The application of pulsed electric field (PEF) technology as a non-thermal cell membrane permeabilization treatment, was widely demonstrated widely to be effective in microbial inactivation studies, ...as well as to increase the rates of heat and mass transfer phenomena in food and biotechnological processes (drying, osmotic treatment, freezing, extraction, and diffusion). Nevertheless, most published papers on the topic do not provide enough information for other researchers to assess results properly. A general rule/guidance in reporting experimental data and most of all exposure conditions, would be to report details to the extent that other researchers will be able to repeat, judge and evaluate experiments and data obtained. This is what is described in the present recommendation paper.
Pulsed electric field (PEF) treatment is a promising technology that has received considerable attention in food and biotechnology related applications food and biotechnology related applications of PEF include:i)“cold” pasteurization of liquid foods and disinfection of wastewater by microbial inactivationii)PEF-assisted processing (drying, extraction or expression)
•PEF treatment is affected by a large number of parameters.•The broad experimental conditions and equipments used limited the comparison of results.•The standardization of experimental procedure is required.•Key information to be reported in microbial inactivation studies.•Key information to be reported in studies of PEF-assisted processing in biotechnology.
The paper presents an approach that reduces several difficulties related to the determination of induced transmembrane voltage (ITV) on irregularly shaped cells. We first describe a method for ...constructing realistic models of irregularly shaped cells based on microscopic imaging. This provides a possibility to determine the ITV on the same cells on which an experiment is carried out, and can be of considerable importance in understanding and interpretation of the data. We also show how the finite-thickness, nonzero-conductivity membrane can be replaced by a boundary condition in which a specific surface conductivity is assigned to the interface between the cell interior (the cytoplasm) and the exterior. We verify the results obtained using this method by a comparison with the analytical solution for an isolated spherical cell and a tilted oblate spheroidal cell, obtaining a very good agreement in both cases. In addition, we compare the ITV computed for a model of two irregularly shaped CHO cells with the ITV measured on the same two cells by means of a potentiometric fluorescent dye, and also with the ITV computed for a simplified model of these two cells.
Cell membrane permeabilization is caused by the application of high intensity electric pulses of short duration. The extent of cell membrane permeabilization depends on electric pulse parameters, ...characteristics of the electropermeabilization media and properties of cells exposed to electric pulses. In the present study, the temperature effect during pulse application on cell membrane fluidity and permeabilization was determined in two different cell lines: V-79 and B16F-1.
While cell membrane fluidity was determined by electron paramagnetic resonance (EPR) method, the cell membrane electropermeabilization was determined by uptake of bleomycin and clonogenic assay. A train of eight rectangular pulses with the amplitude of 500 V/cm, 700 V/cm and 900 V/cm in the duration of 100 μs and with repetition frequency 1 Hz was applied. Immediately after the pulse application, 50 μl droplet of cell suspension was maintained at room temperature in order to allow cell membrane resealing. The cells were then plated for clonogenic assay. The main finding of this study is that the chilling of cell suspension from physiological temperature (of 37 °C) to 4 °C has significant effect on cell membrane electropermeabilization, leading to lower percent of cell membrane permeabilization. The differences are most pronounced when cells are exposed to electric pulse amplitude of 900 V/cm. At the same time with the decreasing of temperature, the cell membranes become less fluid, with higher order parameters in all three types of domains and higher proportion of domain with highest order parameter. Our results indicate that cell membrane fluidity and domain structure influence the electropermeabilization of cells, however it seems that some other factors may have contributing role.
The electroporation effect on tissue can be assessed by measurement of electrical properties of the tissue undergoing electroporation. The most prominent techniques for measuring electrical ...properties of electroporated tissues have been voltage-current measurement of applied pulses and electrical impedance tomography (EIT). However, the electrical conductivity of tissue assessed by means of voltage-current measurement was lacking in information on tissue heterogeneity, while EIT requires numerous additional electrodes and produces results with low spatial resolution and high noise. Magnetic resonance EIT (MREIT) is similar to EIT, as it is also used for reconstruction of conductivity images, though voltage and current measurements are not limited to the boundaries in MREIT, hence it yields conductivity images with better spatial resolution. The aim of this study was to investigate and demonstrate the feasibility of the MREIT technique for assessment of conductivity images of tissues undergoing electroporation. Two objects were investigated: agar phantoms and ex vivo liver tissue. As expected, no significant change of electrical conductivity was detected in agar phantoms exposed to pulses of all used amplitudes, while a considerable increase of conductivity was measured in liver tissue exposed to pulses of different amplitudes.
Electrochemotherapy is a technique where electric pulses in combination with chemotherapeutic agents are applied to tumor cells. In general, patients find electrochemotherapy tolerable, in spite of ...unpleasant sensations associated with contraction of muscles located beneath or in the vicinity of the electrodes. These contractions are due to the intensity of the electric pulses required for effective electropermeabilization of tumor cell membranes. Since a train of eight electric pulses with repetition frequency of 1 Hz is usually applied to the tumors, each pulse in the train excites underlying nerves and provokes muscle contractions. Therefore, for patients involved in electrochemotherapy, the use of pulses with repetition frequency higher than the frequency of tetanic contraction would represent reduced number of muscle contractions and associated unpleasant sensations. Our results of the uptake of Lucifer Yellow into electropermeabilized cells in vitro show that with increased repetition frequency the uptake stays at similar levels even at frequencies up to 8.3 kHz. On the basis of these results the possibilities for the clinical use of pulses with high repetition frequency in electrochemotherapy are considered.
Involuntary muscle contractions and painful sensations during electric pulse delivery are the most unpleasant side effects of electrochemotherapy. The aim of this study was to determine the nature of ...pain caused by the application of electric pulses and to evaluate patients tolerance to the standard electric pulses of 1 Hz repetition frequency and the new 5 kHz protocol. A train of eight electric pulses of 1 Hz and 5 kHz repetition frequencies was delivered to the forearms of 40 healthy volunteers. After the conclusion of each protocol the subjects had to complete the short-form McGill Pain Questionnaire with separate visual analog scales for pain intensity and unpleasantness. All subjects selected at least one superficial and one deep pain descriptor; 85% selected at least two superficial descriptor and 60% at least two deep description. The application of 5 kHz electric pulses was less unpleasant than the standard 1 Hz pulses; however, the pain intensity did not differ between the protocols. Significantly more subjects chose the new 5 kHz protocol as their choice of treatment (P = 0.017). The frequent use of deep descriptors in our study indicates that muscle contractions contribute to the discomfort felt by the subjects during the delivery of electric pulses. The new 5 kHz protocol considerably shortens the treatment session and is also better tolerated. Therefore, the new 5 kHz electrochemotherapy protocol should eventually replace the 1 Hz pulses as new standard.
Electrochemotherapy, a combination of high voltage electric pulses and of an anticancer drug, has been demonstrated to be highly effective in treatment of cutaneous and subcutaneous tumors. Unique ...properties of electrochemotherapy (e.g., high specificity for targeting cancer cells, high degree of localization of treatment effect, capacity for preserving the innate immune response and the structure of the extracellular matrix) are facilitating its wide spread in the clinics. Due to high effectiveness of electrochemotherapy in treatment of cutaneous and subcutaneous tumors regardless of histological origin, there are now attempts to extend its use to treatment of internal tumors. To advance the applicability of electrochemotherapy to treatment of internal solid tumors, new technological developments are needed that will enable treatment of these tumors in daily clinical practice. New electrodes through which electric pulses are delivered to target tissue need to be designed with the aim to access target tissue anywhere in the body. To increase the probability of complete tumor eradication, the electrodes have to be accurately positioned, first to provide an adequate extent of electroporation of all tumor cells and second not to damage critical healthy tissue or organs in its vicinity. This can be achieved by image guided insertion of electrodes that will enable accurate positioning of the electrodes in combination with patient-specific numerical treatment planning or using a predefined geometry of electrodes. In order to be able to use electrochemotherapy safely for treatment of internal tumors located in relative proximity of the heart (e.g., in case of liver metastases), the treatment must be performed without interfering with the heart’s electrical activity. We describe recent technological advances, which allow treatment of liver and bone metastases, soft tissue sarcomas, brain tumors, and colorectal and esophageal tumors. The first clinical experiences in these novel application areas of electrochemotherapy are also described.
In silico experiments (numerical simulations) are a valuable tool for non-invasive research of the influences of tissue properties, electrode placement and electric pulse delivery scenarios in the ...process of electroporation. The work described in this article was aimed at introducing time dependent effects into a finite element model developed specifically for electroporation. Reference measurements were made ex vivo on beef liver samples and experimental data were used both as an initial condition for simulation (applied pulse voltage) and as a reference value for numerical model calibration (measured pulse current). The developed numerical model is able to predict the time evolution of an electric pulse current within a 5% error over a broad range of applied pulse voltages, pulse durations and pulse repetition frequencies. Given the good agreement of the current flowing between the electrodes, we are confident that the results of our numerical model can be used both for detailed in silico research of electroporation mechanisms (giving researchers insight into time domain effects) and better treatment planning algorithms, which predict the outcome of treatment based on both spatial and temporal distributions of applied electric pulses.
Effective conductivity of cell suspensions Pavlin, M.; Slivnik, T.; Miklavcic, D.
IEEE transactions on biomedical engineering,
2002-Jan., 2002, 2002-Jan, 2002-01-00, 20020101, Letnik:
49, Številka:
1
Journal Article
Recenzirano
Using finite-element method (FEM) effective conductivity of a cell suspension was calculated for different cell volume fractions and membrane conductivities. Cells were modeled as spheres having ...equivalent conductivity and were organized in cubic lattices, layers and clusters. The results were compared to different analytical expressions for effective conductivity and they showed that Maxwell theory is valid also for higher volume fractions.
Abstract Background This systematic review has two purposes: to consolidate the current knowledge about clinical effectiveness of electrochemotherapy, a highly effective local therapy for cutaneous ...and subcutaneous tumors; and to investigate the differences in effectiveness of electrochemotherapy with respect to tumor type, chemotherapeutic drug, and route of drug administration. Methods All necessary steps for a systematic review were applied: formulation of research question, systematic search of literature, study selection and data extraction using independent screening process, assessment of risk of bias, and statistical data analysis using two-sided common statistical methods and meta-analysis. Studies were eligible for the review if they provided data about effectiveness of single-session electrochemotherapy of cutaneous or subcutaneous tumors in various treatment conditions. Results In total, 44 studies involving 1894 tumors were included in the review. Data analysis confirmed that electrochemotherapy had significantly ( p < .001) higher effectiveness (by more than 50%) than bleomycin or cisplatin alone. The effectiveness was significantly higher for intratumoral than for intravenous administration of bleomycin ( p < .001 for CR%, p = .028 for OR%). Bleomycin and cisplatin administered intratumorally resulted in equal effectiveness of electrochemotherapy. Electrochemotherapy was more effective in sarcoma than in melanoma or carcinoma tumors. Conclusions The results of this review shed new light on effectiveness of electrochemotherapy and can be used for prediction of tumor response to electrochemotherapy with respect to various treatment conditions and should be taken into account for further refinement of electrochemotherapy protocols.
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