脂双层膜表面结构与稳定性的原子力显微镜研究

用原子力显微镜研究了1,2－二油酸甘油－3－磷酸－1甘油（DOPG）脂双层膜 的表面结构与稳定性。实验结果表明,原子力显微镜的探针与脂双层膜的相互作用 导致脂双层膜表面产生一个永久的损伤。静电相互作用对脂双层膜结构和稳定性的 影响表明,在NaCl溶液中制成的脂质体,随着NaCl浓度的增加,它们的双层膜更稳 定。在低的NaCl浓度则经常被损伤,在1 mol/L NaCl溶液中制备的指双层变得更稳 定。在KCl溶液中结果恰好相反。在高的KCl浓度中经常被损伤,随着KCl浓度的降 低,它们的双层膜更稳定。葡萄糖和蔗糖对脂双层膜结构有稳定作用。     

Effects of synthetic lipids on solubilization and colloid stability of hydrophobic drugs

Aqueous miconazole (MCZ) aggregates were solubilized and/or colloidally stabilized by bilayer-forming synthetic amphiphiles such as dioctadecyldimethylammonium bromide (DODAB) or sodium dihexadecylphosphate (DHP) dispersions. Particle sizing, light absorption and scattering from drug particles, zeta-potential determination, and drug aggregation kinetics from turbidity changes in the presence or absence of lipid dispersions were obtained over a range of drug and lipid concentrations. The very low solubility of MCZ in water made possible the determination of size distributions for drug particles in water and comparison to those in the presence of DODAB or DHP nanosized bilayer fragments or entire and closed bilayer vesicles. Large drug aggregates disappeared upon incubation with nanosized bilayer fragments produced by ultrasonic dispersion with tip. Light-absorption spectra for MCZ in a poor solvent (water), in a good organic solvent (methanol), and in different lipid dispersions showed that solubilization depended on the presence of bilayer fragments. MCZ was poorly soluble in dispersions formed of closed bilayers (vesicles) of DODAB or DHP in the gel state and in phosphatidylcholine (PC) vesicles in the liquid-crystalline state. Increased hydrophobicity at the borders of bilayer fragments explained MCZ solubilization. At [MCZ]>0.4 mM, kinetics of drug aggregation, zeta-potential measurements, and size minimization were obtained upon addition of minute amounts of oppositely charged bilayer fragments ([DHP]=0.05 mM), making possible determination of a remarkable stabilizing effect of drug particles by coverage with anionic bilayer fragments. High drug colloid stability in the presence of charged bilayer fragments was achieved by two different means: (1). at large drug concentrations and small concentrations of bilayer fragments, coverage of large drug particles with bilayer fragments; (2). at large amounts of bilayer fragments, drug solubilization in its monomeric form at the borders of bilayer fragments. Inexpensive, synthetic bilayer fragments offered a large area of hydrophobic nanosurfaces dispersed and electrostatically stabilized in water, opening new prospects for drug solubilization and colloid stabilization of insoluble drug particles.     

Loading of gold nanoparticles inside the DPPC bilayers of liposome and their effects on membrane fluidities

Gold nanoparticles were loaded in the bilayer of dipalmitoylphosphatidylcholine (DPPC) liposomes, named as gold-loaded liposomes. Above the gel to liquid-crystalline phase transition temperature, membrane fluidities of DPPC liposomes were changed by loading the gold nanoparticles. Compared with liposomes without loading the gold nanoparticles, gold-loaded liposomes showed the lower fluorescence anisotropy values. That is, the membrane fluidities of DPPC bilayer were increased by loading the gold nanoparticles. The membrane fluidities were increased as the amount of gold nanoparticles increased. The existence of gold nanoparticles in the DPPC bilayer was observed by transmission electron microscopy. Through the energy dispersive X-ray spectrometer, the particles in DPPC bilayer were confirmed to be gold nanoparticles.     

On the stability of membrane,foam and emulsion bilayers with respect to rupture by hole nucleation

Bilayer membranes and bilayer foam or emulsion films were considered thermodynamically from a unified point of view as two-dimensional phases. The work of fluctuation formation of holes in a bilayer was determined and it was shown that the bilayer can be either stable or unstable with respect to rupture by hole nucleation. When the bilayer is in contact with a solution, its stability with respect to rupture was found to decrease with decreasing concentration of dissolved amphiphiles, and a limiting concentration was shown to exist, below which the bilayer is subject to rupture by hole nucleation. The connection is discussed between existing formulae for the work of fluctuation formation of holes in bilayers.     

Revealing the Preferred Interlayer Orientations and Stackings of Two‐Dimensional Bilayer Gallium Selenide Crystals

Characterizing and controlling the interlayer orientations and stacking orders of two‐dimensional (2D) bilayer crystals and van der Waals (vdW) heterostructures is crucial to optimize their electrical and optoelectronic properties. The four polymorphs of layered gallium selenide (GaSe) crystals that result from different layer stackings provide an ideal platform to study the stacking configurations in 2D bilayer crystals. Through a controllable vapor‐phase deposition method, bilayer GaSe crystals were selectively grown and their two preferred 0° or 60° interlayer rotations were investigated. The commensurate stacking configurations (AA′ and AB stacking) in as‐grown bilayer GaSe crystals are clearly observed at the atomic scale, and the Ga‐terminated edge structure was identified using scanning transmission electron microscopy. Theoretical analysis reveals that the energies of the interlayer coupling are responsible for the preferred orientations among the bilayer GaSe crystals.     

Creating fluid and air-stable solid supported lipid bilayers

Solid supported lipid bilayers are rapidly delaminated when drawn through the air/water interface. We have discovered that a close packed monolayer of specifically bound protein prevents this process. The protection mechanism worked in two ways. First, when protein-protected bilayers were drawn through the air/water interface, a thin bulk water layer was visible over the entire bilayer region, thereby preventing air from contacting the surface. Second, a stream of nitrogen was used to remove all bulk water from a protected bilayer, which remained fully intact as determined by fluorescence microscopy. The condition of this dried bilayer was further probed by fluorescence recovery after photobleaching. It was found that lipids were not two-dimensionally mobile in dry air. However, when the bilayer was placed in a humid environment, 91% of the bleached fluorescence signal was recovered, indicating long-range two-dimensional mobility. The diffusion coefficient of lipids under humid conditions was an order of magnitude slower than the same bilayer under water. Protected bilayers could be rehydrated after drying, and their characteristic diffusion coefficient was reestablished. Insights into the mechanism of bilayer preservation were suggested.     

Sustained-release phenylpropanolamine hydrochloride bilayer caplets containing the hydroxypropylmethylcellulose 2208 matrix. II. Effects of filling order in bilayer compression and manufacturing method of the prolonged-release layer on compactibility of bilayer caplets

The purpose of this study was to establish the manufacturing method of the formulated bilayer caplets containing the hydroxypropylmethylcellulose 2208 (HPMC2208) matrix without lamination. In manufacturing the bilayer caplets containing the HPMC2208 (Metolose 90SH4000) matrix, some bilayer caplets were cracked. We found that cracking of bilayer caplets is not the separation of two layers, but lamination of the prolonged-release layer. It was assumed that Metolose 90SH4000 causes lamination of the prolonged-release layer. Two factors, roller compaction pressure on dry granulation of the prolonged-release layer and filling order of the prolonged-release layer in bilayer compression, were related to lamination of bilayer caplets. The compactibility of the prolonged-release layer decrease with an increase in roller compaction pressure on dry granulation. The compactibility of the prolonged-release layer manufactured by direct compression is superior to that manufactured by dry granulation. The compactibility of the prolonged-release layer in the shape of the second layer, convexo-concave, is superior to that in the shape of the first layer, convexo-convex. This is due to the fact that the density distribution inside the compact in the shape of convexo-concave was more uniform than that in the shape of convexo-convex. The manufacturing method of the formulated bilayer caplets having the prolonged-release layer whose Metolose 90SH4000 content is 60% without lamination is as follows: the prolonged-release layer manufactured by direct compression is fed as the second layer in bilayer compression.     

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.     

Interaction of cytolytic toxin CytB with a supported lipid bilayer: study using an acoustic wave device

An acoustic technique was used to monitor the interaction of the pore-forming cytolytic toxin CytB with a positively charged supported lipid bilayer. The acoustic device, which is based on a waveguide geometry, is sensitive to changes in the mass of the supported bilayer. The specificity of the interaction, rate and extent of the association, reversibility and effect of previous depositions of toxin were investigated. The CytB was found to bind irreversibly to the lipids at all fractional coverages even when the protein-to-lipid ratio was high enough to imply that the protein was associating with the external surface of the bilayer. The CytB formed stable structures with the bilayer at high protein surface concentrations and did not appear to disrupt the bilayer in the manner of a detergent. The rate of association with the bilayer was found to be directly proportional to the solution concentration of CytB at higher concentrations but appeared to be low at a CytB solution concentration of 5 microg mL(-1), leading to relatively low amounts of CytB being associated with the bilayer.     

Detection of membrane biointeractions based on fluorescence superquenching

Assays for biointeractions of molecules with supported lipid bilayers using fluorescence superquenching are described. A conjugated cationic polymer was adsorbed on to silica microspheres, which were then coated with an anionic lipid bilayer. The lipid bilayer attenuated superquenching by acting as a barrier between the conjugated polymer and its quencher. Biointeractions of the lipid bilayer with a membrane lytic peptide, melittin, were detected and quantitated by superquenching of the conjugated polyelectrolyte in flow cytometric and microfluidic bioassays. A higher sensitivity for detecting melittin lysis of the lipid bilayer at lower concentrations and shorter times for melittin action was found using flow cytometry in this study in comparison to other existing methods. This study combined the sensitivity of superquenching and flow cytometry to detect biointeractions with a lipid bilayer, which serves as a platform for developing functional assays for sensor applications, lipid enzymology, and investigations of molecular interactions. In addition, this study demonstrated proof-of-concept for using superquenching detected as a result of lipid bilayer disruption in a microfluidic format.     

硫醇－磷脂混合双层膜的电化学性质及其与蜂毒素的相互 作用研究

报道了硫醇－磷脂混合双层膜的循环伏安和电化学交流阻抗行为研究,并用电化学方法考察了蜂毒素与其相互作用,实验中通过冷冻表面沾有磷脂溶液的硫醇单层膜制备混合双层膜,研究表明双层膜在电极表面形成致密的绝缘层,阻碍了电极表面的电子传递,在双层膜体系上引入的蜂毒素可在膜表面上形成孔洞,破坏膜的绝缘性,降低膜电阻,增加膜电容,使带负电的探针Fe(CN)6^3-的氧化还原反应速度加快。     

Potential-driven structural changes in Langmuir-Blodgett DMPC bilayers determined by in situ spectroelectrochemical PM IRRAS

Combined Langmuir-Blodgett vertical withdrawing and Langmuir-Schaefer horizontal touch (LB-LS) methods were employed to transfer DMPC bilayers onto a Au(111) electrode surface. Charge density measurements and photon polarization modulation infrared reflection absorption spectroscopy were employed to investigate electric field induced changes in the structure of the bilayer. The results show that the physical state and the molecular arrangement found in the monolayer at the air-water interface is to a large extent preserved in the bilayer formed by the LB-LS method. This approach provides an opportunity to produce supported bilayers with a well-designed architecture. The properties of the bilayer formed by the LB-LS method were compared to the properties of the bilayer produced by spontaneous fusion of unilamellar vesicles investigated in an earlier study (Bin, X.; Zawisza, I.; Lipkowski, J. Langmuir 2005, 21, 330-347). The tilt angles of the acyl chains are much smaller in the bilayer formed by the LB-LS method and are closer to the angles observed for vesicles and stacked hydrated bilayers. The tilt angles of the phosphate and choline groups are also smaller and are characteristic of an orientation in which the area per DMPC molecule is small. The electric field induced changes of these angles are also less pronounced in the bilayer formed by the LB-LS method. We have shown that these differences are a result of the higher packing density of the phospholipid molecules in the bilayer formed by the LB-LS method.     

卵磷脂与四乙酰氧基苯基卟啉金属配合物的相互作用

本文用~(31)P.NMR和~1HNMR谱分析了卵磷脂的组分和结构,并以小角X射线散射法(SAXS)研究了所合成的六种四乙酰氧基卟啉金属配合物与卵磷脂的相互作用,发现卟啉分子镶嵌于磷脂双层的疏水链之间,使双分子层间距变大,而金属卟啉分子因其与磷脂的极性头基的静电相互作用,所形成的磷脂双分子层的间距介于纯卵磷脂和含有卟啉分子的卵磷脂所构成的双分子层之间.     

Using bicellar mixtures to form supported and suspended lipid bilayers on silicon chips

Bicellar mixtures, planar lipid bilayer assemblies comprising long- and short-chain phosphatidylcholine lipids in suspension, were used to form supported lipid bilayers on flat silicon substrate and on nanotextured silicon substrates containing arrays of parallel troughs (170 nm wide, 380 nm deep, and 300 nm apart). Confocal fluorescence and atomic force microscopies were used to characterize the resulting lipid bilayer. Formation of a continuous biphasic undulating lipid bilayer membrane, where the crests and troughs corresponded to supported and suspended lipid bilayer regions, is demonstrated. The use of interferometric lithography to fabricate nanotexured substrates provides an advantage over other nanotextured substrates such as nanoporous alumina by offering flexibility in designing different geometries for suspending lipid bilayers.     

DEPOSITION OF METALLIC and SEMICONDUCTING LAYERS ONTO BILAYER LIPID MEMBRANES

In this paper the phenomenon of inorganic substances deposited onto the bilayer lipid membrane is described. Metallic copper and semiconducting compounds such as CuS, FeS, CdS and also AgBr were deposited onto the bilayer lipid membrane. The cyclic voltammetry technique was used for determining electrochemical and photoelectrical properties of coated membranes. Considerable increase of stability and drastic changes of membrane properties were observed. Also investigated were the photoelectrical properties of bilayer lipid membrane with dispersed CdSe or AgBr particles in the absence and presence of pigments.     

Controlled delivery of proteins into bilayer lipid membranes on chip

The study and the exploitation of membrane proteins for drug screening applications requires a controllable and reliable method for their delivery into an artificial suspended membrane platform based on lab-on-a-chip technology. In this work, a polymeric device for forming lipid bilayers suitable for electrophysiology studies and biosensor applications is presented. The chip supports a single bilayer and is configured for controlled protein delivery through on-chip microfluidics. In order to demonstrate the principle of protein delivery, the potassium channel KcsA was reconstituted into proteoliposomes, which were then fused with the suspended bilayer on-chip. Fusion of single proteoliposomes with the membrane was identified electrically. Single channel conductance measurements of KcsA in the on-chip bilayer were recorded and these were compared to previously published data obtained with a conventional planar bilayer system.     

双层类脂膜及其在电化学生物传感器中的应用

详细评述了各种双层类脂膜包括传统的双层类脂膜（ＢＬＭ）、固体载体支撑的自组双层类脂膜（ｓ－ＢＬＭ）、固体载体支撑的混合双层类脂膜（ｅ－ＢＬＭ）的制备方法和特性,比较了其优缺点。介绍了双层类脂膜在电化学生物传感器中的应用,并展望了发展前景。     

Electron paramagnetic resonance studies of an integral membrane peptide inserted into aligned phospholipid bilayer nanotube arrays

This communication reports for the first time the determination of the helical tilt of an integral membrane peptide inserted into aligned phospholipids bilayer nanotube arrays using spin label EPR spectroscopy. Also, we demonstrate herein how the helical tilt of the peptide can be easily calculated using the hyperfine splitting values gleaned from a perpendicularly aligned bilayer in phospholipid bilayer nanotube arrays. EPR spectral simulations were used to verify the method.     

The distribution and conformation of very long-chain plant wax components in a lipid bilayer

Plant wax contains long-chain alkanes and related components which are transported to the surface of the plant by specialized ABC transporter proteins. Here, we determine the distribution and conformation of three wax components, nonacosane, nonacosan-15-one, and nonacosan-15-ol, using unbiased and umbrella sampling molecular dynamics simulations. The molecules all partitioned to the center of the bilayer, with a free-energy difference of -70 kJ/mol between bulk water and the center of the bilayer for the alkane and -55 kJ/mol for the two more-polar molecules. All of the wax molecules were highly mobile in the bilayer, freely moving between opposite leaflets on a time scale of a few nanoseconds. Nonacosan-15-one and nonacosan-15-ol folded double to expose their hydrophilic group to the solvent, whereas nonacosane alternated between orientations spanning the full bilayer and orientations in the center of the bilayer.     

Probing into interesting effects of fractal Ge nanoclusters induced by Pd nanoparticles

Metal/semiconductor thin films are a class of unique materials that have widespread technological applications, particularly in the field of microelectronic devices. New strategies of fractal assessment for Pd/Ge bilayer films formed at various annealing temperatures are of fundamental importance in the development of micro/nanodevices. Herein, Pd/Ge bilayer films with interesting fractal nanoclusters were successfully prepared by evaporation techniques. Temperature-dependent properties of resistance and fractal dimensions in Pd/Ge bilayer films with self-similar Ge fractal nanoclusters were investigated in detail. Experimental results indicated that the fractal crystallization behavior and film resistance in Pd/Ge bilayer films are influenced significantly by annealing temperatures and fractal dimensions. The measurements of film resistance confirmed that there is an evident relationship between the film resistance and the fractal dimension. These phenomena were reasonably explained by the random tunneling junction network mechanism.