Phospholipid Liposomes Research Articles

Fatty Acid Photodecarboxylase Is an Interfacial Enzyme That Binds to Lipid-Water Interfaces to Access Its Insoluble Substrate.

Fatty acid photodecarboxylase (FAP), one of the few natural photoenzymes characterized so far, is a promising biocatalyst for lipid-to-hydrocarbon conversion using light. However, the optimum supramolecular organization under which the fatty acid (FA) substrate should be presented to FAP has not been addressed. Using palmitic acid embedded in phospholipid liposomes, phospholipid-stabilized microemulsions, and mixed micelles, we show that FAP displays a preference for FAs present in liposomes and at the surface of microemulsions. The kinetics of adsorption onto phospholipid and galactolipid monomolecular films further suggests the ability of FAP to bind to and penetrate into membranes, with a higher affinity in the presence of FAs. The FAP structure reveals a potential interfacial recognition site with clusters of hydrophobic and basic residues surrounding the active site entrance. The resulting dipolar moment suggests the orientation of FAP at negatively charged interfaces. These findings provide important clues about the mode of action of FAP and the development of FAP-based bioconversion processes.

Effect of lipopolymer (DSPE-PEG750) on phospholipid monolayers and bilayers differing in the structure of the polar head group

Phospholipid liposomes are the most popular kind of vesicles used in drug delivery due to their high biocompatibility with biomembranes. However, this type of liposomes show some limitations related with their short half-life in bloodstream. This problem can be resolved by the coating of their surface with the application of PEG molecules which form steric and entropic barrier, avoiding recognition of encapsulated liposomes by the reticuloendothelial system (RES). Such modification of liposomes significantly affects the physicochemical characteristics of PEGylated liposomes, which is related to the intermolecular interactions between the lipopolymer molecules and the phospholipid that builds the liposome membrane. In our work we investigated the influence of the PEG-ylated lipid (DSPE-PEG750) on physicochemical properties of phospholipid monolayers and bilayers with different structure of the polar group (DPPC, DPPE, DPPS). The phospholipid monolayers were examined with the application of Langmuir monolayer technique and Brewster angle microscopy (BAM) as well as grazing incidence x-ray diffraction (GIXD). The studies performed on phospholipid bilayer systems were related to dynamic light scattering and zeta potential measurements and the experiments with the calcein release and steady-state fluorescence anisotropy of DPH. The obtained results showed that the type of polar group of phospholipid as well as the addition of PEG-ylated lipid significantly changes the molecular organization of phospholipid monolayers. Moreover, these parameters also influence the properties of phospholipid bilayers such as size, surface charge, stability and permeability.

Effects of DC Magnetic Fields on Magnetoliposomes.

The potential use of magnetic nanoparticles (MNPs) in biomedicine as magnetic resonance, drug delivery, imagenology, hyperthermia, biosensors, and biological separation has been studied in different laboratories. One of the challenges on MNP elaboration for biological applications is the size, biocompatibility, heat efficiency, stabilization in physiological conditions, and surface coating. Magnetoliposome (ML), a lipid bilayer of phospholipids encapsulating MNPs, is a system used to reduce toxicity. Encapsulated MNPs can be used as a potential drug and a gene delivery system, and in the presence of magnetic fields, MLs can be accumulated in a target tissue by a strong gradient magnetic field. Here, we present a study of the effects of DC magnetic fields on encapsulated MNPs inside liposomes. Despite their widespread applications in biotechnology and environmental, biomedical, and materials science, the effects of magnetic fields on MLs are unclear. We use a modified coprecipitation method to synthesize superparamagnetic nanoparticles (SNPs) in aqueous solutions. The SNPs are encapsulated inside phospholipid liposomes to study the interaction between phospholipids and SNPs. Material characterization of SNPs reveals round-shaped nanoparticles with an average size of 12 nm, mainly magnetite. MLs were prepared by the rehydration method. After formation, we found two types of MLs: one type is tense with SNPs encapsulated and the other is a floppy vesicle that does not show the presence of SNPs. To study the response of MLs to an applied DC magnetic field, we used a homemade chamber. Digitalized images show encapsulated SNPs assembled in chain formation when a DC magnetic field is applied. When the magnetic field is switched off, it completely disperses SNPs. Floppy MLs deform along the direction of the external applied magnetic field. Solving the relevant magnetostatic equations, we present a theoretical model to explain the ML deformations by analyzing the forces exerted by the magnetic field over the surface of the spheroidal liposome. Tangential magnetic forces acting on the ML surface result in a press force deforming MLs. The type of deformations will depend on the magnetic properties of the mediums inside and outside the MLs. The model predicts a coexistence region of oblate–prolate deformation in the zone where χ = 1. We can understand the chain formation in terms of a dipole–dipole interaction of SNP.

Anti-inflammatory effects of traditional chinese medicine ingredients based on nano-Strychnos liposomes for external skin application

Strychnos is an effective anti-tumor monomer with anti-inflammatory, analgesic, anti-tumor and enhancing immunity. However, its clinical application is restricted due to its poor water solubility and toxicity. The key to the study of traditional Chinese medicine strychnine is to develop a new low toxicity and high efficiency formulation. Nano liposomes can effectively improve the transdermal absorption rate of the drug, prolong the action time of the drug, enhance the tissue targeting of the drug, so as to achieve the sustained-release effect. Therefore, this paper attempts to explore the anti-inflammatory effect of traditional Chinese medicine ingredients based on the skin topical nano Strychnos liposomes, and provide experimental and theoretical basis for the development of new transdermal formulations of nano Strychnos liposomes. In this paper, Strychnos was used as the research object, and the experimental results showed that Strychnos liposomes with good dispersion stability could be prepared by film ultrasonic method. Although the composite phospholipid liposome only contains 10% hydrogenated phospholipid liposome, its peroxide value is always far lower than that of soybean phospholipid liposome, so the antioxidant capacity of composite phospholipid liposome is higher than that of soybean phospholipid liposome. Nano liposomes have almost no anti-inflammatory effect, while ordinary Strychnos has poor analgesic and anti-inflammatory effects; nano Strychnos liposomes have considerable anti-inflammatory effect.

Epigenetic inhibition assisted chemotherapeutic treatment of lung cancer based on artificial exosomes

We adopted a novel strategy by combining histone deacetylase (HDAC) inhibitors with traditional chemotherapeutics to treat solid tumors. However, chemotherapeutics often have a narrow therapeutic index and need multiple administrations with undesired side effects that lead to the intolerance. To reduce the non-specificity of chemotherapeutics, targeted therapy was introduced to restrict such agents in the tumor with minimum effects on other tissues. We developed bioinspired artificial exosomes (AE), which enabled to deliver chemotherapeutics to the tumors effectively after systemic administration. AE were produced by incorporating membrane proteins from cancer cells into phospholipid liposomes that mimicked the plasma membrane. The synthesized AE were used for the delivery of broad-spectrum chemotherapeutic doxorubicin (DOX) and vorinostat (SAHA), an epigenetic inhibitor. The combination of DOX and SAHA showed synergistic effects on suppressing non-small cell lung cancer cells and xenograft tumors without apparent adverse effects. AE facilitated the delivery of drugs to tumor tissue and extended the retention time of drugs within tumors. Taken together, these studies suggest that the bioengineered artificial exosomes may serve as novel delivery strategy for chemotherapeutics to treat non-small cell lung cancer.

Cell Surface Binding and Lipid Interactions behind Chemotherapy-Drug-Induced Ion Pore Formation in Membranes.

Chemotherapy drugs (CDs) disrupt the lipid membrane’s insulation properties by inducing stable ion pores across bilayer membranes. The underlying molecular mechanisms behind pore formation have been revealed in this study using several methods that confirm molecular interactions and detect associated energetics of drugs on the cell surface in general and in lipid bilayers in particular. Liposome adsorption and cell surface binding of CD colchicine has been demonstrated experimentally. Buffer dissolved CDs were considerably adsorbed in the incubated phospholipid liposomes, measured using the patented ‘direct detection method’. The drug adsorption process is regulated by the membrane environment, demonstrated in cholesterol-containing liposomes. We then detailed the phenomenology and energetics of the low nanoscale dimension cell surface (membrane) drug distribution, using atomic force microscopy (AFM) imaging what addresses the surface morphology and measures adhesion force (reducible to adhesive energy). Liposome adsorption and cell surface binding data helped model the cell surface drug distribution. The underlying molecular interactions behind surface binding energetics of drugs have been addressed in silico numerical computations (NCs) utilizing the screened Coulomb interactions among charges in a drug–drug/lipid cluster. Molecular dynamics (MD) simulations of the CD-lipid complexes detected primarily important CD-lipid electrostatic and van der Waals (vdW) interaction energies. From the energetics point of view, both liposome and cell surface membrane adsorption of drugs are therefore obvious findings. Colchicine treated cell surface AFM images provide a few important phenomenological conclusions, such as drugs bind generally with the cell surface, bind independently as well as in clusters of various sizes in random cell surface locations. The related adhesion energy decreases with increasing drug cluster size before saturating for larger clusters. MD simulation detected electrostatic and vdW and NC-derived charge-based interactions explain molecularly of the cause of cell surface binding of drugs. The membrane binding/association of drugs may help create drug–lipid complexes with specific energetics and statistically lead to the creation of ion channels. We reveal here crucial molecular understanding and features of the pore formation inside lipid membranes that may be applied universally for most of the pore-forming existing agents and novel candidate drugs.

Staphylococcus aureus membrane-damaging activities of four phenolics.

The membrane-damaging activities of four phenolics chosen for their bactericidal activity against Staphylococcus aureus CNRZ3 were investigated: 5,7-dihydroxy-4-phenylcoumarin (DHPC), 5,8-dihydroxy-1,4-naphthoquinone (DHNQ), epigallocatechin gallate (EGCG) and isobutyl 4-hydroxybenzoate (IBHB). Staphylococcusaureus CNRZ3 cells, as well as model liposomes mimicking its membrane phospholipids composition, were treated with each phenolic at its minimal bactericidal concentration. Membrane integrity, intracellular pH and intracellular esterase activity were examined by flow cytometric analysis of S.aureus cells stained with propidium iodide and SYTO® 9, 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein acetoxymethyl ester, and 5(6)-carboxyfluorescein diacetate, respectively. While intracellular pH was affected by the foyr phenolics, only DHNQ and to a lesser extent EGCG, caused a loss of membrane integrity. Flow cytometric analysis of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and DPPC/POPG (2-oleoyl-1-palmitoyl-sn-glycero-3-phosphoglycerol) liposomes stained with Coumarin 6 (which penetrates the lipid bilayer) or 5-N(octadecanoyl)-amino-fluorescein (which binds to the liposome shell) suggested that only EGCG and DHNQ penetrated the bilayer of phospholipids of liposomes. Taken together, these findings support the hypothesis that EGCG and DHNQ bactericidal activity results from their accumulation in the phospholipid bilayer of S.aureus cells membrane causing its disruption.

Surface Plasmon Resonance Monitoring of Mono-Rhamnolipid Interaction with Phospholipid-Based Liposomes.

The interactions of mono-rhamnolipids (mono-RLs) with model membranes were investigated through a biomimetic approach using phospholipid-based liposomes immobilized on a gold substrate and also by the multiparametric surface plasmon resonance (MP-SPR) technique. Biotinylated liposomes were bound onto an SPR gold chip surface coated with a streptavidin layer. The resulting MP-SPR signal proved the efficient binding of the liposomes. The thickness of the liposome layer calculated by modeling the MP-SPR signal was about 80 nm, which matched the average diameter of the liposomes. The mono-RL binding to the film of the phospholipid liposomes was monitored by SPR and the morphological changes of the liposome layer were assessed by modeling the SPR signal. We demonstrated the capacity of the MP-SPR technique to characterize the different steps of the liposome architecture evolution, i.e., from a monolayer of phospholipid liposomes to a single phospholipid bilayer induced by the interaction with mono-RLs. Further washing treatment with Triton X-100 detergent left a monolayer of phospholipid on the surface. As a possible practical application, our method based on a biomimetic membrane coupled to an SPR measurement proved to be a robust and sensitive analytical tool for the detection of mono-RLs with a limit of detection of 2 μg mL-1.

Characterization of milk and soy phospholipid liposomes and inflammation in 3T3-L1 adipocytes

Milk phospholipids (PL) are valuable dairy components that appear to impart human health benefits, including improved cognitive function in infants and adults. The commercial food industry uses primarily plant-based sources of PL, such as soy lecithin. However, it remains unclear whether different compositions of PL from different dietary sources, such as milk, convey the same benefits. We hypothesized that PL derived from bovine milk or soy have differing physiological effects in terms of inflammation due to their differences in composition. The objectives of this study were to characterize milk and soy liposomes by their physicochemical properties and composition and to evaluate their effects in vitro by means of inflammatory gene expression analyses. Milk and soy phospholipid large unilamellar vesicles (MPL-LUV and SPL-LUV, respectively) prepared using thin-film hydration coupled with extrusion were similar in terms of structure, size, and stability; however, they differed significantly in composition. The 3T3-L1 adipocytes were selected for this work because adipocytes are the main site of uptake, synthesis, modification, and breakdown of lipids and are important inflammatory mediators in mammalian systems. In this work, these cells exposed to both liposome varieties showed high biocompatibility and low cytotoxicity up to concentrations of 0.5 mg/mL as measured by colorimetric MTT and lactate dehydrogenase assays. Furthermore, SPL-LUV showed trends toward stimulating inflammation compared with MPL-LUV as measured by expression of 2 proinflammatory cytokines, monocyte chemoattractant protein-1 (MCP-1) and IL-6. Expression of MCP-1 significantly increased 1.82-fold relative to the control upon SPL-LUV treatment, with similar trends for IL-6 (increased 1.59-fold). The MPL-LUV showed relatively no change in cytokine expression. The results obtained in this work suggest that the methodology used to prepare LUV and the composition and proportion of milk PL are important in measuring cell physiology changes and inflammatory status in mammalian cells.

Antiprotozoal Structure-Activity Relationships of Synthetic Leucinostatin Derivatives and Elucidation of their Mode of Action.

Leucinostatin A is one of the most potent antiprotozoal compounds ever described, but little was known on structure–activity relationships (SAR). We used Trypanosoma brucei as a protozoal model organism to test synthetically modified derivatives, resulting in simplified but equally active compounds 2 (ZHAWOC6025) and 4 (ZHAWOC6027), which were subsequently modified in all regions of the molecule to gain an in‐depth SAR understanding. The antiprotozoal SAR matched SAR in phospholipid liposomes, where membrane integrity, leaking, and dynamics were studied. The mode of action is discussed based on a structure–activity analysis of derivatives in efficacy, ultrastructural studies in T. brucei, and artificial membrane models, mimicking membrane stability and membrane potential. The main site of antiprotozoal action of natural and synthetic leucinostatins lies in the destabilization of the inner mitochondrial membrane, as demonstrated by ultrastructural analysis, electron microscopy and mitochondrial staining. Long‐time sublethal exposure of T. brucei (200 passages) and siRNA screening of 12′000 mutants showed no signs of resistance development to the synthetic derivatives.

Piper Nigrum Protects Against Fe (II) Mediated Lipid Peroxidation in Phopholipids Liposomes: Analytical and Biochemical Analysis

Phospholipids liposomes were employed to access the extent of Iron [Fe(II)] induced lipid peroxidation. Our data indicate that Fe(II) caused a significant increase in malondialdehyde (MDA) formation, a lipid peroxidation product. Ethanolic extract of Piper Nigrum (PN) significantly inhibited lipid peroxidation at the highest tested concentrations. The plant with three different extracts (aqueous, ethanolic, and ethyl acetate)was tested for its nutritional value, and a higher amount of carbohydrates (42.66%), fibers (20.96 %), and proteins (14.38 %) were recorded. The mineral content evaluated by atomic absorption spectrophotometry (AAS) showed Ca (30.400 mg/L), Mg (10.475 mg/L), and Mn (2.391 mg/L) in higher concentrations. Quantitative gas chromatographic analysis coupled with a mass spectrometer (GCMS) was used to explore the characteristic biochemical compounds, which confirmed the presence of twenty-four (24) active constituents. D-Limonene (18.59%) was present in higher concentrations, followed by beta-pinene (17.98%), alpha-pinene (12.85%), and caryophyllene (9.91%). Plant extracts and essential oil were further tested against six (6) gram-negative, three (3) gram-positive, and a fungal strain by disc diffusion method. The presence of active compounds in essential oil and crude extracts may be responsible for observed pharmacological efficacies of PN and can be considered as a nutraceutical for the treatment of oxidative stress-related and pathogenic diseases.

Homogeneous and stable (+)-usnic acid loaded liposomes prepared by compressed CO2

The administration of hydrophobic actives and drugs for medical or cosmetical purposes generally requires a formulation that ensures adequate water solubility, which can be achieved through the encapsulation in liposomes. For the vehiculation of (+)-usnic acid (UA), a hydrophobic compound with antioxidant activity, we have prepared liposomes in a one-step process using compressed CO2. The investigated formulations are mainly composed of 1,2-dimyristoyl-sn-glycero-3-phosphocholine and cholesterol, but contain also a small molar fraction (10%) of a synthetic surfactant derived from L-prolinol. In previous investigations liposomes containing L-prolinol derivatives showed a higher efficacy as DNA or drug delivery systems with respect to liposomes of mere phospholipids. As a consequence, they were added to liposomes to make them more suitable UA delivery systems. By testing different surfactant chain lengths and headgroups, we studied how the chemical nature of the surfactant affects the physicochemical vesicle properties and their interaction with UA. Most formulations, especially those containing surfactants with longer alkyl chains (C14 and C16), show a good potentiality as UA delivery systems because they exhibit a higher stability, vesicle-to-vesicle homogeneity and bilayer compaction with respect to analog liposomes prepared by the conventional thin film hydration previously investigated. Our results confirm the advantages of DELOS-SUSP also in the case of mixed liposomes containing phospholipids and synthetic ionic surfactants. Moreover, this study demonstrates that liposomes composed of the same lipids can feature different properties if prepared according to different methodologies. In addition, this investigation points out that also the properties that a solute included in the bilayer show can be affected by the technique used for liposomes preparation.

Synthetic blood and blood products for combat casualty care and beyond.

Synthetic biology adopts an engineering design approach to create innovative treatments that are reliable, scalable, and customizable to individual patients. Interest in substitutes for allogenic blood components, primarily red blood cells and platelets, increased in the 1980s because of concerns over infectious disease transmission. However, only now, with emerging synthetic approaches, are such substitutes showing genuine promise. Affordable alternatives to donated blood would be of enormous benefit worldwide. Several approaches to replacing the oxygen-carrying function of red cells are under advanced investigation. Hemoglobin-based oxygen carriers incorporate modifications to reduce the renal toxicity and nitric oxide scavenging of free hemoglobin. While use of earlier-generation hemoglobin-based oxygen carriers may be limited to circumstances in which blood transfusion is not an option, recent advances in chemical modification of hemoglobin may eventually overcome such problems. Another approach encases hemoglobin molecules in biocompatible synthetic nanoparticles. An alternative is the ex vivo production of red cells in bioreactors, with or without genetic manipulation, that offers the potential of a universal donor product. Various strategies to manufacture synthetic platelets are also underway, ranging from simple phospholipid liposomes encapsulating adenosine diphosphate and decorated with fibrinogen fragments, to more complex capsules with multiple receptor peptide sequences. Ex vivo production of platelets in bioreactors is also possible including, for example, platelets derived from induced pluripotent stem cells that are differentiated into a megakaryocytic lineage. Prior to clinical use, trials assessing synthetic blood components must evaluate meaningful safety and effectiveness outcomes in relatively large numbers of critically ill patients. Overcoming these challenges may be as much a hurdle as product design. This article reviews the state of the science of the synthetic biology approach to developing blood component substitutes.

Comparative Study on Curcumin Loaded in Golden Pompano (Trachinotus blochii) Head Phospholipid and Soybean Lecithin Liposomes: Preparation, Characteristics and Anti-Inflammatory Properties.

In this study, we compared the characteristics and in vitro anti-inflammatory effects of two curcumin liposomes, prepared with golden pompano head phospholipids (GPL) and soybean lecithin (SPC). GPL liposomes (GPL-lipo) and SPC liposomes (SPC-lipo) loaded with curcumin (CUR) were prepared by thin film extrusion, and the differences in particle size, ζ-potential, morphology, and storage stability were investigated. The results show that GPL-lipo and SPC-lipo were monolayer liposomes with a relatively small particle size and excellent encapsulation rates. However, GPL-lipo displayed a larger negative ζ-potential and better storage stability compared to SPC-lipo. Subsequently, the effects of phospholipids in regulating the inflammatory response of macrophages were evaluated in vitro, based on the synergistic effect with CUR. The results showed that both GPL and SPC exerted excellent synergistic effect with CUR in inhibiting the lipopolysaccharide (LPS)-induced secretion of nitric oxide (NO), reactive oxygen species (ROS), and pro-inflammatory genes (tumor necrosis factor (TNF)-α, interleukin 1β (IL-β), and interleukin 6 (IL-6)) in RAW264.7 cells. Interestingly, GPL-lipo displayed superior inhibitory effects, compared to SPC-lipo. The findings provide a new innovative bioactive carrier for development of stable CUR liposomes with good functional properties.

How Can Immuno suppression After Organ Transplantation be Made More Effective and Safer? – A Review on Liposomal Formulations with Consideration of Archaeal Tetraetherlipid

Immune-suppressive agents such as methylprednisolone and cyclosporine exert tremendous side effects, because of high dosage and long-term application required for immune suppression after organ transplantation. Major side effects of methylprednisolone include bleeding of the gastro-intestinal tract, hypertension, and osteoporosis, whereas cyclosporine is nephrotoxic. Liposomes are phospholipid particles that allow delivery of drugs preferentially to the reticuloendothelial system. They can be prepared from phospholipids such as lecithin from soybean or egg yolk, other specific or modified lipids or from membrane-spanning tetraether lipid (TEL), which can be extracted and purified from archaeal cell membranes. One advantage in the use of liposomal application is reduced toxicity of many drugs. We report on various liposomal preparations of cyclosporine, methylprednisolone (L-MPL) and its palmitate derivative (L-MPLP). It has been documented that liposomal cyclosporine A (L-CsA), 1.75 mg/kg/day for seven days has potential for use as an immune-suppressive agent in rats with increased efficacy and decreased nephrotoxicity compared to commercially available forms of intravenous CsA. Liposomal methylprednisolone (L-MPL) 2 mg/kg, intravenously (IV), twice a week shows significantly prolonged cardiac allograft survival in rats and tissue-selective sequestration of the drug in comparison with the same dosage regimen of methylprednisolone in solution, administered daily. We report on organ distribution of L-MPLP in rats after intraperitoneal (IP) administration. Conclusion: Liposomal preparations of immunosuppressants have significantly higher immune-suppressive potential and lower toxicity than non-liposomal preparations. Bipolar TEL can be extracted, fractionated and purified from archaea to form stable liposomes which are extremely resistant, even to gastric fluid. Hence, TEL liposomes allow (besides IV and IP) for oral administration of immuno suppressants after organ transplantation with pharmacological and toxicological advantages as common liposomal phospholipid bilayer preparations.

Amphipol-facilitated elucidation of the functional tetrameric complex of full-length cytochrome P450 CYP2B4 and NADPH-cytochrome P450 oxidoreductase

Interactions of membrane-bound mammalian cytochromes P450 (CYPs) with NADPH-cytochrome P450 oxidoreductase (POR), which are required for metabolism of xenobiotics, are facilitated by membrane lipids. A variety of membrane mimetics, such as phospholipid liposomes and nanodiscs, have been used to simulate the membrane to form catalytically active CYP:POR complexes. However, the exact mechanism(s) of these interactions are unclear because of the absence of structural information of full-length mammalian CYP:POR complexes in membranes. Herein, we report the use of amphipols (APols) to form a fully functional, soluble, homogeneous preparation of full-length CYP:POR complexes amenable to biochemical and structural study. Incorporation of CYP2B4 and POR into APols resulted in a CYP2B4:POR complex with a stoichiometry of 1:1, which was fully functional in demethylating benzphetamine at a turnover rate of 37.7 ± 2.2 min−1, with a coupling efficiency of 40%. Interestingly, the stable complex had a molecular weight (Mw) of 338 ± 22 kDa determined by multiangle light scattering, suggestive of a tetrameric complex of 2CYP2B4:2POR embedded in one APol nanoparticle. Moreover, negative stain electron microscopy (EM) validated the homogeneity of the complex and allowed us to generate a three-dimensional EM map and model consistent with the tetramer observed in solution. This first report of the full-length mammalian CYP:POR complex by transmission EM not only reveals the architecture that facilitates electron transfer but also highlights a potential use of APols in biochemical and structural studies of functional CYP complexes with redox partners.

Unprecedented formation of sterically stabilized phospholipid liposomes of cuboidal morphology.

Sterically stabilized phospholipid liposomes of unprecedented cuboid morphology are formed upon introduction in the bilayer membrane of original polymers, based on polyglycidol bearing a lipid-mimetic residue. Strong hydrogen bonding in the polyglycidol sublayers creates attractive forces, which, facilitated by fluidization of the membrane, bring about the flattening of the bilayers and the formation of cuboid vesicles.

Performance of Cosmetic Ingredients Evaluated by Their Membrane Protection Efficiency

Plant extracts (PEs) are frequently used as ingredients in skin and hair care products to improve protection against the excess of sun exposure. PEs are usually selected based on their antioxidant capacity for reducing the damage in lipids and proteins, which are key molecules in the maintenance of skin homeostasis and hair integrity. Interestingly, biomolecules that do not have any antioxidant capacity can have a protective effect by physically adsorbing and inhibiting leakage of membranes. Our aims are: 1) to develop an experimental protocol that quantifies the level of membrane damage/protection; 2) to recommend an approach to compare the membrane protection efficiency of different PEs; 3) to briefly describe the mechanisms of membrane protection. Evaluation of membrane damage/protection is based on a phospholipid liposome system that signals the membrane rupture caused by photo-induced lipid oxidation. Carboxyfluorescein (CF) emission is monitored as a function of time (120 min) in a controlled experimental setup, in which a certain level of membrane damage is induced by Dimethyl-Methylene Blue (DMMB) photosensitized oxidation during red-light irradiation. Percentage levels of membrane damage/protection are calculated by comparing the CF fluorescence level of DMMB-liposomes in the presence of varying concentrations of membrane-protective ingredients with that of surfactant-dissolved DMMB-liposomes. Membrane protectants reduce the level of CF fluorescence during irradiation of DMMB-liposomes. Gallic acid (GA) and trehalose (Tre) were used as standard compounds to compute the levels of protection by the antioxidant and physical adsorption mechanisms, respectively. The percentage of membrane protection was shown to be linearly proportional to the concentration of the standard protectants. The performance of PEs was measured and compared in terms of GA or Tre membrane protection equivalents. All extracted also had their antioxidant capacities evaluate by the anti-radical DPPH assay. The membrane protection efficiency parallels the anti-radical efficiency for extracts that have antioxidant activity. However, even extracts that are mainly enriched in polysaccharides and that do not have any measurable anti-radical capacity were able to protect membranes. We propose that these extracts protect membranes by the physical adsorption mechanism, in comparison with the protection observed by Tre and also based on the known effect of carbohydrates that can protect against membrane micro-fissures. The membrane damage assay described here can be used to evaluate and compare the performance of cosmetic ingredients with claims of maintaining lipid membrane integrity of skin and hair products.

Design, Function and Applications of Molecular Permeable Polymer Vesicles

Vesicles are formed from the self–assembly of phospholipids, synthetic lipids, and amphiphilic polymers. Among vesicles, polymer vesicles can be introduced various functions such as stimuli–responsivity and have superior mechanical strength. Polymer vesicles also have an inner aqueous phase and can function as enzymatic reactors when enzymes are encapsulated in the inner aqueous phase. Therefore, they have been attracted attention for biomaterials such as nano/microreactors, carriers for drug delivery systems (DDS), and artificial organelle/cell models. However, the bilayers of polymer vesicles are thicker than those of phospholipid liposomes and thus have extremely low molecular permeability, making it difficult to supply molecules from the external environment. This review introduces our efforts to develop molecular permeable polymer vesicles, the functions of the vesicles, and applications for biomaterials as artificial molecular channels and biocatalytic nanoreactors.

DMPC Phospholipid Bilayer as a Potential Interface for Human Cystatin C Oligomerization: Analysis of Protein-Liposome Interactions Using NMR Spectroscopy.

Studies revolving around mechanisms responsible for the development of amyloid-based diseases lay the foundations for the recognition of molecular targets of future to-be-developed treatments. However, the vast number of peptides and proteins known to be responsible for fibril formation, combined with their complexity and complexity of their interactions with various cellular components, renders this task extremely difficult and time-consuming. One of these proteins, human cystatin C (hCC), is a well-known and studied cysteine-protease inhibitor. While being a monomer in physiological conditions, under the necessary stimulus—usually a mutation, it tends to form fibrils, which later participate in the disease development. This process can potentially be regulated (in several ways) by many cellular components and it is being hypothesized that the cell membrane might play a key role in the oligomerization pathway. Studies involving cell membranes pose several difficulties; therefore, an alternative in the form of membrane mimetics is a very attractive solution. Here, we would like to present the first study on hCC oligomerization under the influence of phospholipid liposomes, acting as a membrane mimetic. The protein–mimetic interactions are studied utilizing circular dichroism, nuclear magnetic resonance, and size exclusion chromatography.