Effects of sugar concentration on the electroporation, size distribution and average size of charged giant unilamellar vesicles.

We investigated the effects of sugar concentration on the electroporation, size distribution and average size of giant unilamellar vesicles (GUVs). GUVs were prepared from 40 mol% of 1,2-dioleoyl-sn-glycero-3-phospho-(1'-rac-glycerol) (DOPG) and 60 mol% of 1, 2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) lipids. Pulsed electric field was applied to the 40%DOPG/60%DOPC-GUVs and it induced lateral electric tension (σc) in the membranes of vesicles. The σc-induced probability of rupture (Ppore) and the rate constant of rupture (kp) of GUVs under the sugar concentration, c = 40, 100 and 300 mM, were determined. Both the Ppore and kp increased with the increase of σc, but higher tension was required to generate the same values of Ppore and kp with increasing c. We also investigated average sizes of GUVs from the size distribution of vesicles under various sugar concentrations. With the increase of c, the peak of the size distribution histograms shifted to the region of smaller vesicles. The average size decreased 1.6-fold when c increased from 10 to 300 mM. These investigations help to understand various biomedical, biophysical, and biochemical processes in vesicles and cells. Electroporation, size distribution and average size of charged GUVs were investigated under various sugar concentrations. The sugar concentration influences the electroporation of vesicles and the average size of GUVs.

Effects of cholesterol on the size distribution and bending modulus of lipid vesicles.

The influence of cholesterol fraction in the membranes of giant unilamellar vesicles (GUVs) on their size distributions and bending moduli has been investigated. The membranes of GUVs were synthesized by a mixture of two elements: electrically neutral lipid 1, 2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and cholesterol and also a mixture of three elements: electrically charged lipid 1,2-dioleoyl-sn-glycero-3-phospho-(1'-rac-glycerol) (DOPG), DOPC and cholesterol. The size distributions of GUVs have been presented by a set of histograms. The classical lognormal distribution is well fitted to the histograms, from where the average size of vesicle is obtained. The increase of cholesterol content in the membranes of GUVs increases the average size of vesicles in the population. Using the framework of Helmholtz free energy of the system, the theory developed by us is extended to explain the experimental results. The theory determines the influence of cholesterol on the bending modulus of membranes from the fitting of the proper histograms. The increase of cholesterol in GUVs increases both the average size of vesicles in population and the bending modulus of membranes.

Deformation and poration of giant unilamellar vesicles induced by anionic nanoparticles.

The interaction of anionic magnetite nanoparticles (MNPs) of size 18 nm with negatively charged giant unilamellar vesicles (GUVs) formed from a mixture of neutral dioleoylphosphatidylcholine (DOPC) and negatively charged dioleoylphosphatidylglycerol (DOPG) lipids has been investigated. It has been obtained that NPs induces the deformation of spherical GUVs. The reaction of other GUVs on NPs consists in the appearance of pores in their membranes. We focused the effect of electrostatics on the interaction of charged membranes with MNPs. To study the influence of the surface charge of GUVs on the processes under consideration, we varied the fraction of DOPG in the vesicles from 0 to 100%. We examined the influence of salt concentration in the range of 50-300 mM NaCl concentration. To describe the degree of deformation, a special parameter compactness was introduced. The pore formation in the membranes of GUVs was investigated by the leakage of sucrose. The compactness increases with time and also NPs concentration. The fraction of deformed GUVs increases with the increase of surface charge density of membranes as well as the decrease of salt concentration in buffer. The value of compactness for neutral membrane is 1.25 times higher than that of charged ones. The fraction of deformed GUVs become constant after 20 min, however it increases with NPs concentration. The time taken for stochastic pore formation is less for charged membrane than neutral one. The physical mechanism explaining the experimental results obtained in these investigations.

Electrostatic interaction effects on the size distribution of self-assembled giant unilamellar vesicles.

The influence of electrostatic conditions (salt concentration of the solution and vesicle surface charge density) on the size distribution of self-assembled giant unilamellar vesicles (GUVs) is considered. The membranes of GUVs are synthesized by a mixture of dioleoylphosphatidylglycerol and dioleoylphosphatidylcholine in a physiological buffer using the natural swelling method. The experimental results are presented in the form of a set of histograms. The log-normal distribution is used for statistical treatment of results. It is obtained that the decrease of salt concentration and the increase of vesicle surface charge density of the membranes increase the average size of the GUV population. To explain the experimental results, a theory using the Helmholtz free energy of the system describing the GUV vesiculation is developed. The size distribution histograms and average size of GUVs under various conditions are fitted with the proposed theory. It is shown that the variation of the bending modulus due to changing of electrostatic parameters of the system is the main factor causing a change in the average size of GUVs.

Electrostatic interaction effects on tension-induced pore formation in lipid membranes.

We investigated the effects of electrostatic interactions on the rate constant (k(p)) for tension-induced pore formation in lipid membranes of giant unilamellar vesicles under constant applied tension. A decrease in salt concentration in solution as well as an increase in surface charge density of the membranes increased k(p). These data indicate that k(p) increases as the extent of electrostatic interaction increases. We developed a theory on the effect of the electrostatic interactions on the free energy profile of the membrane containing a prepore and also on the values of k(p); this theory explains the experimental results and fits the experimental data reasonably well in the presence of weak electrostatic interactions. Based on these results, we conclude that a decrease in the free energy barrier of the prepore state due to electrostatic interactions is the main factor causing an increase in k(p).

Rate constant of tension-induced pore formation in lipid membranes.

We investigated the effects of tension induced by micropipet aspiration on giant unilamellar vesicles (GUVs) composed of dioleoylphosphatidylglycerol (DOPG) and dioleoylphosphatidylcholine (DOPC). We analyzed the time course of the fraction of intact GUVs among all of the GUVs under constant tension σ and obtained the rate constants of pore formation kp(σ). To determine kp, we developed an approach using the mean first passage time. The fitting of the theoretical curves of kp versus σ to the experimental data determined the line tension of a prepore, Γ. The value of Γ of a DOPG/DOPC bilayer was smaller than that of a DOPC bilayer.