Preparation of Liposomes from Soy Lecithin Using Liquefied Dimethyl Ether

We investigated a method to prepare liposomes; soy lecithin was dissolved in liquefied dimethyl ether (DME) at 0.56 MPa, which was then injected into warm water. Liposomes can be successfully prepared at warm water temperatures above 45 °C. The transmission electron microscopy (TEM) images of the obtained liposomes, size distribution, ζ-potential measurements by dynamic light scattering and the amount of residual medium were compared by gas chromatography using the conventional medium, diethyl ether. The size of the obtained liposomes was approximately 60–300 nm and the ζ-potential was approximately −57 mV, which was almost the same as that of the conventional medium. Additionally, for the conventional media, a large amount remained in the liposome dispersion even after removal by depressurization and dialysis membrane treatment; however, liquefied DME, owing to its considerably low boiling point, was completely removed by depressurization. Liquefied DME is a very attractive medium for the preparation of liposomes because it does not have the toxicity and residue problems of conventional solvents or the hazards of ethanol addition and high pressure of supercritical carbon dioxide; it is also environmentally friendly.

Continuous-Flow Production of Liposomes with a Millireactor under Varying Fluidic Conditions

Continuous-flow production of liposomes using microfluidic reactors has demonstrated advantages compared to batch methods, including greater control over liposome size and size distribution and reduced reliance on post-production processing steps. However, the use of microfluidic technology for the production of nanoscale vesicular systems (such as liposomes) has not been fully translated to industrial scale yet. This may be due to limitations of microfluidic-based reactors, such as low production rates, limited lifetimes, and high manufacturing costs. In this study, we investigated the potential of millimeter-scale flow reactors (or millireactors) with a serpentine-like architecture, as a scalable and cost-effective route to the production of nanoscale liposomes. The effects on liposome size of varying inlet flow rates, lipid type and concentration, storage conditions, and temperature were investigated. Liposome size (i.e., mean diameter) and size dispersity were characterised by dynamic light scattering (DLS); z-potential measurements and TEM imaging were also carried out on selected liposome batches. It was found that the lipid type and concentration, together with the inlet flow settings, had significant effects on the properties of the resultant liposome dispersion. Notably, the millifluidic reactor was able to generate liposomes with size and dispersity ranging from 54 to 272 nm, and from 0.04 to 0.52 respectively, at operating flow rates between 1 and 10 mL/min. Moreover, when compared to a batch ethanol-injection method, the millireactor generated liposomes with a more therapeutically relevant size and size dispersity.

FEATURES OF THE TECHNOLOGY OF LIPOSOMAL FORMULATION OF A ANALOGUE HYPOTHALAMIC HORMONE SOMATOSTATIN

Background. In connection with the prospect of the use of an analog of the hypothalamic hormone somatostatin synthesized by the laboratory of chemical synthesis Institute of experimental diagnostics and chemotherapy of FSBI «N.N. Blokhin Russian Cancer Research Center» and showed a high anti-tumor activity as a drug arises a need to establish an optimal technology of its receipt. In preliminary studies in a modelformulation for an analog of the hypothalamic hormone somatostatin selected liposome technological process of which has a series of specific steps comprising. Objective. Development of technology for obtaining liposomal formulation hypothalamic hormone somatostatin analogue. Materials and methods. Liposomes analog of the hypothalamic hormone somatostatin obtained by method Bengema in modification for hydrophobic substances. To reduce the diameter of the liposome are used methods extrusion, homogenization and ultrasonic. Analysis of the size of the liposomes was performed by correlation spectroscopy light scattering using nanosizer. The pH of the liposomal dispersion was determined by potentiometry. The quantitative content of the drug substance was determined by spectrophotometry using a standard sample with X (282 ± 3) nm and an alcoholic solution of empty liposomes as a reference solution. Amount of incorporated drug was calculated as the ratio of the concentration of drug in the liposome dispersion after filtration to the concentration of drug in the dispersion after preparation. Results and Conclusion. The hydrophobic nature of the substance causes an analog of the hypothalamic hormone somatostatin technological features of obtaining liposomal formulation. Since the step of forming a film of the lipid substance is dissolved in an organic solvent together with lipids, film is hydrated by a solution of cryoprotectant. Grinding liposomes an analog of the hypothalamic hormone somatostatin appropriate to be carried out using homogenization or extrusion methods, due to the high efficiency of these methods, the preservation stability of the liposomes and a high percentage of inclusion an analog of the hypothalamic hormone somatostatin, included in the liposomal bilayer. At the stage of separating the non-inclusion of substance an analog of the hypothalamic hormone somatostatin due to the insolubility of the substance in the water, you can use the filtering method, without the need for complicated procedures gel filtration, dialysis, etc. Furthermore the process of separating a substance not included can be combined with the sterilization of the liposome dispersion by selecting a particular filter material.

Observations of membrane fusion in a liposome dispersion: the missing fusion intermediate?

Early intermediate structures of liposome-liposome fusion events were captured by freeze-fracture electron microscopic (EM) technique. The images show the morphology of the fusion interface at several different stages of the fusion event. One of the intermediates was captured at a serendipitous stage of two vesicles’ membranes (both leaflets) merging and their contents starting to intermix clearly showing the fusion interface with a previously unseen fusion rim. From the morphological information a hypothetical sequence of the fusion event and corresponding lipid structural arrangements are described.

The influence of tin compounds on the dynamic properties of liposome membranes: A study using the ESR method

The influence of organic and inorganic compounds of tin on the dynamic properties of liposome membranes obtained in the process of dipalmitoylphosphatidylcholine (DPPC) sonication in distilled water was investigated. This was carried out by means of the spin ESR probe method. The probes were selected in such a way as to penetrate different areas of the membrane (a TEMPO probe, 5-DOXYL stearic acid, 16-DOXYL stearic acid). Four compounds of tin were chosen: three organic ones, (CH3)4Sn, (C2H5)4Sn and (C3H7)3SnCl, and one inorganic one, SnCl2. The investigated compounds were added to a liposome dispersion, which was prepared prior to that. The concentration of the admixture was changed within the values from 0 to 10%-mole in proportion to DPPC. The studies indicated that the chlorides of tin display the highest activity in their interaction with liposome membranes. Since these compounds have ionic form in a water solution, the obtained result can mean that this form of admixture has a considerable influence on its activity. Furthermore, it was found that there is a slightly stronger influence of tin compounds with a longer hydrocarbon chain on changes in the probes’ spectroscopic parameters.

Rheological examination of erythrocyte aggregation in the presence of lipid-based fine particles

The development of lipid-based fine particles as potential drug carriers requires a detailed investigation of the possible effects of these carriers on the rheological properties of blood. In this study, we investigated the influence of dynamic conditions on aggregate formation and stability in dispersions of lipid-based fine particles in whole blood under in vitro conditions. The rheological parameters of two concentrations of liposome dispersion and two concentrations of lipid emulsion in blood were examined. A micro-rheological model of aggregating dispersions is proposed in which the apparent viscosity is estimated as the sum of the hydrodynamic and structural parts which are correlated with system structural ordering in the flow. The dynamics of structural ordering of the aggregating system an considered by examining the evolution of the state of the system in phase space depending on the shear rate. The addition of lipid-based particles induced aggregate formation in the blood, which was more pronounced at higher concentrations of lipid-based fine particles. Furthermore, larger and more stable aggregates are formed in liposome dispersions as compared to lipid emulsions in blood.

In vitro evaluation of the controlled release of antibiotics from liposomes

Results of this study have shown significantly prolonged release of selected antibiotic from liposome dispersion as compared to free antibiotic solution of the same initial concentration. Two models of non-steady one-dimensional diffusion were successfully applied to the experimental data and the antibiotic diffusion coefficients were estimated. In addition, the mass transfer resistance of the membrane was shown to be insignificant confirming the suitability of the applied experimental system. Since liposomes are known as systems with slower drug release, then, when it comes to their incorporation in the final form of a preparation for further experiments in vivo, the system of choice would be liposomes with an encapsulated antibiotic drug. The established experimental system could be extended to other liposome formulations with respect to the release rate of the active components.