Transmembrane Potential Of Cells Research Articles

Reconstructing ventricular cardiomyocyte dynamics and parameter estimation using data assimilation

Cardiac ventricular myocyte action potential dynamics are regulated by intricate and nonlinear interactions between the cell transmembrane potential and ionic currents and concentrations. Present technology limits the ability to measure transmembrane potential and multiple ionic currents simultaneously, which narrows the scope of experiments to provide a complete snapshot of the cardiac myocyte state. This limitation presents an obstacle for understanding how perturbations can trigger arrhythmias and more broadly how the myocyte responds to different conditions, such as changes in pacing rate or responses to drug treatment. In this study, we demonstrate that a data-assimilation approach can successfully reconstruct and predict the dynamics of a heterogeneous virtual cardiac ventricular myocyte population in the presence of parameter uncertainty. A population of heterogeneous cardiac ventricular myocytes is generated by varying ionic current conductance parameters, and additional observational uncertainty is mimicked by the addition of Gaussian noise to the transmembrane potential. We demonstrate that the data-assimilation approach accurately reconstructs transmembrane potential, with error less than the magnitude of the noise. Further, the data-assimilation approach successfully estimates the conductances of ionic currents generally with high accuracy and requiring low computational time. As a proof of concept, we apply the data-assimilation approach to reconstruct action potential dynamics from optical mapping experiments in an ex vivo isolated guinea pig heart. Critically, we demonstrate that the ionic conductance parameters estimated from a recording at one pacing frequency can accurately predict action potential dynamics at different rates.

A Many-Shells Model for Cell Transmembrane Potentials for Pulsed Electric Field Applications

A Many-Shells Model for Cell Transmembrane Potentials for Pulsed Electric Field Applications

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.

The Mechanism of Antimicrobial Activity of Conjugated Bile Acids against Lactic Acid Bacilli.

The mechanism underlying antimicrobial activity of conjugated bile acids against strains of lactic acid bacilli is not well understood. The purpose of this study was to investigate two typical conjugated bile acids (glycochenodeoxycholic acid and taurochenodeoxycholic acid) for their mechanisms of antimicrobial activity against four strains of different species of lactic acid bacilli at the physiological pH of the small intestine of humans. The bacterial cell membrane integrity, transmembrane potential, and transmembrane pH gradient were examined using the fluorescence probes SYTO 9 plus propidium iodide, 3,3'-dipropylthiadicarbocyanine iodide, and 5(6)-carboxyfluorescein diacetate N-succinimidyl ester, respectively. The intracellular ATP levels were measured by the firefly luciferase-based bioluminescence method. It was found that the antimicrobial activity of conjugated bile acids against the strains of lactic acid bacilli is strain-specific, and glycochenodeoxycholic acid showed significantly greater antimicrobial activity than taurochenodeoxycholic acid against the strains of lactic acid bacilli. The conjugated bile acids inhibited the growth of strains of lactic acid bacilli by disrupting membrane integrity, dissipating transmembrane potential, reducing the transmembrane pH gradient, and depleting intracellular ATP. In conclusion, the antimicrobial activity of conjugated bile acids against lactic acid bacilli is a multifactorial phenomenon. This study will provide valuable information for developing strategies to improve the ability of lactic acid bacilli to tolerate bile in vivo.

Pulsed Electric Field Ablation of Esophageal Malignancies and Mitigating Damage to Smooth Muscle: An In Vitro Study.

Cancer ablation therapies aim to be efficient while minimizing damage to healthy tissues. Nanosecond pulsed electric field (nsPEF) is a promising ablation modality because of its selectivity against certain cell types and reduced neuromuscular effects. We compared cell killing efficiency by PEF (100 pulses, 200 ns-10 µs duration, 10 Hz) in a panel of human esophageal cells (normal and pre-malignant epithelial and smooth muscle). Normal epithelial cells were less sensitive than the pre-malignant ones to unipolar PEF (15-20% higher LD50, p < 0.05). Smooth muscle cells (SMC) oriented randomly in the electric field were more sensitive, with 30-40% lower LD50 (p < 0.01). Trains of ten, 300-ns pulses at 10 kV/cm caused twofold weaker electroporative uptake of YO-PRO-1 dye in normal epithelial cells than in either pre-malignant cells or in SMC oriented perpendicularly to the field. Aligning SMC with the field reduced the dye uptake fourfold, along with a twofold reduction in Ca2+ transients. A 300-ns pulse induced a twofold smaller transmembrane potential in cells aligned with the field, making them less vulnerable to electroporation. We infer that damage to SMC from nsPEF ablation of esophageal malignancies can be minimized by applying the electric field parallel to the predominant SMC orientation.

Reconstructing action potential and ionic current dynamics of a mammalian ventricular cell using data assimilation.

The dynamics of the cardiac ventricular myocyte action potential (AP) is regulated by intricate and nonlinear interactions between the cell transmembrane potential, and ionic currents and concentrations. Present technology limits the ability to simultaneously measuring multiple ionic currents and concentration during an AP in in vitro single cell experiments, which limits the scope of experimental data to provide a complete snapshot of the cardiac myocyte system state. This limitation presents an obstacle for understanding how cellular perturbations can trigger arrhythmogenic conditions. In this study, we applied a data assimilation approach, which combines limited noisy observations with predictions from a computational model, paired with a parameter estimation to minimize the effects of parameter uncertainty and model error. We performed a series of in silico experiments, in which a forecasting system was tasked with reconstructing AP dynamics of a virtual ventricular myocyte in the presence varying degrees of parameter and model error. With appropriate protocol design, we find that a data assimilation approach can successfully reconstruct and predict the dynamics of not only the directly observed state variables (such as the transmembrane potential and a selected ionic current), but also unobserved state variables, such as unmeasured ionic currents and concentrations. As an extension of the results of these series of in silico experiments, we generated simulations to assess which ionic currents, when measured, offer greater predictive ability using the data assimilation algorithm. This can offer valuable information for the design of single cardiac in vitro experiments, as it is not feasible to measure all ionic currents and concentrations across time within the same cell during an AP. Future work will investigate the longer-term accuracy of this approach predicting potentially pro-arrhythmic perturbations for arrhythmia risk assessment.

Simulation study of cell transmembrane potential and electroporation induced by time-varying magnetic fields

In this work, an improved simulation model was proposed to assess the transmembrane potential (TMP) evolution on the cellular membrane exposed to time-varying magnetic fields (TMFs). Comparatively, we extended the research on TMP induced by TMF to the electroporation phenomenon by introducing the Smoluchowski function, thereby predicting the occurrence of electroporation. The simulation results based on our numerical model showed that with exposure to the sub-microsecond trapezoidal pulsed magnetic field (PMF), the pore density did not reach the conventional electroporation criterion (1014m−2) even if the TMP exceeded the electroporation threshold (1 V); however, with the same energy import, it was easier for the nanosecond pulse to electroporate the membrane evidenced by higher pore density. Further, the capability of predicting the occurrence of electroporation was verified by extending our simulation model to compare experimental results. The comparative analysis showed that our simulation model has predictive and guiding significance for experimental studies and practical applications.

Three-dimensional transistor arrays for intra- and inter-cellular recording.

Electrical impulse generation and its conduction within cells or cellular networks are the cornerstone of electrophysiology. However, the advancement of the field is limited by sensing accuracy and the scalability of current recording technologies. Here we describe a scalable platform that enables accurate recording of transmembrane potentials in electrogenic cells. The platform employs a three-dimensional high-performance field-effect transistor array for minimally invasive cellular interfacing that produces faithful recordings, as validated by the gold standard patch clamp. Leveraging the high spatial and temporal resolutions of the field-effect transistors, we measured the intracellular signal conduction velocity of a cardiomyocyte to be 0.182 m s-1, which is about five times the intercellular velocity. We also demonstrate intracellular recordings in cardiac muscle tissue constructs and reveal the signal conduction paths. This platform could provide new capabilities in probing the electrical behaviours of single cells and cellular networks, which carries broad implications for understanding cellular physiology, pathology and cell-cell interactions.

Cell transmembrane potential in contactless permeabilization by time-varying magnetic fields

BackgroundAlthough experimental results proved the feasibility of using time-varying magnetic field as a contactless cells permeabilization method, the underlying mechanism is still poorly understood. In this study a numerical analysis of the time-dependent transmembrane potential (TMP) at cell membranes during permeabilization by time-varying magnetic fields was proposed, and a first quantification of mechanical stress induced by the magnetic and electric fields, hypothesized to play an important role in the permeabilization mechanism, was carried out. Methods: Starting from the simulation of real in vitro experimental conditions, the analysis was widened quantifying the influence of pulse frequency, cell dimension and distance of the cell from the magnetic field source. The mechanical pressure on cell membrane due to the interaction between free charges and induced electric field and due to the gradient of the magnetic field was quantified in all those conditions in which the TMP values were not high enough to cause membrane permeabilization. Results: TMP values induced by typical in-vitro experimental conditions were far below the values needed for membrane permeabilization, with a strong dependence on pulse frequency and distance of the cell from the coil. Conclusion: The preliminary assessment of the mechanical pressure on cell membrane showed that stress values evaluated in conditions in which TMP values were too low to cause membrane permeabilization were comparable to those known to influence the pores opening mechanisms. Results represent a significant step towards a better comprehension of the mechanism underlying cell membrane permeabilization by time-varying magnetic fields.

Predicting electrotransfer in ultra-high frequency sub-microsecond square wave electric fields

ABSTRACTMeasurement of cell transmembrane potential (TMP) is a complex methodology involving patch-clamp methods or fluorescence-based potentiometric markers, which have limited to no applicability during ultrafast charging and relaxation phenomena. In such a case, analytical methods are applied for evaluation of the voltage potential changes in biological cells. In this work, the TMP-based electrotransfer mechanism during ultra-high frequency (≥1 MHz) electric fields is studied and the phenomenon of rapid membrane charge accumulation, which is non-occurrent during conventional low-frequency electroporation is simulated using finite element method (FEM). The influence of extracellular medium conductivity (0.1, 1.5 S/m) and pulse rise/fall times (10–50 ns) TMP generation are presented. It is shown that the medium conductivity has a dramatic influence on the electroporation process in the high-frequency range of applied pulsed electric fields (PEF). The applied model allowed to grasp the differences in polarization between 100 and 900 ns PEF and enabled successful prediction of the experimental outcome of propidium iodide electrotransfer into CHO-K1 cells and the conductivity-dependent patterns of MHz range PEF-triggered electroporation were determined. The results of this study form recommendations for development and pre-evaluation of future PEF protocols and generators based on ultra-high frequency electroporation for anticancer and gene therapies.

HJH-1, a Broad-Spectrum Antimicrobial Activity and Low Cytotoxicity Antimicrobial Peptide.

With the overuse of antibiotics, multidrug-resistant bacteria pose a significant threat to human health. Antimicrobial peptides (AMPs) are a promising alternative to conventional antibiotics. This study examines the antimicrobial and membrane activity of HJH-1, a cationic peptide derived from the hemoglobin α-subunit of bovine erythrocytes P3. HJH-1 shows potent antimicrobial activity against different bacterial species associated with infection and causes weaker hemolysis of erythrocytes, at least five times the minimum inhibitory concentration (MIC). HJH-1 has good stability to tolerance temperature, pH value, and ionic strength. The anionic membrane potential probe bis-(1,3-dibutylbarbituric acid) trimethine oxonol [DiBAC4(3)] and propidium iodide are used as indicators of membrane integrity. In the presence of HJH-1 (1× MIC), Escherichia coli membranes rapidly depolarise, whereas red blood cells show gradual hyperpolarisation. Scanning electron microscopy and transmission electron micrographs show that HJH-1 (1× MIC) damaged the membranes of Escherichia coli, Staphylococcus aureus, and Candida albicans. In conclusion, HJH-1 damages the integrity of the bacterial membrane, preventing the growth of bacteria. HJH-1 has broad-spectrum antibacterial activity, and these activities are performed by changing the normal cell transmembrane potential and disrupting the integrity of the bacterial membrane.

Abstract 195: Electrochemical treatment produces pH changes in the tumor microenvironment that are toxic to cancer cells

Abstract Background: Locoregional therapy is playing an increasingly central role in the management of hepatocellular carcinoma (HCC), where ablation is distinguished among these techniques by its capacity to effect cure. Electrochemical treatment (EChT), a technique that facilitates necrosis via the application of direct current at low voltages shows promise; however, widespread use has been limited as studies of the underlying biology and the mechanism of action have been limited. Methods: A novel assay for the study of EChT mechanism was designed, wherein HCC cells (2.5e5 cells/mL) were embedded in low-melting temperature agarose (1.5%). EChT was performed on these assays between nitinol cathodes (d = 0.5 mm) and platinum anodes (d = 0.5 mm) in a variety of geometries. Buffering capacity of the encapsulation assay was varied via the addition of HEPES buffer at final concentrations of 10mM, 50mM, or 200mM. Either during or immediately following EChT, the following measurements were recorded: cell viability, pH, ROS burden, transmembrane potential, and temperature. Results: Following EChT, acidic pH was appreciated surrounding the anodes and basic pH surrounding the cathodes in regions where cell death was observed by fluorescence microscopy. Upon increasing the buffering capacity of the assay, the total area of cell death decreased (p &amp;lt; 10-6). There was not sufficient heat generation nor transmembrane potential to account for the observed cell death. A pH-threshold was identified, where a pH &amp;gt; 10.8 or &amp;lt; 4.4 was found to cause cell necrosis in the three different buffer conditions studied. By varying the number of electrodes, their spacing, and length of ablation, it was possible to shape the zone of cell death observed to an arbitrary geometry. Conclusions: The mechanism of EChT-induced cancer cell death is through the spread of acidic and basic species generated by hydrolysis of water upon electron transfer at the electrodes. The diffusion of these species leads to cell death in a predictable, pH dependent manner. These results suggest that an electrochemical ablation device could treat complex HCC tumor geometries with appropriate electrode placement and charge delivery. Citation Format: Nicholas Perkons, Elliot Stein, Chike Nwaezeapu, Joseph Wildenberg, Daniel Ackerman, Gregory Nadolski, Stephen Hunt, Terence Gade. Electrochemical treatment produces pH changes in the tumor microenvironment that are toxic to cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 195.

Insight into the Mode of Action of Celangulin V on the Transmembrane Potential of Midgut Cells in Lepidopteran Larvae.

Celangulin V (CV) is the main insecticidal constituent of Celastrus angulatus. The V-ATPase H subunit of the midgut cells of lepidopteran larvae is the putative target protein of CV. Here, we compared the effects of CV on the midgut membrane potentials of Mythimna separata and Agrotis ipsilon larvae with those of the Cry1Ab toxin from Bacillus thuringiensis and with those of inactive CV-MIA, a synthetic derivative of CV. We investigated the changes in the apical membrane potentials (Vam) and basolateral membrane potentials (Vbm) of the midguts of sixth-instar larvae force-fed with the test toxins. We also measured the Vam and Vbm of larval midguts that were directly incubated with the test toxins. Similar to the effect of Cry1Ab, the Vam of CV-treated midguts rapidly decayed over time in a dose-dependent manner. By contrast, CV-MIA did not influence Vam. Meanwhile, the Vam of A. ipsilon larval midguts directly incubated with CV decayed less than that of M. separata larval midguts, whereas that of larvae force-fed with CV did not significantly change. Similar to Cry1Ab, CV did not affect the Vbm of isolated midguts. CV significantly inhibited V-ATPase activity in a dose-dependent manner. Therefore, CV initially inhibits V-ATPase in the apical membrane and affects intracellular pH, homeostasis, and nutrient transport mechanisms in lepidopteran midgut cells.

Organoruthenium Complexes with C^N Ligands are Highly Potent Cytotoxic Agents that Act by a New Mechanism of Action.

Our study demonstrates that four novel kinetically inert C,N-cyclometalated RuII complexes of the type [Ru(C^N)(N^N)2 ][PF6 ] containing a handle for functionalization on the C^N ligand are very potent cytotoxic agents against several different human cancer cell lines and are up to 400-fold more potent than clinically used cisplatin. In addition, the investigated ruthenium complexes are less cytotoxic in noncancerous cells, and exhibit higher selectivity for cancer cells than conventional platinum anticancer drugs. The high potency of the investigated ruthenium compounds can be attributed to several factors, including enhanced internalization and their capability to change mitochondrial transmembrane potential in cells. The new ruthenium complexes also interfere with protein synthesis with a markedly higher potency than conventional inhibitors of DNA translation. Notably, the latter mechanism has not been hitherto described for other cytotoxic Ru compounds and cisplatin.

High frequency electroporation efficiency is under control of membrane capacitive charging and voltage potential relaxation

The study presents the proof of concept for a possibility to achieve a better electroporation in the MHz pulse repetition frequency (PRF) region compared to the conventional low frequency protocols. The 200ns×10 pulses bursts of 10–14kV/cm have been used to permeabilize Chinese hamster ovary (CHO) cells in a wide range (1Hz–1MHz) of PRF. The permeabilization efficiency was evaluated using fluorescent dye assay (propidium iodide) and flow cytometry. It was determined that a threshold PRF exists when the relaxation of the cell transmembrane potential is longer than the delay between the consequent pulses, which results in accumulation of the charge on the membrane. For the CHO cells and 0.1S/m electroporation medium, this phenomenon is detectable in the 0.5–1MHz range. It was shown that the PRF is an important parameter that could be used for flexible control of electroporation efficiency in the high frequency range.

Theoretical analysis of transmembrane potential of cells exposed to nanosecond pulsed electric field

Purpose: Intracellular electroporation occurs when the cells are exposed to nanosecond pulsed electric field (nsPEF). It is believed the electroporation (formation and extension of pores on the membrane induced by external electric field) is affected significantly by the transmembrane potential. This paper analyzed transmembrane potential induced by nsPEF in the term of pulse frequency spectrum, aiming to provide a theoretical explanation to intracellular bio-effects.Methods: Based on the double-shelled spherical cell model, the frequency dependence of transmembrane potential was obtained by solving Laplace’s equation, while the time course of transmembrane potential was obtained by a method combined with discrete Fourier transform and Laplace transform. First-order Debye equation was used to describe the dielectric relaxation of the cell medium.Results: Frequency-domain analysis showed that when the electric field frequency was higher than 105 Hz, the transmembrane potential on the organelle membrane (ΔΦo) was increasing to exceed the transmembrane potential on the cellular membrane (ΔΦc). In the time-domain analysis, transmembrane potentials induced by four nsPEF (short trapezoid, long trapezoid, bipolar and sine shapes) with the same field strength were compared with each other. It showed that ΔΦo is obviously larger than ΔΦc if the curve of the normalized frequency spectrum of the pulse is more similar with the curve of normalized ΔΦo in frequency domain. Pulses with major frequency components higher than 108 Hz lead to both small ΔΦo and ΔΦc. This may explain why high power pulsed microwave lead to unobvious bio-effects of cells than nsPEF with trapezoid form.Conclusion: Through the pulse frequency spectrum it is clearer to understand the relationship between nsPEF and the transmembrane potential.

Потенциометрические измерения трансмембранного потенциала клеток с использованием проникающих ионов

Потенциометрические измерения трансмембранного потенциала клеток с использованием проникающих ионов

Modelling the Cell Transmembrane Potential Dependence on the Structure of the Pulsed Magnetic Field Coils

During high power pulsed magnetic field treatment of biological samples the cells are subjected to both the high magnetic and induced electric fields. The extent of the influence of each treatment component is poorly studied. The work presents the finite element method analysis of pulsed inductive coils that are used for generation of pulsed magnetic and induced electric fields. The simulated coils, electrical parameters and the output characteristics are evaluated in respect to the induced cell transmembrane potential. The model of the Jurkat T lymphocyte cells is introduced in the analysis. The study includes finite element method analysis of four solenoid coils with different structure and inductance in the range of 2.8 μH to 62 μH. Pulsed magnetic field amplitudes up to 5 T are investigated in this work. DOI: http://dx.doi.org/10.5755/j01.eee.20.8.8432

Numerical analysis of DC-field-induced transmembrane potential of spheroidal cells in axisymmetric orientations

This paper presents the electrostatic analysis of direct-current and steady-state transmembrane potential of non-spherical biological cells. The purpose of this analysis is to clarify the influences of different cell geometries and conductivity of the extracellular medium on transmembrane potential. The cells are modeled as spherical or spheroidal and as having different ratios between the radii in different axial directions. The boundary element method, a numerical method, is applied to the calculation of the transmembrane potential. The calculations show that a decrease in the conductivity affects both magnitude and distribution of transmembrane potential. The cell membrane can be approximated as a perfect dielectric, provided that the conductivity of the extracellular medium is sufficiently high. For the same cell geometries, transmembrane potential is smaller for pairs of cells than for isolated cells, and this potential is more reduced at the contact poles than at the opposite poles. Either different axial lengths or different radii between the cells results in this disparity in transmembrane potential of the cell pair. However, the maximum potential of both cells approaches the same value and is located at the contact poles if the conductivity in the extracellular medium is very low.

Microarray of non-connected gold pads used as high density electric traps for parallelized pairing and fusion of cells

Cell fusion consists of inducing the formation of a hybridoma cell containing the genetic properties of the progenitor cells. Such an operation is usually performed chemically or electrically. The latter method, named electrofusion, is considered as having a strong potential, due to its efficiency and non-toxicity, but deserves further investigations prior to being applicable for key applications like antibody production and cancer immunotherapy. Indeed, to envision such applications, a high amount of hybrid cells is needed. In this context, we present in this paper a device for massive cell pairing and electrofusion, using a microarray of non-connected conductive pads. The electrofusion chamber--or channel--exposes cells to an inhomogeneous electric field, caused by the pads array, enabling the trapping and pairing of cells with dielectrophoresis (DEP) forces prior to electrofusion. Compared to a mechanical trapping, such electric trapping is fully reversible (on/off handling). The DEP force is contactless and thus eases the release of the produced hybridoma. Moreover, the absence of wire connections on the pads permits the high density trapping and electrofusion of cells. In this paper, the electric field mapping, the effect of metallic pads thickness, and the transmembrane potential of cells are studied based on a numerical model to optimize the device. Electric calculations and experiments were conducted to evaluate the trapping force. The structure was finally validated for cell pairing and electrofusion of arrays of cells. We believe that our approach of fully electric trapping with a simple structure is a promising method for massive production of electrofused hybridoma.