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.

In this Letter, we explore how cell electro-deformation and electro-poration are connected. We build a time-domain model of layered concentric shells (a model of biological cells) including their dielectric and elastic properties. We simulate delivery of one trapezoidal voltage pulse to either a single spherical cell or an assembly of three neighboring cells in a specific configuration and calculate cell deformation and pore formation. We describe the qualitative features of the electric field, surface charge density, transmembrane voltage, cell elongation, and pore density distribution at specific times i.e., before, during and after the application of the electric pulse and explore the correlations between them. Our results show that (1) the polarization charge redistribution plays a significant role in the spatial distribution of electrical stresses at μs time scales and (2) the cell deformation and pore density can be correlated with regions of high surface charge density. In future work, our model could be used for understanding basic mechanisms of electro-deformation and electro-poration with high-frequency short bipolar pulses of biological cells in suspension or tissues.

Introduction: Skin electroporation is a promising treatment for transdermal drug delivery, gene electrotransfer, skin rejuvenation, electrochemotherapy, and wound disinfection. Although a considerable amount of in vitro and in vivo studies exists, the translation to clinics is not as fast as one would hope. We hypothesize the reason lies in the inadequate dosimetry, i.e. electrode configurations, pulse parameters and pulse generators used. We suggest adequate dosimetry can be determined by mathematical modeling and would allow comparison of protocols and facilitate translation into clinics. Areas covered: We introduce the mechanisms and applications of skin electroporation, present existing mathematical models and compare the influence of different model parameters. We review electrodes and pulse generators, prototypes as well as commercially available models. Expert opinion: The reasons for slow translation of skin electroporation treatments into clinics lie in uncontrolled and inadequate dosimetry, poor reporting rendering comparisons between studies difficult, and significant differences in animal and human skin morphology, often dismissed in reports. Mathematical models enable comparison of studies, however, when the parameters of the pulses and electrode configuration are not adequately reported, as is often the case, comparisons are difficult, if not impossible. For each skin electroporation treatment, systematic studies determining optimal parameters should be performed and treatment parameters standardized.

Background: Cell membrane permeabilization by pulsed electromagnetic fields (PEMF) is a novel contactless method which results in effects similar to conventional electroporation. The non-invasiveness of the methodology, independence from the biological object homogeneity and electrical conductance introduce high flexibility and potential applicability of the PEMF in biomedicine, food processing, and biotechnology. The inferior effectiveness of the PEMF permeabilization compared to standard electroporation and the lack of clear description of the induced transmembrane transport are currently of major concern. Methods: The PEMF permeabilization experiments have been performed using a 5.5 T, 1.2 J pulse generator with a multilayer inductor as an applicator. We investigated the feasibility to increase membrane permeability of Chinese Hamster Ovary (CHO) cells using short microsecond (15 µs) pulse bursts (100 or 200 pulses) at low frequency (1 Hz) and high dB/dt (>10⁶ T/s). The effectiveness of the treatment was evaluated by fluorescence microscopy and flow cytometry using two different fluorescent dyes: propidium iodide (PI) and YO-PRO®-1 (YP). The results were compared to conventional electroporation (single pulse, 1.2 kV/cm, 100 µs), i.e., positive control. Results: The proposed PEMF protocols (both for 100 and 200 pulses) resulted in increased number of permeable cells (70 ± 11% for PI and 67 ± 9% for YP). Both cell permeabilization assays also showed a significant (8 ± 2% for PI and 35 ± 14% for YP) increase in fluorescence intensity indicating membrane permeabilization. The survival was not affected. Discussion: The obtained results demonstrate the potential of PEMF as a contactless treatment for achieving reversible permeabilization of biological cells. Similar to electroporation, the PEMF permeabilization efficacy is influenced by pulse parameters in a dose-dependent manner.