid ordered phase. Molecular dynamic simulations were carried out to better characterize the distinct behavior of both sterols in a DPPC-membrane. The calculated parameters agreed quite well with the experimental results and a molecular model is proposed to explain differences in the sterols molecules and their effect on the DPPC-bilayer.     

Interfacial interactions between poly[L-lysine]-based branched polypeptides and phospholipid model membranes

The interaction of five poly[L-lysine]-derived branched chain polypeptides of poly[Lys(X(i))] (X(i)K) or poly[Lys(X(i)-DL-Ala(m))] (XAK) with lipid bilayers (DPPC and DPPC/PG, 8:2) was studied by fluorescence polarization techniques. Two fluorescent probes, DPH and TMA-DPH, were utilized to monitor changes of motion in the internal and/or in the polar head regions, respectively. Results indicate that the interaction of polypeptides with neutral (DPPC) bilayers is mainly dependent on the polarity and electrical charge of side chains. The amphoteric E(i)K shows the highest level of interaction. Polycationic polypeptides (H(i)K, P(i)K, TAK) have a relatively small effect on the transition temperature of the lipids, while the polyanionic Succ-EAK has no effect at the alkyl chain region of the bilayer. Data with TMA-DPH indicate the lack of pronounced interaction between the polypeptides and the outer surface of the liposome. Similar tendency was documented for DPPC/PG vesicles. Polypeptides, H(i)K, and P(i)K induce significant changes in the transition temperature, thus indicating their insertion into the hydrophobic core of the bilayer without marked effect on the polar head region. Results suggest that these polypeptides (except E(i)K) have no destabilizing effect on liposomes studied. These properties are considered as beneficial for their use as safe carriers for bioactive molecules.     

Liquid Ordered Phase of Binary Mixtures Containing Dipalmitoylphosphatidylcholine and Sterols

The effect of cholesterol, desmosterol, stigmasterol, sitosterol, ergosterol, and androsterol on the phase behavior of aqueous dispersions of dipalmitoylphosphatidylcholine (DPPC) was studied to understand the role of the side chain in the formation of ordered phases of the type observed in membrane rafts. Thermotropic changes in the structure of mixed dispersions and transition enthalpies were examined by synchrotron X-ray diffraction (XRD) and differential scanning calorimetry (DSC). The observations indicated that cholesterol was more efficient than phytosterols (stigmasterol and sitosterol) or ergosterol in its interaction with DPPC to form the liquid ordered phase (L_o). The L_o induced by cholesterol or desmosterol was stable over a wide temperature range, whereas, the liquid ordered phase containing phytosterols or ergosterol was profoundly dependent on temperature, which should be distinguished as L_oβ and L_oα, representing the phases below and above the main transition temperature. The characteristics in forming ordered structures of cholesterol and other sterols imply that the evolution may have selected cholesterol as the most efficient sterol for animals to form rafts in their cell membranes.     

Phospholipid bilayer free volume analysis employing the thermal ring-closing reaction of merocyanine molecular switches

The free volume properties of phospholipid bilayers have been determined using a new assay that applies the photochromic and solvatochromic properties of merocyanines. The orientation and embedding depth of the merocyanines in the bilayer are controlled using substitution on the merocyanine indole moiety. The free volume changes at the aqueous interface (region 1), the phospholipid headgroup (region 2), and the aliphatic interior (region 3) of the bilayer are compared by analyzing the rate constants for the merocyanine ring-closing reaction. Free volume variations during the P(beta)(')(gel) <--> L(alpha)(liquid) phase transition are observed in region 1, in accordance with large structural rearrangements between the gel and the liquid phases in this region. The largest free volume is found in region 3, and the smallest is found in region 2. This distribution of free volume in the bilayer agrees with computational studies of these systems. Comparison of the free volume in region 2 of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) lipids shows that this method is sensitive to small structural differences between lipids. In region 2, the free volume is found to be approximately 2 times larger in DPPC bilayers, which could be related to different merocyanine interactions with the two phosphatidylcholines. Free volume properties determined on picosecond and second time scales are compared based on an analysis of merocyanine formation and decoloration reactions.     

Effects of silver nanoparticles on the fluidity of bilayer in phospholipid liposome

This paper describes the formation and characterization of liposome entrapping the silver nanoparticles in bilayer. Silver nanoparticles were entrapped in the bilayer of dipalmitoylphosphatidylcholine (DPPC) liposome, named as silver-loaded liposome. Specifically, above the gel to liquid-crystalline phase transition temperature of this lipid (i.e., 41 degrees C), it was observed that membrane fluidities of silver-loaded liposomes were increased, and fluorescence anisotropy values were reduced from 0.114 to 0.097. This might be due to the structural modifications and interactions between DPPC molecules and silver nanoparticles within the bilayer. It was also confirmed that silver nanoparticles were entrapped in hydrophobic region of lipid bilayer with transmission electron microscopy (TEM) and electron energy loss spectroscopy (EELS) measurements.     

Docosahexaenoic acid and eicosapentaenoic acid induce changes in the physical properties of a lipid bilayer model membrane

We investigated the effect of fatty acids such as stearic acid (SA, 18:0), oleic acid (OA, 18:1), eicosapentaenoic acid (EPA, 20:5), and docosahexaenoic acid (DHA, 22:6) on a dipalmitoylphosphatidylcholine (DPPC) bilayer by determining the phase transition temperature, fluorescence anisotropy of 1,6-diphenyl-1,3,5-hexatriene (DPH), and detergent insolubility. Treatment with unsaturated fatty acid broadened and shifted the phase transitions of the DPPC bilayer to a lower temperature. The phase transition temperature and the value of fluorescence anisotropy of DPH at 37 degrees C decreased progressively with increasing treatment amounts of unsaturated fatty acid. A large amount of the DPPC bilayer treated with unsaturated fatty acid was dissolved in Triton X-100, obtaining a low level of detergent insolubility. These modifications of the bilayer physical properties were most pronounced with DHA and EPA treatment. These data show that unsaturated fatty acids, particularly DHA and EPA, induce a marked change in the lipid bilayer structure. The composition of fatty acids in the DPPC bilayer was similar after treatment with various unsaturated fatty acids, suggesting that the different actions of unsaturated fatty acids are attributed to change in the molecular structure (e.g., kinked conformation by double bonds). We further explored the change in physical properties induced by fatty acids dispersed in a water-in-oil-in-water multiple emulsion and found that unsaturated fatty acids acted efficiently on the DPPC bilayer, even when incorporated in emulsion form.