Susceptibility of Legionella gormanii Membrane-Derived Phospholipids to the Peptide Action of Antimicrobial LL-37-Langmuir Monolayer Studies.

LL-37 is the only member of the cathelicidin-type host defense peptide family in humans. It exhibits broad-spectrum bactericidal activity, which represents a distinctive advantage for future therapeutic targets. The presence of choline in the growth medium for bacteria changes the composition and physicochemical properties of their membranes, which affects LL-37's activity as an antimicrobial agent. In this study, the effect of the LL-37 peptide on the phospholipid monolayers at the liquid-air interface imitating the membranes of Legionella gormanii bacteria was determined. The Langmuir monolayer technique was employed to prepare model membranes composed of individual classes of phospholipids-phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylglycerol (PG), cardiolipin (CL)-isolated from L. gormanii bacteria supplemented or non-supplemented with exogenous choline. Compression isotherms were obtained for the monolayers with or without the addition of the peptide to the subphase. Then, penetration tests were carried out for the phospholipid monolayers compressed to a surface pressure of 30 mN/m, followed by the insertion of the peptide into the subphase. Changes in the mean molecular area were observed over time. Our findings demonstrate the diversified effect of LL-37 on the phospholipid monolayers, depending on the bacteria growth conditions. The substantial changes in membrane properties due to its interactions with LL-37 enable us to propose a feasible mechanism of peptide action at a molecular level. This can be associated with the stable incorporation of the peptide inside the monolayer or with the disruption of the membrane leading to the removal (desorption) of molecules into the subphase. Understanding the role of antimicrobial peptides is crucial for the design and development of new strategies and routes for combating resistance to conventional antibiotics.

Insight into the Mechanism of Interactions between the LL-37 Peptide and Model Membranes of Legionella gormanii Bacteria.

Legionella gormanii is a fastidious, Gram-negative bacterium known to be the etiological agent of atypical community-acquired pneumonia. The human cathelicidin LL-37 exhibits a dose-dependent bactericidal effect on L. gormanii. The LL-37 peptide at the concentration of 10 µM causes the bacteria to become viable but not cultured. The antibacterial activity of the peptide is attributed to its effective binding to the bacterial membrane, as demonstrated by the fluorescence lifetime imaging microscopy. In this study, to mimic the L. gormanii membranes and their response to the antimicrobial peptide, Langmuir monolayers were used with the addition of the LL-37 peptide to the subphase of the Langmuir trough to represent the extracellular fluid. The properties of the model membranes (Langmuir monolayers) formed by phospholipids (PL) isolated from the L. gormanii bacteria cultured on the non-supplemented (PL-choline) and choline-supplemented (PL+choline) medium were determined, along with the effect of the LL-37 peptide on the intermolecular interactions, packing, and ordering under the monolayer compression. Penetration tests at the constant surface pressure were carried out to investigate the mechanism of the LL-37 peptide action on the model membranes. The peptide binds to the anionic bacterial membranes preferentially, due to its positive charge. Upon binding, the LL-37 peptide can penetrate into the hydrophobic tails of phospholipids, destabilizing membrane integrity. The above process can entail membrane disruption and ultimately cell death. The ability to evoke such a great membrane destabilization is dependent on the share of electrostatic, hydrogen bonding and Lifshitz-van der Waals LL-37-PL interactions. Thus, the LL-37 peptide action depends on the changes in the lipid membrane composition caused by the utilization of exogenous choline by the L. gormanii.

Physicochemical Characteristics of Model Membranes Composed of Legionella gormanii Lipids.

Legionella gormanii is one of the species belonging to the genus Legionella, which causes atypical community-acquired pneumonia. The most important virulence factors that enable the bacteria to colonize the host organism are associated with the cell surface. Lipids building the cell envelope are crucial not only for the membrane integrity of L. gormanii but also by virtue of being a dynamic site of interactions between the pathogen and the metabolites supplied by its host. The utilization of exogenous choline by the Legionella species results in changes in the lipids' composition, which influences the physicochemical properties of the cell surface. The aim of this study was to characterize the interfacial properties of the phospholipids extracted from L. gormanii cultured with (PL+choline) and without exogenous choline (PL-choline). The Langmuir monolayer technique coupled with the surface potential (SPOT) sensor and the Brewster angle microscope (BAM) made it possible to prepare the lipid monomolecular films (model membranes) and study their properties at the liquid/air interface at 20 °C and 37 °C. The results indicate the effect of the choline addition to the bacterial medium on the properties of the L. gormanii phospholipid membranes. The differences were revealed in the organization of monolayers, their molecular packing and ordering, degree of condensation and changes in the components' miscibility. These findings are the basis for further research on the mechanisms of adaptation of this pathogen, which by changing the native composition and properties of lipids, bypasses the action of antimicrobial compounds and avoids the host immune attack.

Preparation and Surface Characterization of Chitosan-Based Coatings for PET Materials.

Poly(ethylene terephthalate)-PET-is one of the most frequently used polymers in biomedical applications. Due to chemical inertness, PET surface modification is necessary to gain specific properties, making the polymer biocompatible. The aim of this paper is to characterize the multi-component films containing chitosan (Ch), phospholipid 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), immunosuppressant cyclosporine A (CsA) and/or antioxidant lauryl gallate (LG) which can be utilized as a very attractive material for developing the PET coatings. Chitosan was employed owing to its antibacterial activity and also its ability to promote cell adhesion and proliferation favorable for tissue engineering and regeneration purposes. Moreover, the Ch film can be additionally modified with other substances of biological importance (DOPC, CsA and LG). The layers of varying compositions were prepared using the Langmuir-Blodgett (LB) technique on the air plasma-activated PET support. Then their nanostructure, molecular distribution, surface chemistry and wettability were determined by atomic force microscopy (AFM), time-of-flight secondary ion mass spectrometry (TOF-SIMS), X-ray photoelectron spectroscopy (XPS), contact angle (CA) measurements and the surface free energy and its components' determination, respectively. The obtained results show clearly the dependence of the surface properties of the films on the molar ratio of components and allow for a better understanding of the coating organization and mechanisms of interactions at the molecular level both inside the films and between the films and the polar/apolar liquids imitating the environment of different properties. The organized layers of this type can be helpful in gaining control over the surface properties of the biomaterial, thus getting rid of the limitations in favor of increased biocompatibility. This is a good basis for further investigations on the correlation of the immune system response to the presence of biomaterial and its physicochemical properties.

Characteristics of Hybrid Bioglass-Chitosan Coatings on the Plasma Activated PEEK Polymer.

Polyetheretherketone (PEEK) is a biocompatible, chemically and physically stable radiolucent polymer that exhibits a similar elastic modulus to the normal human bone, making it an attractive orthopedic implant material. However, PEEK is biologically inert, preventing strong enough bonding with the surrounding bone tissue when implanted in vivo. Surface modification and composite preparation are the two main strategies for the improvement of the bioactivity of PEEK. In this study, the plasma activated PEEK surfaces with the embedded bioglass, chitosan, and bioglass-chitosan mixed layers applying from the solution dip-coating technique were investigated. The most prominent factors affecting the coating biocompatibility are strictly connected with the composition of its outer surface (its charge and functional groups), hydrophilic-hydrophobic character, wettability and surface free energy, and topography (size of pores/substructures, roughness, stiffness), as well as the personal characteristics of the patient. The obtained surfaces were examined in terms of wettability and surface-free energy changes. Additionally, FTIR (Fourier Transformation Infrared Spectrometry) and SIMS (Secondary Ion Mass Spectrometry) were applied to establish and control the coating composition. Simultaneously the structure of coatings was visualized with the aid of SEM (Scanning Electron Microscopy). Finally, the obtained systems were incubated in SBF (Simulated Body Fluid) to verify the modifications' influence on the bioactivity/biocompatibility of the PEEK surface. Different structures with variable compositions, as well as changes of the wettability, were observed depending on the applied modification. In addition, the incubation in SBF suggested that the bioglass-chitosan ratio influenced the formation of apatite-like structures on the modified PEEK surfaces.

Effect of naproxen on the model lipid membrane formed on the water-chitosan subphase.

Non steroidal anti-inflammatory drugs (NSAIDs) are those of the most common over the counter (OTC) medications widely used by millions of people every day. Unfortunately, despite their popularity those drugs can cause serious side effects in the digestive system (ulcers, bleeding, and pain). These inconveniences are caused by the changes in the structures of the outer phospholipid layers of gastric mucus and mucosa. As a result the H+ ions from the stomach acid can pass easily through these natural protective barriers and damage the epithelial cells which causes ulcers and bleeding. Chitosan as a polysaccharide known for its unique biocompatibility, drug delivery possibilities and wound healing effect has been chosen to examine if it can induce the reduction of undesirable effects of naproxen. This paper focuses on the interactions of the naproxen with a model biological membrane with and without the presence of chitosan. Applying the Langmuir technique coupled with the surface potential measurements and the Brewster angle microscope imaging allowed to characterize successfully examined systems in terms of the monolayer compressibility, thickness, stability, electric properties and morphology. The results proved that the presence of naproxen alters the mechanical and electrical properties of the model membrane depending on its surface pressure. Moreover, the addition of chitosan to the lipid-drug system causes significant changes in the properties of the layer, i.e. a reduction of its compressibility, thickness and morphology modification. Nevertheless, chitosan suppresses some changes induced by naproxen such as alteration of the apparent dipole moment and film stability.

Surface Properties of the Polyethylene Terephthalate (PET) Substrate Modified with the Phospholipid-Polypeptide-Antioxidant Films: Design of Functional Biocoatings.

Surface properties of polyethylene terephthalate (PET) coated with the ternary monolayers of the phospholipid 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), the immunosuppressant cyclosporine A (CsA), and the antioxidant lauryl gallate (LG) were examined. The films were deposited, by means of the Langmuir-Blodgett (LB) technique, on activated by air low temperature plasma PET plates (PETair). Their topography and surface chemistry were determined with the help of atomic force microscopy (AFM) and time-of-flight secondary ion mass spectrometry (TOF-SIMS), respectively, while wettability was evaluated by the contact angle measurements. Then, the surface free energy and its components were calculated from the Lifshitz-van der Waals/Acid-Base (LWAB) approach. The AFM imaging showed that the Langmuir monolayers were transferred effectively and yielded smoothing of the PETair surface. Mass spectrometry confirmed compatibility of the quantitative and qualitative compositions of the monolayers before and after the transfer onto the substrate. Moreover, the molecular arrangement in the LB films and possible mechanisms of DOPC-CsA-LG interactions were determined. The wettability studies provided information on the type and magnitude of the interactions that can occur between the biocoatings and the liquids imitating different environments. It was found that the changes from open to closed conformation of CsA molecules are driven by the hydrophobic environment ensured by the surrounding DOPC and LG molecules. This process is of significance to drug delivery where the CsA molecules can be released directly from the biomaterial surface by passive diffusion. The obtained results showed that the chosen techniques are complementary for the characterization of the molecular organization of multicomponent LB films at the polymer substrate as well as for designing biocompatible coatings with precisely defined wettability.

Characteristics of Phospholipid-Immunosuppressant-Antioxidant Mixed Langmuir-Blodgett Films.

Hemocompatibility is one of the major criteria for the successful cardiovascular applicability of novel biomaterials. In this context, monolayers of certain biomolecules can be used to improve surface biocompatibility. To this end, biocoatings incorporating a phospholipid (1,2-dioleoyl-sn-glycero-3-phosphocholine, DOPC), an immunosuppressant (cyclosporine A, CsA), and an antioxidant material (lauryl gallate, LG) were fabricated by depositing Langmuir films onto gold or mica substrates using the Langmuir-Blodgett (LB) technique. These LB monolayers were thoroughly characterized by means of quartz crystal microbalance (QCM), atomic force microscopy (AFM), cyclic voltammetry (CV), and contact angle (CA) measurements. The obtained results indicate that the properties of these LB films are modulated by the monolayer composition. The presence of LG in the three-component systems (DOPC-CsA-LG) increases the molecular packing and the surface coverage of the substrate, which affects the wettability of the biocoating. From the different compositions studied here, we conclude that DOPC-CsA-LG monolayers with a DOPC/CsA ratio of 1:1 and LG molar fractions of 0.50 and 0.75 exhibit improved surface biocompatible characteristics. These results open up new perspectives on our knowledge and better understanding of phenomena at the biomaterial/host interface.

The effect of chitosan/TiO2/hyaluronic acid subphase on the behaviour of 1,2-dioleoyl-sn-glycero-3-phosphocholine membrane.

The main aim of the study was to determine the effect of two polysaccharides: chitosan (Ch) and hyaluronic acid (HA), and/or titanium dioxide (TiO2) on the structure and behaviour of the 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) membrane. To achieve this goal the surface pressure as a function of the area per molecule (π-A) isotherm for the phospholipid monolayer was recorded. The shape of the π-A isotherms and compression-decompression cycles, as well as the compression modulus values, were analysed in terms of biocompatibility. Besides, morphology and thickness of the phospholipid layers obtained by means of Brewster angle microscope at different π, were determined. The obtained results show that both polysaccharides Ch, HA, as well inorganic TiO2 affect slightly the structure of the DOPC monolayer but do not disrupt it. Their presence brings no typical arrangements of both the polar heads and tails of DOPC molecules at the interface.

The human LL-37 peptide exerts antimicrobial activity against Legionella micdadei interacting with membrane phospholipids.

Legionella micdadei is responsible for community- or nosocomial-acquired pneumonia as well as the influenza-like illness Pontiac fever. The aim of this study was to investigate the ability of L. micdadei to utilize extracellular choline for phosphatidylcholine (PC) synthesis and its consequences for the phospholipid composition of its membrane system and the interaction with the human LL-37 peptide. Comparative analysis of the PC content using isotopic labeling revealed that in presence of exogenous choline 98% of the total PC was synthesized via the Pcs pathway while the remaining 2% were generated via the PE-methylation (PmtA) pathway. PC species were to a greater extent defined by the Pcs pathway in the outer membrane than in the inner membrane. While no major changes in the bacterial lipid content were observed using 31P NMR, indication for utilization of longer acyl chains and slight increase of PG in response to choline addition was observed by a top-down lipidomics screen. The LL-37 peptide inhibited L. micdadei growth in a dose-dependent manner. Bacteria cultured with exogenous choline were more sensitive to the LL-37 peptide when compared to the standard culture condition. Our biophysical investigations show that the peptide perturbs bacterial-derived phospholipid monolayers and this interaction is dependent on the molar portion of PC. This interaction is responsible for the observed changes in the anti-L. micdadei activity of the LL-37 peptide.

The Influence of Polysaccharides/TiO2 on the Model Membranes of Dipalmitoylphosphatidylglycerol and Bacterial Lipids.

The aim of the study was to determine the bactericidal properties of popular medical, pharmaceutical, and cosmetic ingredients, namely chitosan (Ch), hyaluronic acid (HA), and titanium dioxide (TiO2). The characteristics presented in this paper are based on the Langmuir monolayer studies of the model biological membranes formed on subphases with these compounds or their mixtures. To prepare the Langmuir film, 1,2-dipalmitoyl-sn-glycero-3-phospho-rac-(1-glycerol) (DPPG) phospholipid, which is the component of most bacterial membranes, as well as biological material-lipids isolated from bacteria Escherichia coli and Staphylococcus aureus were used. The analysis of the surface pressure-mean molecular area (π-A) isotherms, compression modulus as a function of surface pressure, CS-1 = f(π), relative surface pressure as a function of time, π/π0 = f(t), hysteresis loops, as well as structure visualized using a Brewster angle microscope (BAM) shows clearly that Ch, HA, and TiO2 have antibacterial properties. Ch and TiO2 mostly affect S. aureus monolayer structure during compression. They can enhance the permeability of biological membranes leading to the bacteria cell death. In turn, HA has a greater impact on the thickness of E. coli film.

What affects the biocompatibility of polymers?

In recent decades synthetic polymers have gained increasing popularity, and nowadays they are an integral part of people's daily lives. In addition, owing to their competitive advantage and being susceptible to modification, polymers have stimulated the fast development of innovative technologies in many areas of science. Biopolymers are of particular interest in various branches of medicine, such as implantology of bones, cartilage and skin tissues as well as blood vessels. Biomaterials with such specific applications must have appropriate mechanical and strength characteristics and above all they must be compatible with the surrounding tissues, human blood and its components, i.e. exhibit high hemo- and biocompatibility, low or no thrombo- and carcinogenicity, foreign body response (host response), appropriate osteoconduction, osteoinduction and mineralization. For biocompatibility improvement many surface treatment techniques have been utilized leading to fabricate the polymer biomaterials of required properties, also at nanoscale. This review paper discusses the most important physicochemical and biological factors that affect the biocompatibility, thus the reaction of the living organism after insertion of the polymer-based biomaterials, i.e. surface modification and/or degradation, surface composition (functional groups and charge), size and shapes, hydrophilic-hydrophobic character, wettability and surface free energy, topography (roughness, stiffness), crystalline and amorphous structure, nanostructure, cell adhesion and proliferation, cellular uptake. Particularly, the application of polysaccharides (chitosan, cellulose, starch) in the tissue engineering is emphasized.

Analysis of Molecular Interactions between Components in Phospholipid-Immunosuppressant-Antioxidant Mixed Langmuir Films.

The study of Langmuir monolayers incorporating biomimetic and bioactive substances plays an important role today in assessing the properties and quality of the molecular films for potential biomedical applications. Here, miscibility of binary and ternary monolayers of phospholipid (dioleoyl phosphatidylcholine, DOPC), immunosuppressant (cyclosporine A, CsA), and antioxidant (lauryl gallate, LG) of varying molar fractions was analyzed by means of the Langmuir technique coupled with a surface potential (ΔV) module at the air-water interface. The surface pressure-area per molecule (π-A) isotherms provided information on the physical state of the films at a given surface pressure, the monolayer packing and ordering, and the type and strength of intermolecular interactions. Surface potential-area (ΔV-A) isotherms revealed the molecular orientation changes at the interface upon compression. In addition, the apparent dipole moment of the monolayer-forming molecules was determined from the surface potential isotherms. The obtained results indicated that the film compression provoked subsequent changes of CsA conformation and/or orientation, conferring better affinity for the hydrocarbon environment. The mutual interactions between the components were analyzed here in terms of the excess and total Gibbs energy of mixing, whose values depended on the stoichiometry of the mixed films. The strongest attraction, thus the highest thermodynamic stability, was found for a DOPC-CsA-LG mixture with a 1:1:2 molar ratio. Based on these results, a molecular model for the organization of the molecules within the Langmuir film was proposed. Through this model, we elucidated the significant role of LG in improving the miscibility of CsA in the model DOPC membrane and thus in increasing the stability of self-assembled monolayers by noncovalent interactions, such as H-bonds and Lifshitz-van der Waals forces. The above 1:1:2 combination of three components is revealed as the most promising film composition for the modification of implant device surfaces to improve their biocompatibility. Further insight into mechanisms concerning drug-membrane interactions at the molecular level is provided, which results in great importance for biocoating design and development as well as for drug release at target sites.

Properties of Artificial Phospholipid Membranes Containing Lauryl Gallate or Cholesterol.

Lauryl gallate (LG) is an antioxidant agent. However, it exhibits poor solubility in water. Its interactions with the membrane result in structure evolution thus affecting the membrane functionality. In this paper the Brewster angle microscope coupled with the Langmuir trough was applied to determine the morphology, phase behaviour, thickness and miscibility of ternary Langmuir monolayers with equal mole fractions of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC); 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and an increasing mole fraction of LG. The results were discussed as regards analogous systems where cholesterol (Chol) was the third component. Moreover, the phosphatidylcholine-lauryl gallate (PC-LG) interactions were monitored by the attenuated total reflectance Fourier transform infrared spectroscopy and time-of-flight secondary ion mass spectrometry. Besides lipid composition, the addition of LG was found to be a significant factor to modulate the model membrane properties. The LG molecules adjust themselves to the PC monolayer structure. The hydrophobic fragment is dipped into the membrane interior while the hydroxyl groups of phenolic gallate moiety associate with the polar groups of PC mainly through hydrogen bonding inducing the compacting effect. LG is found to be deeply submerged within DOPC, closer to the double bonds, and its insertion practically does not affect the DPPC/DOPC membrane fluidity. This is crucial for getting more profound insight into the role of LG in stabilizing the non-raft domains, mostly exposed to oxidation in which LG can co-localize and serve its antioxidant function.

Effect of Lauryl Gallate on Wetting Properties of Organized Thin Phospholipid Films on Mica.

To characterize surfaces of phospholipid/lauryl gallate monolayers deposited on mica there were applied numerous methods such as measurements of advancing and receding contact angles and optical profilometry, as well as atomic force microscopy. As a result, there was no found correlation between contact angles (and their hysteresis) or surface roughness. Hence, most monolayer topographical changes at the Ångstrom level accompanied changes in surface chemistry which resulted in the hysteresis of contact angle on thin films. The obtained results indicate that stability and permeability of the binary films are determined by the composition and stoichiometry of the mixed monolayers. These results can be helpful for insight into lauryl gallate behavior in living systems, i.e., in membrane antioxidant protection and pharmacological activities.

Interactions of lauryl gallate with phospholipid components of biological membranes.

The effect of different amounts of lauryl gallate (LG) on properties of the model membranes of phosphatidylcholines (PC), differing in the presence of double bonds in the hydrocarbon chains, and phosphatidylglycerol (PG) was described in terms of phase behaviour of mixtures, interactions between both components, monolayers stability and their organization. The Langmuir monolayer technique was used to monitor the surface thermodynamics (i.e. the excess area and excess Gibbs energy of mixing) on the basis of surface pressure-area per molecule (π-A) isotherms. Simultaneously, morphology of the studied monolayers was visualized by the Brewster angle microscopy (BAM). This allowed evaluating the kind and magnitude of interactions which influence on the phase behaviour and structural properties of the monolayers. The obtained results can be helpful to reveal the mechanism of phospholipid antioxidant protection and important pharmacological (antimicrobial) role of lauryl gallate for production of effective therapeutic substances.

Properties of Langmuir and solid supported lipid films with sphingomyelin.

Biological cell membranes play a crucial role in various biological processes and their functionality to some extent is determined by the hydrophilic/hydrophobic balance. A significant progress in understanding the membrane structure was the discovery of laterally segregated lipid domains, called the lipid rafts. These raft domains are of ordered lamellar liquid-crystalline phase, while rest of the membrane exists in a relatively disordered lamellar liquid-crystalline phase. Moreover, the chemical constitution of the lipid rafts consists of a higher content (up to 50%) of cholesterol (Chol) and sphingomyelin (SM). Sphingomyelin also plays a significant role in the red cells of blood and nerves, in some diseases, as a precursor to ceramides, and other sphingolipid metabolites. In this paper properties of Langmuir and solid supported mixed lipid films of DPPC/SM, DOPC/SM, and Chol/SM are described. Special attention has been paid to wetting properties (hydrophobic/hydrophilic balance) of these films transferred onto a hydrophilic glass surface. To our knowledge such results have not yet been published in the literature. The properties were determined via contact angle measurements and then calculation of the films' apparent surface free energy. The films' wettability and their apparent surface free energy strongly depend on their composition. The energy is affected by both the structure of hydrocarbon chains of glycerophospholipids (DPPC and DOPC) and their interactions with SM. Properties of mixed Chol/SM monolayer depend also on the film stoichiometry. At a low Chol content (XChol=0.25) the interactions between SM and Chol are strong and hence the formation of binary complex is possible. This is accompanied by a decrease in the film surface free energy in comparison to that of pure SM monolayer, contrary to a higher Chol content where the monolayer energy increases. This suggests that cholesterol is excluded from the membrane thus increasing the film hydrophilicity. These results are consistent with the literature data and somehow confirm the hypothesis of lipid raft formation. The roughness of the investigated monolayer surfaces was also determined using optical profilometry. The roughness parameters of the DPPC, SM, and mixed DPPC/SM generally correlate with the changes of their apparent surface free energy, i.e. with the decreasing roughness the apparent surface free energy also decreases. However, this is not the case for mixed DOPC/SM monolayers. Although the roughness increases with SM content the apparent surface free energy decreases. Therefore some other factors, like the presence of unsaturated bonds in the DOPC molecule, influence the film phase state and the energy too. More experiments are needed to explain this hypothesis.

Surface Gibbs energy interaction of phospholipid/cholesterol monolayers deposited on mica with probe liquids.

The mica supported binary monolayers containing phospholipids: 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 2-oleoyl-1-palmitoyl-sn-glycero-3-phosphocholine (POPC), 1,2-dipalmitoyl-sn-glycero-3-phospho-rac-(1-glycerol) (DPPG), and cholesterol (Chol), mixed at different molar fractions, were investigated by measurements of the contact angles of water, formamide and diiodomethane. This allowed calculation of apparent surface Gibbs energy (further in the paper termed as 'surface free energy') of the monolayers according to the theoretical approach developed by Chibowski (contact angle hysteresis model, CAH). Then, based on the surface free energy values, the molar interaction Gibbs energy of the lipid molecules with the given probe liquid was evaluated. These values correlate with the values of excess area, interpreted as an indicator of the condensing effect of cholesterol on phospholipid monolayers at the air-water interface. The results indicate that the thermodynamic parameters of interactions depend on the monolayer composition and the probe liquid used to their determination. Changes of the parameters are discussed in relation to the monolayer packing, ordering, tilting of the molecules, and properties of the probe liquids as well.

Characterization of the binary mixed monolayers of α-tocopherol with phospholipids at the air-water interface.

The mixed Langmuir monolayers composed of model constituents of biological membranes, 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 2-oleoyl-1-palmitoyl-sn-glycero-3-phosphocholine (POPC), and 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), were investigated to provide information on the intermolecular interactions between these membrane components and the physiologically active vitamin E-α-tocopherol (TF), as well as on the phase behavior of these mixed systems. Additionally, topography of these monolayers transferred onto the mica support was investigated by the inverted metallurgical microscope. Morphological characteristics were directly observed by Brewster angle microscopy (BAM). From the surface pressure-area isotherms and the analysis of physicochemical parameters (compressibility and mean molecular area at the maximum compressibility) it was found that depending on the acyl chains saturation degree, TF has different effect on the phospholipids. In the case of DPPC, the addition of TF to the phospholipid film causes destabilization of the ordered hydrocarbon chains, while in the POPC/DOPC-TF systems, the attractive interactions are responsible for the monolayer increased stability. Thus, the results of these studies confirm the hypothesis that α-tocopherol may play a role in the stabilization of biological membranes.

Thermodynamic aspects of cholesterol effect on properties of phospholipid monolayers: Langmuir and Langmuir-Blodgett monolayer study.

Cholesterol is an important component of lipid rafts in mammalian cell membranes. Studies of phospholipid monolayers containing cholesterol provide insight into the role of cholesterol in regulating the properties of animal cells, raft stability, and organization. In this contribution, a study of the characteristics of binary Langmuir monolayers consisting of phospholipids, 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 2-oleoyl-1-palmitoyl-sn-glycero-3-phosphocholine (POPC), 1,2-dipalmitoyl-sn-glycero-3-phospho-rac-(1-glycerol) (DPPG), and cholesterol (Chol), was conducted on the basis of the surface pressure-area per molecule (π-A) isotherms. Analysis of the results obtained provided information on the mean molecular area, the excess Gibbs energy of mixing, and condensation in the monolayer. The mixed monolayers were also deposited onto the mica plates and investigated by the contact angle measurements of water, formamide, and diiodomethane. The contact angles allowed calculating surface free energy of the films from the van Oss et al. approach. It was found that cholesterol determines the molecular packing and ordering of the monolayers closely connected with the kind of phospholipid. This is reflected in the values of surface free energy of the model membranes. From the thermodynamic analysis of phospholipid/cholesterol/liquid interactions, one may draw conclusions about the most favorable composition (stoichiometry) of the binary film which is especially important in view of the lipid rafts formation.