Doxorubicin loaded thermostable nanoarchaeosomes: a next-generation drug carrier for breast cancer therapeutics.

Breast cancer has a poor prognosis due to the toxic side effects associated with high doses of chemotherapy. Liposomal drug encapsulation has resulted in clinical success in enhancing chemotherapy tolerability. However, the formulation faces severe limitations with a lack of colloidal stability, reduced drug efficiency, and difficulties in storage conditions. Nanoarchaeosomes (NA) are a new generation of highly stable nanovesicles composed of the natural ether lipids extracted from archaea. In our study, we synthesized and characterized the NA, evaluated their colloidal stability, drug release potential, and anticancer efficacy. Transmission electron microscopy images have shown that the NA prepared from the hyperthermophilic archaeon Aeropyrum pernix K1 was in the size range of 61 ± 3 nm. The dynamic light scattering result has confirmed that the NA were stable at acidic pH (pH 4) and high temperature (70 °C). The NA exhibited excellent colloidal stability for 50 days with storage conditions at room temperature. The cell viability results have shown that the pure NA did not induce cytotoxicity in NIH 3T3 fibroblast cells and are biocompatible. Then NA were loaded with doxorubicin (NAD), and FTIR and UV-vis spectroscopy results have confirmed high drug loading efficiency of 97 ± 1% with sustained drug release for 48 h. The in vitro cytotoxicity studies in MCF-7 breast cancer cell lines showed that NAD induced cytotoxicity at less than 10 nM concentration. Fluorescence-activated cell sorting (FACS) results confirmed that NAD induced late apoptosis in nearly 92% of MCF-7 cells and necrosis in the remaining cells with cell cycle arrest at the G0/G1 phase. Our results confirmed that the NA could be a potential next-generation carrier with excellent stability, high drug loading efficiency, sustained drug release ability, and increased therapeutic efficacy, thus reducing the side effects of conventional drugs.

Effects of Hydrophobic Gold Nanoparticles on Structure and Fluidity of SOPC Lipid Membranes.

Gold nanoparticles (AuNPs) are promising candidates in various biomedical applications such as sensors, imaging, and cancer therapy. Understanding the influence of AuNPs on lipid membranes is important to assure their safety in the biological environment and to improve their scope in nanomedicine. In this regard, the present study aimed to analyze the effects of different concentrations (0.5, 1, and 2 wt.%) of dodecanethiol functionalized hydrophobic AuNPs on the structure and fluidity of zwitterionic 1-stearoyl-2-oleoyl-sn-glycerol-3-phosphocholine (SOPC) lipid bilayer membranes using Fourier-transform infrared (FTIR) spectroscopy and fluorescent spectroscopy. The size of AuNPs was found to be 2.2 ± 1.1 nm using transmission electron microscopy. FTIR results have shown that the AuNPs induced a slight shift in methylene stretching bands, while the band positions of carbonyl and phosphate group stretching were unaffected. Temperature-dependent fluorescent anisotropy measurements showed that the incorporation of AuNPs up to 2 wt.% did not affect the lipid order in membranes. Overall, these results indicate that the hydrophobic AuNPs in the studied concentration did not cause any significant alterations in the structure and membrane fluidity, which suggests the suitability of these particles to form liposome-AuNP hybrids for diverse biomedical applications including drug delivery and therapy.

Archaeosomes: New Generation of Liposomes Based on Archaeal Lipids for Drug Delivery and Biomedical Applications.

Archaeosomes are a new generation of stable liposomes composed of natural ether lipids extracted from archaea, or synthetic archaeal lipids. Archaea constitute a domain of single-celled microorganisms that are structurally similar to but evolutionarily distinct from bacteria. They synthesize unique membrane lipids with isoprenoid hydrocarbon side chains attached via an ether linkage to the glycerol-phosphate backbone. Compared to the ester linkages found in the lipids of Eukarya and bacteria, the ether linkages in archaeal lipids are more stable in various environmental conditions such as high/low temperatures, acidic or alkaline pH, bile salts, and enzymatic hydrolysis. This feature has intrigued scientists to use archaeal lipids to prepare archaeosomes with superior physicochemical stability and utilize them as effective carriers to deliver various cargos of biomedical importance such as drugs, proteins, peptides, genes, and antioxidants to the target site. Archaeosomes carrying antigens and/or adjuvants are also proven to be better candidates for stimulating antigen-specific, humoral, and cell-mediated immune responses, which broadens their scope in vaccine delivery. These properties associated with excellent biocompatibility and a safety profile provide numerous advantages to the archaeosomes to function as a versatile delivery system. This mini-review will provide an overview of the unique features of archaeal lipids, preparation and characterization of archaeosomes, and emphasize the prospects related to drug delivery and other biomedical applications.

Magneto-mechanical actuation of barium-hexaferrite nanoplatelets for the disruption of phospholipid membranes.

HYPOTHESIS: The magneto-mechanical actuation (MMA) of magnetic nanoparticles with a low-frequency alternating magnetic field (AMF) can be used to destroy cancer cells. So far, MMA was tested on different cells using different nanoparticles and different field characteristics, which makes comparisons and any generalizations about the results of MMA difficult. In this paper we propose the use of giant unilamellar vesicles (GUVs) as a simple model system to study the effect of MMA on a closed lipid bilayer membrane, i.e., a basic building block of any cell. EXPERIMENTS: The GUVs were exposed to barium-hexaferrite nanoplatelets (NPLs, ~50 nm wide and 3 nm thick) with unique magnetic properties dominated by a permanent magnetic moment that is perpendicular to the platelet, at different concentrations (1-50 µg/mL) and pH values (4.2-7.4) of the aqueous suspension. The GUVs were observed with an optical microscope while being exposed to a uniaxial AMF (3-100 Hz, 2.2-10.6 mT). FINDINGS: When the NPLs were electrostatically attached to the GUV membranes, the MMA induced cyclic fluctuations of the GUVs' shape corresponding to the AMF frequency at the low NPL concentration (1 µm/mL), whereas the GUVs were bursting at the higher concentration (10 µg/mL). Theoretical considerations suggested that the bursting of the GUVs is a consequence of the local action of an assembly of several NPLs, rather than a collective effect of all the absorbed NPLs.

Bending elasticity modulus of giant vesicles composed of aeropyrum pernix k1 archaeal lipid.

Thermally induced shape fluctuations were used to study elastic properties of giant vesicles composed of archaeal lipids C25,25-archetidyl (glucosyl) inositol and C25,25-archetidylinositol isolated from lyophilised Aeropyrum pernix K1 cells. Giant vesicles were created by electroformation in pure water environment. Stroboscopic illumination using a xenon flash lamp was implemented to remove the blur effect due to the finite integration time of the camera and to obtain an instant picture of the fluctuating vesicle shape. The mean weighted value of the bending elasticity modulus kc of the archaeal membrane determined from the measurements meeting the entire set of qualification criteria was (1.89 ± 0.18) × 10-19 J, which is similar to the values obtained for a membrane composed of the eukaryotic phospholipids SOPC (1.88 ± 0.17) × 10-19 J and POPC (2.00 ± 0.21) ´ 10-19 J. We conclude that membranes composed of archaeal lipids isolated from Aeropyrum pernix K1 cells have similar elastic properties as membranes composed of eukaryotic lipids. This fact, together with the importance of the elastic properties for the normal circulation through blood system, provides further evidence in favor of expectations that archaeal lipids could be appropriate for the design of drug delivery systems.

Influence of nanoparticle-membrane electrostatic interactions on membrane fluidity and bending elasticity.

The aim of this work is to investigate the effect of electrostatic interactions between the nanoparticles and the membrane lipids on altering the physical properties of the liposomal membrane such as fluidity and bending elasticity. For this purpose, we have used nanoparticles and lipids with different surface charges. Positively charged iron oxide (γ-Fe2O3) nanoparticles, neutral and negatively charged cobalt ferrite (CoFe2O4) nanoparticles were encapsulated in neutral lipid 1-stearoyl-2-oleoyl-sn-glycero-3-phosphocholine and negatively charged 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-L-serine lipid mixture. Membrane fluidity was assessed through the anisotropy measurements using the fluorescent probe 1,6-diphenyl-1,3,5-hexatriene. Though the interaction of both the types of nanoparticles reduced the membrane fluidity, the results were more pronounced in the negatively charged liposomes encapsulated with positively charged iron oxide nanoparticles due to strong electrostatic attractions. X-ray photoelectron spectroscopy results also confirmed the presence of significant quantity of positively charged iron oxide nanoparticles in negatively charged liposomes. Through thermally induced shape fluctuation measurements of the giant liposomes, a considerable reduction in the bending elasticity modulus was observed for cobalt ferrite nanoparticles. The experimental results were supported by the simulation studies using modified Langevin-Poisson-Boltzmann model.

Registration and analysis of the shape fluctuations of nearly spherical lipid vesicles.

The analysis of shape fluctuations of giant nearly spherical lipid vesicles observed via optical microscopy is one of the widely used methods for the determination of the bending elasticity of lipid membranes. Although the method has been used already for three decades, the values of this material constant, obtained by different groups for membranes of the same composition, in identical conditions, differ significantly. The aim of the present work is the development of the method, enabling us to avoid the influence of artifacts on the value of the measured bending modulus. This is achieved by rejection of some images of the vesicle or the whole vesicle when they do not satisfy the requirements (selection criteria) of the applied theory. The bending modulus of 1-stearoyl-2-oleoyl-sn-glycerol-3-phosphocholine lipid membranes is determined via the advanced method described here. The results are compared with the values in the literature and their difference is discussed.

Mechanoformation of neutral giant phospholipid vesicles in high ionic strength solution.

We present new and simple method for formation of giant unilamellar vesicles (GUVs) in high ionic strength solutions, such as phosphate buffered saline (PBS). Mechanoformation method is an alternative method to electroformation method. The advantage of the mechanoformation procedure is that there are no limitations with respect to the ionic strength of the aqueous solutions, because there is no applied electric potential thus no current flow through the formation cell and no electrolysis is induced.

Permeability and the hidden area of lipid bilayers.

The passive water permeability of a lipid vesicle membrane was studied, related to the hydrostatic (not osmotic) pressure difference between the inner and the outer side of the vesicle in a water environment without additives. Each pressure difference was created by sucking a vesicle into a micropipette at a given sucking pressure. The part of the membrane sucked into the micropipette (the projection length) was measured as a function of time. The time dependence can be divided into two intervals. We put forward the idea that smoothing of membrane defects, accompanied by an increase of the membrane area, takes place during the initial time interval, which results in a faster increase of the projection length. In the second time interval the volume of the vesicle decreases due to the permeability of its membrane and the increase of the projection length is slower. The hidden area and the water permeability of a typical lipid bilayer were estimated. The measured permeability, conjugated to the hydrostatic pressure difference, is an order of magnitude higher than the known value of the permeability, conjugated to the osmotic pressure difference. A hypothesis, based on pore formation, is proposed as an explanation of this experimental result.