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Yuchen Zhang, Feihong Dong, Zhengxin Liu, Jinsong Guo, Jue Zhang, Jing Fang
Currently, nanosecond pulsed electric fields (nsPEFs) with short pulse duration and non-thermal effects have various potential applications in medicine and biology, especially in tumor ablation. Additionally, there are a few investigations on its proliferative effects in the normal cell. Clinically, proliferation of endothelial cells can perhaps accelerate the stent endothelialization and reduce the risk of acute thrombosis. To explore the feasibility using nsPEFs to induce proliferation of endothelial cells, in this study, porcine iliac endothelial (PIEC) cell line was cultured and tested by CCK-8 assay after nsPEFs treatment...
2018: PloS One
C Gao, X Zhang, J Chen, J Zhao, Y Liu, J Zhang, J Wang
BACKGROUND: Tongue squamous cell carcinoma is the most common squamous cell carcinoma of the head and neck. Immunotherapy has great potential in the treatment of tongue squamous cell carcinoma because of its unique advantages. However, the efficacy of immunotherapy is limited by the efficiency of antigen phagocytosis by immune cells. MATERIAL AND METHODS: We extracted dendritic cells (DCs) from human peripheral blood. Utilizing a nanosecond pulsed electric field (nsPEF), we deliver the tumour lysate protein into DCs and then incubate the DCs with PBMCs to obtain specific T cells to kill tumour cells...
April 22, 2018: Medicina Oral, Patología Oral y Cirugía Bucal
Xinhua Chen, Ruiqing Zhang, Hao Wen
The impact of ultrashort nanopulse on cellular membrane is of biological significance and thus has been studied intensively. Different from cell study, this ex vivo study aims to investigate the biological effects of nanosecond pulsed electric field (nsPEF) on an independent multimembrane parasite, human hydatid cyst, to observe the unique influence of nanopulse on macromembrane structure, permeabilization, and biochemistry. The 300 ns nsPEF was delivered on an experimental model of single human hydatid cyst ex vivo with eight different parameters...
2018: BioMed Research International
Brittany P Lassiter, Siqi Guo, Stephen J Beebe
Nano-pulse stimulation (NPS), previously called nsPEFs, induced a vaccine-like effect after ablation of orthotopic N1-S1 hepatocellular carcinoma (HCC), protecting rats from subsequent challenges with N1-S1 cells. To determine immunity, immune cell phenotypes were analyzed in naïve, treated and protected rats. NPS provides a positive, post-ablation immuno-therapeutic outcome by alleviating immunosuppressive T regulatory cells (Treg) in the tumor microenvironment (TME), allowing dendritic cell influx and inducing dynamic changes in natural killer cells (NKs), NKT-cells and T-lymphocytes in blood, spleen and liver...
March 13, 2018: Cancers
Akemi Yamaguchi, Hiroshi Ishibashi, Susumu Kono, Midori Iida, Masaya Uchida, Koji Arizono, Nobuaki Tominaga
Herein, we propose using a nanosecond pulsed electric field (nsPEF) technique to assess teratogenicity and embryonic developmental toxicity of estradiol-17β (E2 ) and predict the molecular mechanisms of teratogenicity and embryonic developmental defects caused by E2 on medaka (Oryzias latipes). The 5 hour post-fertilization embryos were exposed to co-treatment with 10 μm E2 and nsPEF for 2 hours and then continuously cultured under non-E2 and nsPEF conditions until hatching. Results documented that the time to hatching of embryos was significantly delayed in comparison to the control group and that typical abnormal embryo development, such as the delay of blood vessel formation, was observed...
May 2018: Journal of Applied Toxicology: JAT
Diganta Dutta, Xavier-Lewis Palmer, Jinhyun Kim, Shizhi Qian, Michael Stacey
The purpose of this study is to map the energy dissipation of Jurkat cells using a single 60 nanosecond pulse electric field (NsPEF), primarily through atomic force microscopy (AFM). The phase shift is generated by the sample elements that do not have a heterogeneous surface. Monitoring and manipulating the phase shift is a powerful way for determining the dissipated energy and plotting the topography. The dissipated energy is a relative value, so the silica wafer and cover slip are given a set reference while the transmission of energy between the tip of the cantilever and cell surfaces is measured...
February 2018: Micron: the International Research and Review Journal for Microscopy
Hanna Hanna, Agnese Denzi, Micaela Liberti, Franck M André, Lluis M Mir
Microsecond pulsed electric fields (μsPEF) permeabilize the plasma membrane (PM) and are widely used in research, medicine and biotechnology. For internal membranes permeabilization, nanosecond pulsed electric fields (nsPEF) are applied but this technology is complex to use. Here we report that the endoplasmic reticulum (ER) membrane can also be electropermeabilized by one 100 µs pulse without affecting the cell viability. Indeed, using Ca2+ as a permeabilization marker, we observed cytosolic Ca2+ peaks in two different cell types after one 100 µs pulse in a medium without Ca2+ ...
October 12, 2017: Scientific Reports
Claudia Muratori, Andrei G Pakhomov, Elena Gianulis, Jade Meads, Maura Casciola, Peter A Mollica, Olga N Pakhomova
Nanosecond pulsed electric fields (nsPEF) are emerging as a novel modality for cell stimulation and tissue ablation. However, the downstream protein effectors responsible for nsPEF bioeffects remain to be established. Here we demonstrate that nsPEF activate TMEM16F (or Anoctamin 6), a protein functioning as a Ca2+ -dependent phospholipid scramblase and Ca2+ -activated chloride channel. Using confocal microscopy and patch clamp recordings, we investigated the relevance of TMEM16F activation for several bioeffects triggered by nsPEF, including phosphatidylserine (PS) externalization, nanopore-conducted currents, membrane blebbing, and cell death...
November 24, 2017: Journal of Biological Chemistry
Gleb P Tolstykh, Gary L Thompson, Hope T Beier, Zachary A Steelman, Bennett L Ibey
Cell swelling and blebbing has been commonly observed following nanosecond pulsed electric field (nsPEF) exposure. The hypothesized origin of these effects is nanoporation of the plasma membrane (PM) followed by transmembrane diffusion of extracellular fluid and disassembly of cortical actin structures. This investigation will provide evidence that shows passive movement of fluid into the cell through nanopores and increase of intracellular osmotic pressure are not solely responsible for this observed phenomena...
March 2017: Biochemistry and Biophysics Reports
Lynn Carr, Sylvia M Bardet, Delia Arnaud-Cormos, Philippe Leveque, Rodney P O'Connor
Cytosolic, synthetic chemical calcium indicators are typically used to visualise the rapid increase in intracellular calcium ion concentration that follows nanosecond pulsed electric field (nsPEF) application. This study looks at the application of genetically encoded calcium indicators (GECIs) to investigate the spatiotemporal nature of nsPEF-induced calcium signals using fluorescent live cell imaging. Calcium responses to 44kV/cm, 10ns pulses were observed in U87-MG cells expressing either a plasma membrane targeted GECI (GCaMP5-G), or one cytosolically expressed (GCaMP6-S), and compared to the response of cells loaded with cytosolic or plasma membrane targeted chemical calcium indicators...
February 2018: Bioelectrochemistry
Sarah D Jensen, Vera A Khorokhorina, Claudia Muratori, Andrei G Pakhomov, Olga N Pakhomova
We demonstrate that conditioning of mammalian cells by electroporation with nanosecond pulsed electric field (nsPEF) facilitates their response to the next nsPEF treatment. The experiments were designed to unambiguously separate the electroporation-induced sensitization and desensitization effects. Electroporation was achieved by bursts of 300-ns, 9 kV/cm pulses (50 Hz, n = 3-100) and quantified by propidium dye uptake within 11 min after the nsPEF exposure. We observed either sensitization to nsPEF or no change (when the conditioning was either too weak or too intense, or when the wait time after conditioning was too short)...
September 8, 2017: Scientific Reports
Maura Casciola, Shu Xiao, Andrei G Pakhomov
Modern technologies enable deep tissue focusing of nanosecond pulsed electric field (nsPEF) for non-invasive nerve and muscle stimulation. However, it is not known if PEF orders of magnitude shorter than the activation time of voltage-gated sodium channels (VGSC) would evoke action potentials (APs). One plausible scenario requires the loss of membrane integrity (electroporation) and resulting depolarization as an intermediate step. We report, for the first time, that the excitation of a peripheral nerve can be accomplished by 12-ns PEF without electroporation...
September 5, 2017: Scientific Reports
Ryan C Burke, Sylvia M Bardet, Lynn Carr, Sergii Romanenko, Delia Arnaud-Cormos, Philippe Leveque, Rodney P O'Connor
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...
October 2017: Biochimica et Biophysica Acta
Keiko Morotomi-Yano, Ken-Ichi Yano
Exposure of cultured human cells to nanosecond pulsed electric fields (nsPEFs) elicits various cellular events, including Ca2+ influx and cell death. Recently, nsPEFs have been regarded as a novel physical treatment useful for biology and medicine, but the underlying mechanism of action remains to be fully elucidated. In this study, we investigated the effect of nsPEFs on transglutaminases (TGs), enzymes that catalyze covalent protein modifications such as protein-protein crosslinking. Cellular TG activity was monitored by conjugation of cellular proteins with biotin-cadaverine, a cell-permeable pseudosubstrate for TGs...
July 2017: FEBS Open Bio
Malak Soueid, Martinus C F Dobbelaar, Sabrina Bentouati, Sylvia M Bardet, Rodney P O'Connor, Delphine Bessières, Jean Paillol, Philippe Leveque, Delia Arnaud-Cormos
In this paper, delivery devices for nanosecond pulsed electric field exposure of biological samples in direct contact with electrodes or isolated are presented and characterized. They are based on a modified electroporation cuvette and two transverse electromagnetic cells (TEM cells). The devices were used to apply pulses with high intensity (4.5 kV) and short durations (3 and 13 ns). The delivery devices were electromagnetically characterized in the frequency and time domains. Field intensities of around 5, 0...
January 2018: Medical & Biological Engineering & Computing
Xinhua Chen, Ruiqing Zhang, Tuerganaili Aji, Yingmei Shao, Yonggang Chen, Hao Wen
The nanosecond pulsed electric field (nsPEF) is investigated as an alternative plan for benign hepatic hydatid cyst. Altogether 72 C57B6 mice were included. Normal group (n = 12) had no parasite injection and the other 60 mice were used to induce hydatid cyst in liver by injecting protoscolices in portal vein. The liver hydatid cysts were exposed to nsPEF with different doses and then follow up. The standard surgery was performed as positive control. The hydatid cyst growth was monitored by ultrasound; the morphology was checked by gross anatomy and pathology was tested by H&E stain...
July 3, 2017: Scientific Reports
Diganta Dutta, Xavier-Lewis Palmer, Anthony Asmar, Michael Stacey, Shizhi Qian
This study reports that the surface charge density changes in Jurkat cells with the application of single 60 nanosecond pulse electric fields, using atomic force microscopy. Using an atomic force microscope tip and Jurkat cells on silica in a 0.01M KCl ionic concentration, we were able to measure the interfacial forces, while also predicting surface charge densities of both Jurkat cell and silica surfaces. The most important finding is that the pulsing conditions varyingly reduced the cells' surface charge density...
September 2017: Micron: the International Research and Review Journal for Microscopy
Xinhua Chen, Yiling Chen, Jianwen Jiang, Liming Wu, Shengyong Yin, Xudong Miao, Robert J Swanson, Shusen Zheng
BACKGROUND: Nanosecond pulsed electric field (nsPEF), which is also termed as nano-pulse stimulation (NPS), has the potential of stimulating immune responses toward cancer cells. The current study investigates its local and systemic antitumor efficacy in vivo in late stage tumors with lung metastasis. METHOD: The 12 canines with spontaneous osteosarcomas and 12 nude mice transplanted with human hepatocellular carcinoma were divided randomly and were given NPS treatment, surgery or no treatment control...
July 4, 2017: Oncotarget
Shengyong Yin, Xudong Miao, Xueming Zhang, Xinhua Chen, Hao Wen
Nanosecond pulsed electric field (nsPEF) is a novel non-thermal tumor ablation technique. However, how nsPEF affect cell physiology at different environmental temperature is still kept unknown. But this issue is of critical clinical practice relevance. This work aim to investigate how nsPEF treated cancer cells react to different environmental temperatures (0, 4, 25, and 37°C). Their cell viability, apoptosis, mitochondrial membrane potential, and reactive oxygen species (ROS) were examined. Lower temperature resulted in higher apoptosis rate, decreased mitochondria membrane potential, and increased ROS levels...
February 2018: Journal of Cellular Physiology
Andrei G Pakhomov, Iurii Semenov, Maura Casciola, Shu Xiao
Electric field pulses of nano- and picosecond duration are a novel modality for neurostimulation, activation of Ca(2+) signaling, and tissue ablation. However it is not known how such brief pulses activate voltage-gated ion channels. We studied excitation and electroporation of hippocampal neurons by 200-ns pulsed electric field (nsPEF), by means of time-lapse imaging of the optical membrane potential (OMP) with FluoVolt dye. Electroporation abruptly shifted OMP to a more depolarized level, which was reached within <1ms...
July 2017: Biochimica et Biophysica Acta
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