Abstract
Electroporation-induced cell sensitization was described as the occurrence of a delayed hypersensitivity to electric pulses caused by pretreating cells with electric pulses. It was achieved by increasing the duration of the electroporation treatment at the same cumulative energy input. It could be exploited in electroporation-based treatments such as electrochemotherapy and tissue ablation with irreversible electroporation. The mechanisms responsible for cell sensitization, however, have not yet been identified. We investigated cell sensitization dynamics in five different electroporation buffers. We split a pulse train into two trains varying the delay between them and measured the propidium uptake by fluorescence microscopy. By fitting the first-order model to the experimental results, we determined the uptake due to each train (i.e. the first and the second) and the corresponding resealing constant. Cell sensitization was observed in the growth medium but not in other tested buffers. The effect of pulse repetition frequency, cell size change, cytoskeleton disruption and calcium influx do not adequately explain cell sensitization. Based on our results, we can conclude that cell sensitization is a sum of several processes and is buffer dependent. Further research is needed to determine its generality and to identify underlying mechanisms.
Highlights
Electroporation is a phenomenon resulting in a transient increase in membrane permeability, which occurs when short high voltage pulses are applied to cells and tissues [1,2]
Electroporation is used in medicine, e.g. electrochemotherapy (ECT) [3,4,5,6], non-thermal irreversible electroporation (IRE) as a method of tissue ablation [7,8,9], gene therapy [10,11], DNA vaccination [12,13] and transdermal drug delivery [14,15,16], as well as in biotechnology [17], and food processing [18,19,20]
Different inter-train delays were tested in the growth medium and the low-conductivity buffer
Summary
Electroporation is a phenomenon resulting in a transient increase in membrane permeability, which occurs when short high voltage pulses are applied to cells and tissues [1,2]. Electroporation is used in medicine, e.g. electrochemotherapy (ECT) [3,4,5,6], non-thermal IRE as a method of tissue ablation [7,8,9], gene therapy [10,11], DNA vaccination [12,13] and transdermal drug delivery [14,15,16], as well as in biotechnology [17], and food processing [18,19,20]. When treating tumors with IRE, a high number of pulses is delivered, which can cause significant Joule heating and thermal damage and complicate the treatment [22,23]. Provided the effect of the electric pulses be enhanced, we can treat larger tumors with
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