High powered, nanosecond duration, pulsed electric fields (nsPEF) cause cell death by a mechanism that is not fully understood and have been proposed as a targeted cancer therapy. Numerous chemotherapeutics work by disrupting microtubules. As microtubules are affected by electrical fields, this study looks at the possibility of disrupting them electrically with nsPEF. Human glioblastoma cells (U87-MG) treated with 100, 10 ns, 44 kV/cm pulses at a frequency of 10 Hz showed a breakdown of their interphase microtubule network that was accompanied by a reduction in the number of growing microtubules. This effect is temporally linked to loss of mitochondrial membrane potential and independent of cellular swelling and calcium influx, two factors that disrupt microtubule growth dynamics. Super-resolution microscopy revealed microtubule buckling and breaking as a result of nsPEF application, suggesting that nsPEF may act directly on microtubules.

Nanosecond pulsed electric fields (nsPEFs) have a variety of applications in the biomedical and biotechnology industries. Cancer treatment has been at the forefront of investigations thus far as nsPEFs permeabilize cellular and intracellular membranes leading to apoptosis and necrosis. nsPEFs may also influence ion channel gating and have the potential to modulate cell physiology without poration of the membrane. This phenomenon was explored using live cell imaging and a sensitive fluorescent probe of transmembrane voltage in the human glioblastoma cell line, U87 MG, known to express a number of voltage-gated ion channels. The specific ion channels involved in the nsPEF response were screened using a membrane potential imaging approach and a combination of pharmacological antagonists and ion substitutions. It was found that a single 10ns pulsed electric field of 34kV/cm depolarizes the transmembrane potential of cells by acting on specific voltage-sensitive ion channels; namely the voltage and Ca2 gated BK potassium channel, L- and T-type calcium channels, and the TRPM8 transient receptor potential channel.

Studying the response of neuronal networks to radiofrequency signals requires the use of a specific device capable of accessing and simultaneously recording neuronal activity during electromagnetic fields (EMF) exposure. In this study, a Microelectrode Array (MEA) that records the spontaneous activity of neurons is coupled to an open transverse electromagnetic (TEM) cell which propagates EMF. We characterize this system both numerically and experimentally at 1.8 GHz. Two MEA versions were compared, for the first time, to determine the impact of their design dissimilarities on the response to EMF. Macroscopic and microscopic measurements using respectively a fiber-optic probe and a temperature-dependent fluorescent dye (Rhodamine-B) were carried out. Results indicate that one MEA shows more stability toward the changes of the surrounding environment compared to the other MEA. Using a fiber-optic thermometer, the measured specific absorption rate (SAR) probe value in the center of the more stable MEA was 5.5±2.3 W/kg. Using a Rhod-B microdosimetry technique, the measured SAR value at the level of the MEA electrodes was 7.0±1.04 W/kg. SAR values are normalized per 1-W incident power. Due to the additional metallic planes and a smaller chip aperture, this new recording chip is steadier in terms of SAR and temperature stability allowing high exposure homogeneity as required during biological experiments. A typical neuronal activity recording under EMF exposure is reported.

Exposing living cells to a certain level of electromagnetic field (EMF) might induce some biological effects including temperature elevation. In this article, we studied two exposure systems at the macro and microscopic levels, allowing the study of the EMF effect on the biological samples exposed to 1.8-GHz signals. The macrosystem was an open transverse electromagnetic (TEM) cell that served as a dosimetry reference for defining limitations and optimal conditions for the temperature calibration using Rhodamine B (RhodB). The microfluidic microsystem was based on the coplanar waveguide (CPW) electrodes. Temperature measurements are carried out with a fluorooptic probe to extract specific absorption rate (SAR) values that are compared with numerical dosimetry, based on an FDTD method. After calibration, the fluorescence fits well with the temperature variation measured by the probe. To investigate dosimetry at a microscopic level, the fluorescence of the temperature-dependent dye RhodB was measured by fluorescence microscopy within the microfluidic channel or the biological cells. Results evidenced that the technique is applicable for RhodB concentrations higher than 1 µm with a value of 50 µm recommended for reliable experiments. For steady detection and SAR assessments, temperature variations of a few tenths of degrees were required.

Despite the biomedical advances of the last century, many cancers including glioblastoma are still resistant to existing therapies leaving patients with poor prognoses. Nanosecond pulsed electric fields (nsPEF) are a promising technology for the treatment of cancer that have thus far been evaluated in vitro and in superficial malignancies. In this paper, we develop a tumor organoid model of glioblastoma and apply intravital multiphoton microscopy to assess their response to nsPEFs. We demonstrate for the first time that a single 10 ns, high voltage electric pulse (35–45 kV/cm), collapses the perfusion of neovasculature, and also alters the diameter of capillaries and larger vessels in normal tissue. These results contribute to the fundamental understanding of nsPEF effects in complex tissue environments, and confirm the potential of nsPEFs to disrupt the microenvironment of solid tumors such as glioblastoma.

Abstract-We propose to analyze the aperture and ITO layer presence of a modified transverse electromagnetic (TEM) cell. This TEM cell can be used to study the potential effects of microwave electromagnetic fields on biological cells. This modified delivery device allows real-time observation of biological cells during exposure. Microscopic observation is achieved through an aperture in the lower wall of the TEM cell that is sealed with a 700-nm film of the transparent conducting material Indium tin oxide (ITO). To determine the device efficiency, numerical and experimental electromagnetic dosimetry was conducted. For assessing the effect of the aperture on the specific absorption rate (SAR) in the exposed sample, a plastic Petri dish containing cell culture medium, full-wave 3-D electromagnetic simulations and temperature measurements were performed. For 1-W input power, the SAR values obtained at 1.8 GHz in the sample exposed in the TEM cell with the sealed or non-sealed aperture of 20-mm diameter were 1.1 W/kg and 23.6 W/kg, respectively. An excellent homogeneity of the SAR distribution was achieved when the aperture was sealed with the ITO layer. The performance of the delivery system was confirmed by microwave exposure and simultaneous observation of living cells.

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