scholarly journals The Good and the Bad of Cell Membrane Electroporation

2021 ◽  
Vol 68 (4) ◽  
pp. 753-764
Author(s):  
Katja Balantič ◽  
Damijan Miklavčič ◽  
Igor Križaj ◽  
Peter Kramar
Keyword(s):  
Cell Membrane ◽  
Lipid Bilayer ◽  
High Voltage ◽  
Food Industry ◽  
Cell Membranes ◽  
Tissue Ablation ◽  
Gene Electrotransfer ◽  
Basic Principles ◽  
Electric Pulses ◽  
Hydrophobic Barrier

Electroporation is used to increase the permeability of the cell membrane through high-voltage electric pulses. Nowadays, it is widely used in different areas, such as medicine, biotechnology, and the food industry. Electroporation induces the formation of hydrophilic pores in the lipid bilayer of cell membranes, to allow the entry or exit of molecules that cannot otherwise cross this hydrophobic barrier. In this article, we critically review the basic principles of electroporation, along with the advantages and drawbacks of this method. We discuss the effects of electroporation on the key components of biological membranes, as well as the main applications of this procedure in medicine, such as electrochemotherapy, gene electrotransfer, and tissue ablation. Finally, we define the most relevant challenges of this romising area of research.

scholarly journals Investigation of Plasmid DNA Delivery and Cell Viability Dynamics for Optimal Cell Electrotransfection In Vitro

2020 ◽  
Vol 10 (17) ◽  
pp. 6070
Author(s):  
Sonam Chopra ◽  
Paulius Ruzgys ◽  
Martynas Maciulevičius ◽  
Milda Jakutavičiūtė ◽  
Saulius Šatkauskas
Keyword(s):  
Cell Membrane ◽  
Cell Viability ◽  
High Voltage ◽  
Plasmid Dna ◽  
Transfection Efficiency ◽  
Dna Delivery ◽  
Output Parameter ◽  
Gene Electrotransfer ◽  
Electric Pulses

Electroporation is an effective method for delivering plasmid DNA molecules into cells. The efficiency of gene electrotransfer depends on several factors. To achieve high transfection efficiency while maintaining cell viability is a tedious task in electroporation. Here, we present a combined study in which the dynamics of both evaluation types of transfection efficiency and the cell viability were evaluated in dependence of plasmid concentration as well as at the different number of high voltage (HV) electric pulses. The results of this study reveal a quantitative sigmoidal (R2 > 0.95) dependence of the transfection efficiency and cell viability on the distance between the cell membrane and the nearest plasmid. We propose this distance value as a new, more accurate output parameter that could be used in further optimization studies as a predictor and a measure of electrotransfection efficiency.

Calcium influx determines the muscular response to electrotransfer

2012 ◽  
Vol 302 (4) ◽  
pp. R446-R453 ◽  
Author(s):  
Pernille Hojman ◽  
Camilla Brolin ◽  
Hanne Gissel
Keyword(s):  
Cell Membrane ◽  
High Voltage ◽  
Voltage Pulse ◽  
Low Voltage ◽  
Calcium Influx ◽  
High Voltage Pulse ◽  
Electric Pulses ◽  
Tnf Α ◽  
Cell Membrane Permeabilization ◽  
The Times

Cell membrane permeabilization by electric pulses (electropermeabilization), results in free exchange of ions across the cell membrane. The role of electrotransfer-mediated Ca2+-influx on muscle signaling pathways involved in degeneration (β-actin and MurF), inflammation (IL-6 and TNF-α), and regeneration (MyoD1, myogenin, and Myf5) was investigated, using pulse parameters of both electrochemotherapy (8 HV) and DNA delivery (HVLV). Three pulsing conditions were used: 8 high-voltage pulses (8 HV), resulting in large permeabilization and ion flux, and a combination of one high-voltage pulse and one low-voltage pulse (HVLV), either alone or in combination with injection of DNA. Mice and rats were anesthetized before pulsing. At the times given, animals were killed, and intact tibialis cranialis muscles were excised for analysis. Uptake of Ca2+ was assessed using 45Ca as a tracer. Using gene expression analyses and histology, we showed a clear association between Ca2+ influx and muscular response. Moderate Ca2+ influx induced by HVLV pulses results in activation of pathways involved in immediate repair and hypertrophy. This response could be attenuated by intramuscular injection of EGTA reducing Ca2+ influx. Larger Ca2+ influx as induced by 8-HV pulses leads to muscle damage and muscle fiber regeneration through recruitment of satellite cells. The extent of Ca2+ influx determines the muscular response to electrotransfer and, thus, the success of a given application. In the case of electrochemotherapy, in which the objective is cell death, a large influx of Ca2+ may be beneficial, whereas for DNA electrotransfer, muscle recovery should occur without myofiber loss to ensure preservation of plasmid DNA.

Feasibility Study for Applying Irreversible Electroporation to the Treatment of Breast Cancer

2009 ◽  
Author(s):  
Robert E. Neal ◽  
Ravi Singh ◽  
Suzy Torti ◽  
Rafael V. Davalos
Keyword(s):  
Breast Cancer ◽  
Immune Response ◽  
Extracellular Matrix ◽  
Numerical Modeling ◽  
Cell Membrane ◽  
Irreversible Electroporation ◽  
Short Length ◽  
Tissue Ablation ◽  
Electric Pulses ◽  
A Cell

Non-thermal irreversible electroporation (IRE) is a new, minimally invasive, localized tissue ablation technique [1]. The procedure uses electrodes to deliver short-length, high voltage electric pulses to destabilize a cell membrane, leading to the creation of nanopores. When the pulses are strong enough, the cell cannot repair the damage and dies [2]. It has been shown that substantial volumes of tissue and cutaneous tumors may be ablated in a non-thermal manner using irreversible electroporation [1, 3]. In addition, this procedure may be predicted by numerical modeling, promotes an immune response, leaves the extracellular matrix intact, does not affect nerves, may be monitored in real-time, and preserves tissue vasculature [2–5].

Irreversible Electroporation (IRE) to Treat Brain Tumors

2008 ◽  
Author(s):  
Paulo A. Garcia ◽  
John Robertson ◽  
John Rossmeisl ◽  
Rafael V. Davalos
Keyword(s):  
Cell Death ◽  
Cell Membrane ◽  
Brain Tumors ◽  
High Voltage ◽  
Electric Pulse ◽  
Irreversible Electroporation ◽  
Transient Effect ◽  
Electric Pulses ◽  
Thermal Cell ◽  
Reversible Electroporation

Electroporation is the phenomenon in which permeability of the cell membrane to ions and macromolecules is increased by exposing the cell to short (microsecond to millisecond) high voltage electric pulses [1]. The application of the electric pulse can have no effect, can have a transient effect known as reversible electroporation, or can cause permanent permeation known as irreversible electroporation (IRE) which leads to non-thermal cell death by necrosis [1, 2].

scholarly journals Evaluation of electroporated area using 2,3,5-triphenyltetrazolium chloride in a potato model

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Seung Jeong ◽  
Hongbae Kim ◽  
Junhyung Park ◽  
Ki Woo Kim ◽  
Sung Bo Sim ◽  
...  
Keyword(s):  
Cell Death ◽  
Cell Membrane ◽  
Irreversible Electroporation ◽  
Tissue Ablation ◽  
Target Tissue ◽  
Triphenyltetrazolium Chloride ◽  
Tissue Model ◽  
Electric Pulses ◽  
Reversible Electroporation ◽  
Over Time

AbstractIrreversible electroporation (IRE) is a tissue ablation method, uses short high electric pulses and results in cell death in target tissue by irreversibly permeabilizing the cell membrane. Potato is commonly used as a tissue model for electroporation experiments. The blackened area that forms 12 h after electric pulsing is regarded as an IRE-ablated area caused by melanin accumulation. Here, the 2,3,5-triphenyltetrazolium chloride (TTC) was used as a dye to assess the IRE-ablated area 3 h after potato model ablation. Comparison between the blackened area and TTC-unstained white area in various voltage conditions showed that TTC staining well delineated the IRE-ablated area. Moreover, whether the ablated area was consistent over time and at different staining times was investigated. In addition, the presumed reversible electroporation (RE) area was formed surrounding the IRE-ablated area. Overall, TTC staining can provide a more rapid and accurate electroporated area evaluation.

Interaction of detergent sclerosants with cell membranes

2014 ◽  
Vol 30 (5) ◽  
pp. 306-315 ◽  
Author(s):  
Kurosh Parsi
Keyword(s):  
Cell Membrane ◽  
Lipid Bilayer ◽  
Cell Membranes ◽  
Membrane Lipid ◽  
Striking Similarity ◽  
Target Cells ◽  
Clotting Factors ◽  
Endothelial Lining ◽  
Ionic Charge ◽  
Vessel Occlusion

Commonly used detergent sclerosants including sodium tetradecyl sulphate (STS) and polidocanol (POL) are clinically used to induce endovascular fibrosis and vessel occlusion. They achieve this by lysing the endothelial lining of target vessels. These agents are surface active (surfactant) molecules that interfere with cell membranes. Surfactants have a striking similarity to the phospholipid molecules of the membrane lipid bilayer. By adsorbing at the cell membrane, surfactants disrupt the normal architecture of the lipid bilayer and reduce the surface tension. The outcome of this interaction is concentration dependent. At high enough concentrations, surfactants solubilise cell membranes resulting in cell lysis. At lower concentrations, these agents can induce a procoagulant negatively charged surface on the external aspect of the cell membrane. The interaction is also influenced by the ionic charge, molecular structure, pH and the chemical nature of the diluent (e.g. saline vs. water). The ionic charge of the surfactant molecule can influence the effect on plasma proteins and the protein contents of cell membranes. STS, an anionic detergent, denatures the tertiary complex of most proteins and in particular the clinically relevant clotting factors. By contrast, POL has no effect on proteins due to its non-ionic structure. These agents therefore exhibit remarkable differences in their interaction with lipid membranes, target cells and circulating proteins with potential implications in a range of clinical applications.

scholarly journals A DISTINCTIVE CELL CONTACT IN THE RAT ADRENAL CORTEX

1972 ◽  
Vol 53 (1) ◽  
pp. 148-163 ◽  
Author(s):  
Daniel S. Friend ◽  
Norton B. Gilula
Keyword(s):  
Cell Membrane ◽  
Adrenal Cortex ◽  
Cell Membranes ◽  
Freeze Fracture ◽  
Typical Feature ◽  
Cell Contact ◽  
Cortical Cells ◽  
Cell Dispersion ◽  
Extensive Cell ◽  
Distinctive Cell

Extensive cell contacts which resemble septate junctions occur between cells in the three major zones of the rat adrenal cortex. Characteristically, they extend between small intercellular canaliculi and the periendothelial space, frequently interrupted by gap junctions and rarely by desmosomes. Zonulae occludentes have not been identified in the adrenal cortex. Along this distinctive cell contact, the cell membranes of apposing cells are separated by 210–300 a bisected by irregularly spaced 100–150-A extracellular particles which are often circular in profile. In lanthanum preparations, these particles appear to form a continuous chain throughout the intercellular space and are visualized as an alveolate structure in sections parallel to the plane of the cell membrane. The cell membrane in the area of septate-like contact does not differ from nonjunctional areas of the cell membrane in freeze-fracture replicas. The cell contact retains its integrity after cell dispersion and after the separation of cell membranes from disrupted cells. The intercellular particles also persist after brief extraction in lipid solvents. Besides adherence, possible functions of this adrenal contact include maintenance of the width of the extracellular space, the provision of channels between intercellular canaliculi and the bloodstream, and utilization as cation depots. Similar structures are also present between adrenal cortical cells of several other species and between interstitial cells of the testis. This type of cell contact may, in fact, be a typical feature of steroid-hormone-secreting tissues in vertebrates.

scholarly journals Pathogenesis of shigella diarrhea. VII. Evidence for a cell membrane toxin receptor involving β1 {arrow} 4-linked N-acetyl-D-glucosamine oligomers

1977 ◽  
Vol 146 (2) ◽  
pp. 535-546 ◽  
Author(s):  
GT Keusch ◽  
M Jacewicz
Keyword(s):  
Cell Membrane ◽  
Cholera Toxin ◽  
Liver Cell ◽  
Hela Cells ◽  
Mammalian Cells ◽  
Cell Membranes ◽  
Proteolytic Enzymes ◽  
Cell Monolayer ◽  
Toxin Receptor ◽  
Toxin Uptake

The binding of ShigeUa dysenteriae 1 cytotoxin to HeLa cells in culture and to isolated rat liver cell membranes was studied by means of an indirect consumption assay of toxicity from the medium, or by determination of cytotoxicity to the HeLa cell monolayer. Both liver cell membranes and HeLa cells removed toxicity from the medium during incubation, in contrast to WI-38 and Y-1 mouse adrenal tumor cells, both of which neither bound nor were affected by the toxin. Uptake of toxin was directly related to concentration of membranes added, time,and temperature, and indirectly related to the ionic strength of the buffer used. The chemical nature of the membrane receptor was characterized by using three principal approaches: (a) enzymatic sensitivity; (b) competitive inhibition and (c) receptor blockade studies. The receptor was destroyed by proteolytic enzymes, phospholipases (which markedly altered the gross appearance of the membrane preparation) and by lysozyme, but not by a variety of other enzymes. Of 28 carbohydrate and glycoprotein haptens studied, including cholera toxin and ganglioside, only the chitin oligosaccharide lysozyme substrates, per N-acetylated chitotriose, chitotetraose, and chitopentaose were effective competitive inhibitors. Greatest inhibition was found with the trimer, N, N', N" triacetyl chitotriose. Of three lectins studied as possible receptor blockers, including phytohemagglutinin, concanavalin A, and wheat germ agglutinin, only the latter, which is known to possess specific binding affinity for N, N', N" triacetyl chitotriose, was able to block toxin uptake. Evidence from all three approaches indicate, therefore, existence of a glycoprotein toxin receptor on mammalian cells, with involvement of oligomeric β1{arrow}4-1inked N-acetyl glucosamine in the receptor. This receptor is clearly distinct from the G(M1) ganglioside thought to be involved in the binding of cholera toxin to the cell membrane of a variety of cell types susceptible to its action.

Ammonia movement and distribution after exercise across white muscle cell membranes in rainbow trout

1996 ◽  
Vol 271 (3) ◽  
pp. R738-R750 ◽  
Author(s):  
Y. Wang ◽  
G. J. Heigenhauser ◽  
C. M. Wood
Keyword(s):  
Membrane Potential ◽  
Cell Membrane ◽  
Intracellular Ph ◽  
Muscle Cell ◽  
Cell Membranes ◽  
White Muscle ◽  
Extracellular Ph ◽  
Posterior Portion ◽  
Arterial Inflow ◽  
Muscle Cell Membrane

Manipulations of pH and electrical gradients in a perfused preparation were used to analyze the factors controlling ammonia distribution and flux in trout white muscle after exercise. Trout were exercised to exhaustion, and then an isolated-perfused white muscle preparation with discrete arterial inflow and venous outflow was made from the posterior portion of the tail. The tail-trunks were perfused with low (7.4)-, medium (7.9)-, and high (8.4)-pH saline, achieved by varying HCO3- concentration ([HCO3-]) at constant Pco2. Intracellular and extracellular pH, ammonia, CO2, K+, Na+, and Cl- were measured. Muscle intracellular pH was not affected by changes in extracellular pH. Increasing extracellular pH caused a decrease in the transmembrane NH3 partial pressure (PNH3) gradient and a decrease in ammonia efflux. When extracellular K+ concentration was increased from 3.5 to 15 mM in the medium-pH group, a depolarization of the muscle cell membrane potential from -92 to -60 mV and a 0.1-unit depression in intracellular pH occurred. Ammonia efflux increased despite a marked reduction in the PNH3 gradient. Amiloride (10(-4) M) had no effect, indicating that Na+/H(+)-NH4+ exchange does not participate in ammonia transport in this system. A comparison of observed intracellular-to-extracellular ammonia distribution ratios with those modeled according to either pH or Nernst potential distributions supports a model in which ammonia distribution across white muscle cell membranes is affected by both pH and electrical gradients, indicating that the membranes are permeable to both NH3 and NH4+. Membrane potential, acting to retain high levels of NH4+ in the intracellular compartment, appears to have the dominant influence during the postexercise period. However, at rest, the pH gradient may be more important, resulting in much lower intracellular ammonia levels and distribution ratios. We speculate that the muscle cell membrane NH3-to-NH4+ permeability ratio in trout may change between the rest and postexercise condition.

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