Dipalmitoylphosphatidylcholine Vesicles Research Articles

Phosphatidylcholine Surface Hydration-Dependent Adsorption to Mucin Enhances Intestinal Mucus Barrier Function.

The crucial role of zwitterionic phosphatidylcholines (PC) within mucus gel is essential for maintaining intestinal homeostasis, while the underlying mechanism remains incompletely understood. Herein, we compared the dynamic interfacial adsorption behavior of saturated dipalmitoylphosphatidylcholine (DPPC) and unsaturated dioleoylphosphatidylcholine (DOPC) to intestinal mucin and their impact on the intestinal mucus barrier function. Results of quartz crystal microbalance with dissipation showed that the highly surface-hydrated DPPC vesicles exhibited significantly faster and more extensive adsorption to purified intestinal mucin than the slightly surface-hydrated DOPC vesicles. Utilizing an intestinal Caco-2/HT29-MTX coculture model, we observed that DPPC vesicles adsorbed much more to the mucus gel compared to DOPC vesicles. Additionally, DPPC vesicle adsorption displayed increased wetting, and converse for DOPC vesicles. Interestingly, both of them exhibited nearly the same protective effects against cell injury induced by peptic-tryptic digests of gliadin (PTG). The partial mechanism involved the binding of PTG to DPPC and DOPC within the mucus gel, thereby restricting PTG contact with the underlying epithelial cells. These findings shed light on the intricate interfacial dynamics of PC adsorption to mucin and their implications for maintaining the integrity of the intestinal mucus barrier.

Structural dynamics of chlorpromazine (CPZ) drug with dipalmitoylphosphatidylcholine (DPPC) lipid: a potential drug for SARS-CoV-2

There is an urgent requirement for drug discovery and more importantly drug repositioning due to infectious new Severe Acute Respiratory Syndrome coronavirus 2. As per the recent report published in the journal L'Encéphale in May 2020, there is a planned ReCoVery Study examining the repurposing the chlorpromazine for the treatment of COVID-19. Here, we apply a combined Raman microspectroscopy and DFT-MD approach to investigate the structural dynamics of the Chlorpromazine (CPZ) drug with dipalmitoylphosphatidylcholine (DPPC) lipid bilayer, identifying the specific position of the drug in the DPPC lipid bilayer. The intensity ratios of the Raman peaks I 2935/I 2880, I 1097/I 1064 and I 1097/I 1129 are representative of the interaction of drugs with lipid alkyl chains and furnish conformation of lipid alkyl chains. Raman imaging microscopy for the study of the distribution of CPZ inside the lipid vesicles is reported. We also investigated the influence of order and disorder ratio in the CPZ on the DPPC liposomes prepared on phase transition temperature. HIGHLIGHTS Drug–membrane interactions using micromolar concentrations of both lipid and drugs. Neuroleptic drug and DPPC vesicles composed of DPPC/drug mixtures reveal qualitative differences between the Raman spectra The temperature-controlled Raman microspectroscopic study has demonstrated that below phase-transition temperature, the fatty acid chains of the phospholipids are stiff and packed in a highly ordered array. DFT and MD simulations to understand molecular interactions, structural dynamics, and Raman spectra. Above phase-transition temperature, the chains are disordered and possess more motional freedom. Communicated by Ramaswamy H. Sarma

Lamellarity and size distributions in mixed DPPC/amphiphilic poly(2-oxazoline) gradient copolymer vesicles and their temperature response

Mixed liposomes of dipalmitoylphosphatidylcholine (DPPC) and gradient (pseudodiblock) poly(2-methyl-2-oxazoline)-grad-poly(2-phenyl-2-oxazoline) (MPOx) copolymers are investigated by small angle neutron scattering (SANS). All experimental data, from different phospholipid-copolymer compositions, concentrations and temperatures are fitted with one model. This model allows the determination of the separate contributions from vesicular populations of different lamellarity and size. MPOx copolymers are proved to modify both the size and lamellarity of DPPC liposomes. The gradient copolymer with higher hydrophilic content induces shrinkage of the uni- and bi-lamellar DPPC vesicles. The copolymer with lower hydrophilic content causes dramatic changes on the lamellarity of DPPC vesicles by the formation of hexa-lamellar vesicles. The tendency of multi-lamellar vesicles to transform into uni-lamellar ones as temperature increases is more pronounced in the presence of the copolymers. These findings may have direct implications on the drug loading and release properties of liposomes and their interactions with cells.

Синтетические меротерпеноиды на основе терпеновых спиртов: синтез, самосборка и мембранотропные свойства

Currently, targeted drug delivery is of great interest in the field of medicine. The study of compounds capable of permeating cell membranes is a major problem in this area. The synthesis of pharmacologically active compounds includes the formation of structures with various combinations of pharmacophore fragments and properties. Amphiphilic compounds tend to exhibit membranotropic activity. From this point of view, the modification of natural products, especially terpenoids, is of particular interest. Terpenoid structures are used as membrane anchors in the development of modulators for membrane-integrated proteins or structures for creating nanocontainers. In this paper we synthesized a number of water-soluble amphiphilic meroterpenoids containing a charged pyridinium fragment on the basis of acyclic terpene alcohols. Residue of terpene alcohols – geraniol (monoterpenol), farnesol (sesquiterpenol), and phytol (diterpenol) – were used as the hydrophobic part of the amphiphilic structure. Linear acyclic alcohols are commercially available reagents and have a structure similar to that of polyprenols in archaeal lipids, which made it possible to obtain synthetic lipid-like meroterpenoids capable of self-assembly in aqueous solutions. The charged pyridinium fragment, which is included in numerous natural compounds, was of interest as a polar component. This meroterpenoids are synthetic analogs of archaeal lipids. It was shown that the studied meroterpenoids form nanosized aggregates in aqueous solutions by the method of dynamic light scattering and the Doppler microelectrophoresis method. Turbidimetric titration on model dipalmitoylphosphatidylcholine vesicles revealed that the synthesized compounds are embedded into the bilayer membrane without destroying it. Self-assembled aggregates of synthesized compounds in water can find application for drug delivery – in the creation of nanocontainers containing membrane anchors capable of interacting with the outer surface of the cell (lipid membrane).

Microfluidic-assisted fabrication of reverse micelle/PLGA hybrid microspheres for sustained vascular endothelial growth factor delivery.

In this study, poly (d, l-lactide-co-glycolide) (PLGA) composite microspheres containing anhydrous reverse micelle (R.M.) dipalmitoylphosphatidylcholine (DPPC) nanoparticles loaded vascular endothelial growth factor (VEGF) were produced using microfluidic platforms. The VEGF-loaded R.M. nanoparticles (VRM) were achieved by initial self-assembly and subsequent lipid inversion of the DPPC vesicles. The fabricated VRMs were encapsulated into the PLGA matrix by flow-focusing geometry microfluidic platforms. The encapsulation efficiency, in vitro release profile, and the bioactivity of the produced composite microspheres were investigated. The release study showed that VEGF was slowly released from the PLGA composite microspheres over 28 days with a reduced initial burst (18 ± 4.17% in the first 24H). The VEGF stability during encapsulation and release period was also investigated, and the results indicated that encapsulated VEGF was well preserved. Also, the bioactivity assay of the PLGA composite microspheres on human umbilical vein endothelial cells was confirmed that the encapsulated VEGF was utterly active. The present monodisperse and controllable VEGF-loaded microspheres with reproducible manner could be widely used in tissue engineering and therapeutic applications.

Effect of C60 nanoparticles on elasticity of small unilamellar vesicles composed of DPPC bilayers**Project supported by the National Natural Science Foundation of China (Grant No. 61475196).

The interaction between C60 nanoparticles and biomembranes has been of great interest in researches over the past decades due to their novel applications as well as potential cytotoxicity. In this work, we study the deformation of the small unilamellar vesicles composed of dipalmitoylphosphatidylcholine (DPPC) lipid bilayers infiltrated with C60 nanoparticles of different molecular concentrations through coarse-grained molecular dynamics simulations. By employing the Helfrich spontaneous curvature model, the bending modulus and the spontaneous curvature of the vesicles with C60 nanoparticles of different concentrations are obtained according to the simulation data. The results show that the bending modulus and the spontaneous curvature of pure DPPC vesicle membranes are approximately 1.6 × 10−19 J and 1.4 nm−1, respectively. Both of them increase linearly approximately as the C60 concentration increases from 0 to 16.3%. The density profiles of vesicles, the order of lipid packing and the diffusion characteristics of DPPC and C60 are also investigated.

Localization of Annexin A6 in Matrix Vesicles During Physiological Mineralization.

Annexin A6 (AnxA6) is the largest member of the annexin family of proteins present in matrix vesicles (MVs). MVs are a special class of extracellular vesicles that serve as a nucleation site during cartilage, bone, and mantle dentin mineralization. In this study, we assessed the localization of AnxA6 in the MV membrane bilayer using native MVs and MV biomimetics. Biochemical analyses revealed that AnxA6 in MVs can be divided into three distinct groups. The first group corresponds to Ca2+-bound AnxA6 interacting with the inner leaflet of the MV membrane. The second group corresponds to AnxA6 localized on the surface of the outer leaflet. The third group corresponds to AnxA6 inserted in the membrane’s hydrophobic bilayer and co-localized with cholesterol (Chol). Using monolayers and proteoliposomes composed of either dipalmitoylphosphatidylcholine (DPPC) to mimic the outer leaflet of the MV membrane bilayer or a 9:1 DPPC:dipalmitoylphosphatidylserine (DPPS) mixture to mimic the inner leaflet, with and without Ca2+, we confirmed that, in agreement with the biochemical data, AnxA6 interacted differently with the MV membrane. Thermodynamic analyses based on the measurement of surface pressure exclusion (πexc), enthalpy (ΔH), and phase transition cooperativity (Δt1/2) showed that AnxA6 interacted with DPPC and 9:1 DPPC:DPPS systems and that this interaction increased in the presence of Chol. The selective recruitment of AnxA6 by Chol was observed in MVs as probed by the addition of methyl-β-cyclodextrin (MβCD). AnxA6-lipid interaction was also Ca2+-dependent, as evidenced by the increase in πexc in negatively charged 9:1 DPPC:DPPS monolayers and the decrease in ΔH in 9:1 DPPC:DPPS proteoliposomes caused by the addition of AnxA6 in the presence of Ca2+ compared to DPPC zwitterionic bilayers. The interaction of AnxA6 with DPPC and 9:1 DPPC:DPPS systems was distinct even in the absence of Ca2+ as observed by the larger change in Δt1/2 in 9:1 DPPC:DPPS vesicles as compared to DPPC vesicles. Protrusions on the surface of DPPC proteoliposomes observed by atomic force microscopy suggested that oligomeric AnxA6 interacted with the vesicle membrane. Further work is needed to delineate possible functions of AnxA6 at its different localizations and ways of interaction with lipids.

A comparative study of the effects of 7β-hydroxycholesterol, 25-hydroxycholesterol, and cholesterol on the structural and thermal phase behavior of multilamellar dipalmitoylphosphatidylcholine bilayer vesicles

The effects of 7β-hydroxycholesterol (7βOH) and 25-hydroxycholesterol (25OH) on the phase behavior and the structural properties of the multilamellar vesicles of dipalmitoylphosphatidylcholine (DPPC) bilayers were investigated with comparing to the effects of cholesterol (Chol). Differential scanning calorimetry (DSC) measurements showed these three sterols have slightly but distinctly different effects on the thermal behavior of DPPC bilayers. X-ray diffraction data analysis on DPPC bilayers containing 30 mol% sterols with the aid of molecular volume data estimated by the neutral buoyancy method indicated that the order of apparent occupied surface area per molecule is DPPC/7βOH > DPPC/25OH > DPPC/Chol at both 25 ℃ and 50 ℃, suggesting that the strength of the condensation effect of these sterols follows inversely this order. Based on the findings in this study, we inferred the molecular orientations of Chol, 7βOH, and 25OH in DPPC bilayers.

Characterization of phospholipid vesicles containing lauric acid: physicochemical basis for process and product development

Lauric acid (LAH) strongly inhibits the growth of acne-causing bacteria. LAH is essentially water-insoluble and the solubility of laurate (LA) salts are medium and temperature dependent. Hence, LAH/LA preparations are difficult to formulate. Here we fully characterized phospholipid vesicles containing up to 50 mol% LAH. Vesicles of dipalmitoylphosphatidylcholine (DPPC) containing LAH, at pHs 7.4 and 5.0, were characterized measuring size, charge, bilayer phase transition temperature (Tm) and permeability of water-soluble probes. Small angle X-ray scattering and cryotransmission electron microscopy showed multilamellar vesicles at low LAH %. Increasing LAH % had a negligible effect on particle size. An internal aqueous compartment in all vesicle's preparations, even at equimolar DPPC: LAH fractions, was demonstrated using water-soluble probes. At pH 5.0, the interaction between DPPC and LAH increased the Tm and phase transition cooperativity showing a single lipid phase formed by hydrogen-bonded DPPC: LAH complexes. At pH 7.4, vesicles containing 50 mol% LAH exhibited distinct phases, ascribed to complex formation between LAH and LA or LAH and DPPC. LAH incorporated in the vesicles minimally permeated a skin preparation at both pHs, indicating that the primary sites of LAH solubilization were the skin layers. These results provide the foundations for developing processes and products containing DPPC: LAH.

Adsorption of Phospholipids at the Air-Water Surface

Phospholipids are ubiquitous components of biomembranes and common biomaterials used in many bioengineering applications. Understanding adsorption of phospholipids at the air-water surface plays an important role in the study of pulmonary surfactants and cell membranes. To date, however, the biophysical mechanisms of phospholipid adsorption are still unknown. It is challenging to reveal the molecular structure of adsorbed phospholipid films. Using combined experiments with constrained drop surfactometry and molecular dynamics simulations, here, we studied the biophysical mechanisms of dipalmitoylphosphatidylcholine (DPPC) adsorption at the air-water surface. It was found that the DPPC film adsorbed from vesicles showed distinct equilibrium surface tensions from the DPPC monolayer spread via organic solvents. Our simulations revealed that only the outer leaflet of the DPPC vesicle is capable of unzipping and spreading at the air-water surface, whereas the inner leaflet remains intact and forms an inverted micelle to the interfacial monolayer. This inverted micelle increases the local curvature of the monolayer, thus leading to a loosely packed monolayer at the air-water surface and hence a higher equilibrium surface tension. These findings provide novel insights, to our knowledge, into the mechanism of the phospholipid and pulmonary surfactant adsorption and may help understand the structure-function correlation in biomembranes.

Coarse-grained molecular dynamics simulations of the effect of edge activators on the skin permeation behavior of transfersomes

Transfersomes (TRS) can provide sustained drug delivery and themselves are biocompatible, biodegradable and nontoxic. Edge activators (EAs) are key factors for increasing the deformability of TRS, and this active deformation mechanism is of commercial interest, especially at the molecular level. Accordingly, in this paper, the deformability of pure dipalmitoyl phosphatidylcholine (DPPC) vesicles, TRS with sodium cholate as an EA, and DPPC vesicles containing pogostone (POG) were compared via umbrella sampling technology. The DPPC conformation and membrane fluidity of these three types of bilayer systems were evaluated, and the changes in the membrane properties of vesicles caused by EAs were studied. EAs could increase the deformability of TRS by decreasing the deformation energy barrier due to their amphiphilic structures, which was similar to those of DPPC molecules. The membrane properties also changed via treatment with EAs including altering the tail chain angle, disturbing the ordered tail chain arrangement and prompting lateral diffusion of DPPC molecules. In addition, the impact of EAs on DPPC bilayers was further demonstrated to be concentration dependent. An ideal concentration was identified for the lowest amount of EA that offered a gel-liquid-crystalline phase transition of DPPC bilayers. Importantly, POG, a lipophobic transdermal drug, can also affect the skin permeation behavior of vesicles but had weaker effects than EA.

Real-Time Monitoring of Interactions between Solid-Supported Lipid Vesicle Layers and Short- and Medium-Chain Length Alcohols: Ethanol and 1-Pentanol.

Lipid bilayers represent the interface between the cell and its environment, serving as model systems for the study of various biological processes. For instance, the addition of small molecules such as alcohols is a well-known process that modulates lipid bilayer properties, being considered as a reference for general anesthetic molecules. A plethora of experimental and simulation studies have focused on alcohol’s effect on lipid bilayers. Nevertheless, most studies have focused on lipid membranes formed in the presence of alcohols, while the effect of n-alcohols on preformed lipid membranes has received much less research interest. Here, we monitor the real-time interaction of short-chain alcohols with solid-supported vesicles of dipalmitoylphosphatidylcholine (DPPC) using quartz crystal microbalance with dissipation monitoring (QCM-D) as a label-free method. Results indicate that the addition of ethanol at different concentrations induces changes in the bilayer organization but preserves the stability of the supported vesicle layer. In turn, the addition of 1-pentanol induces not only changes in the bilayer organization, but also promotes vesicle rupture and inhomogeneous lipid layers at very high concentrations.

Water-soluble meroterpenes containing an aminoglyceride fragment with geraniol residues: synthesis and membranotropic properties

A number of new membrane anchors based on water-soluble aminoglycerides with geraniol fragments have been synthesized. A biomimetic approach was used based on the design of meroterpenes structurally similar to archaeal lipids. Turbidimetry and laser Doppler microelectrophoresis showed that the synthesized compounds were incorporated into unilamellar dipalmitoylphosphatidylcholine (DPPC) vesicles.

Role of oligo(malic acid) on the formation of unilamellar vesicles

Stable unilamellar dipalmitoylphosphatidylcholine vesicles were produced by using oligo(malic acid) and cholesterol. Detailed physico-chemical characterization prove that by using oligo(malic acid) the substitution of PEGylated lipids for sterically stabilization comes possible. The polymer molecules cover the outer surface of spherical-shaped vesicles, and an asymmetrical composition occurs in the two leaflets of the phospholipid bilayer. The oligo(malic-acid) and cholesterol are enriched in the outer side assuring the stabilization of vesicles. Cholesterol plays an important role in the self-assembly of components as it makes the entering of oligomers possible deep into the polar head-region of lipids. The presence of oligo(malic acid) molecules does not induce degradation by hydrolysis of lipid molecules but the vesicle system turns into a sensitive form giving a possibility for pH sensitive targeting. Preliminary investigation on the investigated oligo(malic acid)-stabilized vesicles do not show any toxic effect promising their applicability in the field of liposomal drug delivery.

Soft nanoparticles charge expression within lipid membranes: The case of amino terminated dendrimers in bilayers vesicles.

Interactions of charged nanoparticles with model bio-membranes provide important insights about the soft interaction involved and the physico-chemical parameters that influence lipid bilayers stability, thus providing key features of their cytotoxicity effects onto cellular membranes. With this aim, the self-assembly processes between polyamidoamine dendrimers (generation G = 2.0 and G = 4.0) and dipalmitoylphosphatidylcholine (DPPC) lipids were investigated by means of Zeta potential analysis, x-rays, Raman and quasielastic light scattering experiments. Raman scattering data evidenced that dendrimers penetration produce a perturbation of the DPPC vesicles alkyl chains. A linear increase of liposome zeta-potential with increasing PAMAM concentration evidenced that only a fraction of the dendrimers effective charge contributes to the expression of the charge at the surface of the DPPC liposome. The linear region of the zeta-potential extends toward higher PAMAM concentrations as the dendrimer generation decreases from G = 4.0 to G = 2.0. Further increase in PAMAM concentration, outside of the linear region, causes a perturbation of the bilayer characterized by the loss in multilamellar correlation and the increase of DPPC liposome hydrodynamic radius. The findings of our investigation help to rationalize the effect of nanoparticles electrostatic interaction within lipid vesicles as well as to provide important insights about the perturbation of lipid bilayers membrane induced by nanoparticles inclusion.

Cholesterol affects the interaction between an ionic liquid and phospholipid vesicles. A study by differential scanning calorimetry and nanoplasmonic sensing

The present work aims at studying the interactions between cholesterol-rich phosphatidylcholine-based lipid vesicles and trioctylmethylphosphonium acetate ([P8881][OAc]), a biomass dissolving ionic liquid (IL). The effect of cholesterol was assayed by using differential scanning calorimetry (DSC) and nanoplasmonic sensing (NPS) measurement techniques. Cholesterol-enriched dipalmitoyl-phosphatidylcholine vesicles were exposed to different concentrations of the IL, and the derived membrane perturbation was monitored by DSC. The calorimetric data could suggest that the binding and infiltration of the IL are delayed in the vesicles containing cholesterol. To clarify our findings, NPS was applied to quantitatively follow the resistance of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine incorporating 0, 10, and 50mol% of cholesterol toward the IL exposure over time. The membrane perturbation induced by different concentrations of IL was found to be a concentration dependent process on cholesterol-free lipid vesicles. Moreover, our results showed that lipid depletion in cholesterol-enriched lipid vesicles is inversely proportional to the increasing amount of cholesterol in the vesicles. These findings support that cholesterol-rich lipid bilayers are less susceptible toward membrane disrupting agents as compared to membranes that do not incorporate any sterols. This probably occurs because cholesterol tightens the phospholipid acyl chain packing of the plasma membranes, increasing their resistance and reducing their permeability.

Novel type of isoprenoid membrane anchors: an investigation of binding properties with dipalmitoylphosphatidylcholine vesicles

AbstractIn this work, we present a new type of amphiphilic membrane‐anchoring agents that can be easily obtained by the Diels‐Alder reaction between terpene myrcene and N‐substituted maleimides. The interaction between the compounds and small unilamellar dipalmitoylphosphatidylcholine vesicles was investigated using infrared spectroscopy, microgravimetry, and turbidimetry. The ability of the compounds to embed in the phospholipid membrane was shown to be strongly dependent on the charge of their polar group. The insertion of the compounds studied into the lipid bilayer did not lead to disruption of the dipalmitoylphosphatidylcholine vesicles up to the highest tested drug to lipid molar ratio of 0.5 to 0.6. Low lipid solubilization ability of the compounds as well as their rigid nonplanar structure makes them an interesting alternative to the common membrane‐anchoring structural motifs.

Charge-Mediated Localization of Conjugated Polythiophenes in Zwitterionic Model Cell Membranes.

The selective engineering of conjugated polyelectrolyte (CPE)-phospholipid interfaces is poised to play a key role in the design of advanced biomedical and biotechnological devices. Herein, we report a strategic study to investigate the relationship between the charge of the CPE side group and their association with zwitterionic phospholipid bilayers. The interaction of dipalmitoylphosphatidylcholine (DPPC) phospholipid vesicles with a series of poly(thiophene)s bearing zwitterionic, cationic, or anionic terminal groups (P3Zwit, P3TMAHT and P3Anionic, respectively) has been probed. Although all CPEs showed an affinity for the zwitterionic vesicles, the calculated partition coefficients determined using photoluminescence spectroscopy suggested preferential incorporation within the lipid bilayer in the order P3Zwit > P3Anionic ≫ P3TMAHT. The polarity probe Prodan was used to further qualify the position of the CPE inside the vesicle bilayers via Förster resonance energy transfer (FRET) studies. The varying proximity of the CPEs to Prodan was reflected in the Stern-Volmer quenching constants and decreased in the order P3Anionic > P3TMAHT ≫ P3Zwit. Dynamic light scattering measurements showed an increase in the hydrodynamic diameter of the DPPC vesicles upon addition of each poly(thiophene), but to the greatest extent for P3Anionic. Small-angle neutron scattering studies also revealed that P3Anionic specifically increased the thickness of the headgroup region of the phospholipid bilayer. Epifluorescence and atomic force microscopy imaging showed that P3TMAHT formed amorphous agglomerates on the vesicle surface, P3Zwit was buried throughout the bilayer, and P3Anionic formed a shell of protruding chains around the surface, which promoted vesicle fusion. The global data indicate three distinctive modes of interaction for the poly(thiophene)s within DPPC vesicles, whereby the nature of the association is ultimately controlled by the pendant charge group on each CPE chain. Our results suggest that charge-mediated self-assembly may provide a simple and effective route to design luminescent CPE probes capable of specific localization within phospholipid membranes.

Temperature-dependent bifurcation of cooperative interactions in pure and enriched in β-carotene DPPC liposomes

We examined the influence of temperature on lipid intermolecular interactions and the organization of bilayers within multilamellar dipalmitoylphosphatidylcholine (DPPC) liposomes. We also investigated the effect of 0.5 mol% β-carotene, a non-polar carotenoid, on the adhesive properties of these liposomes. Atomic force microscopy (AFM) and differential scanning calorimetry (DSC) were used to correlate the changes in the physical properties of the liposomal systems with their thermotropic behaviour. Using DSC we detected two transitions in pure DPPC vesicles and in those containing 0.5 mol% β-carotene. In both systems the pretransition occurred at 34.5(1)°C and the main phase transition at 41.4 °C during heating. Upon cooling, the temperatures of the pretransition and the main transition decreased by about 6 °C and 1 °C, respectively. Changes in enthalpy and entropy were also similar in the two investigated systems. Data obtained in parallel AFM force experiments show that the adhesive forces between the liposomal systems and AFM probe strongly depend on the loading rate. Moreover, their characteristic monotonic changes and discontinuities are sensitive to temperature. In the range of temperatures from 27 °C to 31 °C, i.e. below the temperature of phase transition from gel to ripple phase, the adhesive forces measured in a water environment are about an order of magnitude higher in the presence of β-carotene than in pure DPPC liposomes. The observed variable dependence of adhesion on the loading rate suggests that there are changes in the long- and short-range interactions between lipids, and that these may be related to the occurrence of some clustering effects. In addition, the simultaneous existence of different subphases was found in the gel phase of DPPC liposomes. The presence of β-carotene at a level of 0.5 mol% stimulates the structural reorganization of DPPC multilamellar vesicles and enhances the bifurcation phenomenon detected in these systems.

Investigation of Encapsulated Liposomal Antituberculotics and Effects on in Vitro Model Systems

Introduction. Liposomes are widely investigated nanocarriers, which are capable for incorporation of both lipid-soluble and water-soluble drugs. WHO aimed to eradicate TB disease, which is caused by Mycobacterium tuberculosis bacteria. TB kills more than 1 million people every year. The main reasons for death are increasing frequency of MDR (multidrug-resitant) and XDR (extensively drug-resistant) bacteria stems and inappropriate patient compliance.Aim. Preparation and encapsulation of newly synthesized antitubercultic compound into liposomes. To enhance the encapsulation efficiency and to target intracellular bacteria in the macrophages.Methods.We have prepared two types of liposomes.Type I. consists of dipalmitoyl phosphatidylcholine (DPPC).Type II. consists of: dioleoylphosphatidylethanolamine (DOPE), cholesteryl hemisuccinate (CHEMS) and pegylated distearoyl phosphatidylethanolamine (DSPE-PEG).We used thin lipid film technology to prepare multilamellar vesicles (MLV) and henceforward both types were treated with ultrasound or extrusion technique to get small unilamellar vesicles (SUV). The size distribution of liposomes was determined/measured with dynamic light scattering (DLS). The change of diameter of vesicles show the rate of aggregation. We investigated the efficacy of the preparation technology and the stability of liposomes. The used antituberculotic compounds were: isoniazid (INH), TB 504, TB 505 (chemical structure not published yet). Encapsulation efficiency was determined by measuring absorbance after size exclusion chromatography (SEC).Results. DPPC vesicles showed reduced stability than complex liposomes (type II.) both at 4 and 20 °C. By extrusion method we got uniform and more stable vesicles. Encapsulation efficiency was influenced by the diameter of liposome and the physico-chemical properties of antituberculotic compounds. The in vitro activity of liposomal antituberculotic compounds was determined on M. tuberculosis H37Rv culture. Considering that M. tuberculosis is an intracellular pathogen the effect of the compounds was studied on M. tuberculosis H37Rv infected MonoMac6 human monocytes.