纳米脂质体 单层 应用

Liposomes, due to their various forms, require further exploration. These structures can deliver both hydrophilic and hydrophobic drugs for cancer, antibacterial, antifungal, immunomodulation, diagnostics, ophtalmica, vaccines, enzymes and genetic elements. Preparation of liposomes results in different properties for these systems. In addition, based on preparation methods, liposomes types can be unilamellar, multilamellar and giant unilamellar; however, there are many factors and difficulties that affect the development of liposome drug delivery structure. In the present review, we discuss some problems that impact drug delivery by liposomes. In addition, we discuss a new generation of liposomes, which is utilized for decreasing the limitation of the conventional liposomes.

… as a drug delivery system. Until recently, both objectives faced the serious problem that a method for the preparation of a homogeneous and well-characterized population of unilamellar …

… Positively charged unilamellar liposomes enhanced … Liposomal entrapment of drug is prerequisite to enhanced … when liposomes that were preformed in the absence of drug were mixed …

… range of drug release. These results show that suspensions of large unilamellar liposomes can be made to encapsulate a therapeutically useful quantity of drug that is rapidly and …

ABSTRACT The aim of the present investigation was to evaluate the influence of liposome formulation on the ability of vesicles to penetrate a pathological mucus model obtained from COPD affected patients in order to assess the potential of such vesicles for the treatment of chronic respiratory diseases by inhalation. Therefore, Small Unilamellar Liposomes (PLAIN‐LIPOSOMEs), Pluronic® F127‐surface modified liposomes (PF‐LIPOSOMEs) and PEG 2000PE‐surface modified liposomes (PEG‐LIPOSOMEs) were prepared using the micelle‐to‐vesicle transition (MVT) method and beclomethasone dipropionate (BDP) as model drug. The obtained liposomes showed diameters in the range of 40–65nm, PDI values between 0.25 and 0.30 and surface electric charge essentially close to zero. The encapsulation efficiency was found to be dependent on the BDP/lipid ratio used and, furthermore, BDP‐loaded liposomes were stable in size both at 37°C and at 4°C. All liposomes were not cytotoxic on H441 cell line as assessed by the MTT assay. The liposome uptake was evaluated through a cytofluorimetric assay that showed a non‐significant reduction in the internalization of PEG‐LIPOSOMEs as compared with PLAIN‐LIPOSOMEs. The penetration studies of mucus from COPD patients showed that the PEG‐LIPOSOMEs were the most mucus‐penetrating vesicles after 27h. In addition, PEG‐ and PF‐LIPOSOMEs did not cause any effect on bronchoalveolar lavage fluid proteins after aerosol administration in the mouse. The results highlight that PEG‐LIPOSOMEs show the most interesting features in terms of penetration through the pathologic sputum, uptake by airway epithelial cells and safety profile.

… efficiency of hydrophilic drugs in unilamellar liposomes, and will … to compare drug encapsulation efficiencies of liposomes … control of drug encapsulation during liposome …

… Our results have important implications for the use of liposomes as a slow release drug-delivery system within the vasculature, particularly because prolonged circulation times have …

Periocular routes represent a promising approach to chronic eye therapy due to its unique exploitation of scleral structure. Especially, for ophthalmic infection requiring prolonged …

… Here, I describe two major types of liposome preparation: (1) small unilamellar vesicles (SUVs) prepared by “bath sonication” method, (2) large unilamellar vesicles (LUVs) prepared by “…

The purpose of this study was to compare the various methods available to separate non-encapsulated drug from large unilamellar liposomes (LUV). Multilamellar liposomes (MLV) were prepared by thin film hydration using distearoylphosphatidylcholine:cholesterol (2:1 molar ratio). MLVs were passed through a 0.2-μm polycarbonate membrane using an extruder to prepare LUVs. Particle size of liposome preparations was characterized using a submicron particle-size analyser. The non-encapsulated drug was separated by: filtering through Centrifree tubes; passing through gel (Sepharose-4B and Sephadex G-25M); passing through minicolumn; ficoll density gradient; protamine aggregation; or dialysis. The dialysis method was found to be unsuitable for separation of non-encapsulated drug due to equilibration of encapsulated drug as the free drug was dialyzed. The upper limit for lipid concentration was 5 mg mL−1 using the Centrifree method. Separation using gel chromatography led to dilution of liposome preparation. Minicolumn and density gradient techniques did not lead to sample dilution, however the minicolumn method was tedious. The time required for separation of liposomes by protamine aggregation was longer for neutral liposomes. Thus it was concluded that the Centrifree was the fastest method to estimate encapsulation; the density gradient method was ideal to separate non-encapsulated drug; and protamine aggregation was the least expensive method to estimate encapsulation efficiency.

… the interactions of multilamellar with unilamellar liposomes bearing phosphate and … liposomes have been used to predict in vivo drug retention for unilamellar liposomal drug delivery …

… However it is possible that a small unilamellar vesicles (SUVS) coupled to brain drug transport vectors may be transported through the BBB by receptor mediated or absorptive mediated …

… The permeability properties of liposomes are important for understanding the function of … for designing liposomes as selective drug delivery vehicles. Previous investigations of liposome …

… Thus, a 50% reduction in NMR signal means that the liposome preparation is unilamellar and 25% reduction in the intensity of the original NMR signal means there are 2 bilayers in the …

… delivery. The use of liposomes as vehicles for drug delivery, however, was soon found to be … , employed to produce small unilamellar liposomes, affects the sample morphology, we also …

… Delivery of a membrane-bound molecule may be better described by a surface-area … In this work, the size of liposomes was measured in terms of volume, so these liposomes can be …

… Ultrasound is commonly used in the preparation of unilamellar liposome dispersions and is often considered for cell membrane disruption for drug delivery or DNA transfection …

… that result in small unilamellar liposomes is the combination … We speculate that the mechanism of unilamellar liposome … this liposome formation process attractive for use in drug delivery …

Here, we introduce a one-pot method for the bottom-up assembly of complex single- and multicompartment synthetic cells. Cellular components are enclosed within giant unilamellar vesicles (GUVs), produced at the milliliter scale directly from small unilamellar vesicles (SUVs) or proteoliposomes with only basic laboratory equipment within minutes. Toward this end, we layer an aqueous solution, containing SUVs and all biocomponents, on top of an oil–surfactant mix. Manual shaking induces the spontaneous formation of surfactant-stabilized water-in-oil droplets with a spherical supported lipid bilayer at their periphery. Finally, to release GUV-based synthetic cells from the oil and the surfactant shell into the physiological environment, we add an aqueous buffer and a droplet-destabilizing agent. We prove that the obtained GUVs are unilamellar by reconstituting the pore-forming membrane protein α-hemolysin and assess the membrane quality with cryotransmission electron microscopy (cryoTEM), fluorescence recovery after photobleaching (FRAP), and zeta-potential measurements as well as confocal fluorescence imaging. We further demonstrate that our GUV formation method overcomes key challenges of standard techniques, offering high volumes, a flexible choice of lipid compositions and buffer conditions, straightforward coreconstitution of proteins, and a high encapsulation efficiency of biomolecules and even large cargo including cells. We thereby provide a simple, robust, and broadly applicable strategy to mass-produce complex multicomponent GUVs for high-throughput testing in synthetic biology and biomedicine, which can directly be implemented in laboratories around the world.

Among the molecular milieu of the cell, the membrane bilayer stands out as a complex and elusive synthetic target. We report a microfluidic assembly line that produces uniform cellular compartments from droplet, lipid, and oil/water interface starting materials. Droplets form in a lipid-containing oil flow and travel to a junction where the confluence of oil and extracellular aqueous media establishes a flow-patterned interface that is both stable and reproducible. A triangular post mediates phase transfer bilayer assembly by deflecting droplets from oil, through the interface, and into the extracellular aqueous phase to yield a continuous stream of unilamellar phospholipid vesicles with uniform and tunable size. The size of the droplet precursor dictates vesicle size, encapsulation of small-molecule cargo is highly efficient, and the single bilayer promotes functional insertion of a bacterial transmembrane pore.

… approaches for the synthesis of giant unilamellar vesicles as models of synthetic cells, from … of the nascent vesicles. Incorporation of lipids delivered by small unilamellar vesicles (SUVs) …

… If lipid molecules have an effect on GFP synthesis, we would expect a pronounced effect in smaller GUV. Therefore, the vesicle size dependencies of the yield of GFP synthesis were …

… Stable unilamellar vesicles form spontaneously upon mixing aqueous suspensions of long-… or longer) with small amounts (20 mol %) of micellar synthetic short-chain lecithins (fatty …

Creating an artificial cell from the bottom up is a long‐standing challenge and, while significant progress has been made, the full realization of this goal remains elusive. Arguably, one of the biggest hurdles that researchers are facing now is the assembly of different modules of cell function inside a single container. Giant unilamellar vesicles (GUVs) have emerged as a suitable container with many methods available for their production. Well‐studied swelling‐based methods offer a wide range of lipid compositions but at the expense of limited encapsulation efficiency. Emulsion‐based methods, on the other hand, excel at encapsulation but are only effective with a limited set of membrane compositions and may entrap residual additives in the lipid bilayer. Since the ultimate artificial cell will need to comply with both specific membrane and encapsulation requirements, there is still no one‐method‐fits‐all solution for GUV formation available today. This review discusses the state of the art in different GUV production methods and their compatibility with GUV requirements and operational requirements such as reproducibility and ease of use. It concludes by identifying the most pressing issues and proposes potential avenues for future research to bring us one step closer to turning artificial cells into a reality.

Giant unilamellar vesicles (GUVs) have gained great popularity as mimicries for cellular membranes. As their sizes are comfortably above the optical resolution limit, and their lipid composition is easily controlled, they are ideal for quantitative light microscopic investigation of dynamic processes in and on membranes. However, reconstitution of functional proteins into the lumen or the GUV membrane itself has proven technically challenging. In recent years, a selection of techniques has been introduced that tremendously improve GUV-assay development and enable the precise investigation of protein-membrane interactions under well-controlled conditions. Moreover, due to these methodological advances, GUVs are considered important candidates as protocells in bottom-up synthetic biology. In this review, we discuss the state of the art of the most important vesicle production and protein encapsulation methods and highlight some key protein systems whose functional reconstitution has advanced the field. Expected final online publication date for the Annual Review of Biophysics, Volume 50 is May 2021. Please see http://www.annualreviews.org/page/journal/pubdates for revised estimates.

In order to recapitulate complex eukaryotic compartmentalization, synthetic biology aims to recreate cellular membrane-lined compartments from the bottom-up. Many important cellular organelles and cell-produced extracellular vesicles are in the size range of several hundreds of nanometers. Although attaining a fundamental characterization and mimicry of their cellular functions is a compelling goal, the lack of methods for controlled vesicle formation in this size range has hindered full understanding. Here, we show the optimization of a simple and efficient protocol for the production of large unilamellar vesicles (LUVs) with a median diameter in the range of 450–550 nm with high purity. Importantly, we rely on commercial reagents and common laboratory equipment. We thoroughly characterize the influence of different experimental parameters on the concentration and size of the resulting vesicles and assess changes in their lipid composition and surface charge. We provide guidance for researchers to optimize LUV production further to suit specific applications.

Synthesis of giant unilamellar vesicles (GUVs) of charged and uncharged lipids at physiological salt concentration is presented using the starch hydrogel method as an example of the gel assisted synthesis method. The swelling of the gel is assisted by the presence of a high amount of amylopectin in starch and yields giant-sized vesicles, which are unilamellar in nature. This method holds promise since starch is a commonly available cheap bio-compatible material. This work indicates that native starch yields vesicles of better size range as compared to the acid-treated starch. It is demonstrated that contrary to the common belief, pre-hydration of bilayers is not critical to the success of this method. The synthesis of GUVs in physiological salt concentrations is possible since the salt does not produce any osmotic effect on its own. At low starch concentration, the size of the vesicles is found to correlate with the swelling factor. The conjugate effect of the starch concentration and ion leads to the change in the swelling factor of the gel and thereby influence the size and architecture of the vesicles. Also, interactions between starch and lipid play an important role in the formation of the giant vesicles.

… the formation of unilamellar vesicles. Indeed, the most widely used vesicle formation method … During the growing process, small vesicles often fuse into a larger one that may later detach …

The development of greener nano-constructs with noteworthy biological activity is of supreme interest, as a robust choice to minimize the extensive use of synthetic drugs. Essential oils (EOs) and their constituents offer medicinal potentialities because of their extensive biological activity, including the inhibition of fungi species. However, their application as natural antifungal agents are limited due to their volatility, low stability, and restricted administration routes. Nanotechnology is receiving particular attention to overcome the drawbacks of EOs such as volatility, degradation, and high sensitivity to environmental/external factors. For the aforementioned reasons, nanoencapsulation of bioactive compounds, for instance, EOs, facilitates protection and controlled-release attributes. Nanoliposomes are bilayer vesicles, at nanoscale, composed of phospholipids, and can encapsulate hydrophilic and hydrophobic compounds. Considering the above critiques, herein, we report the in-house fabrication and nano-size characterization of bioactive oregano essential oil (Origanum vulgare L.) (OEO) molecules loaded with small unilamellar vesicles (SUV) nanoliposomes. The study was focused on three main points: (1) multi-compositional fabrication nanoliposomes using a thin film hydration–sonication method; (2) nano-size characterization using various analytical and imaging techniques; and (3) antifungal efficacy of as-developed OEO nanoliposomes against Trichophyton rubrum (T. rubrum) by performing the mycelial growth inhibition test (MGI). The mean size of the nanoliposomes was around 77.46 ± 0.66 nm and 110.4 ± 0.98 nm, polydispersity index (PdI) of 0.413 ± 0.015, zeta potential values up to −36.94 ± 0.36 mV were obtained by dynamic light scattering (DLS). and spherical morphology was confirmed by scanning electron microscopy (SEM). The presence of OEO into nanoliposomes was displayed by attenuated total reflection Fourier-transform infrared (ATR-FTIR) spectroscopy. Entrapment efficiency values of 79.55 ± 6.9% were achieved for OEO nanoliposomes. In vitro antifungal activity of nanoliposomes tested against T. rubrum strains revealed that OEO nanoliposomes exhibited the highest MGI, 81.66 ± 0.86%, at a concentration of 1.5 µL/mL compared to the rest of the formulations. In summary, this work showed that bioactive OEO molecules with loaded nanoliposomes could be used as natural antifungal agents for therapeutical purposes against T. rubrum.

… Natural and synthetic phospholipids were purchased from … that one can produce giant vesicles from small-size liposomes … on the platinum electrodes as small drops not exceeding 2 μL (…

… One of the current challenges in synthetic biology is the production of stable membrane … and ATP synthase from Escherichia coli into unilamellar liposomes ranging from 100 nm to 50 …

A simple Giant unilamellar vesicle (sGUV) is a micron-sized spherical vesicle, which is composed of a lipid bilayer that encloses an aqueous solution inside and is suspended in another aqueous medium. As biomimetic models of biological cells, sGUVs are well-established systems for biophysical studies. However, sGUVs primarily mimic anucleate cells, limiting their application as models for non-nucleate cells. We propose compound Giant Unilamellar vesicles (cGUVs) as a more appropriate biomimetic model for nucleated cells. cGUVs are vesicle-in-vesicle structures, where the outer and inner vesicle bilayers can be assumed to represent the cell and the nuclear membrane of a cell, respectively. In this study, we describe a simple method for the synthesis of cGUVs. Briefly, sGUVs were generated using the lipid composition of DMPC and cholesterol within the range of 37-43 mol% in a sucrose medium via the electroformation method. These sGUVs were then subjected to osmotic shock by introducing a hypertonic glucose solution, triggering an immediate transition to a somatocytic shape. This process led to the formation of an intermediate state where the outer and inner vesicles remained connected through a neck, ultimately resulting in the development of vesicle-in-vesicle structure, referred to as cGUVs. Furthermore, it is known that GUVs prepared with higher cholesterol levels in the bilayer exhibit reduced ion permeability. This characteristic allows tuning of conductivity in the annular solution of a cGUV by adjusting the conductivities of the hydrating medium, the inner solution through hypertonic glucose, and the outer solution by adjusting of the dilution or the addition of conductive media, making the resultant cGUVs with controlled conductivities in the outer, annular, and inner regions, relevant for electric field studies. We propose this efficient and straightforward approach for synthesizing compound vesicles that can be used as a biomimetic model for eukaryotic, nucleated cells, advancing their application in biophysical research.

… was integrated into the membrane of smaller vesicles; this finding is quantitatively explained … ratio in smaller vesicles. Finally, we discuss the applications of membrane protein synthesis …

The reconstitution of a cell nucleus in a lipid bilayer-enclosed synthetic cell makes great strides in bottom-up synthetic biology. In this study, we propose a method for assembling a nucleus in giant unilamellar vesicles (GUVs). To induce reconstitution of the nucleus, we utilize interphase egg extract of African clawed frogs Xenopus laevis, known as a biochemically controllable cell-free system capable of transforming an added sperm chromatin into a nucleus in vitro. We enhanced GUV formation efficiency by the inverted emulsion method through incorporating prolonged waiting time and adding chloroform into lipid-dispersed oil, facilitating subsequent nuclear assembly reactions in the GUVs.Characterization of nucleus-like structures formed in the GUVs revealed the presence of dense DNA and accumulated GFP-NLS in the structure, indicative of functional nuclear import. Immunostaining further validated the presence of nuclear pore complexes on the surfaces of these nucleus-like structures. Our approach offers a versatile platform for constructing artificial cellular systems that closely mimic eukaryotic cells. Teaser A cell nucleus is reconstituted in lipid bilayer-enclosed confinements using egg extract from African clawed frogs.

… smaller vesicles by mechanical treatment, the GUVs do not spontaneously reform from the smaller vesicles. … biotechnology: Synthesis of proteins and nucleic acids inside lipid vesicles. J …

A synthetic, amphipathic 30-amino acid peptide with the major repeat unit Glu-Ala-Leu-Ala (GALA) was designed to mimic the behavior of the fusogenic sequences of viral fusion proteins. GALA is a water-soluble peptide with an aperiodic conformation at neutral pH and becomes an amphipathic alpha-helix as the pH is lowered to 5.0 where it interacts with bilayers. Fluorescence energy transfer measurements indicated that GALA induced lipid mixing between phosphatidylcholine small unilamellar vesicles but not large unilamellar vesicles. This lipid mixing occurred only at pH 5.0 and not at neutral pH. Concomitant with lipid mixing, the vesicles increased in diameter from 500 to 750 to 1000 A as measured by dynamic light scattering and internal volume determination. GALA induced leakage of small molecules (Mr 450) at pH 5.0 was too rapid to permit detection of contents mixing. However, retention of larger molecules (Mr 4100) under the same conditions suggests that vesicle fusion is occurring. For a 100/1 lipid/peptide ratio all vesicles fused just once, whereas for a 50/1 ratio higher order fusion products formed. A mass action model gives good simulation of the kinetics of increase in fluorescence intensity and yields rate constants of aggregation and fusion. As the lipid to peptide ratio decreases from 100/1 to 50/1 both rate constants of aggregation and fusion increase, indicating that GALA is a genuine inducer of vesicle fusion. The presence of divalent cations which can alter GALAs conformation at pH 7.5 had little effect on its lipid mixing activity. GALA was modified by altering the sequence while keeping the amino acid composition constant or by shortening the sequence. These peptides did not have any lipid mixing activity nor did they induce an increase in vesicle size. Together, these results indicate that fusion of phosphatidylcholine small unilamellar vesicles induced by GALA requires both a peptide length greater than 16 amino acids as well as a defined topology of the hydrophobic residues.

… giant unilamellar vesicles and … large unilamellar vesicles (LUVs) with diameters of about 100 nm or less, as used for drug delivery applications,17 the internal volume of giant vesicles is …

… to apply as they can involve several days of dialysis. A general and straightforward protocol for production of large unilamellar vesicles is … agents, would produce vesicles of a relatively …

… To illustrate possible applications of this technique, we demonstrate the use of … distribution of large unilamellar vesicles (LUV). By contrast, production of giant unilamellar vesicles (GUV) …

… Vesicle systems containing long-chain saturated lipids have potential in applications where highly ~,table large unilamellar vesicles … the production of large unilameliar vesicles (LUVs) …

… Despite the extensive use and diverse applications of vesicles … Small unilamellar vesicles These vesicles, with sizes ranging … Large unilamellar vesicles These vesicles, ranging from 0.1 …

… application of liposomes and lipid vesicles in biology and medicine has become increasingly widespread in recent years. In biology, lipid vesicles … of large unilamellar vesicles which …

… of time as small unilameilar vesicles (SUVs). Large unilamellar vesicles and GUVs occupy … of biomolecules to organic solvents, severe limitations apply to what kind of molecules can be …

Liposomes and niosomes are currently the most studied lipid vesicles in the nanomedicine field. The system formed by a phospholipid bilayer in aqueous medium allows these vesicles to carry both hydrophilic and lipophilic compounds, providing an increase in solubility of drugs lready used in conventional therapy. The focus on the development of these vesicles should be directed to determining the ideal composition, with low toxicity, biocompatibility and which remains stable for long periods. These characteristics are related to the components used for formulation and the substances that will be encapsulated. Another important point relates to the methods used during formulation, which are important in determining the type of vesicle formed, whether these be large or small, unilamellar or multilamellar. Because of the deliberate actions applied in the development of these vesicles, this review sought to gather updated information regarding the different methods used, including their main components while considering the behavior of each of them when used in different formulations. Also, data showing the importance of formulations in the medical field evidencing studies performed with liposome and niosome vesicles as promising in this area, and others, were included. The approach allows a better understanding of the participation of components in formulations such as cholesterol and non-ionic surfactants, as well as the basis for choosing the ideal components and methods for future research in the development of these vesicles.

Giant vesicles (GVs), which are closed lipid bilayer membranes with a diameter of more than 1 μm, have attracted attention not only as model cell membranes but also for the construction of artificial cells. For encapsulating water-soluble materials and/or water-dispersible particles or functionalizing membrane proteins and/or other synthesized amphiphiles, giant unilamellar vesicles (GUVs) have been applied in various fields, such as supramolecular chemistry, soft matter physics, life sciences, and bioengineering. In this review, we focus on a preparation technique for GUVs that encapsulate water-soluble materials and/or water-dispersible particles. It is based on the centrifugation of a water-in-oil emulsion layered on water and does not require special equipment other than a centrifuge, which makes it the first choice for laboratory use. Furthermore, we review recent studies on GUV-based artificial cells prepared using this technique and discuss their future applications.

In this paper, we innovatively proposed a highly uniform vesicle preparation scheme based on the intervesicle mechanical self-constraint effect of vesicle crowding. By adjusting the spacing of discrete microwell structures, we observed that during the self-assembly of phospholipid molecules in microwells to form giant unilamellar vesicles (GUVs), the scale swelling of the vesicles during the continuous growth process would lead to the crowding of vesicles in adjacent microwells, thus inducing the formation of intervesicle mechanical self-constraint effect. The results of the experiment showed that this paper obtained the optimized discretized microwell structure (micropillar side: 30 μm; pitch: 0 μm), and the corresponding lipid mass was measured and determined, yielding homogeneous giant GUVs of 37.9 ± 2.0 μm. In this paper, homogenized GUVs (∼40 μm) with different cholesterol concentrations (10, 20, and 30%) were obtained by this method, and the above vesicles were subjected to controlled electroporation experiment under external electric fields of 23, 31, and 41 kV/cm, respectively. It showed that the mechanical self-constraint effect of vesicle crowding induced by patterned microstructures during the self-assembly of phospholipid molecules significantly enhances the size homogeneity of GUVs, which would be helpful for the wide applications of GUVs in other areas such as cell-like models and controlled release of drugs.

… small unilamellar vesicles (SUVs). The validity of the protocol has been demonstrated by applying it … However, the large vesicles prepared by hydration methods are found to be mostly …

Model systems such as black lipid membranes or conventional uni- or multilamellar liposomes are commonly used to study membrane properties and structure. However, the construction and dimensions of these models excluded their direct optical microscopic observation. Since the introduction of the simple method of liposome electroformation in alternating electric field giant unilamellar vesicles (GUVs) have become an important model imitating biological membranes. Due to the average diameter of GUVs reaching up to 100 microm, they can be easily observed under a fluorescent or confocal microscope provided that the appropriate fluorescent probe was incorporated into the lipid phase during vesicle formation. GUVs can be formed from different lipid mixtures and they are stable in a wide range of physical conditions such as pH, pressure or temperature. This mini-review presents information about the methods of GUV production and their usage. Particularly, the use of GUVs in studying lipid phase separation and the appearance and behavior of lipid domains (rafts) in membranes is discussed but also other examples of GUVs use in membrane research are given. The experience of the authors in setting up the GUV-forming equipment and production of GUVs is also presented.

The desire to create cell-like models for fundamental science and applications has spurred extensive effort towards creating giant unilamellar vesicles (GUVs). However, a route to selectively self-assemble GUVs in bulk has remained elusive. In bulk solution, membrane-forming molecules such as phospholipids, single-tailed surfactants, and block copolymers typically self-assemble into multilamellar, onion-like structures. So although self-assembly processes can form nanoscale unilamellar vesicles, scaffolding by droplets or surfaces is required to create GUVs. Here we show that it is possible to bulk self-assemble cell-sized GUVs with almost complete selectivity over other vesicle topologies. The seemingly paradoxical pair of features that enables this appears to be having very dynamic molecules at the nanoscale, that create unusually rigid membranes. The resultant self-assembly pathway enables encapsulation of molecules and colloids, and can also generate model primitive cells that can grow and divide.

The purpose of this study was to design ligand-functionalized nanoliposomes that are proficient in providing effective intracellular delivery of an alkaloid drug (galantamine) into PC12 …

… “ordered complexity” of the lipid bilayer and how it impacts the … by mobilizing the phospholipid bilayer into its own defense. … Thus the bilayer has a triple role in this account: a safe space …

Liposomes are widely used in the biological field due to their good biocompatibility and surface modification properties. With the development of biochemistry and material science, many liposome structures and their surface functional components have been modified and optimized one by one, pushing the liposome platform from traditional to functionalized and intelligent, which will better satisfy and expand the needs of scientific research. However, a main limiting factor effecting the efficiency of liposomes is the complicated environmental conditions in the living body. Currently, in order to overcome the above problem, functionalized liposomes have become a very promising strategy. In this paper, binding strategies of liposomes with four main functional elements, namely nucleic acids, antibodies, peptides, and stimuli-responsive motif have been summarized for the first time. In addition, based on the construction characteristics of functionalized liposomes, such as drug-carrying, targeting, long-circulating, and stimulus-responsive properties, a comprehensive overview of their features and respective research progress are presented. Finally, the paper critically presents the limitations of these functionalized liposomes in the current applications and also prospectively suggests the future development directions, aiming to accelerate realization of their industrialization.

… Kalyankar and co-workers14 have individually arrayed intact single liposomes into functionalized microwells (1.2 μm in diameter) etched into a silicon wafer. High-density liposome …

… Herein, rhamnolipid functionalized-nanoliposomes loaded with diacetyl (DT/RL-LPs) were prepared to enhance diacetyl stability and improve the biofilm elimination efficiency. Results …

Aims: This study aims to develop and evaluate oleuropein loaded surface functionalized folate‐targeted – PEG liposomes for the effective management of prostate cancer in an animal model. Materials and methods: Film hydration‐cum‐extrusion technique was used to produce liposomes. Particle size, entrapment efficiency, drug loading, electron microscopy, and drug release study were performed for the characterization. Cell viability and various in vitro studies (phosphatidylserine internalization, TUNEL assay, measurement of mitochondrial membrane potential and caspase‐3 assay) were performed to compare the anticancer and apoptotic effects of developed liposomes against the plain oleuropein. Comparative pharmacokinetic profiling and anticancer efficacy studies including a change in tumor volume, body weight, and survival analysis were performed in mice model. Key findings: The developed liposomes (OL‐FML) showed the particle size of 184.2 ± 9.16 nm, the zeta potential of 1.41 ± 0.24 mV, entrapment efficiency of 63.52 ± 4.15% and drug loading of 21.31 ± 2.37%. OL‐FML showed higher in vitro anti‐proliferative effect and apoptosis on 22Rv1 cells. In vivo pharmacokinetic study revealed a nearly 6 fold increase in the bioavailability of OL‐FML (AUC0→∞ = 641.78 ± 103.764 &mgr;g/mL·hr) as compared to OL solution (AUC0→∞ = 104.11 ± 18.374 &mgr;g/mL·hr) in mice. Increased tumor suppression, weight loss resistance, and survival probability were observed in 22Rv1 induced tumor‐bearing mice with OL‐FML treatment as compared to OL. Significance: The study provides conclusive evidence for the utilization of combining passive and active targeting strategy to enhance the anticancer effect of OL.

… Nanoliposomes incorporating curcumin (curcumin-liposomes) were prepared by adding curcumin in the lipid phase during liposome … into nanoliposomes during their formation. All …

… Two kinds of functionalization of a lipid bilayer vesicle by titania … , a morphologically stable lipid bilayer vesicle having an atomic … The other is the creation of an asymmetric lipid bilayer …

Liposome surface functionalization facilitates enormous potential applications of liposomes, such as enhanced stability, bioactive liposome conjugates, and targeted drug, gene and image agent delivery. Anchoring lipids are needed for grafting ligands of interest and play important roles in ligands grafting density, liposome stability, and liposome chemical and physical characteristics as well. In this report, glyco-functionalized liposome systems based on two kinds of anchoring lipid, phosphatidylethonalamine (PE) and cholesterol (Chol) were prepared by post chemically selective functionalization via Staudinger ligation. The size and stability of the liposomes were confirmed by dynamic light scattering (DLS). Particularly, the impact of anchor lipids on the stability of glyco-functionalized liposomes was investigated by comparing two different anchor lipids, namely Chol-PEG2000-TP and DSPE-PEG2000-TP. In addition, the encapsulation and releasing capacity of the glycosylated liposome based on the two anchoring lipids were investigated by entrapping 5, 6-carboxyfluorescein (CF) dye and monitoring the fluorescence leakage, respectively. Furthermore, the density and accessibility of grafted carbohydrate residues on the liposome surface were evaluated for the two anchoring lipids-derived liposomes with lectin binding, respectively.

… the need for the lipid bilayer. To directly confirm the presence of the liposome coexisting with the cargo, we utilized formulation II, which places a dye in the lipid bilayer. Figures 5 and 6 …

… assembled as part of the lipid bilayer of vesicles, suitable for cell … than the thickness of the lipid bilayer (approximately 4 nm), … study offers more flexibility since functionalized QD up to 50 …

Liposomes loaded with drug–cyclodextrin complexes are widely used as drug delivery systems, especially for species with low aqueous solubility and stability. Investigation of the intimate interactions of macrocycles with liposomes are essential for formulation of efficient and stable drug-in-cyclodextrin-in-liposome carriers. In this work, we reported the preparation of unilamellar vesicles of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) embedded with native β-cyclodextrin and two synthetic derivatives: heptakis(2,3,6-tri-O-methyl)-β-cyclodextrin (TMCD) and heptakis(2,3-di-O-acetyl)-β-cyclodextrin (DACD). We then studied the effect of these macrocycles on the liposomal size, membrane viscosity, and liposomal stability at different temperatures and concentrations. We observed that TMCD and DACD affected vesicle size and the change of size was related to CD concentration. Irrespective of its nature, the macrocycle established interactions with the phospholipidic head groups, preventing cyclodextrins to diffuse into the lipid bilayer, as confirmed by molecular dynamics simulations. Such supramolecular structuring improves liposome stability making these colloid systems promising carriers for biologically active compounds.

… nanoliposomes (NLs) with high affinity for amyloid deposits, was reported [11], [18]. Preliminary results indicate that such nanoliposomes … a distance from the vesicle bilayer, in order not …

Small unilamellar vesicles (SUVs) are indispensable model membranes, organelle mimics, and drug and vaccine carriers. However, the lack of robust techniques to functionalize or organize preformed SUVs limits their applications. Here we use DNA nanostructures to coat, cluster, and pattern sub-100-nm liposomes, generating distance-controlled vesicle networks, strings and dimers, among other configurations. The DNA coating also enables attachment of proteins to liposomes, and temporal control of membrane fusion driven by SNARE protein complexes. Such a convenient and versatile method of engineering premade vesicles both structurally and functionally is highly relevant to bottom-up biology and targeted delivery.

Purpose Previous studies demonstrated the possibility of targeting tumor-angiogenic endothelial cells with positively charged nanocarriers, such as cationic liposomes. We investigated the active targeting potential of positively charged nanoparticles in combination with the heat-induced drug release function of thermosensitive liposomes (TSL). This novel dual-targeted approach via cationic TSL (CTSL) was thoroughly explored using either a novel synthetic phospholipid 1,2-dipalmitoyl-sn-glycero-3-phosphodiglycerol (DPPG2) or a conventional polyethylene glycol (PEG) surface modification. Anionic particles containing either DPPG2 or PEG were also included in the study to highlight difference in tumor enrichment driven by surface charge. With this study, we aim to provide a deep insight into the main differences between DPPG2- and PEG-functionalized liposomes, focusing on the delivery of a well-known cytotoxic drug (doxorubicin; DOX) in combination with local hyperthermia (HT, 41–43°C). Materials and Methods DPPG2- and PEG-based cationic TSLs (PG2-CTSL/PEG-CTSL) were thoroughly analyzed for size, surface charge, and heat-triggered DOX release. Cancer cell targeting and DOX delivery was evaluated by FACS, fluorescence imaging, and HPLC. In vivo particle behavior was analyzed by assessing DOX biodistribution with local HT application in tumor-bearing animals. Results The absence of PEG in PG2-CTSL promoted more efficient liposome–cell interactions, resulting in a higher DOX delivery and cancer cell toxicity compared with PEG-CTSL. By exploiting the dual-targeting function of CTSLs, we were able to selectively trigger DOX release in the intracellular compartment by HT. When tested in vivo, local HT promoted an increase in intratumoral DOX levels for all (C)TSLs tested, with DOX enrichment factors ranging from 3 to 14-fold depending on the type of formulation. Conclusion Cationic particles showed lower hemocompatibility than their anionic counterparts, which was partially mitigated when PEG was grafted on the liposome surface. DPPG2-based anionic TSL showed optimal local drug delivery compared to all other formulations tested, demonstrating the potential advantages of using DPPG2 lipid in designing liposomes for tumor-targeted applications.

… a tail-to-tail fashion similar to the liposome bilayer, but while the outer layer is facing directly … of functionalized liposome surfaces cannot be replicated on SLB systems by direct liposome …

Purpose We investigated the ability of amyloid-β-targeting liposomes, decorated with an anti-transferrin receptor antibody, to cross the blood–brain barrier (BBB), comparing two antibody ligation techniques. Methods Fluorescent or radiolabeled liposomes composed of sphingomyelin/cholesterol and containing phosphatidic acid, known to bind amyloid-β, were further functionalized with the anti-transferrin receptor antibody RI7217. Two different techniques were used to attach RI7217 to the liposomes surface: biotin/streptavidin linkage or thiol–maleimide covalent ligation. Surface plasmon resonance (SPR) and immunoblotting were employed to assess the nanoparticles’ binding performances. Confocal microscopy and radiochemical techniques were used for uptake and permeability studies on an in vitro BBB model made of human brain capillary endothelial cells hCMEC/D3. Results Immunoblotting experiments showed that RI7217-functionalized liposomes bind to transferrin receptor independently of the procedure employed to ligate their surface with the antibody, while SPR experiments showed a slightly higher affinity for covalently functionalized nanoliposomes. The functionalization with RI7217 did not affect the liposomes’ affinity for amyloid-β. The functionalization of liposomes with RI7217, independently of the ligation procedure, gave higher values of uptake and permeability across the barrier model in comparison to the nondecorated ones, without cell monolayer alterations. Of note, the best performing particles were those covalently coupled with the antibody. The ratios of the two radiolabeled lipids (3H-sphingomyelin and 14C-phosphatidic acid) present in the liposome bilayer were found to be similar in the apical and in the basolateral compartments of the barrier model, suggesting that liposomes were transported intact across the cell monolayer. Confocal experiments showed no co-localization of RI7217-liposomes with early/late endosomes or early lysosomes. Conclusion Our results suggest that RI7217 promotes the in vitro barrier crossing of liposomes containing phosphatidic acid, targeting the Alzheimer’s disease amyloid-β peptide. Moreover, for the first time, we prove herein the superior efficiency of covalent coupling of RI7217 versus biotin/streptavidin ligation to facilitate liposomes in overcoming the BBB in vitro.

… We report that treatment with unilamellar phospholipid vesicles (diameter, < 0.1,um) … and substantially increases tumorassociated radioactivity. Vesicles containing aminomannose in the …

… ) across the vesicle membrane. We demonstrate here that the anticancer drugs, adriamycin … rapidly accumulated into egg phosphatidylcholine large unilamellar vesicles in response to a …

… Doxil particles are small (<100 nm) unilamellar vesicles with encapsulated doxorubicin precipitated in the liposomal vesicle by an (NH 4 ) SO 4 gradient [24]. The polyethylene glycol …

Liver cancers are often diagnosed at advanced stages and are the fourth leading cause of cancer death globally. Liver metastases, particularly from colorectal cancer, occur in 66% of patients. Immunotherapies for these cancers are limited by immunosuppressive tumor microenvironments. To address this, phospholipid‐free small unilamellar vesicles (PFSUV) are developed to deliver the toll‐like receptor 7 agonist Imiquimod (IMQ) to hepatocytes. PFSUV consists of 83 mol% cholesterol and 17 mol% Tween80, with IMQ encapsulated in these 75‐nm particles. Intravenous administration of PFSUV‐IMQ sustained liver IFN‐α levels over 24 h while reducing systemic exposure. In a CT26 liver metastasis model, PFSUV‐IMQ combined with Oxaliplatin reduced tumor size, increased CD8+ T cell infiltration, and enhanced tumor apoptosis. In an HCA‐1 liver cancer model, the same treatment decreased tumor burden, increased apoptosis, and reduced lung metastases. Flow cytometry revealed increased CD86+/MHC‐II+ dendritic cells and IFN‐γ+ CD8+ T cells in treated tumors. RNA‐seq shows enrichment of innate immune activation genes after a single dose. These findings suggest that targeted IMQ delivery activates the tumor immune microenvironment, leading to reduced tumor burden in liver cancer and metastasis models.

Localized enzyme-assisted self-assembly (LEASA) has emerged as a powerful tool to generate peptide nanofibers from and within bacteria or cancer cell lines. This approach has led to promising developments in medical imaging, antimicrobial treatments and cancer therapies. Despite these achievements, the features of self-assembly processes localized near the plasma cell membranes and induced by enzymes are not easy to study since biological media compositions are complex and vary over time. From model systems based on giant and small unilamellar vesicles (GUV and SUV respectively) displaying phosphatases at the outer edge of their phospholipid’s membrane, we study the self-assembly of peptides triggered by an enzymatic dephosphorylation. Peptide nanofibers, generated in the close vicinity of GUVs, adsorb all around the membrane where enzymes are located. This process induces the formation of a transient permeability through the membrane without destroying the vesicles, as observed by confocal laser scanning microscopy (CLSM). A cryo-transmission electron microscopy (cryo-TEM) monitoring over time highlights a lag-time before the formation of nano-aggregates located all around SUVs, followed by a rapid formation of short nanofibers near or directly from vesicles. Thanks to the presence of enzymes located on the surface of the vesicles, the micrometer-long nanofibers both adhere and exert such a mechanical force on the spherical shape of vesicles that they deform them. Finally, based on classical molecular dynamics simulations, we propose a mechanism that accounts for all our experimental observations, rationalizing the LEASA process induced on the surface of phospholipid bilayers containing phosphatases.

… The in vivo efficacy of temperature-sensitive unilamellar vesicles encapsulated dacarbazine in … with liposornes prepared from synthetic lipids for use in multimodality cancer therapy. …

The bottom-up construction of enzyme-based artificial cells is generating increasing interest, but achieving artificial cells for "all artificial modules" remains challenging in synthetic biology. Here, we introduce a fully synthetic cell system by integration of biomimetic nanozymes into giant unilamellar vesicles (GUVs). To mimic native peroxidase for free radical generation by taking advantage of Fenton catalysis reactions, we designed and prepared a de novo artificial nanozyme composed of ferritin heavy-chain scaffold protein and catalytic Fe3O4 nanoparticles as the active center. As two examples in bioapplications, we showed this nanozyme-powered GUV system not only mimics intracellular oxidative stress pathways but also induces tumor cell death by sensing and responding to external chemical signals. Specifically, we recreated intracellular biochemical events, including DNA damage and lipid peroxidation, in the compartmentalized GUVs by taking advantage of nanozyme induction of defined catalytic reactions. Additionally, the GUV system also actively induced DNA double-strand breakage and lipid damage of tumor cells, in response to the high expression of H2O2 within the tumor microenvironment. This concept-of-proof study offers a promising option for defining catalysis in biological systems and gives new insights into the de novo creation of artificial cells in a fully synthetic manner.

… The potential of antibody-linked SUVs containing MTX in anticancer therapy was investigated. The … to any Ig, but not toxic to a human kidney cancer line (that did not react with RAMG). …

In advanced drug delivery, versatile liposomal formulations are commonly employed for safer and more accurate therapies. Here we report a method that allows a straightforward production of synthetic monodisperse (~ 100 μm) giant unilamellar vesicles (GUVs) using a microfluidic system. The stability analysis based on the microscopy imaging showed that at ambient conditions the produced GUVs had a half-life of 61 ± 2 h. However, it was observed that ~ 90% of the calcein dye that was loaded into GUVs was transported into a surrounding medium in 24 h, thus indicating that the GUVs may release these small dye molecules without distinguishable membrane disruption. We further demonstrated the feasibility of our method by loading GUVs with larger and very different cargo objects; small soluble fluorescent proteins and larger magnetic microparticles in a suspension. Compared to previously reported microfluidics-based production techniques, the obtained results indicate that our simplified method could be equally harnessed in creating GUVs with less cost, effort and time, which could further benefit studying closed membrane systems.

Disulfiram (DS), an anti-alcoholism medicine, shows strong anti-cancer activity in the laboratory, but the application in clinics for anti-cancer therapy has been limited by its prompt metabolism. Conventional liposomes have shown limited ability to protect DS. Therefore, the aim of this study is to develop PEGylated liposomes of DS for enhanced bio-stability and prolonged circulation. PEGylated liposomes were prepared using ethanol-based proliposome methods. Various ratios of phospholipids, namely: hydrogenated soya phosphatidylcholine (HSPC) or dipalmitoyl phosphatidylcholine (DPPC) and N-(Carbonyl-methoxypolyethylenglycol-2000)-1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE-PEG2000) with cholesterol were used. DS was dissolved in the alcoholic solution in different lipid mol% ratios. The size of the resulting multilamellar liposomes was reduced by high-pressure homogenization. Liposomal formulations were characterized by size analysis, zeta potential, drug loading efficiency and stability in horse serum. Small unilamellar vesicles (SUVs; nanoliposomes) were generated with a size of approximately 80 to 120 nm with a polydispersity index (PDI) in the range of 0.1 to 0.3. Zeta potential values of all vesicles were negative, and the negative surface charge intensity tended to increase by PEGylation. PEGylated liposomes had a smaller size (80–90 nm) and a significantly lower PDI. All liposomes showed similar loading efficiencies regardless of lipid type (HSPC or DPPC) or PEGylations. PEGylated liposomes provided the highest drug biostability amongst all formulations in horse serum. PEGylated DPPC liposomes had t1/2 =77.3 ± 9.6 min compared to 9.7 ± 2.3 min for free DS. In vitro cytotoxicity on wild type and resistant colorectal cancer cell lines was evaluated by MTT assay. All liposomal formulations of DS were cytotoxic to both the wild type and resistant colorectal cancer cell lines and were able to reverse chemoresistance at low nanomolar concentrations. In conclusion, PEGylated liposomes have a greater potential to be used as an anticancer carrier for disulfiram.

… Because optical-trapping does not disrupt the lipid bilayer structures of liposomes, an individual vesicle can be optically trapped and observed for hours, allowing membrane …

The cationic large unilamellar mixed liposomes from 1-palmitoyl-2-… The stability of the liposomal systems was estimated by … and DDAB stabilizes the POPC liposomes. The extent of …

… DPPC Stealth liposomes compared to a … liposomes. The techniques employed in this study can be a valuable tool for the characterization and development of anticancer liposome …

… the bilayer properties of liposomes and showed the nomenclature used to classify the liposomes. … of large single bilayer liposomes by a fast and controlled dialysis. Biochim. Biophys. …

… The deviation from a single exponential function can be characterized by the … of the liposomes prepared by the dialysis method. Fig. 4 shows the gel filtration pattern of bilayer liposomes …

… instrumentation for the scaleup of liposome production. Since no single technique can be applicable … Any liposome with more than one bilayer is defined as being either multilamellar or …

… 5e shows that a small fraction of liposomes with two bilayers hardly changes the scattering curve with respect to the one obtained from single bilayer vesicles (Fig. 5a, dotted line). …

BACKGROUND During past decades, liposomes have emerged as efficient carriers for drugs, diagnostics, vaccines, nutrients and other bioactive agents. Liposomes, the spherical vesicles consisting of phospholipids bilayer have the ability to encapsulate both lipophilic and hydrophilic drugs. Extensive studies have been done in the past for investigating a number of drugs and genes for controlled release with liposomal formulation. Liposomes have also been investigated for their use in cancer treatment. Liposomes offer various advantages because of their biocompatible, biodegradable, nontoxic and non-immunogenic nature. METHODS Liposomes have cell-specific targeting with important applications in the fields of nanotechnology like cancer therapy, diagnosis, gene delivery, cosmetics, agriculture and in food technology. They are prepared by various methods like sonication method, ethanol injection method, lipid film hydration method, micro-emulsion method. CONCLUSION This review will provide an overview of classification, the various formulation methods, characterization, patented formulations and applications of liposomes with future prospects.

The juice exudation of aquatic products oozes out during storage can influence storage quality. Herein, a novel basil essential oil liposome unidirectional water-conducting sustained-release preservation pads (BEOL/UCSP) were prepared with nylon mesh as water-conducting layer, basil essential oil liposome (BEOL) as sustained-release preservation layer, and diatomite and absorbent-cotton as water-absorbing layer. EL/UCSP, β-CL/UCSP, and BEO/UCSP were prepared after BEOL was replaced by eugenol liposome, β-caryophyllene liposome, and BEO. BEOL are microspheres with bilayer structure, had good storage stability, centrifugal stability, thermal stability, embedding capacity, sustained-release, and oxidation resistance, and the main components of preservatives had a synergistic effect on antibacterial properties. The pads without preservative can initially slow down quality deterioration. BEOL/UCSP can directionally absorb exudate and realize long-term sustained-release of preservative, has excellent antibacterial and antioxidant effect, and extended shelf life of Lateolabrax japonicus fillets from 6.0 days to 12.8 days. The BEOL/UCSP can provide technical theoretical support for preservation materials.

… Accurate characterization of the particles, by fluorescence, 31 P-NMR, … a single lipid bilayer around an aqueous cavity. We thus provide the scientific community with a fully characterized …

… , which are vesicles having a phospholipids bilayer … parameters were used to characterize the size of liposomes: … A single dose corresponding to 100.0 mg of each formulation …

… liposome … Liposomes are exceedingly simple; sonicated dispersions of phospholipids produce small vesicles of fairly uniform size (on the order of 3OOA) consisting of a single bilayer …

… for the generation of single-chain Fv (scFv) … liposomes containing nickel-complexed dioleoyl-glycero-succinyl-nitrilotriacetic acid (Ni-NTA-DOGS) as an anchor lipid within the lipid bilayer…

… liposome conjugation, the release of the liposomes by ultrasound, and the internalization of the liposomes, NBD-cholesterol was incorporated into the lipid bilayer of the liposomes. The …

… encapsulation efficiency in single small unilamellar vesicles. The single small unilamellar vesicles … The dye in the membrane is used to determine the vesicle size, and the lumen dye is …

… this approach remained as unilamellar vesicles and no significant change in particle size was observed, they are referred to here as freeze-and-thaw unilamellar vesicles (FAT-ULV). …

… To enhance the yield of giant unilamellar vesicles, we used negatively charged DPPG lipid at 10% concentration of the DPPC lipid, because the presence of DPPG increases the …

… efficiency were obtained when sonication was carried out after the film formation. Vesicle … the effect of charged components on vesicle dimensions, entrapment efficiency and stability. …

… The microencapsulation vesicle (MCV) method is a liposome preparation technique that … sizes with a high encapsulation efficiency. Liposomes encapsulating water-soluble drugs, …

… be considered as a promising vesicular carrier for transdermal … encapsulation efficiency of vesicular systems, which has potential application as a high throughput screening for vesicular …

… Disruption of the lipid-bilayer typically results in vesicle … able to predict the encapsulation efficiency for any unilamellar, … to achieve efficient drug encapsulation into unilamellar, PFE …

In the field of bottom‐up synthetic biology, lipid vesicles provide an important role in the construction of artificial cells. Giant unilamellar vesicles (GUVs), due to their membrane's similarity to natural biomembranes, have been widely used as cellular mimics. So far, several methods exist for the production of GUVs with the possibility to encapsulate biological macromolecules. The inverted emulsion‐based method is one such technique, which has great potential for rapid production of GUVs with high encapsulation efficiencies for large biomolecules. However, the lack of understanding of various parameters that affect production yields has resulted in sparse adaptation within the membrane and bottom‐up synthetic biology research communities. Here, we optimize various parameters of the inverted emulsion‐based method to maximize the production of GUVs. We demonstrate that the density difference between the emulsion droplets, oil phase, and the outer aqueous phase plays a crucial role in vesicle formation. We also investigated the impact that centrifugation speed/time, lipid concentration, pH, temperature, and emulsion droplet volume has on vesicle yield and size. Compared to conventional electroformation, our preparation method was not found to significantly alter the membrane mechanical properties. Finally, we optimize the parameters to minimize the time from workbench to microscope and in this way open up the possibility of time‐sensitive experiments. In conclusion, our findings will promote the usage of the inverted emulsion method for basic membrane biophysics studies as well as the development of GUVs for use as future artificial cells.

… Small unilamellar vesicles (SUVs) were prepared using the method of Pinnaduwage and Bruce22 with modification. Liposome samples were prepared from PC, PC/PG (8:2mol%), or …

… performance. Research involving drug-loaded lipid vesicles entails pharmaceutical and performance … characterize a drug-loaded lipid vesicle system is the entrapment efficiency (EE). …

… yielded one protein per vesicle. The entrapment efficiency was experimentally determined to be about 13%. On the other hand the entrapment efficiency for the small chromate ions was …

Liposomes are attractive vehicles for localized delivery of antibiotics. There exists, however, a gap in knowledge when it comes to achieving high liposomal loading efficiencies for antibiotics. To address this issue, we investigated three antibiotics of clinical relevance against staphylococcal infections with different hydrophilicity and chemical structure, namely, vancomycin hydrochloride, teicoplanin, and rifampin. We categorized the suitability of different encapsulation techniques on the basis of encapsulation efficiency, lipid requirement (important for avoiding lipid toxicity), and mass yield (percentage of mass retained during the preparation process). The moderately hydrophobic (teicoplanin) and highly hydrophobic (rifampin) antibiotics varied significantly in their encapsulation load (max 23.4 and 15.5%, respectively) and mass yield (max 74.1 and 71.8%, respectively), favoring techniques that maximized partition between the aqueous core and the lipid bilayer or those that produce oligolamellar vesicles, whereas vancomycin hydrochloride, a highly hydrophilic molecule, showed little preference to any of the protocols. In addition, we report significant bias introduced by the choice of analytical method adopted to quantify the encapsulation efficiency (underestimation of up to 24% or overestimation by up to 57.9% for vancomycin and underestimation of up to 61.1% for rifampin) and further propose ultrafiltration and bursting by methanol as the method with minimal bias for quantification of encapsulation efficiency in liposomes. The knowledge generated in this work provides critical insight into the more practical, albeit less investigated, aspects of designing vesicles for localized antibiotic delivery and can be extended to other nanovehicles that may suffer from the same biases in analytical protocols.

… We describe the influence of various factors on the encapsulation efficiency of individual … 30 μm unilamellar vesicles (a significant overestimation, since we also observe multilamellar …

The systemic efficacy of the chemotherapeutic agents presently used to treat solid tumors is limited by their low therapeutic index. Previously, our research group improved the in vitro …

… carriers by pegylation is still … PEGylated small unilamellar liposomes (L-GEM) against human PC cell lines and human xenografts as compared to free GEM. The interaction of liposomal …

… of the slower clearance, we believe unilamellar liposomes to be more promising for drug overdose treatment. Thus, only unilamellar liposomes were measured at both 1.45 and 2.9 mg …

… , it is important to examine how addition of TRX changes the liposome structures. This paper examines PEGylated TRX/HSPC liposome in terms of how the addition of TRX affects its …

Cancer is a second leading disease-causing death worldwide that will continuously grow as much as 70% in the next 20 years. Chemotherapy is still becoming a choice for cancer treatment despite its severity of side effects and low success rate due to ineffective delivery of the chemodrugs. Since it was introduced in 1960, significant progress has been achieved in the use of liposomes in drug delivery. The study aims to review relevant literatures on role of PEGylated liposome in enhancing cytotoxic activity of several agents. A systematic literature on the use of PEGylated liposomes in anticancer research via Scopus, Google scholar and PubMed databases was conducted for studies published from 2000 to 2022. A total of 15 articles were selected and reviewed from 312 articles identified covering a variety of anticancer treatments by using PEGylated liposomes. PEGylated liposome which is purposed to achieve steric equilibrium is one of enhanced strategies to deliver anticancer drugs. It has been shown that some improvement of delivery and protection form a harsh gastric environment of several anticancer drugs when they are formulated in a PEGylated liposome. One of the successful drugs that has been clinically used is Doxil®, followed by some other drugs in the pipeline Various drugs (compounds) had been used to enhance the efficacy of PEGylated liposomes for targeted cancer cells in vitro and in vivo. In conclusion, PEGylated liposomes enhance drug activities and have great potential to become efficient anticancer delivery to follow Doxil® in the clinical setting.

Chain-length-dependent membrane changes in PEGylated liposomes induced by doxorubicin loading are revealed using small- and wide-angle X-ray scattering and cryo-EM.

… liposomes. In previous computational works, we performed several studies of the effect of PEGylation … The aim of this study was to explain the effect of liposome PEGylation on their drug-…

… In this paper, liposomes stabilized by PEG-… PEGylated lipid stabilized liposomes, but also significantly improved retention upon dilution. Our findings facilitate new designs of liposome-…

… and the lipid bilayer packing of large unilamellar vesicles (LUV). … Liposome size and specific turbidity decreased with increase … At this PEG–DSPE mol%, an anomalous peak in liposome …

Liposomes are well-established systems for drug delivery and biosensing applications. The design of a liposomal carrier requires careful choice of lipid composition and formulation method. These determine many vesicle properties including lamellarity, which can have a strong effect on both encapsulation efficiency and the efflux rate of encapsulated active compounds. Despite this, a comprehensive study on how the lipid composition and formulation method affect vesicle lamellarity is still lacking. Here, we combine small-angle neutron scattering and cryogenic transmission electron microscopy to study the effect of three different well-established formulation methods followed by extrusion through 100 nm polycarbonate membranes on the resulting vesicle membrane structure. Specifically, we examine vesicles formulated from the commonly used phospholipids 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) via film hydration followed by (i) agitation on a shaker or (ii) freeze–thawing, or (iii) the reverse-phase evaporation vesicle method. After extrusion, up to half of the total lipid content is still assembled into multilamellar structures. However, we achieved unilamellar vesicle populations when as little as 0.1 mol % PEG-modified lipid was included in the vesicle formulation. Interestingly, DPPC with 5 mol % PEGylated lipid produces a combination of cylindrical micelles and vesicles. In conclusion, our results provide important insights into the effect of the formulation method and lipid composition on producing liposomes with a defined membrane structure.

ABSTRACT A commonly held view is that nanocarriers conjugated to polyethylene glycol (PEG) are non-immunogenic. However, many studies have reported that unexpected immune responses have occurred against PEG-conjugated nanocarriers. One unanticipated response is the rapid clearance of PEGylated nanocarriers upon repeat administration, called the accelerated blood clearance (ABC) phenomenon. ABC involves the production of antibodies toward nanocarrier components, including PEG, which reduces the safety and effectiveness of encapsulated therapeutic agents. Another immune response is the hypersensitivity or infusion reaction referred to as complement (C) activation-related pseudoallergy (CARPA). Such immunogenicity and adverse reactivities of PEGylated nanocarriers may be of potential concern for the clinical use of PEGylated therapeutics. Accordingly, screening of the immunogenicity and CARPA reactogenicity of nanocarrier-based therapeutics should be a prerequisite before they can proceed into clinical studies. This review presents PEGylated liposomes, immunogenicity of PEG, the ABC phenomenon, C activation and lipid-induced CARPA from a toxicological point of view, and also addresses the factors that influence these adverse interactions with the immune system. GRAPHICAL ABSTRACT

… was mediated by the molecular recognition of the complementary phosphate and guanidinium groups incorporated in separate unilamellar liposomes. The phosphate group was …

… -liposomes in the Kupffer cells, even already after 15 min, suggesting that the PEG liposomes had … and engineering of PEGylated liposomal formulations for use in multiple drug therapy. …

… The liposomes were found to be small, unilamellar … liposomes. In this study, supercritical fluid technology was successfully used to prepare small, spherical and unilamellar liposomes of …