Effect of D-α-tocopheryl polyethylene glycol 1000 succinate (TPGS) on surfactant monolayers

In the present study, the effects of an amphiphilic polymer, d-α-tocopheryl polyethylene glycol 1000 succinate (TPGS) on model surfactant monolayers dipalmitoylphosphatidylcholine (DPPC), a binary mixture of DPPC with palmitoyloleoyl phosphatidylglycerol (DPPC-POPG) 9:1 (w/w) and binary mixture of DPPC and oleic acid (DPPC-OA) were evaluated. The ability of TPGS to act as an antioxidant adjuvant for pulmonary surfactants was also evaluated. Compression isotherms of surfactant monolayers at 37 °C in a Langmuir-Blodgett trough showed that DPPC and DPPC:TPGS mixed monolayers (1:0.25-1:1, w/w) exhibited low minimum surface tensions (MST) of 1-2 mN/m. Similarly [DPPC:POPG (9:1, w/w)]:TPGS mixed films of 1:0.25-1:1 weight ratios reached 1-2 mN/m MST. DPPC:POPG:TPGS liposomes adsorbed to surface tensions of 29-31 mN/m within 1s. While monolayers of DPPC:OA (1:1, w/w) reached high MST of ～11 mN/m, DPPC:OA:TPGS (1:1:0.25, w/w) film reached near zero MST suggesting that low concentrations of TPGS reverses the effect of OA on DPPC monolayer. Capillary surfactometer studies showed DPPC:TPGS and [DPPC:POPG (9:1, w/w)]:TPGS liposomes maintained 84-95% airway patency. Fluorescence spectroscopy of Laurdan loaded DPPC:TPGS and DPPC:POPG:TPGS liposomes revealed no segregation of lipid domains in the lipid bilayer. Addition of TPGS to soybean liposome significantly reduced thiobarbituric acid reactive substance (TBARS) by 29-39% confirming its antioxidant nature. The results suggest a potential use of TPGS as an adjuvant to improve the surfactant activity as well as act as an antioxidant by scavenging free radicals.     

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.     

Mode of interaction of hydrophobic amphiphilic alpha-helical peptide/dipalmitoylphosphatidylcholine with phosphatidylglycerol or palmitic acid at the air-water interface

Surface pressure (pi)-, surface potential (DeltaV)-, dipole moment (mu( perpendicular))-area (A) isotherms and morphological behavior at the air-water interface were obtained for multicomponent monolayers of two different systems for dipalmitoylphosphatidylcholine (DPPC)/egg-phosphatidylglycerol (PG) (= 68:22, by weight)/Hel 13-5 and DPPC/palmitic acid (PA) (= 90:9, by weight)/Hel 13-5 (Hel 13-5 is a newly designed 18-mer amphiphilic alpha-helical peptide with 13 hydrophobic and 5 hydrophilic amino acid residues). The phase behavior of these model systems was investigated on a subsolution of 0.02 M tris(hydroxymethyl)aminomethane (Tris) buffer (pH 8.4) with 0.13 M NaCl at 298.2 K by employing the Wilhelmy method, the ionizing electrode method, and fluorescence microscopy. Especially, the present study focuses on the interfacial effect of the addition of Hel 13-5 on two binary systems, DPPC/egg-PG and DPPC/PA monolayers, as the substitute for pulmonary surfactant proteins, and on the respective roles of PG and PA for the monolayers in the three-component systems. Constant kink points ( approximately 42 mN m(-1)) clearly appear on the pi-A isotherms, independent of the compositions in the ternary systems, which corresponds to the Hel 13-5 collapse pressure similar to that of SP-B and SP-C as functions in multicomponent monolayers. This implies that Hel 13-5 is squeezed out of ternary monolayers above approximately 42 mN m(-1), resulting in two- to three-dimensional phase transformation. Furthermore, Langmuir isotherms clearly show that Hel 13-5 with egg-PG is squeezed out of the DPPC/egg-PG/Hel 13-5 system, whereas only Hel 13-5 is squeezed out of the DPPC/PA/Hel 13-5 system. Cyclic compression and expansion isotherms of these systems were carried out to confirm the spreading and respreading capacities. In addition, the interfacial behavior of the ternary mixtures has been analyzed by the additivity rule. Morphological examinations and comparisons have verified the interactions of Hel 13-5 with the representative miscible mixture (DPPC/PA system) by fluorescence microscopy. Consequently, distinct morphological variations corresponding to the squeeze-out behavior are observed as a fluorescent contrast recovery. Herein, a new mechanism of the refluorescent phenomenon is proposed by varying the surface composition of Hel 13-5.     

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.     

New protocols for preparing dipalmitoylphosphatidylcholine dispersions and controlling surface tension and competitive adsorption with albumin at the air/aqueous interface

The adsorption behavior of dipalmitoylphosphatidylcholine (DPPC), which is the major component of lung surfactant, at the air/aqueous interface and the competitive adsorption with bovine serum albumin (BSA) were studied with tensiometry, infrared reflection absorption spectroscopy (IRRAS), and ellipsometry. Dynamic surface tensions lower than 1 mN/m were observed for DPPC dispersions, with mostly vesicles, prepared with new protocols, involving extensive sonication above 50 °C. The lipid adsorbs faster and more extensively for DPPC dispersions with vesicles than with liposomes. For DPPC dispersions by a certain preparation procedure at T > T_c, when lipid particles were observed on the surface, dynamic surface tensions as low as 1 mN/m were measured. Moreover, IRRAS intensities and ellipsometric δΔ values were found to be much higher than the values for other DPPC dispersions or spread DPPC monolayers, suggesting that a larger amount of liposomes or vesicles adsorb on the surface. For DPPC/BSA mixtures, the tension behavior is controlled primarily by BSA, which prevents the formation of a dense DPPC monolayer. When BSA is injected into the subphase with a spread DPPC monolayer or into a DPPC dispersion with preadsorbed layers, little or no BSA adsorbs and the DPPC layer remains on the surface. When a DPPC monolayer is spread on a BSA solution at 0.1 wt% at 25 °C, then DPPC lipid can displace the adsorbed BSA molecules. The lack of BSA adsorption, and the expulsion of BSA by DPPC monolayer is probably due to the strong hydrophilicity of the lipid polar headgroup. When a DPPC dispersion is introduced with Trurnit's method or when dispersion drops are sprayed onto the surface of a DPPC/BSA mixture, the surface tension becomes lower and is controlled by DPPC, which can prevent the adsorption of BSA. The results may be important in understanding inhibition of lung surfactants by serum proteins and in designing efficient protocols of surfactant preparation and administration.     

Effect of mycolic acid on surface activity of binary surfactant lipid monolayers

In pulmonary tuberculosis, Mycobacterium tuberculosis lies in close physical proximity to alveolar surfactant. Cell walls of the mycobacteria contain loosely bound, detachable surface-active lipids. In this study, the effect of mycolic acid (MA), the most abundant mycobacterial cell wall lipid, on the surface activity of phospholipid mixtures from lung surfactant was investigated using Langmuir monolayers and atomic force microscopy (AFM). In the presence of mycolic acid, all the surfactant lipid mixtures attained high minimum surface tensions (between 20 and 40 mN/m) and decreased surface compressibility moduli <50 mN/m. AFM images showed that the smooth surface topography of surfactant lipid monolayers was altered with addition of MA. Aggregates with diverse heights of at least two layer thicknesses were found in the presence of mycolic acid. Mycolic acids could aggregate within surfactant lipid monolayers and result in disturbed monolayer surface activity. The extent of the effect of mycolic acid depended on the initial state of the monolayer, with fluid films of DPPC-POPC and DPPC-CHOL being least affected. The results imply inhibitory effects of mycolic acid toward lung surfactant lipids and could be a mechanism of lung surfactant dysfunction in pulmonary tuberculosis.     

Dynamic tension and adsorption behavior of aqueous lung surfactants

The dynamic tension behavior, at constant or at pulsating area conditions, of two commercial lung surfactants in saline is reported. The bubble method, at constant or pulsating area, at 37°C and the pendant drop method at 23°C were used. For Exosurf, a commercial synthetic lung surfactant consisting of dissolved tyloxapol and dispersed dipalmitoylphosphatidylcholine (or DPPC) and hexadecanol (H), the equilibrium and dynamic tensions are high (over 30 mN m⁻¹) and similar to those of tyloxapol alone. Aqueous DPPC/H mixtures have lower tensions than Exosurf. Survanta, a commercial lung surfactant replacement drug consisting of DPPC, other lipids, and two hydrophobic lung surfactant proteins, produces dynamic surface tensions that are substantially lower than those of Exosurf. Diluted 10-fold, Survanta produces under pulsating area (at 20 cycles min⁻¹) lower minimum tensions than undiluted Survanta (6 vs. 12 mN m⁻¹), but higher maximum tensions. In addition, Survanta tension behavior is unusual, having three local maxima and three local minima per cycle, suggesting major variations of its surface composition in each cycle. Monolayer pressure-area isotherms and Fourier transform infrared-attenuated total reflection (FTIR-ATR) spectroscopy results on deposited Langmuir–Blodgett films support this suggestion. They also provide direct evidence of the presence of phospholipids (DPPC or others) on the surface, but only indirect evidence of the presence of other components, on the surface of aqueous Exosurf or Survanta.     

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.     

Interactions between hematological derivatives and dipalmitoyl phosphatidyl choline: implications for adult respiratory distress syndrome

Blood and its components flood the alveoli in adult respiratory distress syndrome (ARDS) and may be responsible for the inhibition of lung surfactant in this syndrome. We have evaluated the surface properties of dipalmitoyl phosphatidylcholine (DPPC) monolayers-the main component of lung surfactant, in the presence of blood and its components. Experiments were performed using a Langmuir-Blodgett trough at physiological temperature (37 degrees C), pH 7.0 and using 0.9% saline as the sub-phase. Whole blood (WB), membranes obtained from whole blood cells (Mem), lysed blood (LB), homogenized blood clot (CLOT), serum (SER), platelet rich plasma (PRP), platelet poor plasma (PPP) and individual plasma proteins (albumin, fibrinogen) were added to the sub-phase in the equivalent proportion of 10 parts of DPPC per million parts (w/v) of hematological inhibitor. Cell membranes were found to be the most inhibitory agent for DPPC surface activity as evidenced by an increase in the minimum surface tension (from 0.818 +/- 0.219 to 7.373 +/- 0.854 mN/m) and percentage area change required to reduce the surface tension from 30 to 10 mN/m (from 21.24 +/- 0.99 to 66.83 +/- 4.44). The inhibitory potential of pure plasma proteins differed from those of more complex blood derivatives like platelet rich plasma and serum. Whole blood and platelet poor plasma were non-inhibitory, but serum, platelet rich plasma and clot significantly increased the minimum surface tension of DPPC to 6.819 +/- 0.925, 6.625 +/- 2.261 and 6.060 +/- 0.640 mN/m, respectively. These results were statistically significant with one-way analysis of variance and Newman-Keul's test (P < 0.05). The present study suggests that, not only the presence of the individual blood component(s) in the lung alveoli but also their interactions decide their inhibitory capability. Pure plasma proteins are not representative of the inhibitory effects of blood derivatives like serum, platelet rich plasma and blood cell membranes which would be more relevant for inhibitory models of ARDS.     

Relationships between equilibrium spreading pressure and phase equilibria of phospholipid bilayers and monolayers at the air-water interface

The intricate interplay between the bilayer and monolayer properties of phosphatidylcholine (PC), phosphatidylglycerol (PG), and phosphatidylethanolamine (PE) phospholipids, in relation to their polar headgroup properties, and the effects of chain permutations on those polar headgroup properties have been demonstrated for the first time with a set of time-independent bilayer-monolayer equilibria studies. Bilayer and monolayer phase behavior for PE is quite different than that observed for PC and PG. This difference is attributed to the characteristic biophysical PE polar headgroup property of favorable intermolecular hydrogen-bonding and electrostatic interactions in both the bilayer and monolayer states. This characteristic hydrogen-bonding ability of the PE polar headgroup is reflected in the condensed nature of PE monolayers and a decrease in equilibrium monolayer collapse pressure at temperatures below the monolayer critical temperature, T(c) (whether above or below the monolayer triple point temperature, T(t)). This interesting phenomena is compared to equilibrated PC and PG monolayers which collapse to form bilayers at 45 mN/m at temperatures both above and below monolayer T(c). Additionally, it has been demonstrated by measurements of the equilibrium spreading pressure, pie, that at temperatures above the bilayer main gel-to-liquid-crystalline phase-transition temperature, T(m), all liquid-crystalline phospholipid bilayers spread to form monolayers with pie around 45 mN/m, and spread liquid-expanded equilibrated monolayers collapse at 45 mN/m to form their respective thermodynamically stable liquid-crystalline bilayers. At temperatures below bilayer T(m), PC and PG gel bilayers exhibit a drop in bilayer pi(e) values < or =0.2 mN/m forming gaseous monolayers, whereas the value of pic of spread monolayers remains around 45 mN/m. This suggests that spread equilibrated PC and PG monolayers collapse to a metastable liquid-crystalline bilayer structure at temperatures below bilayer T(m) (where the thermodynamically stable bilayer liquid-crystalline phase does not exist) and with a surface pressure of 45 mN/m, a surface chemical property characteristically observed at temperatures above bilayer T(m) (monolayer T(c)). In contrast, PE gel bilayers, which exist at temperatures below bilayer T(m) but above bilayer T(s) (bilayer crystal-to-gel phase-transition temperature), exhibit gel bilayer spreading to form equilibrated monolayers with intermediate pie values in the range of 30-40 mN/m; however, bilayer pie and monolayer pic values remain equal in value to one another. Contrastingly, at temperatures below bilayer T(s), PE crystalline bilayers exhibit bilayer pie values < or =0.2 mN/m forming equilibrated gaseous monolayers, whereas spread monolayers collapse at a value of pic remaining around 30 mN/m, indicative of metastable gel bilayer formation.     

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.     

Penetration behaviour of alkylbetainate chlorides into lipid monolayers

In this paper, the penetration behaviour of the alkylbetainate chloride surfactants (C(n)BC, n=10-16) into lipid monolayers of dipalmitoylphosphatidylserine (DPPS), dipalmitoylphosphatidic acid (DPPA), dipalmitoylphosphatidylethanolamine (DPPE), palmitoyoleoylphosphatidylcholine (POPC) and cholesterol (CHOL) is investigated using the Langmuir trough technique. The penetration of C(n)BC is followed by measurement of the surface pressure increase (Δπ) at a constant surface area after the injection of C(n)BC into the aqueous phase, underneath the lipid monolayer previously spread at the air-water interface at 25°C and at different initial surface pressures (π(i)). The influence of both the lipid head group and the surfactant hydrocarbon chain length on the effectiveness of C(n)BC penetration into these monolayers is discussed. The results have shown that C(n)BC adsorb at the air-water interface giving evidence of their surface-active properties. The adsorption kinetics of C16BC into different lipid monolayers are lipid head charge and lipid head volume-dependent. The magnitude of the surface pressure increase (Δπ) arises in the following order: DPPA>DPPS?CHOL≈DPPE>POPC. C(n)BC penetration into negatively-charged (DPPS and DPPA) monolayers does not seem to depend on surfactant alkyl-chain length compared to uncharged (CHOL) and zwitterionic (DPPE and POPC) monolayers for which Δπ increases with a larger alkyl-chain length. Electrostatic interactions are mainly involved in the affinity of C(n)BC with monolayers but the hydrophobic effect plays also a role.     

Interfacial behaviour and mechanical properties of spread lung surfactant protein/lipid layers

The surface behaviour of spread dipalmitoyl phosphatidyl choline (DPPC), lung surfactant protein C (SP-C), and their mixtures were characterised using a captive bubble surfactometer. The surface tension was determined by using axisymmetric bubble shape analysis. Surface dilatational rheological behaviour was characterised by sinusoidal oscillation of the bubble volume and at frequencies 0.006-0.025 Hz. The pi/A isotherms of DPPC, SP-C, and their mixtures were described with a generalised equation of state. Monolayer cycling of mixed DPPC/SP-C layers yields isotherms with a plateau in the range of 50-53 mN/m. When the surface pressure becomes higher SP-C is squeezed out of the film, but it re-enters the film upon expansion. Surface dilatational elasticities of DPPC films had a maximum at about 30 mN/m. At higher surface pressures, the films became brittle and the elasticity decreased. A slightly pronounced maximum was found at a surface pressure exceeding 55 mN/m. The dilatational viscosity had two distinct maxima, corresponding with those in the elasticity curves, i.e. one before the minimum area demand, and one in the range of over-compression. This was explained by the formation of a second ordered complex structure in the range of film over-compression. SP-C films show continuously increasing dilatational elasticities and viscosities with a maximum at f approximately 0.02 Hz. Mixed monolayers, DPPC+2 mol% SP-C, had dilatational elasticities increasing with surface pressure. In contrast to DPPC alone, an elasticity maximum appeared in the range of the squeeze out plateau. The dilatational viscosity had two distinct maxima as observed for DPPC, whereas the maximum before the squeeze out plateau is very broad like that of SP-C. The viscosity decreased for frequencies higher 0.02 Hz favouring elastic properties of the film. Our data provide experimental evidence that SP-C mixed with DPPC yield higher elasticities and viscosities as compared with films formed by the single components. This behaviour is likely to support breathing cycles, especially for the turn from inspiration to expiration and vice versa.     

Effect of temperature on surface properties of cervical tissue homogenate and organic phase monolayers

The temperature dependence of Langmuir monolayers of normal and cancerous human cervical tissues and their organic phases between temperatures of 37 and 45 degrees C was evaluated. Analysis of the surface pressure-area isotherms revealed significantly different increase in fluidity of the cancerous cervical tissue monolayer at 42 degrees C as opposed to the normal cervical tissue monolayers (p<0.05). Similarly, in the case of cervical cancerous organic phase monolayers significant increase of fluidity was observed at 40 degrees C whereas no such change was observed in the normal cervical organic phase monolayers. The effect of temperature was found to be different in cancerous and normal cervical tissues and this may be due to the different lipid profiles in them. Cancerous cervical tissues had 1.8-fold higher total lipids as compared to the normals. Similarly, the PC, PE, PI, PG, SM and PS levels in cancerous cervical tissues were 3.6, 2.0, 2.3, 4.7, 1.7 and 2.2 times higher than those of normal cervical tissues, respectively. Significant cancer-normal difference in minimum surface tension and hysteresis area was found at all temperatures studied for both tissue homogenates and organic phases. For example, cancerous tissue homogenates showed minimum surface tensions of 51.9+/-4.6, 54.4+/-5.9, 57.6+/-6.0 and 51.9+/-5.6mN/m at temperatures 37, 40, 42 and 45 degrees C whereas the corresponding values for normal cervical tissue homogenates were 39.3+/-3.6, 39.2+/-3.7, 39.2+/-3.8 and 39.1+/-3.6, respectively. The fluidity change at hyperthermic range of temperature can be correlated to the increased efficiency of drug on combination therapy with hyperthermia. These results may have implications in manipulating the fluidity of cervical cancer tissue membranes for better permeability thereby leading to better therapeutic strategies for cervical cancer.     

Influence of hydrophobic alkylated gold nanoparticles on the phase behavior of monolayers of DPPC and clinical lung surfactant

The effect of hydrophobic alkylated gold nanoparticles (Au NPs) on the phase behavior and structure of Langmuir monolayers of dipalmitoylphosphatidylcholine (DPPC) and Survanta, a naturally derived commercial pulmonary surfactant that contains DPPC as the main lipid component and hydrophobic surfactant proteins SP-B and SP-C, has been investigated in connection with the potential implication of inorganic NPs in pulmonary surfactant dysfunction. Hexadecanethiolate-capped Au NPs (C(16)SAu NPs) with an average core diameter of 2 nm have been incorporated into DPPC monolayers in concentrations ranging from 0.1 to 0.5 mol %. Concentrations of up to 0.2 mol % in DPPC and 16 wt % in Survanta do not affect the monolayer phase behavior at 20 °C, as evidenced by surface pressure-area (π-A) and ellipsometric isotherms. The monolayer structure at the air/water interface was imaged as a function of the surface pressure by Brewster angle microscopy (BAM). In the liquid-expanded/liquid-condensed phase coexistence region of DPPC, the presence of 0.2 mol % C(16)SAu NPs causes the formation of many small, circular, condensed lipid domains, in contrast to the characteristic larger multilobes formed by pure lipid. Condensed domains of similar size and shape to those of DPPC with 0.2 mol % C(16)SAu NPs are formed by compressing Survanta, and these are not affected by the C(16)SAu NPs. Atomic force microscopy images of Langmuir-Schaefer-deposited films support the BAM observations and reveal, moreover, that at high surface pressures (i.e., 35 and 45 mN m(-1)) the C(16)SAu NPs form honeycomb-like aggregates around the polygonal condensed DPPC domains. In the Survanta monolayers, the C(16)SAu NPs were found to accumulate together with the proteins in the liquid-expanded phase around the circular condensed lipid domains. In conclusion, the presence of hydrophobic C(16)SAu NPs in amounts that do not influence the π-A isotherm alters the nucleation, growth, and morphology of the condensed domains in monolayers of DPPC but not of those of Survanta. Systematic investigations of the effect of the interaction of chemically defined NPs with the lipid and protein components of lung surfactant on the physicochemical properties of surfactant films are pertinent to understanding how inhaled NPs impact pulmonary function.     

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.     

Membrane properties of mixed ganglioside GM1/phosphatidylcholine monolayers

The interaction between ganglioside G_M1 (GM1) and --dipalmitoylphosphatidylcholine (DPPC) in mixed monolayers was investigated using surface pressure measurements and atomic force microscopy (AFM), and the effects of GM1, surface pressure and temperature on the properties of the membranes were examined. Mixed GM1/DPPC monolayers were deposited on mica using the Langmuir–Blodgett (LB) technique for AFM. GM1 and DPPC were miscible below the 0.2 mole fraction of GM1 and there was attractive interaction between GM1 and DPPC. The AFM images for the GM1/DPPC monolayers (X_GM1 < 0.2) at 30 mN m⁻¹ and 25 °C indicated a percolation pattern which means a micro phase separation: namely, the mixed film composed of GM1 and DPPC phase-separated from the DPPC liquid-condensed film. The AFM images for the mixed monolayers at 33 mN m⁻¹ indicated a specific morphology when the surface pressure was varied from 30 to 40 mN m⁻¹. The percolation pattern in the AFM image at 25 °C came to be destroyed with increasing temperature and completely disappeared at 45 °C. The change in the morphology of mixed GM1/DPPC monolayers on varying the surface pressure and temperature is thought to be related to signal transduction and a preventive mechanism against viral infections in the human body.     

A monolayer study on interactions of docetaxel with model lipid membranes

Docetaxel (DCT) is an antineoplastic drug for the treatment of a wide spectrum of cancers. DCT surface properties as well as miscibility studies with l-alpha-dipalmitoyl phosphatidylcholine (DPPC), which constitutes the main component of biological membranes, are comprehensively described in this contribution. Penetration studies have revealed that when DCT is injected under DPPC monolayers compressed to different surface pressures, it penetrates into the lipid monolayer promoting an increase in the surface pressure. DCT is a surface active molecule able to decrease the surface tension of water and to form insoluble films when spread on aqueous subphases. The maximum surface pressure reached after compression of a DCT Langmuir film was 13 mN/m. Miscibility of DPPC and DCT in Langmuir films has been studied by means of thermodynamic properties as well as by Brewster angle microscopy (BAM) analysis of the mixed films at the air-water interface, concluding that DPPC and DCT are miscible and they form non-ideally mixed monolayers at the air-water interface. Helmholtz energies of mixing revealed that no phase separation occurs. In addition, Helmholtz energies of mixing become more negative with decreasing areas per molecule, which suggests that the stability of the mixed monolayers increases as the monolayers become more condensed. Compressibility values together with BAM images indicate that DCT has a fluidizing effect on DPPC monolayers.     

Langmuir monolayer of artificial pulmonary surfactant mixtures with an amphiphilic peptide at the air/water interface: comparison of new preparations with surfacten (Surfactant TA)

Interfacial behavior was studied on the pulmonary lipid mixture containing a newly designed amphiphilic alpha-helical peptide (Hel 13-5) that consists of 13 hydrophobic and 5 hydrophilic amino acid residues. Moreover, the data obtained were compared with those of commercially available Surfacten (Surfactant TA) which has been clinically used for neonatal respiratory distress syndrome (NRDS) in Japan. Surface pressure (pi)-A and surface potential (DeltaV)-area (A) isotherms were measured for our synthetic preparations and Surfacten. Herein, a mixture of dipalmitoylphosphatidylcholine (DPPC)/egg-phosphatidylglycerol (PG)/palmitic acid (PA) (68:22:9 by weight) was used as the constituent of basic preparations. Monolayers were spread on 0.02 M Tris buffer (pH 7.4) with 0.13 M NaCl at the air/liquid interface, and the surface behavior was investigated by employing the Wilhelmy method, an ionizing electrode method, and fluorescence microscopy (FM). Cyclic compression and expansion isotherms of the prepared materials (or products) (DPPC/PG/PA/Hel 13-5) were examined to confirm the spreading and respreading ability. For the prepared products, a plateau region exists on pi-A and DeltaV-A isotherms at approximately 42 mN m(-1), indicating that Hel 13-5 is squeezed out of surface monolayers together with fluid components (PG) upon lateral compression. That is, the squeeze-out phenomenon induces a 2D-3D phase transformation. In particular, the inclination of the pi-A isotherms at X(Hel 13-5) = 0.1 in the plateau region was almost zero irrespective of the molecular area. As proposed in the earlier report (Nakahara, H.; Lee, S.; Sugihara, G.; Shibata, O. Langmuir 2006, 22, 5792-5803), an observed refluorescence phenomenon was discussed for FM measurements. This phenomenon provides evidence of the squeeze-out motion with fluid molecules. Furthermore, the cyclic pi-A and DeltaV-A isotherms show larger hysteresis areas and better respreading abilities in comparison with the previous ternary systems (DPPC/PG/Hel 13-5 and DPPC/PA/Hel 13-5) that are very important properties in pulmonary functions. FM photographs and the temperature dependence of pi-A and DeltaV-A isotherms suggest that the phase behavior of the present preparation product is very similar to that of Surfacten in terms of the domain size and in parameters such as collapse pressures, maximum DeltaV values, and so on. These results demonstrate that PG and PA even in the present preparations work well for compression-expansion cycling as is the case in the previous ternary systems, and the present preparations show comparable properties to Surfacten in vitro.     

Interaction of hexadecylbetainate chloride with biological relevant lipids

The present work investigates the interaction of hexadecylbetainate chloride (C(16)BC), a glycine betaine-based ester with palmitoyl-oleoyl-phosphatidylcholine (POPC), sphingomyelin (SM), and cholesterol (CHOL), three biological relevant lipids present in the outer leaflet of the mammalian plasma membrane. The binding affinity and the mixing behavior between the lipids and C(16)BC are discussed based on experimental (isothermal titration calorimetry (ITC) and Langmuir film balance) and molecular modeling studies. The results show that the interaction between C(16)BC and each lipid is thermodynamically favorable and does not affect the integrity of the lipid vesicles. The primary adsorption of C(16)BC into the lipid film is mainly governed by a hydrophobic effect. Once C(16)BC is inserted in the lipid film, the polar component of the interaction energy between C(16)BC and the lipid becomes predominant. Presence of CHOL increases the affinity of C(16)BC for membrane. This result can be explained by the optimal matching between C(16)BC and CHOL within the film rather by a change of membrane fluidity due to the presence of CHOL. The interaction between C(16)BC and SM is also favorable and gives rise to highly stable monolayers probably due to hydrogen bonds between their hydrophilic groups. The interaction of C(16)BC with POPC is less favorable but does not destabilize the mixed monolayer from a thermodynamic point of view. Interestingly, for all the monolayers investigated, the exclusion surface pressures are above the presumed lateral pressure of the plasma membranes suggesting that C(16)BC would be able to penetrate into mammalian plasma membranes in vivo. These results may serve as a useful basis in understanding the interaction of C(16)BC with real membranes.