Reversible Electroporation Research Articles

Analysis of magnetic resonance contrast agent entrapment following reversible electroporation in vitro.

Administering gadolinium-based contrast agent before electroporation allows the contrast agent to enter the cells and enables MRI assessment of reversibly electroporated regions. The aim of this study was evaluation of contrast agent entrapment in Chinese hamster ovary (CHO) cells and comparison of these results with those determined by standard in vitro methods for assessing cell membrane permeability, cell membrane integrity and cell survival following electroporation. Cell membrane permeabilization and cell membrane integrity experiments were performed using YO-PRO-1 dye and propidium iodide, respectively. Cell survival experiments were performed by assessing metabolic activity of cells using MTS assay. The entrapment of gadolinium-based contrast agent gadobutrol inside the cells was evaluated using T1 relaxometry of cell suspensions 25 min and 24 h after electroporation and confirmed by inductively coupled plasma mass spectrometry. Contrast agent was detected 25 min and 24 h after the delivery of electric pulses in cells that were reversibly electroporated. In addition, contrast agent was present in irreversibly electroporated cells 25 min after the delivery of electric pulses but was no longer detected in irreversibly electroporated cells after 24 h. Inductively coupled plasma mass spectrometry showed a proportional decrease in gadolinium content per cell with shortening of T1 relaxation time (R 2 = 0.88 and p = 0.0191). Our results demonstrate that the contrast agent is entrapped in cells exposed to reversible electroporation but exits from cells exposed to irreversible electroporation within 24 h, thus confirming the hypothesis on which detection experiments in vivo were based.

Modeling methods in overlapping electroporation treatments: Pulse number effects on tissue conductivity and ablation area

Irreversible electroporation (IRE) is a non-thermal tissue ablation technique that utilizes high-voltage pulses between electrode pairs to destroy tissue. Numerical models are essential for predicting treatment outcomes and aiding in treatment planning. This paper studies a nonlinear conductivity model (NC) and a multiparametric nonlinear conductivity model (MPNC) to investigate overlapping electroporation treatments using a three-needle electrode configuration. The NC model accounts for the effects of the electric field and temperature on tissue conductivity during multiple pulsed electric field applications. The MPNC model incorporates the influence of pulse number, alongside electric field and temperature, on tissue conductivity. Results indicate that in the MPNC model, tissue conductivity is significantly higher in regions where the electric field intensity is below the threshold for irreversible electroporation, with the maximum difference reaching 0.4S/m. In contrast, tissue conductivity is slightly lower in regions where the electric field intensity exceeds the IRE threshold. Additionally, the MPNC model predicts a smaller area of irreversible electroporation and a larger area of reversible electroporation, with these differences becoming more pronounced as the distance between electrode needles increases. These findings underscore the importance of considering pulse number effects in numerical models to enhance the accuracy of treatment planning for irreversible electroporation-based therapies.

Effect of Pulse Width and Intensity on Cell Death in Reversible Electroporation of Cancerous Cells

Electroporation (EP) is the process of increasing the permeability of a biological cell or tissue by applying a short-term and sufficient external electric field. The utilization of proper pulse settings is required for EP-based treatments to be successful. Our aim in this study is to examine the effect of different electrical pulse widths and strength on EP efficiency. Human osteosarcoma cells (U20S) were used in the study. Eight-square-pulses with a frequency of 1Hz at 10µs, 1ms, 5ms, 10ms, and 20ms widths with low electric fields (20-500V/cm) were applied to U20S cells. 10-15 minutes after the applications, the cells were incubated in 96-well plates with 10 thousand cells in each well for 24 hours. Efficiency of pulses of different intensity and width was evaluated by MTT analysis method. The percent inhibition of U20S cancer cells elevated as the pulse width increased in almost all electric field values. The highest cell inhibition (%) occurred in pulses with an electric field of 500 V/cm and a width of 20ms (inhibition ratio: 76.25%). No inhibition was observed in the cells at 10µs, 1ms, 5ms, 10ms width pulses with 20 V/cm electric field and 10µs, 1ms width pulses with 50V/cm electric field. In conclusion, our findings show that the electric field intensity and pulse width used in electroporation play an important role in U20S cancer cell death. According to our results, it may be more appropriate to use high-voltage short-width pulses or low-voltage long-width pulses in reversible EP studies.

A Model for Reversible Electroporation to Deliver Drugs into Diseased Tissues.

Drug delivery through electroporation could be highly beneficial for the treatment of different types of diseased tissues within the human body. In this work, a mathematical model of reversible tissue electroporation is presented for injecting drug into the diseased cells. The model emphasizes the tissue boundary where the drug is injected as a point source. In addition, the effect of drug loss at tissue boundaries through extracellular space is studied elaborately. Multiple pulses are applied to deliver a sufficient amount of drug into the targeted cells. The set of differential equations that model the physical circumstances are solved numerically. This model obtains a mass transfer coefficient (MTC), in terms of pore fraction coefficient and drug permeability that controls the drug transport from extracellular to intracellular space. The drug penetration throughout the tissue is captured for the application of different pulses. The boundary effects on drug concentration are highlighted in this study. The advocated model is able to perform homogeneous drug transport into the cells so that the affected tissue is treated completely. This model can be applied to optimize clinical experiments by avoiding the lengthy and costly in vivo and in vitro experiments.

Electrochemotherapy with intravenous, intratumoral, or combined administration of bleomycin in the treatment of colorectal hepatic metastases in a rat model

Electrochemotherapy (ECT) combines the reversible electroporation (rEP) with intravenous (i.v.) or intratumoral (i.t.) administration of chemotherapeutic drugs. We conducted this study to compare the efficacy of i.v., i.t., and i.v. + i.t. injection of bleomycin (BLM) in ECT treatment of colorectal hepatic metastases in a rat model. WAG/Rij rats were randomized into three groups and underwent ECT with i.v., i.t., or i.v. + i.t. injection of BLM. Tumor volumes and oxygenation were measured by means of ultrasound and photoacoustic imaging. Moreover, liver and tumor tissue were analyzed by histology and immunohistochemistry. The i.v. and i.v. + i.t. groups exhibited a 44.0% and 46.6% reduction in oxygen saturation of the tumor tissue when compared to pretreatment values, whereas the i.t. group only showed a reduction of 35.2%. The extent of tumor tissue necrosis did not statistically differ between the groups. However, the i.t. group showed a tendency towards a lower necrosis rate. Cell proliferation, apoptotic cell death, vascularization, and immune cell infiltration were comparable in the treated tumors of the three groups. ECT with i.v. administration of BLM should be preferred in clinical practice, as the combined i.v. + i.t. therapy did not show superior oncological outcomes in the present study.

Ultrasonic-responsive piezoelectric stimulation enhances sonodynamic therapy for HER2-positive breast cancer

IntroductionBreast cancer ranks second as the most common malignancy globally, after lung cancer. Among the various subtypes of breast cancer, HER2 positive breast cancer (HER2 BC)poses a particularly challenging prognosis due to its heightened invasiveness and metastatic potential. The objective of this study was to construct a composite piezoelectric nanoparticle based on poly(vinylidene fluoride-trifluoroethylene) (P(VDF-TrFE)) for imaging and treatment of HER2 BC.MethodBy reshaping the crystal structure of P(VDF-TrFE) piezoelectric nanoparticles, improving hydrophilicity, and incorporating imaging capabilities, we developed piezoelectric composite nanoparticles (PGd@tNBs) that integrate imaging and therapeutic functions. The in vitro characterization encompassed the assessment of piezoelectric properties, hydrophilicity, imaging performance, and therapeutic efficacy of these particles. The targeting and therapeutic effectiveness of PGd@tNBs particles were further validated in the SK-BR3 cell line and subsequently confirmed in HER2-positive tumor-bearing mice.ResultsThe nanoparticle demonstrated excellent biocompatibility and impressive multimodal imaging performance. Magnetic resonance imaging (MRI) observations revealed significant accumulation of PGd@tNBs particles in the HER2 positive tumor, exhibiting superior contrast-enhanced ultrasound performance compared to traditional ultrasound contrast agents, and small animal in vivo imaging showed that PGd@tNBs particles were primarily excreted through respiration and urinary metabolism. Piezoforce Microscopy characterization highlighted the outstanding piezoelectric properties of PGd@tNBs particles. Upon targeted binding to HER2-BC, ultrasound stimulation influenced the cell membrane potential, leading to reversible electroporation. This, in turn, affected the balance of calcium ions inside and outside the cells and the mitochondrial membrane potential. Following ingestion by cells, PGd@tNBs, when exposed to ultrasound, triggered the generation of reactive oxygen species (ROS), resulting in the consumption of glutathione and superoxide dismutase and achieving sonodynamic therapy. Notably, repeated ultrasound stimulation, post PGd@tNBs particles binding and entry into cells, increased ROS production and elevated the apoptosis rate by approximately 45%.ConclusionIn conclusion, the PGd@tNBs particles developed exhibit outstanding imaging and therapeutic efficacy, holding potential for precise diagnosis and personalized treatment of HER2 BC.

Cell-inspired, massive electromodulation of friction via transmembrane fields across lipid bilayers.

Transient electric fields across cell bilayer membranes can lead to electroporation and cell fusion, effects crucial to cell viability whose biological implications have been extensively studied. However, little is known about these behaviours in a materials context. Here we find that transmembrane electric fields can lead to a massive, reversible modulation of the sliding friction between surfaces coated with lipid-bilayer membranes-a 200-fold variation, up to two orders of magnitude greater than that achieved to date. Atomistic simulations reveal that the transverse fields, resembling those at cell membranes, lead to fully reversible electroporation of the confined bilayers and the formation of inter-bilayer bridges analogous to the stalks preceding intermembrane fusion. These increase the interfacial dissipation through reduced hydration at the slip plane, forcing it to revert in part from the low-dissipation, hydrated lipid-headgroup plane to the intra-bilayer, high-dissipation acyl tail interface. Our results demonstrate that lipid bilayers under transmembrane electric fields can have striking materials modification properties.

Changes in Mitochondrial Membrane Potential in In Vitro Electroporation with Nano- and Microsecond Pulses.

With the introduction of nanosecond (ns) pulses, it was suggested that such pulses could be used to permeabilize intracellular membranes, including the mitochondrial membrane. The results presented thus far, however, are not conclusive. Interestingly, the effect of longer microsecond (μs) pulses on changes in mitochondria has never been investigated. We, therefore, investigated the changes in mitochondrial membrane permeability through changes in mitochondrial membrane potential (MMP) in CHO and H9c2 cells after electroporation with 4 ns, 200 ns, and 100 μs pulses. In the range of reversible electroporation, the decrease in MMP generally depended on the cell line. In CHO, ns pulses decreased MMP at lower electroporation intensities than μs. In H9c2, ns and μs were equally effective. In the range of irreversible electroporation, MMP decreased even further, regardless of pulse duration and cell type. The analysis at different time points showed that the changes in MMP within the first hour after pulse treatment are dynamic. Our results on the efficacy of ns pulses are consistent with published data, but with this study we show that μs pulses cause similar changes in MMP as ns pulses, demonstrating that electroporation affects MMP regardless of pulse duration. At the same time, however, differences in MMP changes were observed between different cell lines, indicating some dependence of MMP changes on cell type.

Mechanism study on the influences of buffer osmotic pressure on microfluidic chip-based cell electrofusion.

Cell electrofusion is a key process in many research fields, such as genetics, immunology, and cross-breeding. The electrofusion efficiency is highly dependent on the buffer osmotic pressure properties. However, the mechanism by which the buffer osmotic pressure affects cell electrofusion has not been theoretically or numerically understood. In order to explore the mechanism, the microfluidic structure with paired arc micro-cavities was first evaluated based on the numerical analysis of the transmembrane potential and the electroporation induced on biological cells when the electrofusion was performed on this structure. Then, the numerical model was used to analyze the effect of three buffer osmotic pressures on the on-chip electrofusion in terms of membrane tension and cell size. Compared to hypertonic and isotonic buffers, hypotonic buffer not only increased the reversible electroporation area in the cell-cell contact zone by 1.7 times by inducing a higher membrane tension, but also significantly reduced the applied voltage required for cell electroporation by increasing the cell size. Finally, the microfluidic chip with arc micro-cavitieswas fabricated and tested for electrofusion of SP2/0 cells. The results showed that no cell fusion occurred in the hypertonic buffer. The fusion efficiency in the isotonic buffer was about 7%. In the hypotonic buffer, the fusion efficiency was about 60%, which was significantly higher compared to hypertonic and isotonic buffers. The experimental results were in good agreement with the numerical analysis results.

In-silico research on the effects of variable voltage pulse protocols on the cell survival and permeabilization fractions in electroporated cancerous tissues

Reversible electroporation is used to increase the cell membrane permeability by the application of short duration, high voltage electric pulses with electrodes strategically positioned to induce localized drug concentration in the tumor tissue. However, inadequate selection of pulses’ characteristics can lead to relevant loss of cell viability or lack of cell permeabilization, affecting aversively the results of the therapy. Accordingly, the balance between cell membrane electro-permeabilization and cell death is crucial, and the recursive modification of electroporation parameters can favor this balance to increase the drug uptake into viable cells. Particularly, the controlled diminution of the voltage level as the number of pulses increases can reduce the cell damage without compromising the membrane electro-permeabilization. In this work, a previously developed in-silico tool, based on the Global Method of Approximate Particular Solutions (GMAPS), is calibrated, validated, and used to study the differences between constant voltage and variable voltage pulse protocols (CVP and VVP) in terms of the magnitude and distribution of cell survival and permeabilization fractions along the tissue domain, and the internalization efficiency of the therapy. In VVP protocols, the reduction of the applied voltage with the number of pulses is numerically estimated to reach a target volume-averaged survival fraction in the most sensitive zone of the tissue. Numerical results indicate that differences between the pulse protocol types (CVP and VVP) depend on the initial voltage level ∅0 and number of pulses K, with the VVP protocol bringing about superior results for the larger values of ∅0 and K considered here.

Electroporation in Cancer Therapy: A Simplified Model Derived from the Hodgkin-Huxley Model.

Cancer remains a global health challenge, necessitating effective treatments with fewer side effects. Traditional methods such as chemotherapy and surgery often have complications. Pulsed electric fields and electroporation have emerged as promising approaches to mitigate these challenges. This study presents a comprehensive overview of electroporation as an innovative tool in cancer therapy, encompassing critical elements such as pulse generators and delivery devices. Furthermore, it introduces a simplified reversible electroporation model grounded in the Hodgkin-Huxley model. This model ensures resting potential stability by regulating ionic currents. When membrane charges reach the electroporation threshold, the model swiftly increases the fraction of open pores, resulting in a rapid rise in electroporation current. Conversely, as the transmembrane potential drops below the threshold, the model gradually reduces the fraction of open pores, leading to a gradual decline in electroporation current, indicating pore resealing. This model contributes to easier modeling and implementation of reversible electroporation dynamics, providing a valuable tool for further exploration of electroporation for cancer therapy.

A microdosimetry analysis of reversible electroporation in scattered, overlapping, and cancerous cervical cells

We present a numerical method for studying reversible electroporation on normal and cancerous cervical cells. This microdosimetry analysis builds on a unique approach for extracting contours of free and overlapping cervical cells in the cluster from the Extended Depth of Field (EDF) images. The algorithm used for extracting the contours is a joint optimization of multiple-level set function along with the Gaussian mixture model and Maximally Stable Extremal Regions. These contours are then exported to a multi-physics domain solver, where a variable frequency pulsed electric field is applied. The trans-Membrane voltage (TMV) developed across the cell membrane is computed using the Maxwell equation coupled with a statistical approach, employing the asymptotic Smoluchowski equation. The numerical model was validated by successful replication of existing experimental configurations that employed low-frequency uni-polar pulses on the overlapping cells to obtain reversible electroporation, wherein, several overlapping clumps of cervical cells were targeted. For high-frequency calculation, a combination of normal and cancerous cells is introduced to the computational domain. The cells are assumed to be dispersive and the Debye dispersion equation is used for further calculations. We also present the resulting strength-duration relationship for achieving the threshold value of electroporation between the normal and cancerous cervical cells due to their size and conductivity differences. The dye uptake modulation during the high-frequency electric field electroporation is further advocated by a mathematical model.

Utilizing Human-Induced Pluripotent Stem Cells to Study Cardiac Electroporation Pulsed-Field Ablation.

Electroporation is a promising nonthermal ablation method for cardiac arrhythmia treatment. Although initial clinical studies found electroporation pulsed-field ablation (PFA) both safe and efficacious, there are significant knowledge gaps concerning the mechanistic nature and electrophysiological consequences of cardiomyocyte electroporation, contributed by the paucity of suitable human in vitro models. Here, we aimed to establish and characterize a functional in vitro model based on human-induced pluripotent stem cells (hiPSCs)-derived cardiac tissue, and to study the fundamentals of cardiac PFA. hiPSC-derived cardiomyocytes were seeded as circular cell sheets and subjected to different PFA protocols. Detailed optical mapping, cellular, and molecular characterizations were performed to study PFA mechanisms and electrophysiological outcomes. PFA generated electrically silenced lesions within the hiPSC-derived cardiac circular cell sheets, resulting in areas of conduction block. Both reversible and irreversible electroporation components were identified. Significant electroporation reversibility was documented within 5 to 15-minutes post-PFA. Irreversibly electroporated regions persisted at 24-hours post-PFA. Per single pulse, high-frequency PFA was less efficacious than standard (monophasic) PFA, whereas increasing pulse-number augmented lesion size and diminished reversible electroporation. PFA augmentation could also be achieved by increasing extracellular Ca2+ levels. Flow-cytometry experiments revealed that regulated cell death played an important role following PFA. Assessing for PFA antiarrhythmic properties, sustainable lines of conduction block could be generated using PFA, which could either terminate or isolate arrhythmic activity in the hiPSC-derived cardiac circular cell sheets. Cardiac electroporation may be studied using hiPSC-derived cardiac tissue, providing novel insights into PFA temporal and electrophysiological characteristics, facilitating electroporation protocol optimization, screening for potential PFA-sensitizers, and investigating the mechanistic nature of PFA antiarrhythmic properties.

Skin electroporation for transdermal drug delivery: Electrical measurements, numerical model and molecule delivery

Skin electroporation for drug delivery involves the application of Pulsed Electric Fields (PEFs) on the skin to disrupt its barrier function in a temporary and non-invasive manner, increasing the uptake of drugs. It represents a potential alternative to delivery methods that are invasive (e.g. injections) or limited. We have developed a drug delivery system comprising nanocomposite hydrogels which act as a reservoir for the drug and an electrode for applying electric pulses on the skin. In this study, we employed a multi-scale approach to investigate the drug delivery system on a mouse skin model, through electrical measurements, numerical modeling and fluorescence microscopy. The Electrical properties indicated a highly non-linear skin conductivity behavior and were used to fine-tune the simulations and study skin recovery after electroporation. Simulation of electric field distribution in the skin showed amplitudes in the range of reversible tissue electroporation (400–1200 V/cm), for 300 V PEF. Fluorescence microscopy revealed increased uptake of fluorescent molecules compared to the non-pulsed control. We reported two reversible electroporation domains for our configuration: (1) at 100 V PEF the first local transport regions appear in the extracellular lipids of the stratum corneum, demonstrated by a rapid increase in the skin's conductivity and an increased uptake of lucifer yellow, a small hydrophilic fluorophore and (2) at 300 V PEF, the first permeabilization of nucleated cells occurred, evidenced by the increased fluorescence of propidium iodide, a membrane-impermeable, DNA intercalating agent.

Engineering of Multivalent Membrane-Anchored DNA Frameworks for Precise Profiling of Variable Membrane Permeability During Reversible Electroporation.

Electroporation techniques have emerged as attractive tools for intracellular delivery, rendering promising prospects towards clinical therapies. Transient disruption of membrane permeability is the critical process for efficient electroporation-based cargo delivery. However, smart nanotools for precise characterization of transient membrane changes induced by strong electric pulses are extremely limited. Herein, multivalent membrane-anchored fluorescent nanoprobes (MMFNPs) that take advantages of flexible functionalization and spatial arrangement of DNA frameworks are developed for in situ evaluation of electric field-induced membrane permeability during reversible electroporation . Single-molecule fluorescence imaging techniques are adopted to precisely verify the excellent analytical performance of the engineered MMFNPs. Benefited from tight membrane anchoring and sensitive adenosine triphosphate (ATP)profiling, varying degrees of membrane disturbances are visually exhibited under different intensities of the microsecond pulse electric field (µsPEF). Significantly, the dynamic process of membrane repair during reversible electroporation is well demonstrated via ATP fluctuations monitored by the designed MMFNPs. Furthermore, molecular dynamics (MD) simulations are performed for accurate verification of electroporation-driven dynamic cargo entry via membrane nanopores. This work provides an avenue for effectively capturing transient fluctuations of membrane permeability under external stimuli, offering valuable guidance for developing efficient and safe electroporation-driven delivery strategies for clinical diagnosis and therapeutics.

In silico study about the influence of electroporation parameters on the cellular internalization, spatial uniformity, and cytotoxic effects of chemotherapeutic drugs using the Method of Fundamental Solutions.

Reversible electroporation is a suitable technique to aid the internalization of medicaments in cancer tissues without inducing permanent cellular damage, allowing the enhancement of cytotoxic effects without incurring in electric-driven necrotic or apoptotic processes by the presence of non-reversible aqueous pores. An adequate selection of electroporation parameters acquires relevance to reach these goals and avoid opposite effects. This work applies the Method of Fundamental Solutions (MFS) for drug transport simulations in electroporated cancer tissues, using a continuum tumor cord approach and considering both electro-permeabilization and vasoconstriction effects. The MFS algorithm is validated with published results, obtaining satisfactory accuracy and convergence. Then, MFS simulations are executed to study the influence of electric field magnitude [Formula: see text], number of electroporation treatments [Formula: see text], and electroporation time [Formula: see text] on three assessment parameters of electrochemotherapy: the internationalization efficacy accounting for the ability of the therapy to introduce moles into viable cells, cell-kill capacity indicating the faculty to reduce the survival fraction of cancer cells, and distribution uniformity specifying the competence to supply drug homogeneously through the whole tissue domain. According to numerical results, when [Formula: see text] is the reversibility threshold, a positive influence on the first two parameters is only possible once specific values of [Formula: see text] and [Formula: see text] have been exceeded; when [Formula: see text] is just the irreversibility threshold, any combination of [Formula: see text] and [Formula: see text] is beneficial. On the other hand, the drug distribution uniformity is always adversely affected by the application of electric pulses, being this more noticeable as [Formula: see text], [Formula: see text], and [Formula: see text] increases.

Nitrogen removal performance of aerobic denitrifying bacteria enhanced by an iron-anode pulsed electric field.

Pulsed electric field (PEF) technology has attracted considerable attention because it can efficiently treat pollutants that are difficult to degrade. In this study, a PEF system using iron as the electrode was constructed to investigate the effect of PEF-Fe on the growth and metabolism of aerobic denitrifying bacteria and the effectiveness of wastewater nitrogen removal. The chemical oxygen demand, NO3--N and nitrate removal rates were 98.93%, 97.60% and 24.40 mg·L-1·h-1, respectively, under optimal conditions. As confirmed in this study, PEF-Fe could improve the key enzyme activities of W207-14. Scanning electron microscopy revealed that the surface of PEF-Fe-treated W207-14 was intact and smooth without any irreversible deformation. Flow cytometry combined with fluorescence staining analysis also confirmed reversible electroporation on the cell membrane surface of PEF-Fe-treated W207-14. Differentially expressed gene enrichment analysis showed that PEF-Fe activated the transmembrane transport function of ATP-binding cassette transporte (ABC) transport proteins and enhanced the cell membrane permeability of aerobic denitrifying bacteria. The significant differential expression of iron-sulphur cluster proteins facilitated the regulation of electron transport and maintenance of the dynamic balance of iron ions within the PEF-Fe system.

Enhanced Cell Viability and Migration of Primary Bovine Annular Fibrosus Fibroblast-like Cells Induced by Microsecond Pulsed Electric Field Exposure.

This study is the first to report the enhancement of cell migration and proliferation induced by in vitro microsecond pulsed electric field (μsPEF) exposure of primary bovine annulus fibrosus (AF) fibroblast-like cells. AF primary cells isolated from fresh bovine intervertebral disks (IVDs) are exposed to 10 and 100 μsPEFs with different numbers of pulses and applied electric field strengths. The results indicate that 10 μs-duration pulses induce reversible electroporation, while 100 μs pulses induce irreversible electroporation of the cells. Additionally, μsPEF exposure increased AF cell proliferation up to 150% while increasing the average migration speed by 0.08 μm/min over 24 h. The findings suggest that the effects of PEF exposure on cells are multifactorial-depending on the duration, intensity, and number of pulses used in the stimulation. This highlights the importance of optimizing the μsPEF parameters for specific cell types and applications. For instance, if the goal is to induce cell death for cancer treatment, then high numbers of pulses can be used to maximize the lethal effects. On the other hand, if the goal is to enhance cell proliferation, a combination of the number of pulses and the applied electric field strength can be tuned to achieve the desired outcome. The information gleaned from this study can be applied in the future to in vitro cell culture expansion and tissue regeneration.

Characterizing reversible, irreversible, and calcium electroporation to generate a burst-dependent dynamic conductivity curve

The relationships between burst number, reversible, irreversible, and calcium electroporation have not been comprehensively evaluated in tumor tissue-mimics. Our findings indicate that electroporation effects saturate with a rate constant (τ) of 20 bursts for both conventional and high frequency waveforms (R2 > 0.88), with the separation between reversible and irreversible electroporation thresholds converging at 50 bursts. We find the lethal thresholds for calcium electroporation are statistically similar to reversible electroporation (R2 > 0.99). We then develop a burst-dependent dynamic conductivity curve that now incorporates electroporation effects due to both the electric field magnitude and burst number. Simulated ablation and thermal damage volumes vary significantly between finite element models using either the conventional or new burst-dependent dynamic conductivity curve (p < 0.05). Lastly, for clinically relevant protocols, thermal damage is indicated to not begin until 50 bursts, with maximum nonthermal ablation volumes at 100 bursts (1.5–13% thermal damage by volume). We find that >100 bursts generated negligible increases in ablation volumes with 40–70% thermal damage by volume at 300 bursts. Our results illustrate the need for considering burst number in minimizing thermal damage, choosing adjuvant therapies, and in modeling electroporation effects at low burst numbers.

Reversible Pulsed Electrical Fields as an In Vivo Tool to Study Cardiac Electrophysiology: The Advent of Pulsed Field Mapping.

During electrophysiological mapping of tachycardias, putative target sites are often only truly confirmed to be vital after observing the effect of ablation. This lack of mapping specificity potentiates inadvertent ablation of innocent cardiac tissue not relevant to the arrhythmia. But if myocardial excitability could be transiently suppressed at critical regions, their suitability as targets could be conclusively determined before delivering tissue-destructive ablation lesions. We studied whether reversible pulsed electric fields (PFREV) could transiently suppress electrical conduction, thereby providing a means to dissect tachycardia circuits in vivo. PFREV energy was delivered from a 9-mm lattice-tip catheter to the atria of 12 swine and 9 patients, followed by serial electrogram assessments. The effects on electrical conduction were explored in 5 additional animals by applying PFREV to the atrioventricular node: 17 low-dose (PFREV-LOW) and 10 high-dose (PFREV-HIGH) applications. Finally, in 3 patients manifesting spontaneous tachycardias, PFREV was applied at putative critical sites. In animals, the immediate post-PFREV electrogram amplitudes diminished by 74%, followed by 78% recovery by 5 minutes. Similarly, in patients, a 69.9% amplitude reduction was followed by 84% recovery by 3 minutes. Histology revealed only minimal to no focal, superficial fibrosis. PFREV-LOW at the atrioventricular node resulted in transient PR prolongation and transient AV block in 59% and 6%, while PFREV-HIGH caused transient PR prolongation and transient AV block in 30% and 50%, respectively. The 3 tachycardia patients had atypical atrial flutters (n=2) and atrioventricular nodal reentrant tachycardia. PFREV at putative critical sites reproducibly terminated the tachycardias; ablation rendered the tachycardias noninducible and without recurrence during 1-year follow-up. Reversible electroporation pulses can be applied to myocardial tissue to transiently block electrical conduction. This technique of pulsed field mapping may represent a novel electrophysiological tool to help identify the critical isthmus of tachycardia circuits.