ABSTRACT
The outer membrane (OM) of Gram-negative bacteria is a complex and asymmetric bilayer that antimicrobial peptides must disrupt in order to provoke the cell lysis. The inner and external leaflets of the OM are mainly composed of phospholipids (PL), and lipopolysaccharide (LPS), respectively. Supported lipid bilayers are interesting model systems to mimic the lipid asymmetric scaffold of the OM and determine the quantitative and mechanistic effect of antimicrobial agents, using complementary physicochemical techniques. We report the formation of asymmetric PL/LPS bilayers using the Langmuir-Blodgett/Langmuir-Schaefer technique on two different surfaces (sapphire and mica) with synthetic phospholipids constituting the inner leaflet and bacteria-extracted mutant LPS making up the outer one. The combination of neutron reflectometry and atomic force microscopy techniques allowed the examination of the asymmetric scaffold structure along the normal to the interface and its surface morphology in buffer conditions. Our results allow discrimination of two structurally related peptides, one neutral and inactive, and the other cationic and active. The active cationic plasticin PTCDA1-KF disrupts the asymmetric OM at relevant concentrations through a carpeting scenario characterized by a dramatic removal of lipid molecules from the surface.
Subject(s)
Gram-Negative Bacteria , Biomimetic Materials , Cell Membrane , Eye Proteins , Lipid Bilayers , Lipopolysaccharides , Nerve Tissue ProteinsABSTRACT
Bacterial resistance to antibiotics has become today a major public health issue. In the development of new anti-infectious therapies, antimicrobial peptides appear as promising candidates. However, their mechanisms of action against bacterial membranes are still poorly understood. We describe for the first time the interaction and penetration of plasticins into lipid monolayers and bilayers modeling the two leaflets of the asymmetrical outer membrane of Gram-negative bacteria. The lipid composition of these monolayers mimics that of each leaflet: mixtures of LPS Re 595 mutant and wild type S-form from Salmonella enterica for the external leaflet, and SOPE/SOPG/cardiolipin (80/15/5) for the inner one. The analysis of the interfacial behavior of native (PTCDA1) and modified (PTCDA1-KF) antimicrobial plasticins showed that PTCDA1-KF exhibited better surface properties than its unmodified counterpart. Both peptides could penetrate into the model monolayers at concentrations higher than 0.1 µM. The penetration was particularly enhanced for PTCDA1-KF into the mixed LPS monolayer, due to attractive electrostatic interactions. Grazing X-ray diffraction and atomic force microscopy studies revealed the changes in LPS monolayers organization upon peptide insertion. The interaction of plasticins with liposomes was also monitored by light scattering and circular dichroism techniques. Only the cationic plasticin achieved full disaggregation and structuration in α helices, whereas the native one remained aggregated and unstructured. The main steps of the penetration mechanism of the two plasticins into lipid models of the external leaflet of the outer membrane of Gram-negative bacteria have been established.
Subject(s)
Cell Membrane/chemistry , Eye Proteins/chemistry , Gram-Negative Bacteria/chemistry , Lipopolysaccharides/chemistry , Nerve Tissue Proteins/chemistry , Cell Membrane/metabolism , Circular Dichroism , Eye Proteins/metabolism , Gram-Negative Bacteria/metabolism , Lipopolysaccharides/metabolism , Liposomes/chemistry , Liposomes/metabolism , Membrane Lipids/chemistry , Membrane Lipids/metabolism , Microscopy, Atomic Force , Nerve Tissue Proteins/metabolism , Phospholipids/chemistry , Phospholipids/metabolism , Protein Binding , Salmonella enterica/chemistry , Salmonella enterica/metabolism , Static Electricity , X-Ray DiffractionABSTRACT
INTRODUCTION: Porphyrins are photosensitizers usable in photodynamic therapy. Although these molecules are clinically effective, their low water solubility and their lack of specificity are major drawbacks to their development. Our study was aimed at analysing the interfacial behaviour of glycoconjugated tetraphenylporphyrins newly synthesized at the Curie Institute, and their interaction with model membranes bearing a specific lectin mimicking a mannose membrane receptor in retinoblastoma. MATERIAL AND METHODS: The interfacial behaviour of the porphyrins was analysed by surface pressure measurements, and their specific interaction with the lectin, by dynamic light scattering (liposomes) and the quartz crystal microbalance technique (supported bilayers). RESULTS: All porphyrin derivatives were able to organize at the air/liquid interface. The dendrimeric compounds formed more stable monolayers than the others, and generally showed good mixing properties with the phospholipid used for liposome preparation. In the presence of concanavalin A, the porphyrin bearing-liposomes behaved differently depending on the nature (mannosylated or not) of the porphyrins. DISCUSSION: The interfacial behaviour of the tetraphenylporphyrins is directly related to the orientation of the tetrapyrrolic macrocycle controlled by the grafted groups. Incorporated into a liposome bilayer, glycodendrimeric porphyrins expose their sugar moieties at the vesicle surface. The spacer length plays a crucial role by increasing sugars freedom and enhancing glycosylated liposomes interaction with the lectin. CONCLUSION: Compared to the other studied compounds, the glycodendrimeric porphyrins seem very promising compounds and are now evaluated on cell cultures.
Subject(s)
Biomimetics , Cell Membrane/drug effects , Glycoconjugates/pharmacology , Photosensitizing Agents/pharmacology , Porphyrins/pharmacology , Cell Line, Tumor , Concanavalin A/chemistry , Dendrimers , Glycoconjugates/chemistry , Humans , Lectins , Membranes, Artificial , Phospholipids/chemistry , Photochemotherapy , Photosensitizing Agents/chemistry , Porphyrins/chemistry , Retinal Neoplasms/metabolism , Retinoblastoma/metabolism , Surface PropertiesABSTRACT
Photodynamic therapy is a potentially efficient treatment for various solid tumours, among which retinoblastoma. Its efficacy depends on the preferential accumulation of photosensitizers in the malignant tissues and their accessibility to light. The specificity of drugs for retinoblastoma cells can be improved by targeting a mannose receptor overexpressed at their surface. With the aim of assessing the recognition of newly synthesized glycodendrimeric porphyrins by such receptors, we have built and characterized an original synthetic biomimetic membrane having similar lipidic composition to that of the retinal cell membranes and bearing Concanavalin A, as a model of the mannose receptor. The interaction of the porphyrin derivatives with liposomes and supported planar bilayers has been studied by dynamic light scattering and quartz crystal microbalance with dissipation monitoring (QCM-D). Only mannosylated porphyrins interacted significantly with the membrane model. The methodology used proved to be efficient for the selection of potentially active compounds.
Subject(s)
Biomimetics , Cell Membrane/metabolism , Lectins/metabolism , Lipid Bilayers/metabolism , Mannose/metabolism , Porphyrins/metabolism , Concanavalin A/metabolism , Humans , Liposomes , Quartz Crystal Microbalance Techniques , Retinoblastoma , Tumor Cells, CulturedABSTRACT
In photodynamic therapy, the specificity of a photosensitizer and its penetration into tumor cells are crucial. We have analyzed the ability of newly synthesized meso-(tetraphenyl)porphyrins to be recognized by a model of mannose-specific proteins overexpressed at the surface of retinoblastoma cells. The specific interaction of porphyrin with Con A was studied by surface pressure measurements, fluorescence spectroscopy, dynamic light scattering, and QCM-D. The extent of porphyrins binding to Con A was highly dependent upon their chemical structure. Glycodendrimeric porphyrins showed the higher binding constant to Con A. The length of the spacer separating the sugar from the tetrapyrrolic ring appeared to be crucial in controlling the interaction of the compounds with the lectin in solution or immobilized onto a solid substrate. The methodology used proved to be efficient for the selection of potentially active compounds. The glycodendrimeric porphyrins, especially the derivative having the longer spacer, interacted more significantly with the lectin than the compound devoid of any sugar.
Subject(s)
Dendrimers/chemical synthesis , Liposomes/chemistry , Photosensitizing Agents/chemistry , Porphyrins/chemistry , Spectrometry, Fluorescence/methods , Concanavalin A/chemistry , Models, Chemical , Surface PropertiesABSTRACT
Photodynamic therapy (PDT) is considered one efficient treatment against retinoblastoma. The specificity of a photosensitizer and its penetration into cancerous cells are crucial for achieving tumor necrosis. The selection of photosensitizers such as porphyrin derivatives by tumor cells thus depends to a large extent on their ability to interact with the biological membrane. In this work, we have studied by surface pressure measurements and fluorescence spectroscopy the interaction between three newly synthesized dendrimeric phenylporphyrins and monolayers or liposomes with increasing cholesterol content mimicking the retinoblastoma cell membrane. The morphology of phospholipid-cholesterol-porphyrin mixed monolayers was also analyzed by Brewster angle microscopy. The results showed that the increase in cholesterol content in the model membranes had almost no effect on the effective penetration of the drugs into the lipid layers. Conversely, the chemical structure of the glycodendrimeric phenylporphyrins and the presence of sugar moieties especially appeared to play a crucial role. Although the non-glycoconjugated phenylporphyrin penetrated to a greater extent than glycodendrimeric ones into the liposome membrane, this could be achieved at a high lipid/porphyrin ratio only. Glycodendrimeric porphyrins exhibited improved surface properties compared to the non-glycoconjugated derivative and could penetrate into lipid layers even at low lipid/porphyrin ratios and high surface pressures. Our work highlights the role in the passive diffusion of porphyrins into biomimetic cancer cell membranes, of complex interactions among the lipid molecules, the sugar moieties, and the hydrophobic macrocycle of the porphyrins.
Subject(s)
Carbohydrates/chemistry , Cell Membrane/chemistry , Cholesterol/chemistry , Liposomes/chemistry , Photosensitizing Agents/chemistry , Porphyrins/chemistry , Cell Line, Tumor , Humans , Photochemotherapy , Retinoblastoma/drug therapy , Spectrometry, FluorescenceABSTRACT
In multiple emulsion systems, oily or aqueous transfers may occur between the dispersed droplets through the continuous phase. These transfers are controlled by both the surfactant system (micellar transport), and the partial solubility of one phase in another (molecular transport). The latter could be anticipated from the knowledge of oil polarity, if this information could easily be obtained. In this work, the relative polarity of eight oils used for various purposes has been evaluated from the comparison of their dielectric requirement for solubilization, their interfacial tension and chromatographic analysis. The results showed the complementarities of HPLC analysis and interfacial tension measurements and their superiority over the solubilization method for classifying oils as a function of their polarity.
Subject(s)
Oils/chemistry , Solvents/chemistry , Water/chemistry , Alkanes/chemistry , Chromatography, High Pressure Liquid/methods , Dioxanes/chemistry , Emulsions/chemistry , Fatty Alcohols/chemistry , Mineral Oil/chemistry , Palmitates/chemistry , Phase Transition , Reproducibility of Results , Solubility , Static Electricity , Surface Properties , Surface Tension , Triglycerides/chemistryABSTRACT
New nanoassemblies were instantaneously prepared by mixing two aqueous solutions, one containing a beta-cyclodextrin polymer (pbetaCD), and the other a hydrophobically modified by alkyl chains dextran (MD). The formation mechanism and the inner structure of these nanoassemblies were analysed using surface tension measurements and (1)H NMR spectroscopy. The effect of a hydrophobic guest molecule, such as benzophenone (BZ), on the formation and stability of the nanoassemblies was also evaluated. MD exhibited the typical behaviour of a soluble amphiphilic molecule and adsorbed at the air/water interface. Whereas the injection of native beta-CDs in the solution beneath the adsorbed MD monolayer did not produce any change in the surface tension, that of the pbetaCD resulted in an increase in the surface tension, indicating the desorption of the polymer from the interface. This result accounts for a cooperative effect of beta-CDs linked together in the pbetaCD polymer on dextran desorption. The presence of benzophenone in the system hindered the sequestration of dextran alkyl moieties by beta-CD in the polymer without impeding the formation of associative nanoassemblies of 100-200 nm. (1)H NMR investigations demonstrated that, in the BZ-loaded nanoassemblies, the hydrophobic molecule was mainly located into the cyclodextrin cavities.
Subject(s)
Benzophenones/chemistry , Dextrans/chemistry , Nanoparticles/chemistry , Propylene Glycols/chemistry , beta-Cyclodextrins/chemistry , Hydrophobic and Hydrophilic Interactions , Magnetic Resonance SpectroscopyABSTRACT
The surface properties of the neoglycolipid (GlcNAcE(3)G(28)) and of its PEO-lipid (E(3)G(28)) moiety mixed with phospholipids (dipalmitoylphosphatidylcholine, DPPC; distearoylphosphatidylcholine, DSPC; diarachidoylphosphatidylcholine, DAPC; and dibehenoylphosphatidylcholine, DBPC) were studied in Langmuir monolayers at various mixture compositions and surface pressures. The pi-A isotherms of the pure compounds revealed that because of the presence of the sugar group in its molecule, GlcNAcE(3)G(28) collapsed at a higher surface pressure and occupied a larger molecular area than the PEO-lipid moiety. It was also observed that the presence of the PEO-lipid (E(3)G(28)) in the mixtures triggered a strong alteration of both phospholipid pi-A isotherm profiles and surface diffraction spectra, an indication that the disordering of the initially structured phospholipid monolayers took place. Unlike E(3)G(28), GlcNAcE(3)G(28) did not disorganize phospholipid monolayers but generated a partial segregation of the film-forming components. The calculated excess free energies of mixing (DeltaG(exc)) for GlcNAcE(3)G(28)-phospholipid mixtures enabled us to predict the stability of such systems.
Subject(s)
Phospholipids , Surface Properties , PressureABSTRACT
The serotonergic system plays a critical role in a wide variety of physiological and behavioral processes. Dysregulation of the tightly controlled extracellular concentration of serotonin (5-hydroxytryptamine, 5-HT) appears to be at the origin of a host of metabolic and psychiatric disorders. Since the plasma membrane 5-HT transporter (SERT) is the major protagonist in regulating extracellular 5-HT concentration, SERT is the target of most drugs interacting with the serotonergic system. Unfortunately, some of the drugs towards SERT (e.g. amphetamine derivatives) interfere with cell homeostasis leading to cell toxicity. Developing new SERT ligands devoid of any side-effect represents a major priority in the treatment of 5-HT-associated pathologies. Here, we report structure-activity relationships (SAR) and three-dimensional QSAR (3D-QSAR) studies of a library of 121 compounds including 5-HT analogs, harmanes, benzothiazoles, indanones, amphetamine derivatives and substrate-type 5-HT releasers, with the goal of identifying the structural determinants crucial for SERT uptake. In the absence of data about the bioactive form of 5-HT, conformational analysis of 5-HT was performed using quantum chemistry calculations. This led to three 5-HT stable conformers with anti, -gauche and +gauche side-chain conformation. These conformers, used as templates for superimposition with all the library compounds, enabled the design of a reliable 6-points pharmacophore representative of SERT uptake activity. Molecular dynamics (MD) simulations performed with compounds that are efficiently, moderately, poorly or not transported by SERT allowed to assess the validity of our pharmacophore. Altogether, our data provide for the first time a reliable pharmacophore of SERT uptake activity, which may help to the design of new drugs targeting SERT.
Subject(s)
Blood Platelets/metabolism , Models, Molecular , Serotonin/pharmacokinetics , Adult , Drug Design , Humans , Quantitative Structure-Activity Relationship , Serotonin/analogs & derivatives , Serotonin/chemistryABSTRACT
We have recently described how oligonucleotide (ON) stability and release from O/W cationic emulsions are governed by the lipid composition. The aim of the present paper was to investigate the properties of the ON/lipid complexes through fluorescence resonance energy transfer (FRET), size, surface tension measurements and cryomicroscopy. Starting from a typical emulsion containing stearylamine as a cationic lipid, the influence of the lipid structure (monocationic molecules bearing mono or diacyl chains, or polycations) as well as of the presence of PEGylated lipids, were studied. The presence of a positive charge on the droplet surface clearly contributed to enhance the ON interaction with lipid monolayers and to bring the ON molecules closer to the interface. Hydrophobic interactions through the acyl chains were shown to further enhance the anchorage of the ON/lipid complexes. In contrast, the incorporation of PEGylated lipids acted as a barrier against the establishment of electrostatic bindings, the polyethyleneglycol chains acting themselves as interaction sites for the ON leading to hydrophilic complexes. Similar features were observed for the polycationic lipid, and cryomicroscopy revealed the existence of bridges of various intensities between the droplets of the emulsion containing either PEG or the polycation, probably because of the configuration of the ON at the interface.
Subject(s)
Lipids/chemistry , Oligonucleotides/chemistry , Polyethylene Glycols/chemistry , Algorithms , Amines/chemistry , Cations , Chemical Phenomena , Chemistry, Physical , Coloring Agents , Cryoelectron Microscopy , Emulsions , Energy Transfer , Excipients , Fluorescence , Surface TensionABSTRACT
The serotonin transporter (SERT) is the target of several important antidepressant and psychostimulant drugs. It has been shown that under defined conditions, the transporter spread at the air/water interface was able to bind its specific ligands. In this paper, the interfacial organization of the protein has been assessed from dynamic surface pressure and ellipsometric measurements. For areas comprising between 10,400 and 7,100 A(2)/molecule, ellipsometric measurements reveal an important change in the thickness of the SERT film. This change was attributed to the reorientation of the transporter molecules from a horizontal to their natural predictive transmembrane orientation. The thickness of the SERT film at 7,100 A(2)/molecule was found to be approximately equal to 84 A and coincided well with the theoretical value estimated from the calculations based on the dimensions of alpha-helices containing membrane proteins. These data suggest that the three-dimensional arrangement of the SERT may be represented as a box with lengths d(z)=83--85 A and d(y) or d(x)=41--47 A.
Subject(s)
Carrier Proteins/chemistry , Membrane Glycoproteins/chemistry , Membrane Transport Proteins , Nerve Tissue Proteins , Air , Blood Platelets/chemistry , Humans , Pressure , Serotonin Plasma Membrane Transport Proteins , Water/chemistryABSTRACT
We recently described a positively charged O-W emulsion as a delivery system for oligonucleotides (ON) [Teixeira et al., Pharm. Res. 16 (1999) 30-36]. The present paper investigates the role of the main formulation parameters that may have an influence on the release-rate of a model ON in a protein-containing medium, i.e. the nature of the oily core, the presence of pegylated lipids, the lipid phase transition temperature, and the cationic lipid structure. The use of cationic lipids bearing diacyl chains (and especially polycations) appeared as the only efficient strategy to reduce the ON release rate. In order to have a better insight on the nature of the interactions between the ON and the interfacial lipids, adsorption isotherms at the air-water interface, fluorescence resonance energy transfer and zeta-potential measurements have been performed. Electrostatic interactions were found to play a crucial role. In contrast, the incorporation of PEG-phospholipids acted as a barrier and maintained the ON molecules distant from the interface, leading to a more rapid release. Finally, ON integrity was assessed by a competitive hybridization assay. The results suggest the existence of a transient ion-pair (ON-cationic lipids) protecting ON against nuclease degradation even after its release from the emulsions.
Subject(s)
Drug Delivery Systems , Oligonucleotides/administration & dosage , Cells, Cultured , Drug Stability , Emulsions , Fluorescence , Oligonucleotides/chemistry , Solubility , Surface TensionABSTRACT
The interfacial behavior of the neoglycolipids formed of Guerbet alcohol (G(28)) bound to a triethylene glycol spacer (E(3)) and to a sugar moiety (alpha- and beta-fucose) spread at the air/water interface has been studied under dynamic conditions of compression. Although the alpha (alpha-FucE3G28)- and beta-fucose (beta-FucE3G28) derivatives possessed the same chemical structure, the positioning of the sugar moiety relative to the whole molecule had a significant influence on the organization of neoglycolipid molecules in the spread monolayers. Thus, beta-fucose molecules exhibited higher compressibilities and larger molecular areas than a alpha/beta (84/16%) mixture (alpha(84)-FucE3G28). The comparison of the compressional behavior of the fucose derivatives with that of Guerbet alcohol in the absence and in the presence of the triethylene glycol spacer shows that the presence of the E(3) chain is necessary to stabilize the lipid at the interface and that the incorporation of a sugar moiety into the molecule resulted in an important expansion of a monolayer. Despite their different interfacial behaviors, the two sugar derivatives formed ideal mixtures when cospread at the air/water interface. Conversely, in the presence of a phospholipid, such as DMPC, repulsive interactions were observed and appeared to be stronger for DMPC/alpha(84)-FucE3G28 mixed monolayers. The membrane fluidity of DMPC liposomes bearing the studied amphiphilic molecules was assessed by fluorescence depolarization measurements. The results reveal that whereas G(28) was deeply inserted into the liposome bilayers, the presence of a E(3) chain and of a sugar moiety in these bilayers induced a transfer of the amphiphilic derivatives from the hydrophobic core towards polar headgroups of phospholipid molecules.
Subject(s)
Fucose/chemistry , Glycolipids/chemical synthesis , Phospholipids/chemistry , Glycolipids/chemistry , Liposomes , Membrane Fluidity , Surface PropertiesABSTRACT
The cytoplasmic delivery of drugs encapsulated into pH-sensitive liposomes is under the control of a lamellar-to-hexagonal transition. In a previous study, under anhydrous conditions, oligonucleotides (ODN) encapsulated in pH-sensitive liposomes composed of dioleoylphosphatidylethanolamine (DOPE)/oleic acid (OA)/cholesterol (CHOL) were shown to modify the phase behaviour of DOPE. In the present study, the lipid/ODN interactions were evaluated in fully hydrated samples by surface tension measurements, differential scanning calorimetry, X-ray diffraction and turbidimetry. Concerning the lipids, it was shown that OA provoked a disorganisation of DOPE lamellar phases and led to the complete disappearance of hexagonal transition along with heating. The addition of CHOL further decreased the lipid packing in the bilayers. Concerning ODN, these molecules provoked an increase in the surface pressure of a DOPE/OA/CHOL monolayer, indicating the existence of molecular interactions with the lipids. At a supramolecular level, ODN induced a more ordered organisation of DOPE molecules in the lamellar and hexagonal phases, and completely abolished the disorganisational effect of OA and CHOL.
Subject(s)
Glycerophospholipids/chemistry , Liposomes/chemistry , Oligodeoxyribonucleotides/chemistry , Phosphatidylethanolamines , Calorimetry, Differential Scanning , Chemical Phenomena , Chemistry, Physical , Cholesterol/chemistry , Drug Carriers/chemistry , Hydrogen-Ion Concentration , In Vitro Techniques , Lipid Bilayers/chemistry , Macromolecular Substances , Oleic Acid/chemistry , Scattering, Radiation , Surface Tension , Water , X-RaysABSTRACT
The purified serotonin transporter (SERT) was spread at the air/water interface and the effects both of its surface density and of the temperature on its interfacial behavior were studied. The recorded isotherms evidenced the existence of a stable monolayer undergoing a lengthy rearrangement. SERT/ligand interactions appeared to be dependent on the nature of the studied molecules. Whereas an unrelated drug (chlorcyclizine) did not bind to the spread SERT, it interacted with its specific ligands. Compared to heterocyclic drugs, for which binding appeared to be concentration-dependent, a 'two-site' mechanism was evidenced for pinoline and imipramine.
Subject(s)
Carrier Proteins/metabolism , Membrane Glycoproteins/metabolism , Membrane Transport Proteins , Nerve Tissue Proteins , Air , Binding Sites , Biological Transport , Carrier Proteins/chemistry , Dose-Response Relationship, Drug , Humans , Ligands , Membrane Glycoproteins/chemistry , Serotonin Plasma Membrane Transport Proteins , Solutions , Surface Tension , Temperament , Water/chemistryABSTRACT
Covalent modification of glucose oxidase from Aspergillus niger by the palmitic acid ester of N-hydroxysuccinimide at a molar ratio ester:protein of 56:1 results in the formation of the enzyme derivative with 11 attached palmitic chains. Surface hydrophobicity measurements by a fluorescent probe, 8-anilino-1-naphthalenesulfonate, indicate a drastic increase in the hydrophobicity index of glucose oxidase after such a modification. The modified glucose oxidase displays a much higher adsorption affinity for hydrophilic (silica) as well as for hydrophobic (silica coated by phosphatidyl choline and cholesterol monolayers and polystyrene latex beads) surfaces, and forms more compact surface layers compared to the native glucose oxidase. Such a difference results from a spontaneous formation of micelle-like aggregates (clusters) of the hydrophobized enzyme molecules (average size 500 nm), which come into contact with a surface. A possible structure of the glucose oxidase surface layers and the nature of the forces determining the adsorption of the enzyme on various adsorbents are discussed. Copyright 1999 Academic Press.
ABSTRACT
The penetrant ability of the native glucose oxidase, GOx, and of the hydrophobically modified enzyme GO(mod) realized by grafting to its lysine residues alkyl C16 chains, into phosphatidylcholine dibehenoyl (DBPC), phosphatidylcholine dipalmitoyl (DPPC), phosphatidyl-ethanolamine dipalmitoyl (DPPE), phosphatidyl-serine dipalmitoyl (DPPS), and cholesterol (CHOL) monolayers was assessed by surface pressure measurements at constant area by enzyme injection to the aqueous phase beneath spread monolayers. As revealed by the magnitude of surface pressure increments (DeltaPi), both the quantities and the rates of penetration of the enzymes into these monolayers were lipid chemical nature and enzyme concentration dependent. When compared with GOx, GO(mod) displayed an enhanced penetrant ability into all the studied monolayers that resulted in rapidly attained DeltaPi plateau values, characteristic of stable systems. The influence of lipid hydrocarbon chain length and of the polar headgroup charge on the efficiency and effectiveness of GOx and GO(mod) penetration into these monolayers is discussed. Copyright 1999 Academic Press.
ABSTRACT
PURPOSE: To study the interactions between a polymeric matrix and a drug during storage at a temperature lower than the glass transition temperature of the polymers. METHODS: Poly(lactide-co-glycolide) microspheres loaded with different progesterone ratios were stored at 4. 20 and 40 degrees C. DSC-scans were recorded at regular intervals, depending on the storage temperature. RESULTS: The physical aging of the polymeric matrix, as monitored by the amplitude of the endotherm associated with the glass transition, is slowed down by crystalline progesterone. The development of the progesterone polymorphic depends on the interface/volume ratio of the crystals. CONCLUSIONS: For polymeric drug delivery systems, the determination of all studies parameters must take into account an effect of dispersed drugs which are more sensitive as the storage temperature is lower than the glass transition temperature of the matrix.