Molecular interactions of cord factor with dipalmitoylphosphatidylcholine monolayers: implications for lung surfactant dysfunction in pulmonary tuberculosis

Molecular interactions between mycobacterial cell wall lipid, cord factor (CF) and the abundant surfactant lipid, dipalmitoylphosphatidylcholine (DPPC) were investigated using Langmuir monolayers at physiological temperatures (37 degrees C). Surface topography of the films was visualized by atomic force microscopy (AFM). Thermodynamic behavior of the mixed monolayers was evaluated by investigating the molecular area excess, excess Gibbs free energy of mixing and maximum compressibility modulus (SCM(max)). Cord factor formed immiscible and thermodynamically unstable monolayers with DPPC. Monolayer presence of cord factor altered the physical state of DPPC monolayers from liquid condensed to liquid expanded with the lowering of SCM(max) from 160 to 40 mN/m, respectively. AFM imaging exhibited smooth homogenous surface topography of DPPC films which in the presence of cord factor was markedly altered with the appearance of aggregates and increased surface roughness. The results highlight the capacity of cord factor to disturb DPPC monolayer organization and structure. Interfacial presence of cord factor results in DPPC monolayer fluidization. Lung surfactant function is attributed to its ability to form well packed low compressibility films. Such molecular interactions suggest a dysfunction of lung surfactant in pulmonary tuberculosis due to surfactant monolayer fluidization.     

Thermodynamic characteristics and Langmuir-Blodgett deposition behavior of mixed DPPA/DPPC monolayers at air/liquid interfaces

The role of dipalmitoylphosphatic acid (DPPA) as a transfer promoter to enhance the Langmuir-Blodgett (LB) deposition of a dipalmitoylphosphatidylcholine (DPPC) monolayer at air/liquid interfaces was investigated, and the effects of Ca2+ ions in the subphase were discussed. The miscibility of the two components at air/liquid interfaces was evaluated by surface pressure-area per molecule isotherms, thermodynamic analysis, and by the direct observation of Brewster angle microscopy (BAM). Multilayer LB deposition behavior of the mixed DPPA/DPPC monolayers was then studied by transferring the monolayers onto hydrophilic glass plates at a surface pressure of 30 mN/m. The results showed that the two components, DPPA and DPPC, were miscible in a monolayer on both subphases of pure water and 0.2 mM CaCl2 solution. However, an exception occurs between X(DPPA)=0.2 and 0.5 at air/CaCl2-solution interface, where a partially miscible monolayer with phase separation may occur. Negative deviations in the excess area analysis were found for the mixed monolayer system, indicating the existence of attractive interactions between DPPA and DPPC molecules in the monolayers. The monolayers were stable at the surface pressure of 30 mN/m for the following LB deposition as evaluated from the area relaxation behavior. It was found that the presence of Ca2+ ions had a stabilization effect for DPPA-rich monolayers, probably due to the association of negatively charged DPPA molecules with Ca2+ ions. Moreover, the Ca2+ ions may enhance the adhesion of DPPA polar groups to a glass surface and the interactions between DPPA polar groups in the multilayer LB film structure. As a result, Y-type multilayer LB films containing DPPC could be fabricated from the mixed DPPA/DPPC monolayers with the presence of Ca2+ ions.     

Sum frequency generation spectroscopic study of the condensation effect of cholesterol on a lipid monolayer

Sum frequency generation (SFG) spectra and surface pressure–molecular area (π–A) isotherms have been obtained for mixed cholesterol–DPPC monolayers with cholesterol mole fractions, x(chol.), from 0 to 1.0, at the air–water interface, under same conditions, at 22 °C. Analysis of the spectra indicated that incorporation of cholesterol into the monolayers at 3 mN m⁻¹ greatly increases the conformational and orientational order of the alkyl chains of DPPC, maximizing these properties at x(chol.)=0.4. Analysis also indicated that order in the mixed monolayers at 15 and 35 mN m⁻¹ is not affected by incorporation of cholesterol. The π–A isotherms measured at 3 mN m⁻¹ for the mixed monolayer with x(chol.)=0.4 have the largest negative deviation of the molecular area relative to those of ideal mixtures (the so-called “condensation effect” of cholesterol), indicating the most thermodynamically stable state. Comparison of results from SFG spectra and π–A isotherms explicitly proved that the condensation effect can be interpreted in terms of conformational and orientational ordering of the alkyl chains of DPPC.     

Thermodynamic and infrared analyses of the interaction of chlorpromazine with phospholipid monolayers

An investigation has been made of the interaction between chlorpromazine (CPZ) and monolayers of 1,2-dipalmitoyl-sn-3-glycerophosphatidylcholine (DPPC) and 1,2-dipalmitoyl-sn-3-glycero[phospho-rac-(1-glycerol)] (DPPG), both at the air/water interface and in transferred Langmuir-Blodgett films. The Gibbs free energy, DeltaG, and the compressibility modulus (C(S)(-1)), obtained from the surface pressure isotherms, indicated changes in the in-plane interactions of CPZ/DPPG mixed monolayers, with positive values of DeltaG. The arrangement of CPZ in the zwitterionic DPPC monolayers causes a weaker interaction in CPZ/DPPC mixed monolayers, with the DeltaG fluctuating around zero. IR measurements in transferred monolayers showed that CPZ did not affect the conformational order of the acyl chains, its effects being limited to the bands corresponding to the headgroups. Furthermore, since no shift was observed for the acyl chain bands, the phase transition induced by CPZ is not a liquid expanded (LE) to liquid condensed (LC) transition, as the latter is associated with chain ordering. Taken together, the IR and compressibility results demonstrate that the effect from CPZ cannot be correlated with temperature changes in the subphase for pure monolayers, in contrast to models proposed by other authors.     

Excess fibrinogen adsorption to monolayers of mixed lipids

Adsorption of fibrinogen to the monolayers of mixed lipids, dipalmitoyl phosphatidyl choline (DPPC) and eicosylamine (EA) was measured at a surface pressure of 20 mN/m by an in situ surface plasmon resonance technique. Pressure–area isotherms of DPPC + EA mixtures on water and buffer subphases indicated good lipid miscibility and some contraction of the monolayers at intermediate and higher surface pressures. Surface electric potential of the DPPC + EA monolayers showed excess values for intermediate DPPC:EA ratios. Fibrinogen adsorption and its adsorption rates from a dilute solution (0.03 mg/ml) were proportional to the fraction of EA in the monolayer indicating that protein binding was primarily driven by electrostatic interactions between positive EA charges in the monolayer and a net negative protein charge. At a higher protein concentration (0.06 mg/ml) both the fibrinogen adsorbed amount and its maximum adsorption rate showed excess values relative to the pure EA for 1:1, 2:1 and 3:1 DPPC + EA monolayers. This excess adsorption could be explained, in part, by the contraction of the monolayers with intermediate DPPC:EA ratios which resulted in an excess surface electric potential.     

Structural characteristics of dipalmitoylphosphatidylcholine and hydrolysates from sunflower protein isolate mixed monolayers spread at the air-water interface

In this work, surface film balance and Brewster angle microscopy techniques have been used to analyze the structural characteristics (structure, topography, reflectivity, thickness, miscibility, and interactions) of hydrolysates from sunflower protein isolate (SPI) and dipalmitoylphosphatidylcholine (DPPC) mixed monolayers spread on the air-water interface. The degree of hydrolysis (DH) of SPI, low (5.62%), medium (23.5%), and high (46.3%), and the protein/DPPC mass fraction were analyzed as variables. The structural characteristics of the mixed monolayers deduced from the surface pressure (pi)-area (A) isotherms depend on the interfacial composition and degree of hydrolysis. At surface pressures lower than the equilibrium surface pressure of SPI hydrolysate (pi(e)(SPI hydrolysate)), both DPPC and protein are present in the mixed monolayer. At higher surface pressures (at pi > pi(e)(SPI hydrolysate)), collapsed protein residues may be displaced from the interface by DPPC molecules. The differences observed between pure SPI hydrolysates and DPPC in reflectivity (I) and monolayer thickness during monolayer compression have been used to analyze the topographical characteristics of SPI hydrolysates and DPPC mixed monolayers at the air-water interface. The topography, reflectivity, and thickness of mixed monolayers confirm at microscopic and nanoscopic levels the structural characteristics deduced from the pi-A isotherms.     

Mixed DPPC/DPTAP monolayers at the air/water interface: influence of indolilo-3-acetic acid and selenate ions on the monolayer morphology

The interactions of mixed monolayers of two lipids, zwitterionic 1,2-dipalmitoyl-phosphatidylcholine (DPPC) and positively charged 1,2-dipalmitoyl-3-trimethylammonium-propane (DPTAP), with phytohormone indolilo-3-acetic acid (IAA) and selenate anions in the aqueous subphase were studied. For this purpose, isotherms of the surface pressure versus the mean molecular area were recorded. Domain formation was investigated by using Brewster angle microscopy (BAM). The method of grazing incidence X-ray diffraction (GIXD) was also applied for the characterization of the organization of lipid molecules in condensed monolayers. It was found that selenate ions contribute to monolayer condensation by neutralizing the positive net charge of mixed monolayers whereas IAA molecules penetrated the lipid monolayer, causing its expansion/fluidization. When both solutes were introduced into the subphase, a competition between them for interaction with the positively charged lipids in the monolayer was observed.     

Effects of albumin and erythrocyte membranes on spread monolayers of lung surfactant lipids

Dipalmitoyl phosphatidylcholine (DPPC), one of the main constituents of lung surfactant is mainly responsible for reduction of surface tension to near 0 mN/m during expiration, resisting alveolar collapse. Other unsaturated phospholipids like palmitoyloleoyl phosphatidylglycerol (PG), palmitoyloleoyl phosphatidylcholine (POPC) and neutral lipids help in adsorption of lung surfactant to the air-aqueous interface. Lung surfactant lipids may interact with plasma proteins and hematological agents flooding the alveoli in diseased states. In this study, we evaluated the effects of albumin and erythrocyte membranes on spread films of DPPC alone and mixtures of DPPC with each of PG, POPC, palmitoyloleoyl phosphatidylethanolamine (PE), cholesterol (CHOL) and palmitic acid (PA) in 9:1 molar ratios. Surface tension-area isotherms were recorded using a Langmuir-Blodgett (LB) trough at 37 degrees C with 0.9% saline as the sub-phase. In the presence of erythrocyte membranes, DPPC and DPPC+PA monolayers reached minimum surface tensions of 7.3+/-0.9 and 9.6+/-1.4 mN/m, respectively. Other lipid combinations reached significantly higher minimum surface tensions >18 mN/m in presence of membranes (Newman Keul's test, p<0.05). The relative susceptibility to membrane inhibition was [(DPPC+PG, 7:3)=(DPPC+PG, 9:1)=(DPPC+POPC)=(DPPC+PE)=(DPPC+CHOL)]>[(DPPC+PA)=(DPPC)]. The differential response was more pronounced in case of albumin with DPPC and DPPC+PA monolayers reaching minimum surface tensions less than 2.4 mN/m in presence of albumin, whereas DPPC+PG and DPPC+POPC reached minimum surface tensions of around 20 mN/m in presence of albumin. Descending order of susceptibility of the spread monolayers of lipid mixtures to albumin destabilization was as follows: [(DPPC+PG, 7:3)=(DPPC+PG, 9:1)=(DPPC+POPC)]>[(DPPC+PE)=(DPPC+CHOL)]>[(DPPC+PA)=(DPPC)] The increase in minimum surface tension in presence of albumin and erythrocyte membranes was accompanied by sudden increases in compressibility at surface tensions of 15-30 mN/m. This suggests a monolayer destabilization and could be indicative of phase transitions in the mixed lipid films due to the presence of the hydrophobic constituents of erythrocyte membranes.     

Lung surfactant dysfunction in tuberculosis: effect of mycobacterial tubercular lipids on dipalmitoylphosphatidylcholine surface activity

In pulmonary tuberculosis, Mycobacterium tuberculosis bacteria reside in the alveoli and are in close proximity with the alveolar surfactant. Mycolic acid in its free form and as cord factor, constitute the major lipids of the mycobacterial cell wall. They can detach from the bacteria easily and are known to be moderately surface active. We hypothesize that these surface-active mycobacterial cell wall lipids could interact with the pulmonary surfactant and result in lung surfactant dysfunction. In this study, the major phospholipid of the lung surfactant, dipalmitoylphosphatidylcholine (DPPC) and binary mixtures of DPPC:phosphatidylglycerol (PG) in 9:1 and 7:3 ratios were modelled as lung surfactant monolayers and the inhibitory potential of mycolic acid and cord factor on the surface activity of DPPC and DPPC:PG mixtures was evaluated using Langmuir monolayers. The mycobacterial lipids caused common profile changes in all the isotherms: increase in minimum surface tension, compressibility and percentage area change required for change in surface tension from 30 to 10 mN/m. Higher minimum surface tension values were achieved in the presence of mycolic acid (18.2 ± 0.7 mN/m) and cord factor (13.28 ± 1.2 mN/m) as compared to 0 mN/m, achieved by pure DPPC film. Similarly higher values of compressibility (0.375 ± 0.005 m/mN for mycolic acid:DPPC and 0.197 ± 0.003 m/mN for cord factor:DPPC monolayers) were obtained in presence of mycolic acid and cord factor. Thus, mycolic acid and cord factor were said to be inhibitory towards lung surfactant phospholipids. Higher surface tension and compressibility values in presence of tubercular lipids are suggestive of an unstable and fluid surfactant film, which will fail to achieve low surface tensions and can contribute to alveolar collapse in patients suffering from pulmonary tuberculosis. In conclusion a biophysical inhibition of lung surfactant may play a role in the pathogenesis of tuberculosis and may serve as a target for the development of new drug loaded surfactants for this condition.     

Monolayer behavior of binary systems of betulinic acid and cardiolipin: thermodynamic analyses of Langmuir monolayers and AFM study of Langmuir-Blodgett monolayers

Betulinic acid (BA, a natural pentacyclic triterpene) can induce mitochondrial membrane damage and trigger the mitochondrial pathway of apoptosis in tumor cells. The monolayer behavior of binary systems of BA and cardiolipin (CL, a unique phospholipid found only in mitochondria membrane in animals) was studied by surface pressure-area (π-A) measurements and analyses and Atomic force microscopy (AFM) observation. The miscibility analysis presents that in mixed monolayers BA takes both tilted and nearly perpendicular orientations at surface pressure below 30 mN/m but only nearly perpendicular orientation at 30 mN/m. The thermodynamic stability analysis indicates that phase separation and repulsion occur in mixed BA/CL monolayers. The compressibility analysis shows that at 30 mN/m, 20% addition of BA does markedly translate the liquid-condensed CL monolayer to mixed BA/CL monolayer with the coexistence of liquid-condensed and liquid-expanded phases. The AFM images of supported monolayers give direct evidence of the conclusions obtained from the analyses of π-A isotherms. These results confirm that at high surface pressure near to real biologic situations, BA orients nearly perpendicularly with hydroxyl group toward water, causes phase separation and changes the permeability of CL film, which correlates with the mitochondrial membrane damage induced by BA.     

Langmur monolayers of cerebroside with different head groups originated from sea cucumber: Binary systems with dipalmitoylphosphatidylcholine (DPPC)

Surface properties (Langmuir monolayer) of two different cerebrosides which are extracted from the sea cucumber (Bohadschia argus) were investigated. A main difference in chemical structure of cerebroside between BAC-2a and BAC-4 is their head groups (glucose and galactose, respectively). Furthermore, miscibility and interaction between dipalmitoylphosphatidylcholine (DPPC) and cerebrosides (BAC-2a and BAC-4) in the monolayer have been systematically examined. The surface pressure (π)−area (A), the surface potential (ΔV)−A, and the dipole moment (μ)−A isotherms for monolayers of DPPC, cerebrosides, and their binary combinations have been measured using the Wilhelmy method and the ionizing electrode method. BAC-4 forms a stable liquid-expanded (LE) monolayer, whereas BAC-2a has a first-order phase transition from the LE phase to the liquid-condensed (LC) state on 0.15 M NaCl at 298.2 K. The fundamental properties for each cerebroside monolayer were elucidated in terms of the surface dipole moment based on the three-layer model [R.J. Demchak, T. Fort Jr., J. Colloid Interface Sci. 46 (1974) 191–202] for both cerebrosides and the apparent molar quantity change (Δs^γ, Δh^γ, and Δu^γ) for BAC-2a. In addition, their miscibility with DPPC was examined by the variation of the molecular areas and the surface potentials as a function of cerebroside mole fractions, the additivity rule. The miscibility was also confirmed by constructing the two-dimensional phase diagrams. The phase diagrams for the both binary systems were of negative azeotropic type. That is, the two-component DPPC/BAC-2a and DPPC/BAC-4 monolayers are miscible. Furthermore, the Joos equation for the analysis of the collapse pressure of binary monolayers allowed calculation of the interaction parameter and the interaction energy between the DPPC and cerebroside monolayers. The miscibility in the monolayer state was also confirmed by the morphological observation with Brewster angle microscopy (BAM), fluorescence microscopy (FM), and atomic force microscopy (AFM).     

Effects of Fullerenes on Phospholipid Membranes: A Langmuir Monolayer Study

We investigate two‐component Langmuir monolayers of dipalmitoylphosphatidylcholine (DPPC)/C₆₀ by recording surface pressure/area (π/A) and surface potential/area (ΔV/A) isotherms and by direct Brewster angle microscopy (BAM) imaging. Atomic force microscopy (AFM) is employed to study morphologies of the mixed monolayers transferred to a solid substrate by the Langmuir–Blodgett technique. C₆₀ is shown to have little influence on isotherms of the DPPC/C₆₀ monolayers even at a molar fraction as high as X_C60=0.3. The elastic modulus ( ) versus π curves of the DPPC/C₆₀ monolayers almost overlay each other, as well as that of pure DPPC, that is, the elasticities of pure DPPC monolayers and DPPC/C₆₀ monolayers are remarkably similar. AFM studies reveal that fullerene flocs form at low surface pressures (π≤15 mN m^?1), are gradually disaggregated and dispersed in the DPPC monolayer with increasing surface pressure up to 35 mN m^?1, and are then progressively squeezed out to form protruded islands as the surface pressure increases up to 65 mN m^?1. Our work provides experimental support to the computational result that C₆₀ can dissolve in lipid bilayers without significantly compromising their mechanical properties, a finding which has important implications for the toxicity and development of drug vehicles from fullerene materials.     

Structural studies of the monolayers and bilayers formed by a novel cholesterol-phospholipid chimera

Langmuir isotherm, neutron reflectivity, and small angle neutron scattering studies have been conducted to characterize the monolayers and vesicular bilayers formed by a novel chimeric phospholipid, ChemPPC, that incorporates a cholesteryl moeity and a C-16 aliphatic chain, each covalently linked via a glycerol backbone to phosphatidylcholine. The structures of the ChemPPC monolayers and bilayers are compared against those formed from pure dipalmitoylphoshatidylcholine (DPPC) and those formed from a 60:40 mol % mixture of DPPC and cholesterol. In accord with previous findings showing that very similar macroscopic properties were exhibited by ChemPPC and 60:40 mol % DPPC/cholesterol vesicles, it is found here that the chimeric lipid and lipid/sterol mixture have very similar monolayer structures (each having a monolayer thickness of ～26 ?), and they also form vesicles with similar lamellar structure, each having a bilayer thickness of ～50 ? and exhibiting a repeat spacing of ～65 ?. The interfacial area of ChemPPC, however, is around 10 ?(2) greater than that of the combined DPPC/cholesterol unit in the mixed lipid monolayer (viz., 57 ± 1 vs 46 ± 1 ?(2), at 35 mN·m(-1)), and this difference in area is attributed to the succinyl linkage which joins the ChemPPC steroid and glyceryl moieties. The larger area of the ChemPPC is reflected in a slightly thicker monolayer solvent distribution width (9.5 vs 9 ? for the DPPC/cholesterol system) and by a marginal increase in the level of lipid headgroup hydration (16 vs 13 H(2)O per lipid, at 35 mN·m(-1)).     

Topography of dipalmitoyl-phosphatidyl-choline monolayers penetrated by β-casein

In this work we have analyzed the topography by atomic force microscopy (AFM) of dipalmitoyl-phosphatidyl-choline (DPPC) monolayers previously spread at the air–water interface and penetrated by β-casein. AFM images of β-casein–DPPC monolayers were taken from Langmuir–Blodgett films deposited onto hydrophilic mica substrates at different initial surface pressures (π_i) and after the compression of the mixed films. The monolayer topography depends on the initial structure of the phospholipid:liquid expanded (LE) at 3 mN/m, coexistence between LE and liquid condensed (LC) structures at 7 mN/m, at the end of the LE–LC transition at 10 mN/m, and with a LC structure at 15 mN/m. The area occupied by DPPC domains in the mixed film increases with the π_i value, especially for DPPC with a LC structure at 15 mN/m. At this surface pressure the thickness of the film is at a maximum. After the film compression at 25 mN/m, which is above the equilibrium spreading pressure of β-casein (), this protein is displaced from the interface by DPPC and the topography of the mixed monolayer depends on the initial structure of the DPPC monolayer. A notable feature of the topography of these mixed monolayers is the presence of multilayers of β-casein and DPPC of high thickness (50–70 nm) at the lower π_i values. Although the film is dominated by DPPC at the highest surface pressures (at 25 mN/m), β-casein is not displaced totally from the interface and coexists as β-casein collapsed domains within the network of the DPPC structure.     

Properties of two-component Langmuir monolayer of single chain perfluorinated carboxylic acids with dipalmitoylphosphatidylcholine (DPPC)

The surface pressure (π)– and the surface potential (ΔV)–area (A) isotherms were obtained for two-component monolayers of four different perfluorocarboxylic acids (FCns; perfluorododecanoic acid: FC12, perfluorotetradecanoic acid: FC14, perfluorohexadecanoic acid: FC16, perfluorooctadecanoic acid: FC18) with dipalmitoylphosphatidylcholine (DPPC) on substrate solution of 0.15 M NaCl (pH 2.0) at 298.2 K as a function of compositions in the mixtures by employing the Wilhelmy method, the ionizing electrode method, the fluorescence microscopy, and the atomic force microscopy. The data for the two-component monolayers on these systems were analyzed in terms of the additivity rule. Assuming a regular surface mixture, the Joos equation which allows one to describe the collapse pressure of a two-component monolayer with miscible components was used to declare the miscibility of the monolayer state, and an interaction parameter and an interaction energy were calculated. The new finding was that FCns and DPPC are miscible or immiscible depending on chain length increment of fluorocarbon part. That is, FC12/DPPC monolayer was perfectly miscible, and FC14/DPPC, and FC16/DPPC (0 ≤ X_FC16 ≤ 0.3) monolayers were partially miscible. While FC16/DPPC (0.3 < X_FC16 < 1) and FC18/DPPC systems are immiscible in the monolayer state. Furthermore, the mean molecular area, the surface dipole moment, and the phase diagrams enabled us to estimate the molecular orientation of four different perfluorocarboxylic acids/DPPC in the two-component monolayer state. One type of phase diagrams was obtained and classified into the positive azeotropic type. The miscibility of FCns and DPPC in the monolayer was also supported by fluorescence microscopy and atomic force microscopy. FC12/DPPC, FC14/DPPC and FC16/DPPC (0 ≤ X_FC16 ≤ 0.3) two-component monolayers on 0.15 M NaCl (pH 2) showed that FC12, FC14 and FC16 (0 ≤ X_FC16 ≤ 0.3) can dissolve or partially dissolve the ordered solid DPPC domains formed upon compression. This indicates that these fluorinated amphiphiles soften or harden the lipid depending on their chain length.     

Condensing effect of palmitic acid on DPPC in mixed Langmuir monolayers

The interaction between deuterated dipalmitoylphosphatidylcholine (DPPC-d62) and palmitic acid (PA) in mixed Langmuir monolayers is studied using vibrational sum frequency generation (VSFG) spectroscopy. Palmitic acid is an additive in exogenous lung surfactant preparations such as Survanta and Surfaxin. The effect of PA on the chain conformation and orientation of DPPC in the liquid-expanded and condensed phases is explored. A condensing effect of PA on DPPC is observed with VSFG. At 12 mN/m, DPPC-d62 alone is in the liquid-expanded phase. Adding PA increases the conformational ordering of DPPC chains and causes DPPC to transition from the expanded phase into the condensed phase. At 42 mN/m, DPPC-d62 and PA form a mixed structure in the condensed phase. The presence of PA decreases the chain tilt angle of DPPC, increasing the orientational ordering of DPPC chains. At 42 mN/m, there is also evidence from the frequency red shift of the PO2- symmetric stretch that the carboxyl group of PA forms a hydrogen bond with the phosphate group of DPPC in the condensed phase. From this work the effect of PA on DPPC is 2-fold: (1) PA increases the chain ordering of DPPC and promotes the LE and TC phase separation and (2) due to the miscibility between DPPC and PA in the condensed phase, PA decreases the collapse pressure.     

Thermodynamic and structural characterization of a mixed perfluorocarbon-phospholipid ternary monolayer surfactant system

Pulmonary lung surfactant is a mixture of surfactants that reduces surface tension during respiration. Perfluorinated surfactants have potential applications for artificial lung surfactant formulations, but the interactions that exist between these compounds and phospholipids in surfactant monolayer mixtures are poorly understood. We report here, for the first time, a detailed thermodynamic and structural characterization of a minimal pulmonary lung surfactant model system that is based on a ternary phospholipid-perfluorocarbon mixture. Langmuir and Langmuir-Blodgett monolayers of binary and ternary mixtures of the surfactants 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dipalmitoyl-sn-glycero-3-phosphoglycerol (DPPG) and perfluorooctadecanoic acid (C18F) have been studied in terms of miscibility, elasticity and film structure. The extent of surfactant miscibility and elasticity has been evaluated via Gibbs excess free energies of mixing and isothermal compressibilities. Film structure has been studied by a combination of atomic force microscopy and fluorescence microscopy. Combined thermodynamic and microscopy data indicate that the ternary monolayer films were fully miscible, with the mixed films being more stable than their pure individual components alone, and that film compressibility is minimally improved by the addition of perfluorocarbons to the phospholipids. The importance of these results is discussed in context of these mixtures' potential applications in pulmonary lung surfactant formulations.     

Halofantrine-phospholipid interactions: monolayer studies.

The antimalarial agent halofantrine penetrates dipalmitolylphosphatidylcholine (DPPC) monolayers resulting in an increase in surface pressure and an expansion in area occupied by the lipid components of the monolayer. This phenomenon is observed at concentrations (0.05-0.2 microm) of halofantrine that have no surface activity. Penetration increases with drug concentration and is greatest at low initial surface pressures of the monolayer. A critical surface pressure of the DPPC monolayer has been determined from constant area and constant pressure conditions. The magnitude of these values support the hypothesis that halofantrine readily penetrates the DPPC monolayers. The presence of cholesterol in the DPPC monolayer hampers penetration and a lower critical surface pressure is obtained under such conditions. Even then, a slower rate of penetration is observed only in monolayers maintained at high initial surface pressures (10, 15 mN/m), corresponding to the liquid condensed phase of the monolayer, and not at low surface pressures (2.5, 5.0 mN/m). These results help to give a better understanding of the dynamics of the halofantrine-phospholipid interaction as well as the pharmacodynamic character of the drug.     

Stability of binary model membranes--prediction of the liposome stability by the Langmuir monolayer study

In this paper, usefulness of the Langmuir monolayer study is demonstrated for predictions of the stability of liposomes composed of dipalmitoyl phosphatidylcholine (DPPC) and cholesterol (Chol). Thermodynamic analysis of the surface pressure (π)-area (A) isotherms of the DPPC/Chol systems was performed, which allowed for concluding on miscibility of the components, their molecular packing, and the interactions between molecules. It was found that the most stable system, in which the strongest interactions between molecules occured, was DPPC/Chol at x(Chol)=0.25. The stability of liposomes of the same composition as that in the Langmuir monolayers was analyzed by determining the size distribution of vesicles and the polydispersity as a function of time. The changes of these parameters confirmed that the system of the greatest stability is that with low Chol content.     

Thermodynamic behavior and relaxation processes of mixed DPPC/cholesterol monolayers at the air/water interface

This study investigated the thermodynamic behavior and relaxation processes of mixed DPPC/cholesterol monolayers at the air/water interface at 37°C. Surface pressure–area isotherms and relaxation curves for the mixed monolayers were obtained by using a computer-controlled film balance. In the thermodynamic analysis of the mixed monolayers, the areas of monolayers exhibited negative deviations from the ideal values at all compositions for lower surface pressures. However, at higher surface pressures, distinctively positive deviations from ideality were observed at lower DPPC contents. Excess free energies of mixing had been calculated and the most stable state of the mixed monolayer with x_DPPC=0.5 or 0.6 was found. Moreover, the relaxation kinetics of the mixed monolayers was investigated by measuring the surface area as a function of time at a constant surface pressure of 40 mN m⁻¹. It was shown that the relaxation processes could be described by the models considering nucleation and growth mechanisms.