Nanosecond Pulsed Electric Fields Research Articles

Relationship between Apoptosis Induction in Cancer Cells and the Amount of Electrical Stimulation on Frequency Analysis of Nanosecond Pulsed Electric Field

Relationship between Apoptosis Induction in Cancer Cells and the Amount of Electrical Stimulation on Frequency Analysis of Nanosecond Pulsed Electric Field

Considerations for Exploring Nanosecond Pulsed Electric Fields (nsPEFs) for Treatments of Cancer, Benign Skin Diseases, Atrial Fibrillation, and for New Mechanistic Understandings

Pulsed power includes acquiring electrical energy, compressing it, and releasing it in instantaneous bursts that are low in energy but very high in power. When the pulse duration is near the plasma membrane charging time constant, which is the time during which the cell interior is exposed to the applied pulsed electric field, it affects intracellular structures and functions. The technology is called nanosecond Pulsed Electric Fields (nsPEFs), nanosecond electric pulses (nsEP), or Nanopulse Stimulation (NPSTM) according to Pulse Biosciences, Inc., a company taking the technology to the market. Initial studies showed the elimination of tumor cells in vitro by apoptosis, and other regulated cell death mechanisms, elimination of rodent and canine osteosarcoma, and a basal cell carcinoma clinical trial. In the rat liver and mouse breast cancers, tumor-free animals were in situ vaccinated (ISV), preventing the recurrence of the treated cancers. The technology has also focused on treating benign skin diseases, with some advantages over cryoablation. More recently, the same technology called nanosecond pulsed-field ablation (nsPFA) has been used to treat cardiac arrhythmias like Atrial Fabulation (AFib) with catheters in humans. In pre-clinical studies and now in humans, this technology is showing advantages over radiofrequency ablation and cryoablation. On a new mechanistic landscape, nonlethal nsPEFs modulation of electron transport in the plasma membrane and the mitochondria show potential for controlling redox homeostasis and metabolism. Furthermore, different nsPEF waveforms have different effects on cells; waveforms can differ by pulse duration, rise time, electric field, and/or post-pulse features. So, for nsPEFs, there is a lethal side used for ablation as with NPS and nsPFA and a more recently recognized nonlethal side indicating new possibilities to differentially modify cell physiology depending on the different nsPEF waveforms.

New Developments in Powder and Powder Metallurgy Processes Using Nanosecond Pulsed Electric Fields

New Developments in Powder and Powder Metallurgy Processes Using Nanosecond Pulsed Electric Fields

Anti-tumor effects of nanosecond pulsed electric fields in a murine model of pancreatic cancer

Nanosecond Pulsed Electric Fields (nsPEFs) treatment has demonstrated anti-tumor effects on various cancer cell lines. However, the use of this treatment in pancreatic cancer is limited. This study demonstrated that nsPEFs treatment effectively suppressed the proliferation and metastasis of pancreatic cancer cells, while also inducing DNA damage. Meanwhile, animal experiments have shown that nsPEFs effectively suppressed the growth of pancreatic cancer, even in cases where the tumor volume exceeded 500–600 mm3 at the initiation of treatment. Notably, a single treatment session was found to significantly inhibit tumor growth, while also showing no adverse effects on the main organs of the mice. RNA sequencing and bioinformatics revealed that seven key genes (CDK1, CENPA, UBE2C, CCNB2, PLK1, CCNA2, and CCNB14) were significantly correlated with the overall survival rate of patients with pancreatic cancer. Through the application of the competing endogenous RNA (ceRNA) hypothesis, two miRNAs (has-let-7b-5p and hsa-miR-193b-3p) and four lncRNAs (MIR4435-2HG, ZNF436-AS1, LINC01089, and MIR4435-2HG) were identified as significantly impacting the overall survival of pancreatic cancer patients. We have effectively developed an mRNA-miRNA-lncRNA network that has the potential to stimulate further investigation into the underlying mechanisms of nsPEFs on pancreatic cancer.

Nanosecond pulse electric field treatment initiates mitochondrial apoptosis pathway by inducing mitochondrial morphological changes in myocardial cells.

As an emerging myocardial ablation technique, the mechanism of nanosecond pulse electric field (nsPEF) ablation is currently less studied. Mitochondria are one of the important membrane structure organelles in cells, participating in numerous life activities within the cell. This study aimed to explore the morphological changes of mitochondria in living cells following nsPEF treatment. Myocardial cells were treated with a self-made solid-state LTD high-voltage nanosecond pulse generator with a pulse width of 100ns for 80 times. The changes in mitochondrial membrane potential and cell apoptosis in rat myocardial cells after nsPEFs were investigated using JC-1 assay kit, apoptosis double staining assay kit, and mitochondrial fluorescence probe. The results showed that after nsPEF treatment, the mitochondrial membrane potential decreased, apoptosis increased, and the average mitochondrial area decreased from 0.48 µm2 in live myocardial cells to 0.16 µm2. The average circumference ranges from 3.17µm dropped to 1.60µm. The shape factor decreased from 1.92 to 1.41. The aspect ratio has decreased from 2.16 to 1.59. nsPEF treatment induces changes in the morphology of myocardial cell mitochondria. Based on the results of mitochondrial membrane potential and apoptosis, it can be inferred that under this equipment and parameter conditions, nsPEF treatment first causes changes in mitochondrial morphology, and then initiates the mitochondrial apoptosis pathway, which may provide experimental basis for investigating the potential mechanism of nsPEF ablation of myocardial cells.

Effects of buffer composition and plasmid toxicity on electroporation-based non-viral gene delivery in mammalian cells using bursts of nanosecond and microsecond pulses.

Gene electrotransfer (GET) is non-viral gene delivery technique, also known as electroporation-mediated gene delivery or electrotransfection. GET is a method used to introduce foreign genetic material (such as DNA or RNA) into cells by applying external pulsed electric fields (PEFs) to create temporary pores in the cell membrane. This study was undertaken to examine the impact of buffer composition on the efficiency of GET in mammalian cells Also, we specifically compared the effectiveness of high-frequency nanosecond (ns) pulses with standard microsecond (µs) pulses. For the assessment of cell transfection efficiency and viability, flow cytometric analysis, luminescent assays, and measurements of metabolic activity were conducted. The efficiency of electrotransfection was evaluated using two different proteins encoding plasmids (pEGFP-N1 and Luciferase-pcDNA3). The investigation revealed that the composition of the electroporation buffer significantly influences the efficacy of GET in CHO-K1 cell line. The different susceptibility of cell lines to the electric field and the plasmid cytotoxicity were reported. It was also shown that electroporation with nanosecond duration PEF protocols ensured equivalent or even better transfection efficiency than standard µsPEF. Additionally, we successfully performed long-term transfection of the murine 4T1 cell line using high-frequency nanosecond PEFs and confirmed its' applicability in an in vivo model. The findings from the study can be applied to optimize electrotransfection conditions.

Enhanced Cellular Doxorubicin Uptake via Delayed Exposure Following Nanosecond Pulsed Electric Field Treatment: An In Vitro Study.

The combination of nanosecond Pulsed Electric Field (nsPEF) with pharmaceuticals is a pioneering therapeutic method capable of enhancing drug uptake efficacy in cells. Utilizing nsPEFs configured at 400 pulses, an electric field strength of 15 kV/cm, a pulse duration of 100 ns, and a repetition rate of 10 pulses per second (PPS), we combined the nsPEF with a low dose of doxorubicin (DOX) at 0.5 μM. Upon verifying that cells could continuously internalize DOX from the surrounding medium within 1 h post nsPEF exposure, we set the DOX exposure period to 10 min and contrasted the outcomes of varying sequences of DOX and nsPEF administration: pulsing followed by DOX, DOX followed by pulsing, and DOX applied 40 min after pulsing. Flow cytometry, CCK-8 assays, and transmission electron microscopy (TEM) were employed to examine intracellular DOX accumulation, cell viability, apoptosis, cell cycle, and ultrastructural transformations. Our findings demonstrate that exposing cells to DOX 40 min subsequent to nsPEF treatment can effectively elevate intracellular DOX levels, decrease cell viability, and inhibit the cell cycle. This research work presents a novel approach to enhance DOX uptake efficiency with moderate conditions of both DOX and nsPEF.

Efficacy of a surgical cardiac ablation clamp using nanosecond pulsed electric fields: An acute porcine model

ObjectiveTo examine the effectiveness of a recently developed nonthermal technology, nanosecond pulse-field ablation (nsPFA), for surgical ablation of the atria in a beating heart porcine model. MethodsSix pigs underwent sternotomy and ablation using an nsPFA parallel clamp. The ablation electrodes (53 mm long) were embedded in the jaws of the clamp. Nine lesions per pig were created in locations chosen to be representative of the Cox-maze procedure. Four lesions were intended to electrically isolate parts of the atrium: the right atrial appendage, left atrial appendage, right pulmonary veins, and left pulmonary veins. For these lesions, exit block testing was performed both after ablation and before euthanasia; the time between the 2 tests was 3.3 ± 0.5 hours (range, 2-4 hours). Using purse string sutures, 5 more lesions were created up to the superior vena cava, down to the inferior vena cava, across the right atrial free wall, and at 2 distinct locations on the left atrial free wall. The clamp delivered a train of nanosecond duration pulses, with a total duration of 2.5 seconds, independent of tissue thickness. The heart tissue was stained with 1% triphenyltetrazolium chloride after a dwelling period of 2 hours. Subsequently, each lesion was cross sectioned at 5-mm intervals to assess the ablation depth and transmurality. In some sections, transmurality could not be established on the basis of triphenyltetrazolium chloride staining alone; for these lesions, Gomori-trichrome stains were used, and the histologic sections were evaluated for transmurality. ResultsThe ablation time was 2.5 seconds per lesion, for a total of only 22.5 seconds ablation time to create 9 lesions. A total of 53 lesions were created, resulting in 388 separate histologic sections. Transmurality was established in 386 sections (99.5%). Mean tissue thickness was 3.1 ± 1.5 mm (range, 0.2-8.6 mm). Exit block was confirmed in 23 of the 24 lesions (96%) postablation and 23 of 24 (96%) before the animals were humanely killed. Over the course of the procedure, neither pulse-induced arrhythmias nor any other complications were noted. ConclusionsThe novel nsPFA clamp device was effective in creating acute conduction block and transmural lesions in both the right and left atria in an acute porcine model. This nonthermal energy source has great potential to both shorten procedural time and enable effective ablation in the beating heart.

Pulsed electric field induces exocytosis and overexpression of MAGE antigens in melanoma

Nanosecond pulsed electric field (nsPEF) has emerged as a promising approach for inducing cell death in melanoma, either as a standalone treatment or in combination with chemotherapeutics. However, to date, there has been a shortage of studies exploring the impact of nsPEF on the expression of cancer-specific molecules. In this investigation, we sought to assess the effects of nsPEF on melanoma-specific MAGE (Melanoma Antigen Gene Protein Family) expression. To achieve this, melanoma cells were exposed to nsPEF with parameters set at 8 kV/cm, 200 ns duration, 100 pulses, and a frequency of 10 kHz. We also aimed to comprehensively describe the consequences of this electric field on melanoma cells' invasion and proliferation potential. Our findings reveal that following exposure to nsPEF, melanoma cells release microvesicles containing MAGE antigens, leading to a simultaneous increase in the expression and mRNA content of membrane-associated antigens such as MAGE-A1. Notably, we observed an unexpected increase in the expression of PD-1 as well. While we did not observe significant differences in the cells' proliferation or invasion potential, a remarkable alteration in the cells' metabolomic and lipidomic profiles towards a less aggressive phenotype was evident. Furthermore, we validated these results using ex vivo tissue cultures and 3D melanoma culture models. Our study demonstrates that nsPEF can elevate the expression of membrane-associated proteins, including melanoma-specific antigens. The mechanism underlying the overexpression of MAGE antigens involves the initial release of microvesicles containing MAGE antigens, followed by a gradual increase in mRNA levels, ultimately resulting in elevated expression of MAGE antigens post-experiment. These findings shed light on a novel method for modulating cancer cells to overexpress cancer-specific molecules, thereby potentially enhancing their sensitivity to targeted anticancer therapy.

Nano-Pulse Stimulation Therapy Initiates Regulated Cell Death in Skin, Unlike Bovie Radiofrequency Ablation and Cryoablation.

This study describes a unique new bioelectric approach for clearing skin lesions and illustrates the clinical and histological differences between this new method and the standards of cryoablation and Bovie® radiofrequency ablation (RFA). To determine the advantage of stimulating regulated cell death with nanosecond pulsed electric fields over the necrosis response elicited by thermal ablation modalities. Human abdominal skin was treated with cryoablation, Bovie® RFA, and nano-pulse stimulation (NPS) therapy four times before an abdominoplasty procedure was performed to collect skin for histology. The clinical appearance and corresponding histology of each treatment were documented over time and compared. NPS therapy triggered regulated cell death as indicated by the appearance of activated Caspase-3 at 2 h post treatment and the absence of nuclear staining 1 day post treatment. Epidermal regeneration follows without impacting the noncellular dermis in contrast to cryoablation and Bovie® RFA which trigger necrosis and often cause scarring, inflammation, or permanent pigmentary changes. The main differences between NPS therapy and other ablation modalities are the level of fibrosis, amount of scarring, elastic fiber concentration, and inflammation. An analysis of the skin thickness 30 days after the treatment indicates that NPS-treated skin is the most similar to untreated skin but cryoablated and RF-ablated skin were 2- and 3.5-fold thicker, respectively, suggesting that they initiate necrosis rather than regulated cell death. We conclude that NPS therapy is a unique nonthermal modality that may be applied for clearing benign skin lesions by initiating the skin's own programmed cell death pathway instead of necrosis as used by cryoablation and Bovie® RFA.

Ablating Myocardium Using Nanosecond Pulsed Electric Fields: Preclinical Assessment of Feasibility, Safety, and Durability.

Unlike conventional microsecond pulsed electrical fields that primarily target the cell membranes, nanosecond pulses are thought to primarily electroporate intracellular organelles. We conducted a comprehensive preclinical assessment of catheter-based endocardial nanosecond pulsed field ablation in swine. A novel endocardial nanosecond pulsed field ablation system was evaluated in a total of 25 swine. Using either a low-dose (5-second duration) or high-dose (15-second duration) strategy, thoracic veins and discrete atrial and ventricular sites were ablated. Predetermined survival periods were <1 (n=1), ≈2 (n=7), ≈7 (n=6), 14 (n=2), or ≈28 (n=9) days, and venous isolation was assessed before euthanasia. Safety assessments included evaluation of esophageal effects, phrenic nerve function, and changes in venous caliber. All tissues were subject to careful gross pathological and histopathologic examination. All (100%) veins (13 low-dose, 34 high-dose) were acutely isolated, and all reassessed veins (6 low-dose, 15 high-dose) were durably isolated. All examined vein lesions (10 low-dose, 22 high-dose) were transmural. Vein diameters (n=15) were not significantly changed. Of the animals assessed for phrenic palsy (n=9), 3 (33%) demonstrated only transient palsy. There were no differences between dosing strategies. Thirteen mitral isthmus lesions were analyzed, and all 13 (100%) were transmural (depth, 6.4±0.4 mm). Ventricular lesions were 14.7±4.5 mm wide and 7.1±1.3 mm deep, with high-dose lesions deeper than low-dose (7.9±1.2 versus 6.2±0.8 mm; P=0.007). The esophagus revealed nontransmural adventitial surface lesions in 5 of 5 (100%) animals euthanized early (2 days) post-ablation. In the 10 animals euthanized later (14-28 days), all animals demonstrated significant esophageal healing-8 with complete resolution, and 2 with only trace fibrosis. A novel, endocardial nanosecond pulsed field ablation system provides acute and durable venous isolation and linear lesions. Transient phrenic injury and nontransmural esophageal lesions can occur with worst-case assessments suggesting limits to pulsed field ablation tissue selectivity and the need for dedicated assessments during clinical studies.

Regulating Schwann Cell Growth by Nanosecond Pulsed Electric Field for Peripheral Nerve Regeneration In Vitro.

Schwann cells (SCs) are myelinating cells of the peripheral nervous system, playing a crucial role in peripheral nerve regeneration. Nanosecond Pulse Electric Field (nsPEF) is an emerging method applicable in nerve electrical stimulation that has been demonstrated to be effective in stimulating cell proliferation and other biological processes. Aiming to assess whether SCs undergo significant changes under nsPEF and help explore the potential for new peripheral nerve regeneration methods, cultured RSC96 cells were subjected to nsPEF stimulation at 5 kV and 10 kV, followed by continued cultivation for 3-4 days. Subsequently, some relevant factors expressed by SCs were assessed to demonstrate the successful stimulation, including the specific marker protein, neurotrophic factor, transcription factor, and myelination regulator. The representative results showed that nsPEF significantly enhanced the proliferation and migration of SCs and the ability to synthesize relevant factors that contribute positively to the regeneration of peripheral nerves. Simultaneously, lower expression of GFAP indicated the benign prognosis of peripheral nerve injuries. All these outcomes show that nsPEF has great potential as an efficient treatment method for peripheral nerve injuries by stimulating SCs.

Integrated Time Nanosecond Pulse Irreversible Electroporation (INSPIRE): Assessment of Dose, Temperature, and Voltage on Experimental and Clinical Treatment Outcomes.

This study sought to investigate a novel strategy using temperature-controlled delivery of nanosecond pulsed electric fields as an alternative to the 50-100 microsecond pulses used for irreversible electroporation. INSPIRE treatments were carried out at two temperatures in 3D tumor models using doses between 0.001 s and 0.1 s. The resulting treatment zones were quantified using viability staining and lethal electric field intensities were determined numerically. Computational modeling was then used to determine parameters necessary for INSPIRE treatments to achieve equivalent treatment zones to clinical electroporation treatments and evaluate the potential for these treatments to induce deleterious thermal damage. Lethal thresholds between 1109 and 709 V/cm were found for nominal 0.01 s treatments with pulses between 350 ns and 2000 ns at physiological temperatures. Further increases in dose resulted in significant decreases in lethal thresholds. Given these experimental results, treatment zones comparable to clinical electroporation are possible by increasing the dose and voltage used with nanosecond duration pulses. Temperature-controlled simulations indicate minimal thermal cell death while achieving equivalent treatment volumes to clinical electroporation. Nanosecond electrical pulses can achieve comparable outcomes to traditional electroporation provided sufficient electrical doses or voltages are applied. The use of temperature-controlled delivery may minimize thermal damage during treatment. Intense muscle stimulation and the need for cardiac gating have limited irreversible electroporation. Nanosecond pulses can alleviate these challenges, but traditionally have produced significantly smaller treatment zones. This study suggests that larger ablation volumes may be possible with the INSPIRE approach and that future in vivo studies are warranted.

Electric field dependence of the electron drift velocity in n-type InxGa1-xAs1-yBiy epilayer

The effect of Bi incorporation into InxGa1-xAs lattice on the nanosecond pulsed electric field dependence of the drift velocity of electrons in n-type InxGa1-xAs1-yBiy alloys with various doping densities is investigated at room temperature. The electronic band structure of the alloys is calculated by Finite Element Methods (FEM). The electron temperature is determined from analytical modeling of the power dissipation mechanism. The drift velocity of the electrons (vdrift) does not saturate up to 5.4 kV/cm electric field for the lowest doping density sample. The sample is burned out at a higher electric field due to the impact ionization mechanism. However, the vdrift saturates at a high electric field region in the samples with higher doping densities, and the saturation of electric field values of vdrift depends on the doping density for these samples. The highest electron drift mobility (μdrift) is obtained as 6236 cm2/Vs for the sample with the lowest electron density. The heating of the only electrons through the applied electric field is not enough to initiate inter-valley transfer because of scatterings during transport. Hence, the electric field dependence on electron transport occurs only in the central valley. The effect of the scatterings on the electron transport is identified by considering ionized impurity and phonon scatterings.

Effects of Pulse Width and Electrical Energy of Low‐Voltage Nanosecond Pulsed Electric Fields on Mitochondria in Cancer Cells

Nanosecond pulsed electric fields (nsPEFs) have recently shown promise as a cancer therapy. Although nsPEF‐induced apoptotic responses have been observed, the stress on individual cell components has not been quantified. Therefore, the authors built a cell equivalent circuit, including mitochondria, to quantitatively calculate the voltage, electric field, and electrical energy applied to the cell components in the frequency domain. Additionally, pulse width and electrical energy changed in mitochondrial membrane potential over time. The results indicated that the change of pulsed voltage switched the cell stimulation pathway, resulting in different mitochondrial membrane potentials over time. The frequency response analysis confirmed that the potential on mitochondrial membranes increased under shorter pulse width conditions. This suggested that the frequency analysis provided in this paper is useful for relating the site of cellular stimulation to induce physiological effects. © 2024 Institute of Electrical Engineers of Japan and Wiley Periodicals LLC.

Cardiomyocytes Permeabilization and Electrotransfection by Unipolar and Bipolar Asymmetric Electric Field Pulses.

Short electric field pulses represent a novel potential approach for achieving uniform electroporation within tissue containing elongated cells oriented in various directions, such as electroporation-based cardiac ablation procedures. In this study, we investigated how electroporation with nanosecond pulses with respect to different pulse shapes (unipolar, bipolar, and asymmetric) influences cardiomyocyte permeabilization and gene transfer. For this purpose, rat cardiomyocytes (H9c2) were used. The efficacy of the pulsed electric field protocols was assessed by flow cytometry and electrogene transfer by fluorescent and holotomographic microscopy. The response of the cells was assessed by the metabolic activity (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide[MTT] assay), F-actin distribution in cells by confocal microscopy, and muscle atrophy F-box (MAFbx) marker. We show nano- and microsecond pulse protocols, which are not cytotoxic for cardiac muscle cells and can be efficiently used for gene electrotransfection. Asymmetric nanosecond pulsed electric fields were similarly efficient in plasmid delivery as microsecond and millisecond protocols. However, the millisecond protocol induced a higher MAFbx expression in H9c2 cells.

Evaluation of genetic response of mesenchymal stem cells to nanosecond pulsed electric fields by whole transcriptome sequencing.

Mesenchymal stem cells (MSCs) modulated by various exogenous signals have been applied extensively in regenerative medicine research. Notably, nanosecond pulsed electric fields (nsPEFs), characterized by short duration and high strength, significantly influence cell phenotypes and regulate MSCs differentiation via multiple pathways. Consequently, we used transcriptomics to study changes in messenger RNA (mRNA), long noncoding RNA (lncRNA), microRNA (miRNA), and circular RNA expression during nsPEFs application. To explore gene expression profiles and potential transcriptional regulatory mechanisms in MSCs pretreated with nsPEFs. The impact of nsPEFs on the MSCs transcriptome was investigated through whole transcriptome sequencing. MSCs were pretreated with 5-pulse nsPEFs (100 ns at 10 kV/cm, 1 Hz), followed by total RNA isolation. Each transcript was normalized by fragments per kilobase per million. Fold change and difference significance were applied to screen the differentially expressed genes (DEGs). Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses were performed to elucidate gene functions, complemented by quantitative polymerase chain reaction verification. In total, 263 DEGs were discovered, with 92 upregulated and 171 downregulated. DEGs were predominantly enriched in epithelial cell proliferation, osteoblast differentiation, mesenchymal cell differentiation, nuclear division, and wound healing. Regarding cellular components, DEGs are primarily involved in condensed chromosome, chromosomal region, actin cytoskeleton, and kinetochore. From aspect of molecular functions, DEGs are mainly involved in glycosaminoglycan binding, integrin binding, nuclear steroid receptor activity, cytoskeletal motor activity, and steroid binding. Quantitative real-time polymerase chain reaction confirmed targeted transcript regulation. Our systematic investigation of the wide-ranging transcriptional pattern modulated by nsPEFs revealed the differential expression of 263 mRNAs, 2 miRNAs, and 65 lncRNAs. Our study demonstrates that nsPEFs may affect stem cells through several signaling pathways, which are involved in vesicular transport, calcium ion transport, cytoskeleton, and cell differentiation.

An integrated nanofiller spray and nanosecond pulse electrically-assisted method for synergistically interlaminar toughening and in-situ damage monitoring of CFRP composites

Nanomaterials have already been explored their potential for interlaminar toughening and crack-monitoring capabilities within the field of composite materials. However, efficient enhancement of interlaminar fracture toughness remains an unresolved challenge. Herein, a novel integrated filler spray and nanosecond pulsed electric field (nsPEF)-assisted curing technique is proposed for the fabrication of high-performance composite material system for the first time. The spraying technique proves to be an effective approach for depositing fillers, including carbon nanotubes (CNTs), graphene nanoplatelets (GNPs), and carbon nanofibers (CNFs), onto carbon fiber prepreg. Meanwhile, the nsPEF-assisted technique is utilized to enhance the wettability of resin to fiber, thus reducing void defects. The efficacy of this integrated technique is evidenced by the particularly low content of fillers (0.047 wt%). The experimental results show a substantial reduction of porosity in the composite by 49.36% (from 1.56% to 0.79%). Meanwhile, the mode I initiation and steady-state toughness of samples with CNTs and GNPs are remarkably improved by up to 47% and 82%, respectively, compared to the baseline. This represents the most significant improvement reported to date for non-functionalized CNTs and GNPs at such low concentrations. Moreover, the relative electrical resistance variation (ΔR/R0%) of CNT/GNP sensors used for in-situ damage sensing increased by ∼ 14%. This work pioneers the investigation of the synergistic effects of two promising techniques for preparing composite materials, offering advantages in terms of simplicity, multifunctionality, and the potential for industrial scalability.

Synergistic effects of nanosecond pulsed plasma and electric field on inactivation of pancreatic cancer cells in vitro

Nanosecond pulsed atmospheric pressure plasma jets (ns-APPJs) produce reactive plasma species, including charged particles and reactive oxygen and nitrogen species (RONS), which can induce oxidative stress in biological cells. Nanosecond pulsed electric field (nsPEF) has also been found to cause permeabilization of cell membranes and induce apoptosis or cell death. Combining the treatment of ns-APPJ and nsPEF may enhance the effectiveness of cancer cell inactivation with only moderate doses of both treatments. Employing ns-APPJ powered by 9 kV, 200 ns pulses at 2 kHz and 60-nsPEF of 50 kV/cm at 1 Hz, the synergistic effects on pancreatic cancer cells (Pan02) in vitro were evaluated on the metabolic activities of cells and transcellular electrical resistance (TER). It was observed that treatment with ns-APPJ for > 2 min disrupts Pan02 cell stability and resulted in over 30% cell death. Similarly, applying nsPEF alone, > 20 pulses resulted in over 15% cell death. While the inactivation activity from the individual treatment is moderate, combined treatments resulted in 80% cell death, approximately 3-to-fivefold increase compared to the individual treatment. In addition, reactive oxygen species such as OH and O were identified at the plasma-liquid interface. The gas temperature of the plasma and the temperature of the cell solution during treatments were determined to be near room temperature.

Macrophages enhanced with nanosecond pulsed electric fields and TGF-β1 strengthened the recovery of spinal cord injury.

Macrophages enhanced with nanosecond pulsed electric fields and TGF-β1 strengthened the recovery of spinal cord injury.