The interaction of antimicrobial peptides with membranes.

The interaction of antimicrobial peptides (AMPs) with biological membranes is in the focus of research since several years, and the most important features and modes of action of AMPs are described in this review. Different model systems can be used to understand such interactions on a molecular level. As a special example, we use 2D and 3D model membranes to investigate the interaction of the natural cyclic (Ar-1) and the synthetic linear molecule arenicin with selected amphiphiles and phospholipids. A panoply of sophisticated methods has been used to analyze these interactions on a molecular level. As a general trend, one observes that cationic antimicrobial peptides do not interact with cationic amphiphiles due to electrostatic repulsion, whereas with non-ionic amphiphiles, the peptide interacts only with aggregated systems and not with monomers. The interaction is weak (hydrophobic interaction) and requires an aggregated state with a large surface (cylindrical micelles). Anionic amphiphiles (as monomers or micelles) exhibit strong electrostatic interactions with the AMPs leading to changes in the peptide conformation. Both types of peptides interact strongly with anionic phospholipid monolayers with a preference for fluid layers. The interaction with a zwitterionic layer is almost absent for the linear derivative but measurable for the cyclic arenicin Ar-1. This is in accordance with biological experiments showing that Ar-1 forms well defined stable pores in phospholipid and lipopolysaccharide (LPS) membranes (cytotoxicity). The synthetic linear arenicin, which is less cytotoxic, does not affect the mammalian lipids to such an extent. The interaction of arenicin with bacterial membrane lipids is dominated by hydrogen bonding together with electrostatic and hydrophobic interactions.

From Langmuir Monolayers to Multilayer Films.

This feature article is intended to describe a route from Langmuir monolayers as the most suitable and well-defined models to polyelectrolyte multilayers. The latter are structurally controlled not with angstrom but with nanometer precision; however, they are very modular with regard to building blocks and function and are robust, therefore promising many diverse applications. There have been many methods developed to structurally characterize Langmuir monolayers; therefore, they serve as models in membrane biophysics and materials science as well as in general physics as two-dimensional model systems. Many of these methods as well as ideas to control interfaces could be taken over to study polyelectrolyte multilayers with their extended internal interfaces. Finally, as an outlook we try to sketch various aspects to transit toward systems with higher structural hierarchy, enabling the coupling of different functions and arriving at responsive three-dimensional systems.

The evaporation behavior of sessile droplets from aqueous saline solutions.

Quantitative experiments on the evaporation from sessile droplets of aqueous saline (NaCl) solutions show a strong dependence on salt concentration and droplet shape. The experiments were performed with seven decades of initial NaCl concentrations, with various droplet sizes and with different contact angles. The evaporation rate is significantly lower for high salt concentrations and small contact angles than what is expected from the well-accepted diffusion-controlled evaporation scenario for sessile droplets, even if the change of the vapor pressure due to the salt is taken into account. Particle tracking velocimetry reveals that this modification of the evaporation behavior is caused by marangoni flows that are induced by surface tension gradients originating from the local evaporative peripheral salt enrichment. In addition it is found that already very low salt concentrations lead to a pinning of the three phase contact line. Whereas droplets with concentration ≥10(-6) M NaCl are pinned as soon as evaporation starts, droplets with lower salt concentration do evaporate in a constant contact angle mode. Aside from new, fundamental insights the findings are also relevant for a better understanding of the widespread phenomenon of corrosion initiated by sessile droplets.

Prospects for plasmonic hot spots in single molecule SERS towards the chemical imaging of live cells.

Single molecule surface enhanced Raman scattering (SM-SERS) is a highly local effect occurring at sharp edges, interparticle junctions and crevices or other geometries with a sharp nanoroughness of plasmonic nanostructures ("hot spots"). The emission of an individual molecule at SM-SERS conditions depends on the local enhancement field of the hot spots, as well as the binding affinity and positioning at a hot spot region. In this regard, the stability of near-field nano-optics at hot spots is critical, particularly in a biological milieu. In this perspective review, we address recent advances in the experimental and theoretical approaches for the successful development of SM-SERS. Significant progress in the understanding of the interaction between the excitation electromagnetic field and the surface plasmon modes at the metallic or metallic/dielectric interface of various curvatures are described. New knowledge on methodological strategies for positioning the analytes for SM-SERS and Raman-assisted SERS or the SERS imaging of live cells has been acquired and displayed. In the framework of the extensive development of SM-SERS as an advancing diagnostic analytical technique, the real-time SERS chemical imaging of intracellular compartments and tracing of individual analytes has been achieved. In this context, we highlight the tremendous potential of SERS chemical imaging as a future prospect in SERS and SM-SERS for the prediction and diagnosis of diseases.

Microcontact-printing-assisted access of graphitic carbon nitride films with favorable textures toward photoelectrochemical application.

An "ink" (cyanamide) infiltrated anodic aluminum oxide (AAO) stamp is found capable of printing carbon nitride films featuring regular microstructures of the stamp onto the substrates via in situ "chemical vapor deposition". A photocurrent density of 30.2 µA cm(-2 --) at 1.23 VRHE is achieved for a film on a conductive substrate, which is so far the highest value for pure carbon nitride based photoelectrochemical devices.

Highly effective hot spots for SERS signatures of live fibroblasts.

Pre-formed silver-boron nanoparticles of 22 nm form pearl-like necklace nanostructures with interparticle junctions of less than 10 nm length in the matrix of polyethylene glycol (8000 Da). The silver necklace nanostructure is stable at 37 °C or 70 °C and also inside a live cell medium. A polyethylene glycol matrix with a shorter chain length (1000 Da) does not protect the nanoparticles against attraction, and random aggregates are formed. Silver necklace nanostructures exhibit strong Raman enhancement by more than ∼10(9) which is much higher than for silver-citrate or random silver-boron aggregates. The polymeric matrix of 8000 Da contributes strongly to the electromagnetic field enhancement and removes the chemical contribution to the surface Raman scattering increase. The stable interparticle junctions act as local hot spots for strong Raman scattering signals collected from live fibroblasts and allow systematic in situ studies.

Preparation of multifunctional polysaccharide microcontainers for lipophilic bioactive agents.

Chitosan/xanthan gum microcontainers with a core-shell structure formed due to chemical interactions between polysaccharide chains induced by ultrasonication are presented. Containers were prepared by sonication of water-immiscible (oil-like) liquids in the solution of polysaccharides. One-step fabrication of the container permanent shell is possible, because of the contribution of ultrasonically caused formation of hydrogen bonds and amide linkages. We synthesized containers in a wide size range from 350 nm to 7500 nm, varying in oil/water ratio. The microcontainers were modified with oppositely charged polyelectrolytes and microparticles, which could be used to impart the specified properties to the system. The biocide 4,5-dichloro-2-n-octyl-4-isothiazoline-3-one (DCOIT) was loaded into the proposed containers by utilizing its solution as an oil phase. The following incorporation of the DCOIT containers into the polymer coating demonstrated more sustained antimicrobial activity (∼30%) of the biocide in the encapsulated state, compared to its non-encapsulated form.

Why fluorination of the polar heads reverses the positive sign of the dipole potential of Langmuir monolayers: a vibrational sum frequency spectroscopic study.

Natural nonionic amphiphiles forming monolayers, bilayers, micelles, or biomembranes create a positive dipole potential at the boundary with water. In a series of papers we have reported on Langmuir monolayers with CF3 terminals of the polar heads, which show a negative surface dipole potential ΔV (Petrov , J. G.; Andreeva, T. D.; Kurt, D. K.; Möhwald, H. J. Phys. Chem. B 2005, 109, 14102). Here we use vibrational sum frequency spectroscopy (SF) to study the origin of the opposite ΔV signs of Langmuir films of CH3(CH2)20COCH2CH3 (ethyl ether, EE) and CH3(CH2)20COCH2CF3 (fluorinated ethyl ether, FEE). The vibrational sum frequency spectra are recorded at the same film density of the S-phase of the EE and FEE monolayers and analyzed in the spectral regions of OH, COC, CH3, and CF3 stretching vibrations because these functional groups could be responsible for the different dipole potentials. We compare the rearrangement of the pure water surface by EE and FEE monolayers and the conformations of EE and FEE polar heads. The analysis is performed according to the three-capacitor model of the dipole potential of Langmuir monolayers (Demchak, R. T.; Fort, T., Jr. J. Colloid Interface Sci. 1974, 46, 191). The results show that reversal of the ΔV sign caused by fluorination of the polar heads originates from the upward-oriented CF3 terminals of the FEE heads, whose negative normal dipole moment component determines the negative dipole potential of the FEE monolayer.

Nanoparticle modification by weak polyelectrolytes for pH-sensitive pickering emulsions.

The affinity of weak polyelectrolyte coated oxide particles to the oil-water interface can be controlled by the degree of dissociation and the thickness of the weak polyelectrolyte layer. Thereby the oil in water (o/w) emulsification ability of the particles can be enabled. We selected the weak polyacid poly(methacrylic acid sodium salt) and the weak polybase poly(allylamine hydrochloride) for the surface modification of oppositely charged alumina and silica colloids, respectively. The isoelectric point and the pH range of colloidal stability of both particle-polyelectrolyte composites depend on the thickness of the weak polyelectrolyte layer. The pH-dependent wettability of a weak polyelectrolyte-coated oxide surface is characterized by contact angle measurements. The o/w emulsification properties of both particles for the nonpolar oil dodecane and the more polar oil diethylphthalate are investigated by measurements of the droplet size distributions. Highly stable emulsions can be obtained when the degree of dissociation of the weak polyelectrolyte is below 80%. Here the average droplet size depends on the degree of dissociation, and a minimum can be found when 15 to 45% of the monomer units are dissociated. The thickness of the adsorbed polyelectrolyte layer strongly influences the droplet size of dodecane/water emulsion droplets but has a less pronounced impact on the diethylphthalate/water droplets. We explain the dependency of the droplet size on the emulsion pH value and the polyelectrolyte coating thickness with arguments based on the particle-wetting properties, the particle aggregation state, and the oil phase polarity. Cryo-SEM visualization shows that the regularity of the densely packed particles on the oil-water interface correlates with the degree of dissociation of the corresponding polyelectrolyte.

Dipolar interactions and miscibility in binary Langmuir monolayers with opposite dipole moments of the hydrophilic heads.

We investigate unusual binary Langmuir monolayers with the same long CH3(CH2)21 hydrocarbon chains and fluorinated -O-CH2CF3 (FEE) versus nonfluorinated -O-CH2CH3 (EE) hydrophilic heads, whose opposite dipoles assist miscibility, in contrast to the equally oriented polar head dipoles of almost all natural or synthetic amphiphiles that minister to phase separation. Although two-component bulk micelles, lipid bilayers, and monolayers with fluorinated and nonfluorinated chains, which also have opposite dipoles, often show phase separation, we find complete miscibility and nonideality of the FEE-EE mixtures demonstrated via deviation of the composition dependencies of the mean molecular area at fixed surface pressure from the additivity rule. The composition dependencies of the excess molecular areas exhibit minima and maxima which show specific structural changes at particular compositions. They originate from the dipolar and steric interactions between the polar heads, because the interactions between the same chains of FEE and EE do not vary. The pi/A isotherms and the pi/X(FEE) phase diagram reveal that mixtures with molar fractions X(FEE) > or = 0.3 exist in an upright solid phase even in uncompressed state. This result is confirmed by the compressibility values and via Brewster angle microscopy, which does not show optical anisotropy at X(FEE) > or = 0.3. Comparison of the collapse and phase-transition molecular areas with literature data suggests that the upright architecture corresponds to LS-phase or S-phase with more defects as the S-phase in the pure monolayers. The mixtures with X(FEE) < 0.3 exist in tilted L2' phase at low surface pressures. Their mean molecular areas are smaller than the corresponding values in the EE film, which manifests reduction of the tilt of the EE chains with increasing FEE content. We ascribe the chain erection to partial dehydration of the EE heads caused by dipolar attraction between the EE and FEE heads. The excess free energy of mixing deltaG(exc)pi is positive but much smaller than the negative total free energy of mixing AG mix(pi) showing a spontaneous miscibility at all compositions due to an entropy increase. The analysis of the conflict between the deltaG(mix)pi minimum at molar fraction X(FEE) = 0.5 and the minimum and negative value of the excess molecular area A(pi,exc) at X(FEE) = 0.8 shows that the A(pi,exc)/X(FEE) minimum has not an electrostatic but a short-range structural origin.

Structure of the Langmuir monolayers with fluorinated ethyl amide and ethyl ester polar heads creating dipole potentials of opposite sign.

This study experimentally checks our previous hypothesis (Petrov, J. G.; Polymeropoulos, E. E.; Moehwald, H. Langmuir 2007, 23, 2623) that different conformations of the fluorinated heads of RCONHCH(2)CF(3) and RCOOCH(2)CF(3) monolayers cause the opposite signs and the striking difference of 1.480 V between their surface potentials Delta V. In situ X-ray diffraction at grazing incidence (GIXD) shows that both monolayers form orthorhombic lattices with closely packed chains tilted to the next-nearest neighbors in the RCONHCH(2)CF(3) film and upright in the RCOOCH(2)CF(3) monolayer. The packing of the chains in the plane perpendicular to them, which excludes the effect of the tilt, shows the same distance between the next-nearest neighbors, but significantly closer nearest neighbors in the RCONHCH(2)CF(3) film. This difference implies a specific anisotropic attraction between the adjacent amide heads. IR reflection absorption spectroscopy (IRRAS) shows that the -CONHCH(2)CF(3) heads have trans conformation and participate in H-bonds forming a -NH...O=C- lateral network. We speculate that such structure hinders the energetically optimal orientation of the hydrophobic -CH(2)CF(3) terminals toward air, so that the (delta+)C-(F (delta-))(3) dipoles at the monolayer/water boundary yield a strong positive contribution to Delta V. In contrast, most of the unbounded by H-bonds -COOCH(2)CF(3) heads statistically orient their hydrophobic (delta+)C-(F (delta-))(3) dipoles toward air, yielding a negative average dipole moment at the monolayer/water boundary and negative surface dipole potential.

Composite multilayered biocompatible polyelectrolyte films with intact liposomes: stability and temperature triggered dye release.

The design of new quasi-2D biocompatible films able to release a drug in a controlled manner through the application of physical stimuli is of outstanding interest in biomaterials science. Herein, construction of composite nanofilms with multiple strata of stabilized large unilamellar liposomes is developed. The film has a multilayered architecture formed by the layer-by-layer (LbL) technique utilising two biocompatible polyelectrolytes, hyaluronic acid and poly-l-lysine (HA and PLL), onto which phospholipid liposome "interlayers" are adsorbed and subsequently embedded by further polyelectrolyte adsorption. First of all the morphology of the film surface is characterised and compared to the morphology obtained for a film containing the same polyelectrolytes but stiff polystyrene (PS) latex particles instead of the soft phospholipid vesicles. Both morphologies appear to be similar suggesting that the embedded vesicles keep their spherical shape. As a second step, it has been shown that carboxyfluorescein (CF) encapsulated in the vesicles remains inside without sustained release at room temperature indicating that the liposomes stay intact upon LbL immobilization. A substantial fraction (about 70%) of the vesicles absorbed onto the polyelectrolyte film are desorbed upon further polyelectrolyte adsorption to reach the embedded state. Hence to achieve high loading of the film, multiple vesicle depositions have to be performed. It is shown that the total number of deposited vesicles on the film is proportional to the number m of vesicle deposition steps (up to m = 3). The encapsulated dye can be released in a controlled manner by a temperature increase above the main phase transition temperature of the lipid mixture used to prepare the vesicles. Release kinetics is faster for film-entrapped vesicles than for those in solution due to destabilization of the lipidic bilayer by the polyelectrolyte environment. The developed composite films represent a new surface biocoating able to preserve, in native state, or to release in a controlled way, surface immobilized materials under external stimuli.