BackgroundThe cell membrane acts as a barrier that hinders free entrance of most hydrophilic molecules into the cell. Due to numerous applications in medicine, biology and biotechnology, the introduction of impermeant molecules into biological cells has drawn considerable attention in the past years. One of the most famous methods in this field is electroporation, in which electric pulses with high intensity and short duration are applied to the cells. The aim of our study was to investigate the effect of time-varying magnetic field with different parameters on transmembrane molecular transport.Materials and methods.‘Moreover, a comparison was made between the uptake results due to magnetic pulse exposure and electroporation mediated uptake.’ at the end of Background part. The Chinese hamster ovary (CHO) cells were exposed to magnetic pulses of 2.2 T peak strength and 250 μs duration delivered by Magstim stimulator and double 70 mm coil. Three different frequencies of 0.25, 1 and 10 Hz pulses with 112, 56 and 28 number of pulses were applied (altogether nine experimental groups) and Lucifer Yellow uptake was measured in each group. Moreover, maximum uptake of Lucifer Yellow obtained by magnetic pulses was compared to the measured uptake due to electroporation with typical parameters of 8 pulses of 100 μs, repetition frequency of 1 Hz and electric field intensities of 200 to 600 V/cm.Results and conclusions.Our results show that time-varying magnetic field exposure increases transmembrane molecular transport and this uptake is greater for lower frequencies and larger number of pulses. Besides, the comparison shows that electroporation is more effective than pulsed magnetic field, but the observed uptake enhancement due to magnetic exposure is still considerable.

The precise flow characteristics that promote different atherosclerotic plaque types remain unclear. We previously developed a blood flow-modifying cuff for ApoE−/− mice that induces the development of advanced plaques with vulnerable and stable features upstream and downstream of the cuff, respectively. Herein, we sought to test the hypothesis that changes in flow magnitude promote formation of the upstream (vulnerable) plaque, whereas altered flow direction is important for development of the downstream (stable) plaque. We instrumented ApoE−/− mice (n = 7) with a cuff around the left carotid artery and imaged them with micro-CT (39.6 µm resolution) eight to nine weeks after cuff placement. Computational fluid dynamics was then performed to compute six metrics that describe different aspects of atherogenic flow in terms of wall shear stress magnitude and/or direction. In a subset of four imaged animals, we performed histology to confirm the presence of advanced plaques and measure plaque length in each segment. Relative to the control artery, the region upstream of the cuff exhibited changes in shear stress magnitude only (p < 0.05), whereas the region downstream of the cuff exhibited changes in shear stress magnitude and direction (p < 0.05). These data suggest that shear stress magnitude contributes to the formation of advanced plaques with a vulnerable phenotype, whereas variations in both magnitude and direction promote the formation of plaques with stable features.

BackgroundIn order to reduce the side-effects of chemotherapy, combined chemotherapy-electroporation (electrochemotherapy) has been suggested. Electroporation, application of appropriate electric pulses to biological cells, can significantly enhance molecular uptake of cells due to formation of transient pores in the cell membrane. It was experimentally demonstrated that the efficiency of electroporation is under the control of electric pulse parameters. However, the theoretical basis for these experimental results is not fully explained. In order to predict the outcome of experiments and optimize the efficiency of electroporation before each treatment, we developed a model to investigate the effect of pulse shape on efficiency of electroporation.ResultsOur model is based on a developed chemical-kinetics scheme and trapezium barrier model, while self-consistency was taken into account. This model is further supplemented with a molecular transport model to acquire the molecular uptake of cells. The investigated pulse shapes in this study were unipolar rectangular pulses with different rise and fall times, triangular, sinusoidal and bipolar rectangular pulses and also sinusoidal modulated unipolar pulses with different percentages of modulation. The obtained results from our modelling and simulations are in good agreement with previously published experimental results.ConclusionsWe therefore conclude that this model can be used to predict the effects of arbitrarily shaped electroporation pulses on cell membrane conductivity and molecular transport across the cell membrane.

This review provides an overview of the effect of blood flow on endothelial cell (EC) signalling pathways, applying microarray technologies to cultured cells, and in vivo studies of normal and atherosclerotic animals. It is found that in cultured ECs, 5-10% of genes are up- or down-regulated in response to fluid flow, whereas only 3-6% of genes are regulated by varying levels of fluid flow. Of all genes, 90% are regulated by the steady part of fluid flow and 10% by pulsatile components. The associated gene profiles show high variability from experiment to experiment depending on experimental conditions, and importantly, the bioinformatical methods used to analyse the data. Despite this high variability, the current data sets can be summarized with the concept of endothelial priming. In this concept, fluid flows confer protection by an up-regulation of anti-atherogenic, anti-thrombotic, and anti-inflammatory gene signatures. Consequently, predilection sites of atherosclerosis, which are associated with low-shear stress, confer low protection for atherosclerosis and are, therefore, more sensitive to high cholesterol levels. Recent studies in intact non-atherosclerotic animals confirmed these in vitro studies, and suggest that a spatial component might be present. Despite the large variability, a few signalling pathways were consistently present in the majority of studies. These were the MAPK, the nuclear factor-κB, and the endothelial nitric oxide synthase-NO pathways.

We present a study of the variability of the minimal transmembrane voltage resulting in detectable electroporation of the plasma membrane of spherical and irregularly shaped CHO cells (we denote this voltage by ITVc). Electroporation was detected by monitoring the influx of Ca(2+), and the transmembrane voltage was computed on a 3D finite-elements model of each cell constructed from its cross-section images. We found that ITVc was highly variable, particularly in irregularly shaped cells, where it ranged from 512-1028 mV. We show that this range is much too large to be an artifact due to numerical errors and experimental inaccuracies, implying that for cells of the same type and exposed to the same number of pulses with the same duration, the value of ITVc can differ considerably from one cell to another. We also observed that larger cells are in many cases characterized by a higher ITVc than a smaller one. This is in qualitative agreement with the reports that higher membrane curvature facilitates electroporation, but quantitative considerations suggest that the observed variability of ITVc cannot be attributed entirely to the differences in membrane curvature.