This review explores the emerging integration of natural agents with electroporation (EP) techniques in oncology. Natural compounds derived from plants, microbes, and marine organisms hold promise for cancer treatment due to their multifaceted mechanisms of action, including apoptosis induction and inhibition of tumor progression. EP, a method that enhances cell membrane permeability, facilitates the efficient intracellular delivery of these bioactive compounds. This synergistic approach has demonstrated significant efficacy in preclinical studies and is progressing toward clinical application, particularly for cancers resistant to conventional therapies. We provide an in-depth analysis of this innovative therapeutic strategy’s mechanisms, preclinical evidence, and clinical progress, highlighting its potential to transform cancer treatment paradigms. Future research directions are proposed to optimize and personalize these integrative therapies.
Bioelectric signaling regulates development, regeneration, and cancer suppression. This computational study introduces a minimal control-logic framework bridging ion channel pharmacology to multicell bioelectric patterning in regeneration (e.g., Xenopus limbs, planarian polarity). Using dimensionless ordinary differential equations anchored to pharmacological timescales (e.g., propranolol PK, hours), we modeled three ion channel drugs (amiloride, propranolol, ivermectin) in single cells, embedded them in gap-junction-coupled 10 × 10 tissue lattices, and applied DEAP evolutionary optimization to recover target Vmem patterns. Multiple (k 1 , k 2 , k 3 ) parameter sets yielded identical tissue-scale Vmem patterns, despite perturbations such as ivermectin-induced 2 × k 3 Cl⁻ scaling. This parameter robustness positions tissues as active controllers maintaining patterning goals amid variability. Simulated legacy drugs controlled tissue-level bioelectric/voltage goal states, and different microscopic configurations were able to reach a target voltage state following in silico evolutionary optimization of a few parameters. New predictions include depolarizing Na + blockade inducing ectopic posteriorization and hyperpolarizing Cl⁻ activation restoring anterior–posterior Vmem gradients in planarian regeneration. This is falsifiable with CC2-DMPE dyes. These simulations could reframe legacy ion-channel drugs as bioelectric goal-state controllers beyond single-target inhibition, providing an extensible quantitative systems pharmacology pipeline (GitHub: Cameron-99/bioelectric-qsp-model) for multi-physics models and bioelectricity research.
A widespread energy crisis is the result of the current global situation, which is characterized by high-energy needs and finite resources. Nonrenewable energy sources are fast depleting while green energy options are still being ignored. Alternative energy generation techniques are desperately needed. Recently, there has been a lot of interest in microbial fuel cells (MFCs) due to their favorable operating conditions and the affordability of a variety of eco-friendly fuel substrates. Bioelectricity is created by microorganisms actively breaking down substrates, providing a long-term answer to the growing energy problems. Numerous investigations have yielded fresh insights into MFCs, demonstrating that a broad range of microorganisms can be effectively employed with a broad range of carbon sources, including waste materials. Therefore, the microbial transformation of waste through advanced bioremediation processes such as MFCs, which provide a potentially attractive alternative to conventional methods utilized in wastewater treatment, enables the immediate generation of bioelectricity. This not only keeps up with the latest technological advancements but also reduces overall process expenses. The current status of MFCs, their setup and mode of operation, factors for optimizing MFCs, the range of waste kinds, and microorganisms suitable for generating bioelectricity are all objectively assessed in this review. The review comprehensively focuses on MFC’s processes as applied in bioelectricity generation and environmental sustainability.
Atherosclerosis (AS) remains the primary pathological driver of cardiovascular diseases (CVDs), which account for ∼32% of global deaths. Early diagnosis of AS is critical for reducing mortality, but conventional diagnostic tools fail to dynamically assess oxidative stress—a key pathogenic factor in AS progression. Reactive oxygen species (ROS) are abnormally elevated in early AS plaques, making them a promising biomarker for early detection. However, existing ROS-responsive probes suffer from poor targeting specificity and insufficient sensitivity. RGD-targeted ROS-responsive dual-functional molecular probes, which integrate integrin-binding specificity and ROS-triggered signal activation, have emerged as a novel solution to these limitations. This review comprehensively summarizes the design principles, synthesis strategies, and preclinical evaluation ( in vitro and in vivo ) of RGD-targeted ROS-responsive probes for AS imaging. We focus on probe optimization strategies, including structural modification of RGD ligands, selection of ROS-sensitive moieties, and integration of fluorophores with optimal optical properties. Additionally, we discuss the current challenges in clinical translation and future directions for improving probe performance, with a dedicated focus on the potential applications of these probes in bioelectricity research—an emerging intersection of oxidative stress and vascular physiology. A systematic analysis of PubMed-listed literature (2020–2024, 77 articles; literature screening criteria: keywords including “atherosclerosis”, “RGD”, “ROS-responsive”, “molecular probe”, “targeted imaging”; excluding reviews, case reports, and studies with incomplete data) highlights the potential of these dual-functional probes to address unmet clinical needs in early AS diagnosis and bioelectricity-related research, providing insights for researchers in the fields of molecular imaging, cardiovascular medicine, and bioelectricity.
Growing insight into bioelectric regulation of cell behavior has driven extensive use of electrical stimulation in in vitro studies over the last decade, notably in the enhancement of stem cell maturation. This has prompted the development of electrical stimulation bioreactors to provide the stimulation parameters required for such studies. However, biological outcomes are highly sensitive to stimulation parameters, electrode geometry, and electrode-electrolyte interface. These are factors that current commercial and custom systems handle differently; commercial platforms are robust but often cost-prohibitive, whereas custom open-source systems are cheaper but typically lack validated, user-friendly hardware and standardization. In this perspective, we argue for openly documented electrical stimulation platforms that are not only published but also made available through open-source science manufacturers. Coupling transparent designs with distributed assembly and open validation data offers the potential to lower current barriers to entry, enabling wider adoption of pre-characterized, affordable systems that improve reproducibility and mechanistic clarity.
Objective: Anaerobic digestate is rich in nitrogen, and its treatment poses substantial challenges to conventional processes due to its ammonium concentration, high alkalinity, and low biodegradable carbon content. In this study, microbial fuel cells (MFCs) were investigated for the treatment of digestate. Methods: An orthogonal experimental design was employed to evaluate the effects of influent ammonium concentration (100–300 mg/L), pH (8–10), and external resistance (100–1000 Ω) on nitrogen removal and electricity generation. Results: The MFC consistently achieved high nitrate removal efficiencies (>97%), whereas ammonium removal was strongly condition-dependent. The highest ammonium removal efficiency (66.6%) and total nitrogen removal efficiency (84.8%) were obtained at 300 mg/L NH 4 + , pH 10, and an external resistance of 1,000 Ω. Although pH was identified as the dominant factor influencing performance, microbial community analysis revealed the enrichment of Paracoccus and Thauera , suggesting that nitrogen removal was mainly governed by the coupling of heterotrophic nitrification–aerobic denitrification with bioelectrochemical processes. Conclusions: This work demonstrates the feasibility of using MFCs for the treatment of high-strength digestate and provides valuable insights into the optimization of nitrogen-rich wastewater management.
Microbial fuel cells (MFCs) are bioelectric devices that use bacterial metabolic activity to convert chemical energy to electricity, which offers a potential way to green energy production. Exoelectrogenic bacteria that can transfer their metabolic electrons outside their cells are the primary contributing organisms in this process. This study aims to screen and identify exoelectrogenic bacteria for electricity generation in MFCs. Seawater samples were collected from Kuakata, Bangladesh, and a dual-chambered MFC was used to detect the presence of exoelectrogenic bacteria in the sample. The exoelectrogenic bacterium was screened and isolated from the anode biofilm of the MFC using an MnO 2 -supplemented agar medium. The ability of electricity generation by the isolated bacteria was also determined in a dual-chambered MFC. Then, the isolated exoelectrogen was identified via biochemical assay and 16S rRNA gene sequencing. The isolated exoelectrogenic bacteria generated a voltage of up to 0.341 volts, a current density of 0.044 mA/cm 2 , and a maximum power density of 0.015 mW/cm 2 in the MFC. Biochemical assay and sequencing analysis reported the bacteria as Pseudomonas aeruginosa (similarity index: 99.45%), which demonstrates potential for bioelectricity and sustainable energy production, contributing to novel, eco-friendly resources for biofuel production in Bangladesh.
Spinal cord injury (SCI) disrupts neurological pathways controlling movement, sensation, and autonomic processes, commonly causing long-term impairments. The restoration of damaged spinal tissue continues to pose significant therapeutic challenges. The RISEUP project explores an innovative combinatorial therapy that integrates stem cell transplantation with & micro;sPEFs delivered by an Electro-Pulsed Bio-Hybrid (EPB) device. This in silico study faithfully reproduces the in vivo experimental setup that will be applied within the project to assess EPB functionality and stimulation effects. By positioning the EPB numerical model over the thoracic vertebral region of interest in the ViZOO Neurorat anatomical rat model, and by implementing laminectomy and SCI, a virtual replica of the in vivo experiments is proposed, aiming to inform, in a prospective way, whether electric (E-)field intensities estimated by in vitro studies and microdosimetry and shown to be suitable for electroporation could be reached in vivo, in the complex and evolving environment of the injured spinal cord. To reflect the biological evolution of SCI over the stimulation time, two lesion phases were simulated. First, acute SCI represents the immediate post-injury environment, while chronic models capture the later phase characterized by altered conductivity and structural reorganization. This dual-phase approach allows for a more accurate assessment of the experiments. Finally, dosimetric assessment was computed and evaluated in terms of induced E-field distribution, and peripheral nerves response. Peak E-fields beneath the EPB ranged from 20 to 70 kV/m, sufficient for transient membrane permeabilization and stem cell activation. Field penetration improved in chronic lesions due to increased conductivity, with deeper median fields (similar to 2 kV/m) compared to acute models. Neurofunctional safety was evaluated by coupling the E-field output to axonal electrophysiological models of intercostal nerves. The findings demonstrate that the EPB device can deliver effective, localized stimulation with minimal off-target neural activation. Furthermore, temporal lesion evolution influences field distribution, underscoring the importance of phase-specific assessment.
Low-energy micro- and millisecond electric pulses (EPs) charge and depolarize the cellular plasma membrane (PM) below the electroporation threshold. Conversely, individual nanosecond EPs (NSEPs) are too brief to initiate PM depolarization by charging the cells. It is hypothesized that a single NSEP induces cell depolarization via PM electroporation upon application of high-power EPs. However, low-energy NSEP bursts with a very high pulse repetition frequency may prevent electroporation. This method, a temporal summation of NSEPs, results in subsequent PM charging and depolarization. To visualize PM voltage changes during low-energy EPs, we employed optical measurements using FluoVolt™ (an organic fluorescent reporter of membrane potential [MP]) and a custom-made streak imaging system. Ultra-fast streak kymographs depicting MP changes were obtained after exposure to a ∼0.2 kV/cm single 200 µs EP and 5 MHz trains of 1000 and 2000 NSEPs with 100 ns duration. Immediately following exposure, a small FluoVolt™ response (up to ∼7% fluorescence change) was observed in the PM areas facing electrodes. The response duration directly correlated with the pulse width (PW) or duration of the NSEPs burst interval. The single 200 µs EP was more effective at charging the PM than an equivalent-energy 5 MHz burst of 2000 NSEPs of 100 ns duration. Furthermore, similar amplitudes of PM fluorescence changes between ∼0.2 kV/cm bursts of 1000 and 2000 NSEPs suggest that increasing either PW or applied voltage is necessary to enhance the extent of PM depolarization. Nonetheless, the modest depolarization effect reported herein was sufficient to open voltage-gated Ca2+ channels in neurons. These findings indicate that a 5 MHz burst of low-energy NSEPs and single µs EPs effectively induce PM depolarization and Ca2+ responses without causing any cellular damage.
Background: All cells, including bacteria, maintain an ion gradient across the plasma membrane. The resulting membrane potential is important for cell growth, division, and health. Our current research shows that a subset of Bacillus subtilis (B. subtilis) cells exhibit spontaneous, rapid (similar to 2 min) "spikes" in the membrane potential. The subset of cells that spike increases, and the spikes become more frequent, when cells are exposed to ethanol, an environmental stressor. Our goal is to highlight the association between membrane potential and cell elongation. Methods: To achieve this, membrane potential was measured using tetramethyl rhodamine methyl ester, a voltage-sensing fluorescent dye. Cell fluorescence and elongation were measured simultaneously over a 30-min period using fluorescence and bright field microscopy. Images were analyzed using a combination of ImageJ and custom MATLAB software. Results: We find that the population of cells that spike have slower elongation rates than those with no spikes in both the untreated and ethanol-treated cells over the 30 min of observation.
Surgeons face serious challenges removing cancer fully during surgery, with incomplete cancer removal posing significant risks to patients. To address this problem, Lucell Diagnostics Inc. is developing an innovative cancer detection platform called membrane voltage profiling (MVPro; patent pending). This groundbreaking method exploits the discovery that cancer cells exhibit a physiological biomarker, depolarization, as revealed by fluorescent voltage-sensitive dyes. Cancer cells fluoresce in specific patterns and intensities that differ from normal cells, allowing precise identification. We present here our preliminary results on the feasibility of using voltage to locate cancer cells. The aims were: to perform controls showing whether MVPro affects the normal pathology process; to optimize tissue transfer for margin cell collection; to confirm that the VSD DiBAC(4)(3) is appropriate for MVPro of skin cells; to determine whether MVPro finds cancer in the same specimens as pathological analysis. These studies on nonmelanoma skin cancer specimens reveal that MVPro is low risk to the patient, integrates with existing surgical protocols, finds cancer in the same specimens as does pathology, and presents no complications for pathological analysis. Once development of this methodology is complete, MVPro will yield an annotated, 2D heat map covering the entire surgical margin, indicating the location of cells with a high likelihood of being cancer. This will empower surgeons to confirm a negative surgical margin before closing. This simple type of intraoperative imaging, performed in or near the operating theater, has the potential to improve surgical outcome, cut health care costs, and enhance post-surgical quality of life by preserving healthy tissue. Because the setup costs are relatively small and the reagents are inexpensive, we believe MVPro could be of great benefit to underserved areas. Indeed, MVPro could benefit health care systems globally, from cutting-edge hospitals to small clinics in underserved regions.
This study combines microdosimetry techniques and computational models to investigate the effects of ultrashort pulsed electric fields (PEFs) on cellular membranes. It focuses on identifying optimal stimulation protocols to meet RISEUP project goals, where an implantable electro pulsed bio-hybrid (EPB) device is under development for spinal cord injury neurogenesis. The EPB employs PEFs stimulation to modulate intracellular calcium fluxes, promoting stem cell proliferation and differentiation, by targeting plasma and endoplasmic reticulum (ER) membranes via electroporation. This approach integrates cutting-edge research to advance neurogenesis using mesenchymal stem cells (MSCs) and induced neuronal stem cells (iNSCs). In this work, starting from high-resolution confocal microscopy images, a semi-automatic reconstruction procedure is employed to generate 3D virtual digital twins of iNSCs and MSCs, incorporating their subcellular structures. Microdosimetric simulations are conducted to model the effects of various bipolar pulse intensities (9, 12, 15 V) and durations (10, 100, 1000 & micro;s) on a mixture of virtual stem cells within the EPB device. At 12 V, a 10 & micro;s-bipolar pulse is estimated able to porate plasma membranes, whereas increasing the pulse duration to 1000 & micro;s results in ER electropermeabilization, showing that, at given pulse intensities, adjusting the pulse duration allows poration of both plasma and ER membranes. This strategy is particularly important when voltage cannot be increased, such as in RISEUP, where the fixed onboard power source limits voltage modulation. In such cases, pulse duration becomes a key parameter for achieving the desired membrane poration effects. Furthermore, advanced 3D virtual cells are undeniable in microdosimetry to optimize innovative stimulation protocols aimed at targeting specific cellular compartments.
RISEUP, short for Regeneration of Injured Spinal Cord by Electro Pulsed Bio-Hybrid Implant, is a European project aimed at offering a real possibility of recovery after spinal cord injury. This approach leverages highly intense microsecond pulsed electric fields (& micro;sPEFs) to transiently permeabilize cellular membranes, thereby guiding stem cells proliferation and differentiation pathways. In vitro experiments are underway to develop an optimized stimulation protocol. An initial experimental phase employed faced electrodes (FE), while a second phase uses cylindrical electrodes (CE), corresponding to electro pulsed bio-hybrid (EPB) device - Prototype 0. Since the two configurations generate different electric (E-) field distributions, a numerical calibration of the voltage applied to the CE system was required. The objective of this calibration is to ensure that the E-field intensities generated between the CE electrodes match those previously shown to induce electroporation responses in stem cells with the FE setup. Numerical models of both FE and CE were implemented in COMSOL Multiphysics, and a calibration procedure was fine-tuned to ensure that the CE setup produced an average electric field comparable to the 30 kV/m value achieved by the FE system. The calibration process adjusted the applied voltage in the CE system from the nominal 15 V to 35.4 V to achieve a median electric field of 30 kV/m within the extracellular matrix volume that will host the stem cells. Following this, a microdosimetric study using advanced 3D virtual stem cells, including intracellular compartments, was performed to validate the calibration. The study confirmed that the two systems produced comparable electroporation effects, as evidenced by similar transmembrane potentials and pore densities on both plasma and intracellular membranes. The developed calibration methodology effectively harmonizes the stimulation conditions between different electrodes technologies employed in the RISEUP project, providing crucial support for optimizing in vitro protocols during experimental phases.
Background and Objectives: Electroporation ablation is a promising nonsurgical and minimally invasive technique for tumor ablation; however, no monitoring is currently available. In this article, we present recent advances in the numerical workflow toward a peroperative numerical evaluation of clinical irreversible electroporation (IRE) procedures of liver tumors. The objective of this study is to propose an updated numerical workflow for the digital twin of electroporation ablation, to provide relevant information to physicians performing IRE for hepatocellular carcinoma (HCC). Methods: The workflow consists of four main steps: (1) an image registration algorithm to align the contrast-enhanced cone beam computed tomography (CBCT), where the region of interest are visible, with the lower-quality CBCT acquired after needle insertion; (2) extraction of needles position by manual selection directly on the CBCT containing the needles; (3) accurate and efficient numerical computation of the electric field (EF) distribution, using a static linear model and the finite difference method to simulate the EF at the maximum voltage applied between each electrode pair; and (4) numerical assessment of the tumor coverage by the 3D EF. Results: We propose a criterion for electrical heterogeneity of the medium near the electrode thanks to the measurements provided by the Nanoknife IRE device. The full protocol was tested on three representative patients with nodular HCCs <5 cm. The complete numerical workflow, from image registration and needle detection to the computation, requires at most less than 15 min following image acquisitions, making it suitable for clinical use. Interestingly, the number of finite difference computations increases linearly with the number of needles N, despite the number of electrode pairs increasing as N ( N - 1 ) / 2 . In addition, the pulse amplitude can be modified without the need for full recomputation, enabling online adjustment of the treatment plan.
