ABSTRACT
High-speed imaging of giant unilamellar vesicles (GUVs) in recent years has shown significant shape deformation of these vesicles under electroporating direct current (DC) pulsed electric fields, possibly altering the surface distribution of transmembrane potential (TMP) and, thereby, the location and extent of electroporation on the bilayer membrane. The development of TMP, the corresponding shape deformation, and the extent of electroporation depend upon the waveform of the applied electric field. In this work, the deformation of vesicles was carried out under a high-intensity, single cycle of a sinusoidal pulsed electric field (SSPEF) and a square wave pulsed electric field (SWPEF). The cylindrical shape deformations of vesicles were observed for both SSPEF and SWPEF and were dependent upon the ratio of conductivity of the inner medium to the outer medium, α. For α = 1 and α > 1, the vesicles deformed into prolate cylinders as a result of Maxwell stress, whereas they were compressed into oblate cylinders for α < 1. Vesicles subjected to a SSPEF relaxed following either the pore closure dominated t2 or the efflux and lipid loss dominated, slow t3 mechanism depending upon the value of α. For α = 1 and α < 1, the relaxation of the vesicles was found to be predominantly dependent upon pore closure. On the other hand, a majority of vesicles gained excess area during poration when α > 1, which can be attributed to a higher TMP and faster charging of the membrane. The predictions of the approximate model for the deformation of vesicles agreed with the experiment, with deviations between the two as a result of the simplicity of the model. Moreover, the degree of deformation of vesicles [measured by the aspect ratio (AR)] and shape deformations of vesicles were found to be dependent upon the pulse width (TP) and amplitude (E0) of the SSPEF. The specific temporal variation of pore-forming tendencies of SSPEF and SWPEF, with their associated peculiarities, can be judiciously used for controlling electroporation in cells and vesicles.
ABSTRACT
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.