| Autor: |
Murauskas A; Faculty of Electronics, Vilnius Gediminas Technical University, Vilnius, Lithuania., Staigvila G; Faculty of Electronics, Vilnius Gediminas Technical University, Vilnius, Lithuania., Girkontaitė I; Department of Immunology, State Research Institute Centre for Innovative Medicine, Vilnius, Lithuania., Zinkevičienė A; Department of Immunology, State Research Institute Centre for Innovative Medicine, Vilnius, Lithuania., Ruzgys P; Biophysics Group, Vytautas Magnus University, Kaunas, Lithuania., Šatkauskas S; Biophysics Group, Vytautas Magnus University, Kaunas, Lithuania., Novickij J; Faculty of Electronics, Vilnius Gediminas Technical University, Vilnius, Lithuania., Novickij V; Faculty of Electronics, Vilnius Gediminas Technical University, Vilnius, Lithuania. |
| Abstrakt: |
Measurement of cell transmembrane potential (TMP) is a complex methodology involving patch-clamp methods or fluorescence-based potentiometric markers, which have limited to no applicability during ultrafast charging and relaxation phenomena. In such a case, analytical methods are applied for evaluation of the voltage potential changes in biological cells. In this work, the TMP-based electrotransfer mechanism during ultra-high frequency (≥1 MHz) electric fields is studied and the phenomenon of rapid membrane charge accumulation, which is non-occurrent during conventional low-frequency electroporation is simulated using finite element method (FEM). The influence of extracellular medium conductivity (0.1, 1.5 S/m) and pulse rise/fall times (10-50 ns) TMP generation are presented. It is shown that the medium conductivity has a dramatic influence on the electroporation process in the high-frequency range of applied pulsed electric fields (PEF). The applied model allowed to grasp the differences in polarization between 100 and 900 ns PEF and enabled successful prediction of the experimental outcome of propidium iodide electrotransfer into CHO-K1 cells and the conductivity-dependent patterns of MHz range PEF-triggered electroporation were determined. The results of this study form recommendations for development and pre-evaluation of future PEF protocols and generators based on ultra-high frequency electroporation for anticancer and gene therapies. |
