Search Results (23)

The electric field orientation method effectively promotes the orientation and arrangement of BN nanosheets, forming a thermal conduction network and enhancing the thermal conductivity of the composite material. In this study, microsecond pulsed electric field and direct current electric field were applied to induce the orientation and arrangement of BN nanosheets and improve the thermal conductivity of epoxy resin composites. Under a microsecond pulsed electric field of 50 Hz, 1.5 μs, and 8 kV/mm, the average orientation angle of BN nanosheets increased by 147.7%, and the thermal conductivity of the composite reached 0.352 W/(m·K), which is 1.84 times that of pure epoxy resin. In contrast, under a DC electric field of 70 V/mm, the average orientation angle of BN nanosheets increased by only 57.9%, while the thermal conductivity of the composite reached 0.364 W/(m·K), 1.91 times that of pure epoxy resin. The results indicate that the microsecond pulsed electric field primarily enhances the local orientation of the fillers to improve thermal conductivity, whereas the DC electric field mainly enhances the global arrangement of the fillers to achieve a similar effect. Additionally, thermogravimetric analysis and differential scanning calorimetry were conducted to evaluate the thermal properties of the composites. The results demonstrate that after BN nanosheets orientation and arrangement within the epoxy resin induced by both microsecond pulsed and DC electric fields, the composites exhibited a higher glass transition temperature and improved thermal stability. This study systematically explores the effects of microsecond pulsed and DC electric fields on filler orientation and arrangement, providing valuable insights for the fabrication of electric field-oriented composites. Full article

In the tissue regeneration field, stem cell transplantation represents a promising therapeutic strategy. To favor their implantation, proliferation and differentiation need to be controlled. Several studies have demonstrated that stem cell fate can be controlled by applying continuous electric field stimulation. This study aims to characterize the effect of a specific microsecond electric pulse stimulation (bipolar pulses of 100 µs + 100 µs, delivered for 30 min at an intensity of 250 V/cm) to induce an increase in cell proliferation on mesenchymal stem cells (MSCs) and induced neural stem cells (iNSCs). The effect was evaluated in terms of (i) cell counting, (ii) cell cycle, (iii) gene expression, and (iv) apoptosis. The results show that 24 h after the stimulation, cell proliferation, cell cycle, and apoptosis are not affected, but variation in the expression of specific genes involved in these processes is observed. These results led us to investigate cell proliferation until 72 h from the stimulation, observing an increase in the iNSCs number at this time point. The main outcome of this study is that the microsecond electric pulses can modulate stem cell proliferation. Full article

Background/Objectives: Electrochemotherapy (ECT) is a safe and efficient method of targeted drug delivery using pulsed electric fields (PEF), one that is based on the phenomenon of electroporation. However, the problems of electric field homogeneity within a tumor can cause a diminishing of the treatment efficacy, resulting only in partial response to the procedure. This work used gold nano-particles for electric field amplification, introducing the capability to improve available elec-trochemotherapy methods and solve problems associated with field non-homogeneity. Methods: We characterized the potential use of gold nanoparticles of 13 nm diameter (AuNPs: 13 nm) in combination with microsecond (0.6–1.5 kV/cm × 100 μs × 8 (1 Hz)) and nanosecond (6 kV/cm × 300–700 ns × 100 (1, 10, 100 kHz and 1 MHz)) electric field pulses. Finally, we tested the most prominent protocols (microsecond and nanosecond) in the context of bleomycin-based electrochemotherapy (4T1 mammary cancer cell line). Results: In the nano-pulse range, the synergistic effects (improved permeabilization and electrotransfer) were profound, with increased pulse burst frequency. Addi-tionally, AuNPs not only reduced the permeabilization thresholds but also affected pore resealing. It was shown that a saturated cytotoxic response with AuNPs can be triggered at significantly lower electric fields and that the AuNPs themselves are non-toxic for the cells either separately or in combination with bleomycin. Conclusions: The used electric fields are considered sub-threshold and/or not applicable for electrochemotherapy, however, when combined with AuNPs results in successful ECT, indicating the methodology’s prospective applicability as an anticancer treatment method. Full article

The phenomenon known as bipolar cancellation is observed when biphasic nanosecond electric field pulses are used, which results in reduced electroporation efficiency when compared to unipolar pulses of the same parameters. Basically, the negative phase of the bipolar pulse diminishes the effect of the positive phase. Our study aimed to investigate how bipolar cancellation affects Ca²⁺ electrochemotherapy and cellular response under varying electric field intensities and pulse durations (3–7 kV/cm, 100, 300, and 500 ns bipolar 1 MHz repetition frequency pulse bursts, n = 100). As a reference, standard microsecond range parametric protocols were used (100 µs × 8 pulses). We have shown that the cancellation effect is extremely strong when the pulses are closely spaced (1 MHz frequency), which results in a lack of cell membrane permeabilization and consequent failure of electrochemotherapy in vitro. To validate the observations, we have performed a pilot in vivo study where we compared the efficacy of monophasic (5 kV/cm × ↑500 ns × 100) and biphasic sequences (5 kV/cm × ↑500 ns + ↓500 ns × 100) delivered at 1 MHz frequency in the context of Ca²⁺ electrochemotherapy (B16-F10 cell line, C57BL/6 mice, n = 24). Mice treated with bipolar pulses did not exhibit prolonged survival when compared to the untreated control (tumor-bearing mice); therefore, the bipolar cancellation phenomenon was also occurrent in vivo, significantly impairing electrochemotherapy. At the same time, the efficacy of monophasic nanosecond pulses was comparable to 1.4 kV/cm × 100 µs × 8 pulses sequence, resulting in tumor reduction following the treatment and prolonged survival of the animals. Full article

Pulsed power equipment is often characterized by high energy density and field intensity. In the presence of strong electric field intensity, charge accumulation within insulators exacerbates electric field non-uniformity, leading to potential insulation breakdown, thereby posing a significant threat to the safe operation of pulsed power equipment. In this manuscript, we introduce nonlinear composite materials with field-dependent conductivity and permittivity to adaptively regulate the distribution of the pulsed electric field in insulation equipment. Finite-element modeling and analysis of the needle-plate electrodes and high-voltage bushing are carried out to comprehensively investigate the non-uniformity of the distribution of the electric field and the homogenization effect of various nonlinear materials in the presence of pulsed excitations of different timescales. Numerical results indicate that the involvement of nonlinear composite materials significantly improves the electric field distribution under pulse excitations. In addition, variations in the rising time of the pulses affect the maximum electric field intensity within the insulators considerably, but for pulses of nanosecond and microsecond scales, the tendencies are the opposite. Finally, via the simulations of the bushing, we illustrate that some measures proposed for improving the uniformity of the electric field under low frequencies, e.g., increasing the length of the electric field equalization layer and the distance of the underside of the electric field equalization layer from the grounding screen, are still effective for the homogenization of pulsed electric field. Full article

Pulsed Field Ablation (PFA) is the latest and most intriguing technology for catheter ablation of atrial fibrillation, due to its capability to generate irreversible and cardiomyocytes-selective electroporation of cell membranes by delivering microsecond-lasting high-voltage electrical fields, leading to high expectations. The first trials to assess the clinical success of PFA, reported an arrhythmia-free survival at 1-year of 78.5%, while other trials showed less enthusiastic results: 66.2% in paroxysmal and 55.1% in persistent AF. Nevertheless, real world data are encouraging. The isolation of pulmonary veins with PFA is easily achieved with 100% acute success. Systematic invasive remapping showed a high prevalence of durable pulmonary vein isolation at 75 and 90 days (range 84–96%), which were significatively lower in redo procedures (64.3%). The advent of PFA is prompting a reconsideration of the role of the autonomic nervous system in AF ablation, as PFA-related sparing of the ganglionated plexi could lead to the still undetermined effect on late arrhythmias’ recurrences. Moreover, a new concept of a blanking period could be formulated with PFA, according to its different mechanism of myocardial injury, with less inflammation and less chronic fibrosis. Finally, in this review, we also compare PFA with thermal energy. Full article

In the last few years, pulsed electric fields have emerged as promising clinical tools for tumor treatments. This study highlights the distinct impact of a specific pulsed electric field protocol, PEF-5 (0.3 MV/m, 40 μs, 5 pulses), on astrocytes (NHA) and medulloblastoma (D283) and glioblastoma (U87 NS) cancer stem-like cells (CSCs). We pursued this goal by performing ultrastructural analyses corroborated by molecular/omics approaches to understand the vulnerability or resistance mechanisms triggered by PEF-5 exposure in the different cell types. Electron microscopic analyses showed that, independently of exposed cells, the main targets of PEF-5 were the cell membrane and the cytoskeleton, causing membrane filopodium-like protrusion disappearance on the cell surface, here observed for the first time, accompanied by rapid cell swelling. PEF-5 induced different modifications in cell mitochondria. A complete mitochondrial dysfunction was demonstrated in D283, while a mild or negligible perturbation was observed in mitochondria of U87 NS cells and NHAs, respectively, not sufficient to impair their cell functions. Altogether, these results suggest the possibility of using PEF-based technology as a novel strategy to target selectively mitochondria of brain CSCs, preserving healthy cells. Full article

Gene delivery by the pulsed electric field is a promising alternative technology for nonviral transfection; however, the application of short pulses (i.e., nanosecond) is extremely limited. In this work, we aimed to show the capability to improve gene delivery using MHz frequency bursts of nanosecond pulses and characterize the potential use of gold nanoparticles (AuNPs: 9, 13, 14, and 22 nm) in this context. We have used bursts of MHz pulses 3/5/7 kV/cm × 300 ns × 100 and compared the efficacy of the parametric protocols to conventional microsecond protocols (100 µs × 8, 1 Hz) separately and in combination with nanoparticles. Furthermore, the effects of pulses and AuNPs on the generation of reactive oxygen species (ROS) were analyzed. It was shown that gene delivery using microsecond protocols could be significantly improved with AuNPs; however, the efficacy is strongly dependent on the surface charge of AuNPs and their size. The capability of local field amplification using AuNPs was also confirmed by finite element method simulation. Finally, it was shown that AuNPs are not effective with nanosecond protocols. However, MHz protocols are still competitive in the context of gene delivery, resulting in low ROS generation, preserved viability, and easier procedure to trigger comparable efficacy. Full article

The most likely intentional high-power electromagnetic (EM) interference, threatening the operation of technologically advanced electronic infrastructure, will have the forms of sub- and nanosecond ultra-wideband (UWB) pulses, several hundred nanosecond pulses of attenuated sinusoids, and sub- and microsecond sinusoidal pulses. The protection of electronic objects against high-power EM pulses is provided by different types of metal enclosure shields with technological apertures, inside which sensitive electronic objects can be placed. These technological apertures allow external EM interference to penetrate into the enclosure, making the EM shielding imperfect. The EM protection against the EM pulses has been mainly assessed based on the so-called shielding effectiveness (SE) parameters. The SE parameters (SE^e and SE^m for the electric and magnetic fields, respectively) are useful for designing and comparing EM shields. In relatively small shielding enclosures, which have recently become the subject of interest, the SE parameters have been studied for relatively long transient EM interference, longer than 150 ns, i.e., for the EM pulses whose duration is much longer than the time that the pulse takes to pass the small enclosure. In this work, we dealt with an ultrashort transient interference pulse, the duration of which was much shorter than the pulse transit time through the enclosure. The intentional high-power EM subnanosecond UWB pulse is an example of such a pulse. For such an ultrashort pulse, we studied the EM shielding performance of a small size enclosure numerically (W:H:D = 455 mm:50 mm:463 mm) with aperture (W:H = 80 mm:30 mm). The ultrashort EM interference pulse of a Gaussian distribution of the electric and magnetic fields with amplitudes of 10⁶ V/m and 2.68·10³ A/m, respectively, applied in this study, had a duration of 0.0804 ns (FWHM). This means that the high-power EM interference pulse was about 18 times shorter than the time that it takes to pass the enclosure (equal to about 1.5 ns). Our numerical simulations of the subnanosecond high-power EM interference of the interior of the enclosure with aperture were performed in the time domain using the commercial code CST Microwave Studio. First of all, the time-domain simulations resulted in 2D and 3D images and 2D vector maps of the electric and magnetic fields, which visualized the temporal and spatial development of the EM field in the enclosure with aperture caused by the incident subnanosecond high-power EM interference. The development of the associated electric and magnetic fields proceeded in two phases: first in the form of EM waves and later as an interference pattern, traveling forth and back between the front and rear enclosure walls. Due to the energy loss through the aperture, suffered by the traveling EM field and the tendency of the EM field to be evenly distributed over time throughout the entire enclosure, the amplitudes of the EM field decreased about 30 times within 90 ns. Despite the energy loss, the EM field developed in the enclosure existed at least 1000 times longer than the subnanosecond duration of the incident EM pulse (i.e., at least 90 ns as demonstrated by the numerical calculation). Apart from the EM field development visualization, the time-domain simulation enabled easy tracking of the temporal behavior of the EM field in selected points in the enclosure. Such tracking showed that each point in the enclosure was passed by a series of subnanosecond EM pulses, called internal EM pulses, over a relatively long time (at least over the simulation duration of 90 ns). This means that over 90 ns, the points in the enclosure were repeatedly influenced by the series of about 500 subnanosecond internal EM pulses. The amplitudes of many of these pulses were only (3–5) times lower than that of the incident EM pulse. Despite the lower amplitudes, these internal pulses may cause severe EM interference inside the enclosure. This shows a substantial change in the nature of the EM interference caused by a subnanosecond high-power EM plane wave when a given point is not shielded (a single interference of the subnanosecond strong EM pulse) and when a given point is shielded by the enclosure with aperture (a repetitive interference of subnanosecond weaker EM pulses). With the time dependence of the EM field amplitudes obtained from the time-domain calculation at a selected point in the enclosure, it is easy to determine the SE^e and SE^m at that point as a function of time. In this way, evaluation of the local SE^e and SE^m (for selected points in the enclosure) can be performed. Moreover, the 2D and 3D images and 2D vector maps calculated in the time domain for a given time enabled easy calculation of the SE^e and SE^m maps for various times. Such maps, shown for the first time in this paper, give a more global view of the shielding properties of the enclosure with an aperture. This all shows the advantages of the use of the time-domain approach for studying EM shielding in the case of ultrashort EM interferences. Full article

Electrochemotherapy (ECT) with bleomycin has been effectively used in recent years to treat various skin tumors. Microsecond electric pulses significantly improve bleomycin (BLM) delivery and its anticancer potential. Up to now, we can determine electric field distribution in the targeted tissue, however, the distribution of the injected drug is still not well known. In this study, we propose the combination of indocyanine green (ICG) with bleomycin as a practical approach for ECT, enabling drug distribution control and detection. Normal skeletal muscle (L6) and fibrosarcoma (WEHI-164) cells were used for the viability evaluation by MTT assay after 24 and 72 h. Cells were exposed to the ESOPE protocol alone and in combination with drugs. Additionally, visualization of the uptake of ICG and ICG + BLM supported by electroporation was performed by confocal microscopy. The mast cell tumor (MCTs) was diagnosed in the feline case. The mixture of ICG + BLM was injected into the tumor, and ECT was performed under near-infrared fluorescence imaging (NIRF). The obtained results indicate the safety of the used procedure in vitro and in vivo. ICG does not affect ECT protocols in vitro. No significant cell viability decrease was noted only in the case of WEHI-164 cells post-ECT. Moreover, it does not adversely affect the procedure; in the case of in vivo surgery, it helps to control the drug distribution before and after ECT and identify the sentinel lymph node. Full article

In modern oncology, therapies are based on combining monotherapies to overcome treatment resistance and increase therapy precision. The application of microsecond-pulsed electric fields (PEF) is approved to enhance local chemotherapeutic drug uptake within combination electrochemotherapy regimens. Reactive oxygen species (ROS) have been implicated in anticancer effects, and cold physical plasma produces vast amounts of ROS, which have recently been shown to benefit head and neck cancer patients. PEF and cold plasma technology have been linked to immunogenic cell death (ICD) induction, a regulated cell death accompanied by sterile inflammation that promotes antitumor immunity. To this end, we investigated the combined effect of both treatments regarding their intracellular ROS accumulation, toxicity, ICD-related marker expression, and optimal exposure sequence in a leukemia model cell line. The combination treatment substantially increased ROS and intracellular glutathione levels, leading to additive cytotoxic effects accompanied by a significantly increased expression of ICD markers, such as the eat-me signal calreticulin (CRT). Preconditioned treatment with cold plasma followed by PEF exposure was the most potent treatment sequence. The results indicate additive effects of cold plasma and PEF, motivating further studies in skin and breast tumor models for the future improvement of ECT in such patients. Full article

As the initial antibody technology, the preparation of hybridoma cells has been widely used in discovering antibody drugs and is still in use. Various antibody drugs obtained through this technology have been approved for treating human diseases. However, the key to producing hybridoma cells is efficient cell fusion. High-voltage microsecond pulsed electric fields (μsHVPEFs) are currently one of the most common methods used for cell electrofusion. Nevertheless, the membrane potential induced by the external microsecond pulse is proportional to the diameter of the cell, making it difficult to fuse cells of different sizes. Although nanosecond pulsed electric fields (nsPEFs) can achieve the fusion of cells of different sizes, due to the limitation of pore size, deoxyribonucleic acid (DNA) cannot efficiently pass through the cell pores produced by nsPEFs. This directly causes the significant loss of the target gene and reduces the proportion of positive cells after fusion. To achieve an electric field environment independent of cell size and enable efficient cell fusion, we propose a combination of nanosecond pulsed electric fields and low-voltage microsecond pulsed electric fields (ns/μsLVPEFs) to balance the advantages and disadvantages of the two techniques. The results of fluorescence experiments and hybridoma culture experiments showed that after lymphocytes and myeloma cells were stimulated by a pulse (ns/μsLVPEF, μsHVPEF, and control), compared with μsHVPEF, applying ns/μsLVPEF at the same energy could increase the cell fusion efficiency by 1.5–3.0 times. Thus far, we have combined nanosecond and microsecond pulses and provided a practical solution that can significantly increase cell fusion efficiency. This efficient cell fusion method may contribute to the further development of hybridoma technology in electrofusion. Full article

Gene transfer into primary immune cells as well as into cell lines is essential for scientific and therapeutical applications. One of the methods used for gene transfer is electroporation (EP). EP is a method where a pulsed electric field (PEF) causes a highly transient permeability of the targeted cell membrane. In this work, we present the electrotransfection of CHO-K1, 4T1 cell lines, and primary murine DCs with detectable protein-encoding plasmids in the sub-microsecond range. Microsecond (µs)- and nanosecond (ns)-range pulsed electric field transfection protocols were used. The efficiency of electrotransfection was evaluated using green fluorescent protein (GFP)-encoding plasmids (4.7 kbp; p-EGFP-N1) and plasmids expressing a firefly luciferase and red fluorescent protein (tdTomato) (8.5 kbp; pcDNA3.1(+)/Luc2 = tdT)). It was shown that the used nsPEFs protocol (7 kV/cm × 300 ns × 100, 1 MHz) ensured a better transfection efficiency than µsPEFs (1.2 kV/cm × 100 µs × 8, 1 Hz). Plasmid size and concentration had a strong impact on the cell transfection efficiency too. We also showed that there were no significant differences in transfection efficiency between immature and mature DCs. Finally, the nsPEF protocols were successfully applied for the stable transfection of the CHO-K1 cell line with the linearized pcDNA3.1(+)/Luc2 = tdT plasmid. The results of the study are applicable in gene therapy and DNA vaccination studies for the derivation of optimal electrotransfection conditions. Full article

The possibility to artificially adjust and fine-tune gene expression is one of the key milestones in bioengineering, synthetic biology, and advanced medicine. Since the effects of proteins or other transgene products depend on the dosage, controlled gene expression is required for any applications, where even slight fluctuations of the transgene product impact its function or other critical cell parameters. In this context, physical techniques demonstrate optimistic perspectives, and pulsed electric field technology is a potential candidate for a noninvasive, biophysical gene regulator, exploiting an easily adjustable pulse generating device. We exposed mammalian cells, transfected with a NF-κB pathway-controlled transcription system, to a range of microsecond-duration pulsed electric field parameters. To prevent toxicity, we used protocols that would generate relatively mild physical stimulation. The present study, for the first time, proves the principle that microsecond-duration pulsed electric fields can alter single-gene expression in plasmid context in mammalian cells without significant damage to cell integrity or viability. Gene expression might be upregulated or downregulated depending on the cell line and parameters applied. This noninvasive, ligand-, cofactor-, nanoparticle-free approach enables easily controlled direct electrostimulation of the construct carrying the gene of interest; the discovery may contribute towards the path of simplification of the complexity of physical systems in gene regulation and create further synergies between electronics, synthetic biology, and medicine. Full article

Pulsed electric fields in the sub-microsecond range are being increasingly used in biomedical and biotechnology applications, where the demand for high-voltage and high-frequency pulse generators with enhanced performance and pulse flexibility is pushing the limits of pulse power solid state technology. In the scope of this article, a new pulsed generator, which includes four independent MOSFET based Marx modulators, operating individually or combined, controlled from a computer user interface, is described. The generator is capable of applying different pulse shapes, from unipolar to bipolar pulses into biological loads, in symmetric and asymmetric modes, with voltages up to 6.5 kV and currents up to 65 A, in pulse widths from 100 ns to 100 µs, including short-circuit protection, current and voltage monitoring. This new scientific tool can open new research possibility due to the flexibility it provides in pulse generation, particularly in adjusting pulse width, polarity, and amplitude from pulse-to-pulse. It also permits operating in burst mode up to 5 MHz in four independent channels, for example in the application of synchronized asymmetric bipolar pulses, which is shown together with other characteristics of the generator. Full article