Electroporation Of Cells Research Articles

Pulsed field ablation using additional mapping and substrate modification to treat atrial fibrillation - one year outcome comparison

Abstract Background Pulsed field ablation (PFA) is a relatively new non-thermal form of ablation employing irreversible cell electroporation as an alternative to radio-frequency ablation. Conventional use of PFA relies on fluoroscopy. Electro-anatomical and voltages mapping (EAM) may potentially provide refinement of PFA via ablation of non-PV targets and accurate localisation of catheter placement. Currently there is limited data on the efficacy of the use of voltages criteria of EAM in conjunction PFA in the treatment of atrial fibrillation. Objective The aim of our study was to determine the effect on atrial arrhythmia recurrence with the use of EAM with PFA in patients requiring atrial fibrillation ablation. Methods This single centre retrospective observational analysis included one hundred patients with symptomatic AF (paroxysmal and persistent) undergoing ablation for the first time. Fifty patients underwent PFA-PVI alone and fifty patients underwent PFA-PVI with EAM. Data were obtained on patient characteristics, outcomes, procedure details and complications up to one year. A blanking period of three months was used post-ablation. For rhythm monitoring we utilised ECGs and holters at three, six, nine and twelve months and twenty-four hour holter monitoring at six and nine months. We also used remote monitoring in between these periods to record any symptomatic episodes atrial arrhythmias. Results Baseline patient characteristics are represented in Table 1. Despite having no significant difference in category of AF between groups (Paroxysmal, persistent, longstanding persistent) the mapping group were significantly older (68.2 years ± 10.2 vs 60.1 ± 11.11; p=0.01), longer duration of documented atrial fibrillation (45.9 ± 50 vs 28.6 ± 38 months; p=0.038), higher risk for CV events with CHA2DS2-VASc score (2.2 ± 1.5 vs 1.42 ± 1.2; p=0.04), however, procedure times were greater in the PVI and mapping group (72.8 ± 6.2 vs 41.1 ± 17 minutes; p=0.03). Successful PVI was achieved acutely in all patients upon voltage guided mapping validation and most importantly 1 year follow up revealed comparable outcome in both groups (Fig 1). Conclusion Patients who underwent PFA with mapping were older, greater co-morbidity and longer duration of AF burden. Yet, the addition of EAM with PFA did demonstrate equivalent achievement of sinus rhythm and was relatively high as higher than 80 % at one year follow up. This study may suggest a preference to using EAM in patients who may be deemed to be older and have a greater co-morbidity. It emerges as a great hope to safer approach and successful intervention to reduce AF burden.mapping vs PVI onlyGraph showing one year follow up

Efficient electroporation in primary cells with PEDOT:PSS electrodes.

Precise and efficient delivery of macromolecules into cells enhances basic biology research and therapeutic applications in cell therapies, drug delivery, and personalized medicine. While pulsed electric field electroporation effectively permeabilizes cell membranes to deliver payloads without the need for toxic chemical or viral transduction agents, conventional bulk electroporation devices face major challenges with cell viability and heterogeneity due to variations in fields generated across cells and electrochemistry at the electrode-electrolyte interface. Here, we introduce the use of microfabricated electrodes based on the conducting polymer poly(3,4-ethylenedioxythiophene) doped with polystyrene sulfonate (PEDOT:PSS), which substantially increases cell viability and transfection efficiency. As a proof of concept, we demonstrate the enhanced delivery of Cas9 protein, guide RNA, and plasmid DNA into cell lines and primary cells. This use of PEDOT:PSS enables rapid modification of difficult-to-transfect cell types to accelerate their study and use as therapeutic platforms.

Multi-stage electroporation dynamics induced by ionization wave–substrate interaction in cold plasma jet

The reactive species-independent nature of cold plasma's electric field is pivotal in biomedical applications. This work proposes to connect the plasma fluid model and the asymptotic Smoluchowski model for electroporation, providing a unified framework to investigate the evolution of the electric field in the biological substrate and the multi-stage electroporation response of the human cell. Two common substrates with distinct dielectric properties, namely, the cultivation medium and epidermis, are selected to report three stages of ionization wave (IW)–substrate interaction. The three-stage streamer discharge dynamics (restrike, axial-radial transition, and radial expansion of IW) induce three-stage cell electroporation dynamics (slow charging, fast charging, and electroporation), though the two processes are asynchronous. Specifically, the inner membrane covered the cell nucleus with ultra-short charging time that undergoes only the first two discharge stages in both substrates. Whether the cell membrane is exposed to the third stage of discharge depends on the permittivity of the substrate. The asynchrony can be attributed to the difference in the charging time of the cell membranes and substrates affected by the substrate permittivity. The presented model can provide quantitative insights into the cell electroporation induced by the IW–substrate interaction and theoretical guidance for plasma biomedical applications.

Genetic Modification of Mice Using Prime Editing.

Genetically modifying mice traditionally involved complex methods of designing and validating targeting constructs, embryonic stem cell electroporation and selection, blastocyst injection, and breeding chimeras for germline transmission. Such arduous steps were best carried out by specialized gene targeting cores in academia or through expensive commercial vendors. Further, the time from initiation to completion of a project often took at least 1 year and, in some cases, much longer (or never), with no guarantees of success. The RNA-programmable CRISPR system of gene editing has greatly streamlined the generation of gene modifications (e.g., small substitutions, insertions, and deletions) in the mouse with high rates of success. Several editing platforms exist for gene/genome targeting in mice and other animal models previously difficult or impossible to alter. Here, we provide a simplified method of generating genetically modified mice using the prime editing platform. © 2024 Wiley Periodicals LLC. Basic Protocol 1: Design, cloning, and synthesis of engineered pegRNA (epegRNA) Basic Protocol 2: Microinjection of PE2 components into mouse zygote Basic Protocol 3: Genotyping founder mice and breeding for germline transmission.

Survival advantage of native and engineered T cells is acquired by mitochondrial transfer from mesenchymal stem cells

BackgroundApoptosis, a form of programmed cell death, is critical for the development and homeostasis of the immune system. Chimeric antigen receptor T (CAR-T) cell therapy, approved for hematologic cancers, retains several limitations and challenges associated with ex vivo manipulation, including CAR T-cell susceptibility to apoptosis. Therefore, strategies to improve T-cell survival and persistence are required. Mesenchymal stem/stromal cells (MSCs) exhibit immunoregulatory and tissue-restoring potential. We have previously shown that the transfer of umbilical cord MSC (UC-MSC)-derived mitochondrial (MitoT) prompts the genetic reprogramming of CD3+ T cells towards a Treg cell lineage. The potency of T cells plays an important role in effective immunotherapy, underscoring the need for improving their metabolic fitness. In the present work, we evaluate the effect of MitoT on apoptotis of native T lymphocytes and engineered CAR-T cells.MethodsWe used a cell-free approach using artificial MitoT (Mitoception) of UC-MSC derived MT to peripheral blood mononuclear cells (PBMCs) followed by RNA-seq analysis of CD3+ MitoTpos and MitoTneg sorted cells. Target cell apoptosis was induced with Staurosporine (STS), and cell viability was evaluated with Annexin V/7AAD and TUNEL assays. Changes in apoptotic regulators were assessed by flow cytometry, western blot, and qRT-PCR. The effect of MitoT on 19BBz CAR T-cell apoptosis in response to electroporation with a non-viral transposon-based vector was assessed with Annexin V/7AAD.ResultsGene expression related to apoptosis, cell death and/or responses to different stimuli was modified in CD3+ T cells after Mitoception. CD3+MitoTpos cells were resistant to STS-induced apoptosis compared to MitoTneg cells, showing a decreased percentage in apoptotic T cells as well as in TUNEL+ cells. Additionally, MitoT prevented the STS-induced collapse of the mitochondrial membrane potential (MMP) levels, decreased caspase-3 cleavage, increased BCL2 transcript levels and BCL-2-related BARD1 expression in FACS-sorted CD3+ T cells. Furthermore, UC-MSC-derived MitoT reduced both early and late apoptosis in CAR-T cells following electroporation, and exhibited an increasing trend in cytotoxic activity levels.ConclusionsArtificial MitoT prevents STS-induced apoptosis of human CD3+ T cells by interfering with the caspase pathway. Furthermore, we observed that MitoT confers protection to apoptosis induced by electroporation in MitoTpos CAR T-engineered cells, potentially improving their metabolic fitness and resistance to environmental stress. These results widen the physiological perspective of organelle-based therapies in immune conditions while offering potential avenues to enhance CAR-T treatment outcomes where their viability is compromised.

Generation of RAB4A homozygous knockout induced pluripotent stem cell (iPSC) line

The RAB4A gene is a member of the largest group in the Ras superfamily of small GTPases, which regulate membrane trafficking. The encoded protein is associated with early endosomes and is involved in sorting and recycling. In this study, we generated induced pluripotent stem cells (iPSC) from a healthy individual by electroporation of peripheral blood mononuclear cells. We generated a RAB4A homozygous knockout human iPSC line via CRISPR/Cas9 gene editing. The iPSCs-RAB4A−/− had a normal karyotype, expressed pluripotency markers, and maintained trilineage differentiation potential.

Enhanced non-viral gene delivery via calcium phosphate/DNA co-precipitates with low-voltage pulse electroporation in NK-92 cells for immunocellular therapy.

Achieving high cell transfection efficiency is essential for various cell types in numerous disease applications. However, the efficient introduction of genes into natural killer (NK) cells remains a challenge. In this study, we proposed a design strategy for delivering exogenous genes into the NK cell line, NK-92, using a modified non-viral gene transfection method. Calcium phosphate/DNA nanoparticles (pDNA-CaP NPs) were prepared using co-precipitation methods and combined with low-voltage pulse electroporation to facilitate NK-92 transfection. The results demonstrated that the developed pDNA-CaP NPs exhibited a uniform diameter of approximately 393.9 nm, a DNA entrapment efficiency of 65.8%, and a loading capacity of 15.9%. Furthermore, at three days post-transfection, both the transfection efficiency and cell viability of NK-92 were significantly improved compared to standalone plasmid DNA (pDNA) electroporation or solely relying on the endocytosis pathway of pDNA-CaP NPs. This study provides valuable insights into a novel approach that combines calcium phosphate nanoparticles with low-voltage electroporation for gene delivery into NK-92 cells, offering potential advancements in cell therapy.

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.

Integrating electromagnetic cancer stress with immunotherapy: a therapeutic paradigm.

An array of published cell-based and small animal studies have demonstrated a variety of exposures of cancer cells or experimental carcinomas to electromagnetic (EM) wave platforms that are non-ionizing and non-thermal. Overall effects appear to be inhibitory, inducing cancer cell stress or death as well as inhibition in tumor growth in experimental models. A variety of physical input variables, including discrete frequencies, amplitudes, and exposure times, have been tested, but drawing methodologic rationale and mechanistic conclusions across studies is challenging. Nevertheless, outputs such as tumor cytotoxicity, apoptosis, tumor membrane electroporation and leak, and reactive oxygen species generation are intriguing. Early EM platforms in humans employ pulsed electric fields applied either externally or using interventional tumor contact to induce tumor cell electroporation with stromal, vascular, and immunologic sparing. It is also possible that direct or external exposures to non-thermal EM waves or pulsed magnetic fields may generate electromotive forces to engage with unique tumor cell properties, including tumor glycocalyx to induce carcinoma membrane disruption and stress, providing novel avenues to augment tumor antigen release, cross-presentation by tumor-resident immune cells, and anti-tumor immunity. Integration with existing checkpoint inhibitor strategies to boost immunotherapeutic effects in carcinomas may also emerge as a broadly effective strategy, but little has been considered or tested in this area. Unlike the use of chemo/radiation and/or targeted therapies in cancer, EM platforms may allow for the survival of tumor-associated immunologic cells, including naïve and sensitized anti-tumor T cells. Moreover, EM-induced cancer cell stress and apoptosis may potentiate endogenous tumor antigen-specific anti-tumor immunity. Clinical studies examining a few of these combined EM-platform approaches are in their infancy, and a greater thrust in research (including basic, clinical, and translational work) in understanding how EM platforms may integrate with immunotherapy will be critical in driving advances in cancer outcomes under this promising combination.

Advancements in irreversible electroporation ablation technology for treating atrial fibrillation

Pulsed field ablation (PFA), an emerging treatment method for atrial fibrillation, has demonstrated significant potential in arrhythmia therapy. PFA employs high-intensity, short-duration electric fields to induce irreversible electroporation in myocardial cells, disrupting abnormal cardiac rhythms, and restoring normal heart function. This technique exhibits high success rates and low recurrence rates in animal studies and early clinical trials, offering advantages over traditional methods by reducing damage to adjacent structures such as the esophagus, phrenic nerve, and pulmonary veins. This review examines PFA’s application mechanisms, benefits, key operational parameters and the design and safety of related ablation devices. It emphasizes PFA’s potential to enhance both the efficacy and safety of atrial fibrillation treatment and explores future research directions and technological developments.

Analysis of In Situ Electroporation Utilizing Induced Electric Field at a Wireless Janus Microelectrode.

In situ electroporation, a non-invasive technique for enhancing the permeability of cell membranes, has emerged as a powerful tool for intracellular delivery and manipulation. This method allows for the precise introduction of therapeutic agents, such as nucleic acids, drugs, and proteins, directly into target cells within their native tissue environment. Herein, we introduce an innovative electroporation strategy that employs a Janus particle (JP)-based microelectrode to generate a localized and controllable electric field within a microfluidic chip. The microfluidic device is engineered with an indium tin oxide (ITO)-sandwiched microchannel, where the electric field is applied, and suspended JP microelectrodes that induce a stronger localized electric field. The corresponding simulation model is developed to better understand the dynamic electroporation process. Numerical simulations for both single-cell and chain-assembled cell electroporation have been successfully conducted. The effects of various parameters, including pulse voltage, duration medium conductivity, and radius of Janus microelectrode, on cell membrane permeabilization are systematically investigated. Our findings indicate that the enhanced electric intensity near the poles of the JP microelectrode significantly contributes to the electroporation process. In addition, the distribution for both transmembrane voltage and the resultant nanopores can be altered by conveniently adjusting the relative position of the JP microelectrode, demonstrating a selective and in situ electroporation technique for spatial control over the delivery area. Moreover, the obtained differences in the distribution of electroporation between chain cells can offer insightful directives for the electroporation of tissues or cell populations, enabling the precise and targeted modulation of specific cell populations. As a proof of concept, this work can provide a robust alternative technique for the study of complex and personalized cellular processes.

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.

Considering Joule heating in coupled electroporation and electrodeformation modeling of glioblastoma cells

Cells exposed to a pulsed electric field undergo electroporation(EP) and electrodeformation(ED) under electric field stress, and a coupled model of EP and ED of glioblastoma(GBM) taking into account Joule heating is proposed. The model geometry is extracted from real cell boundaries, and the effects of Joule heating-induced temperature rise on the EP and ED processes are considered. The results show that the temperature rise will increase the cell's local conductivity, leading to a decrease in the transmembrane potential(TMP). The temperature rise also causes a decrease in the dynamic Young's modulus of the cell membrane, making the cell less resistant to deformation. In addition, GBM cells are more susceptible to EP in the middle portion of the cell and ED in the three tentacle portions under pulsed electric fields, and the cells undergo significant positional shifts. The ED of the nucleus is similar to spherical cells, but the degree of ED is smaller.

Microfluidic Electroporation Arrays for Investigating Electroporation-Induced Cellular Rupture Dynamics.

Electroporation is pivotal in bioelectrochemistry for cellular manipulation, with prominent applications in drug delivery and cell membrane studies. A comprehensive understanding of pore generation requires an in-depth analysis of the critical pore size and the corresponding energy barrier at the onset of cell rupture. However, many studies have been limited to basic models such as artificial membranes or theoretical simulations. Challenging this paradigm, our study pioneers using a microfluidic electroporation chip array. This tool subjects live breast cancer cell species to a diverse spectrum of alternating current electric field conditions, driving electroporation-induced cell rupture. We conclusively determined the rupture voltages across varying applied voltage loading rates, enabling an unprecedented characterization of electric cell rupture dynamics encompassing critical pore radius and energy barrier. Further bolstering our investigation, we probed cells subjected to cholesterol depletion via methyl-β-cyclodextrin and revealed a strong correlation with electroporation. This work not only elucidates the dynamics of electric rupture in live cell membranes but also sets a robust foundation for future explorations into the mechanisms and energetics of live cell electroporation.

The Advancement and Utilization of Marx Electric Field Generator for Protein Extraction and Inducing Structural Alterations

This study introduces an innovative two-range, 12-stage Marx pulse generator employing thyristor switches designed specifically for the electroporation of biological cells. The generator consists of two module capacitors of different capacitances (1 μF and 0.25 μF), which enable the generation of electrical pulses with different durations and amplitudes of up to 25 kV. Safety aspects, including overcurrent and overvoltage protection mechanisms, are implemented in both the software and the hardware. In the experimental section, the tests of the Marx generator with resistive load are described in detail, and the results for the voltage fluctuations, pulse duration, and output characteristics of the generator are presented. The advantages of the design, including the high output voltage, the wide range of repetition rates, and the flexibility of the pulse parameters, are emphasized. Additionally, the research showcases the utilization of the devised generator for industrial purposes. Hence, an investigation into the efficiency of protein extraction from microalgae (Chlorella vulgaris) and the impacts of pulsed electric fields (PEFs) on the structural characteristics of casein micelles (CSMs) was chosen as an illustrative example. The obtained results provide valuable insights into the application of PEF in food processing and biotechnology and underline the potential of the developed generator for sustainable and environmentally friendly practices.

The comparison of the dynamics of Ca2+ and bleomycin intracellular delivery after cell sonoporation and electroporation in vitro

Ca2+, in combination with SP or EP, induces cell cytotoxicity much faster compared to BLM. The application of BLM in combination with, SP or EP, reaches the level of cell death, induced by similar combination with Ca2+, only after 72 h. The methods of SP and EP were calibrated according to the level of differential cytotoxicity, determined after 6 days (using cell clonogenic assay). The combination of Ca2+ SP induces cell death faster than Ca2+ EP – after Ca2+ SP it increases to a maximum level after 15 min and remains constant for up to 6 days, while the cytotoxic efficiency after Ca2+ EP increases to the level of Ca2+ SP only after 72 h. The combination of BLM SP shows a very similar dynamics to BLM EP – both reach maximal level of cytotoxicity after 48–72 h.Ca2+ and BLM in combination with SP have shown similar levels of cytotoxicity at higher acoustic pressures (≥250 kPa); therefore, Ca2+ SP can be used to induce immediate and maximal level of cytotoxic effect. The faster cytotoxic efficiency of Ca2+ in combination with SP than EP was determined to be due to the involvement of microbubble inertial cavitation.

Modeling of Cell Electroporation With Emphasis on Ions Transport-Induced Dynamic Alterations in Membrane Conductivity

Modeling of Cell Electroporation With Emphasis on Ions Transport-Induced Dynamic Alterations in Membrane Conductivity

Generation of KCNH2 heterozygous knockout induced pluripotent stem cell (iPSC) line (Long and Short QT Syndrome)

KCNH2 (Potassium Voltage-Gated Channel Subfamily H Member) encodes a voltage-activated potassium channel role as rapidly activating-delayed rectifier potassium channel that plays an essential role in the final repolarization of the ventricular action potential. Mutations in this gene can cause long QT syndrome and short QT syndrome. Transcript variants encoding distinct isoforms were also identified. In this study, we generated induced pluripotent stem cells (iPSC) from a healthy individual by electroporation of peripheral blood mononuclear cells and generated a KCNH2 heterozygous knockout human iPSC line via CRISPR/Cas9 gene editing. The resulting iPSCs had a normal karyotype, were free of genomically integrated epitomal plasmids, expressed pluripotency markers, and maintained trilineage differentiation potential.

Abstract 6640: Zap! Irreversible electroporation in pancreatic cancer cells induces proinflammatory cell death and improves progression free survival

Abstract Pancreatic cancer is the third leading cause of cancer-related death due to its late diagnosis, few treatment options, and extremely low survival rates due to its immunosuppressive microenvironment. Irreversible electroporation (IRE),is a tumor non-thermal ablation technique that applies high-voltage electrical pulses to permeabilize the cell membrane that is irreversible and leads to necrotic cell death. IRE has had strong success in clinical trials nevertheless, little is known about the effect of IRE beyond the initiation of cell death in the tumor tissue. We hypothesize that IRE can not only induce cell death by membrane disruption to facilitate a proinflammatory microenvironment but also causes death via cell cycle arrest. We utilized in-vitro, and immunocompetent in-vivo murine models to investigate changes to pancreatic cancer and the tumor microenvironment. Our findings show that treating the cells with an increasing voltage of IRE induces more cell death. We observed cell cycle arrest in the G1 phase which may have been caused by DNA damage. There was upregulation of DNA damage repair machinery in the treated samples. Our histopathology data suggests increased necrosis on the tumor treated with IRE, as also PCR array analysis showed more proinflammatory cell death caused by pyroptosis. Finally, we demonstrated a reduction in the tumor size and a better survival rate aftertreatment. Our data suggest that Irreversible electroporation has a promising future in inducing proinflammatory type of cell death in the tumor and benefits increased patient survival. More investigation is needed to define ideal parameters for the electrical pulses and device development for tumor ablation in larger animals like porcine models. Citation Format: Manali Powar, Khan Mohammad Imran, Irving C. Allen, Rafael V. Davalos, Zaid Salamah, Kenneth Aycock, Rebecca M. Brock, Sheryl Coutermarsh-Ott, Katie Orr. Zap! Irreversible electroporation in pancreatic cancer cells induces proinflammatory cell death and improves progression free survival [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6640.

Quantitative evaluation method of triboelectric nano-electroporation based on electrical impedance spectroscopy

Electroporation refers to a biophysical process wherein cell permeability is altered by inducing pores of the cell membrane through application of an electric field. It has found extensive applications in cancer therapy and gene delivery. The alteration of electrical impedance during electroporation was investigated by synergistically employing a self-powered triboelectric nanogenerator (TENG) and electrical impedance spectroscopy (EIS), thus serving as a theoretical foundation for real-time efficacy assessment of electroporation in clinical settings. A self-powered electroporation experimental system was established, and the findings demonstrated that upon reaching the threshold level of cell transmembrane potential, the application time of electric field led to electroporation of the cells, resulting in a decline in the impedance value of the blood. As the electric field persists, the impedance value gradually diminishes; when the blood impedance value changes insignificantly, the degree of cell electroporation is saturated. During the process of tumor cell electroporation, the relationships between HeLa cell impedance Z* and time T and cell concentration C is Z* = 7×10−11T5- 4×10−8T4+9×10−6T3-8×10−4T2+0.0117T-9.3587 C+124.2. Simultaneously, the occurrence and reversibility of electroporation were verified through Propidium Iodide (PI) fluorescence staining. The numerical calculations presented in this paper are based on the extended HANAI theory. Furthermore, the simulations were conducted through finite element analysis and perform equivalent circuit fitting on the experimental data. The numerical and simulation results are consistent with the experimental findings, validating the study's establishment of a quantifiable relationship between electroporation degree and cell parameter Z*、Rc、Xc and RRBC. As evidenced, the alteration in electrical impedance spectra can effectively indicate the extent of electroporation. Therefore, it is expected that employing the EIS approach will become an effective means to measure the therapeutic efficacy of electroporation in future clinical applications.