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A special issue of Applied Sciences (ISSN 2076-3417). This special issue belongs to the section "Applied Biosciences and Bioengineering".

Deadline for manuscript submissions: closed (20 January 2026) | Viewed by 20005

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Prof. Dr. Luis Redondo

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Guest Editor

Unit for Innovation and Research in Engineering, Lisbon School of Engineering, UnIRE/ISEL, 1959-007 Lisbon, Portugal
Interests: pulsed power technology and applications; semiconductor based pulsed power generators; pulsed electric field applications
Special Issues, Collections and Topics in MDPI journals

Prof. Dr. Bucur Novac

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Guest Editor

Wolfson School of MEME, Loughborough University, Loughborough, UK
Interests: pulsed power physics and technology

Special Issue Information

Dear Colleagues,

Over the past decade, advances related to pulsed electric field (PEF) techniques have matured considerably, contributing to a broad range of applications in the medical, environmental, food, energy, and biotechnological fields.

This Special Issue will provide an excellent opportunity for sharing the latest results related to the development of technologies associated with the generation of PEFs, from pulsed power generators to treatment chambers, as well as their applications in various domains. In addition, there will also be works presented that describe PEF techniques applied for cancer treatment and tumor ablation, disinfection, decontamination, hygiene, cell and tissue stimulation, wound healing, biomass processing and biofuel generation, food safety and food preservation, agricultural crops, and farming as well as biomedical applications, together with their characteristic diagnostics and analytics.

This Special Issue of Applied Sciences, “Applications of Pulsed Electric Field (PEF) Interactions with Biological Cells”, aims to attract novel contributions covering a wide range of research and applications of the experimental techniques and effects of PEFs on biological cells.

Prof. Dr. Luis Redondo
Prof. Dr. Bucur Novac
Guest Editors

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Keywords

pulsed electric field (PEF) generation
diagnostic and analytics
PEF applications in the medical, environmental, food, energy, and biotechnological fields
experimental techniques and effects from using PEFs in biological cells

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Published Papers (9 papers)

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Research

14 pages, 1704 KB

Open AccessArticle

Electron Paramagnetic Resonance Study of Pulsed Electric Field-Assisted Internalization of Spin Probes into Yeast Cells

by Tomasz Kubiak and Bernadeta Dobosz

Appl. Sci. 2026, 16(4), 1973; https://doi.org/10.3390/app16041973 - 16 Feb 2026

Viewed by 294

Abstract

Efficient internalization of chemical entities into cells across the plasma membrane barrier is crucial for most biomedical applications. Among the physical factors that can enhance the delivery of active substances, the electric field plays an important role. In our experiments, electron paramagnetic resonance (EPR) spectroscopy was used to monitor the process of electric field-assisted endocytosis of a spin probe TEMPO (2,2,6,6-tetramethylpiperidine 1-oxyl) into yeast cells (Saccharomyces cerevisiae). It was found that a long train of low-amplitude voltage pulses (0.5–20 V_pp) can affect the uptake of nitroxide by cells. In addition to the pulse amplitude, the key parameters influencing the internalization process are: frequency, duty cycle and time of exposure to the electric field. The sodium chloride content in the medium in which the cells are suspended is also of great importance. For higher salt concentrations, starting from 3.5% w/w, a significant decrease in cell survival was observed after exposure to an electrical stimulus, which also translated into a decreasing spin probe uptake. Our studies confirm the usefulness of EPR spectroscopy in the study of internalization processes and show that the parameters of electric field pulses and the environmental salinity have a significant impact on the course of electroendocytosis. Full article

(This article belongs to the Special Issue Applications of Pulsed Electric Field (PEF) Interactions with Biological Cells)

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14 pages, 1626 KB

Open AccessArticle

Experimental Evaluation of Pulse Width Effects Under Equal-Dose Pulsed Electric Field Treatment on A375 Cells

by Hongyu Kou, Feiyu Wu, Kai Chen, Shupeng Wang, Runze Liang and Chenguo Yao

Appl. Sci. 2026, 16(2), 1086; https://doi.org/10.3390/app16021086 - 21 Jan 2026

Viewed by 280

Abstract

Pulsed electric fields (PEFs) are widely recognized as a non-thermal, selective physical therapy with wide clinical application in tumor ablation. The pulse width determines how electrical energy is distributed across plasma membrane to intracellular organelles. However, under an engineering-defined equal-dose condition (N·E²·tp), which serves as a practical control parameter rather than a measure of true cellular energy absorption, systematic and comparable experimental characterization of cellular and subcellular responses across pulse widths from the microsecond to nanosecond range remains limited. In this study, PEFs with pulse widths ranging from 100 μs to 50 ns were applied under equal-dose constraints, and cellular responses were evaluated using transmission electron microscopy (TEM), multi-organelle fluorescence imaging, and flow cytometry. The results indicate that pulse-width-dependent effects were observed under a fixed pulse-number, dose-equalized framework in which electric field strength varied across conditions. Structural and functional changes were observed in multiple organelles, including the nucleus, mitochondria, endoplasmic reticulum, and Golgi apparatus. Notably, nanosecond pulses were more effective in inducing mitochondrial membrane potential loss and increasing the proportion of apoptotic or non-viable cells. These findings demonstrate that, under equal-dose conditions, pulse width is a key temporal parameter governing PEF-induced biological effects, indicating that identical dose constraints do not necessarily result in equivalent biological responses. This work provides experimental foundation for parameter selection and optimization in PEF-based biomedical applications. Full article

(This article belongs to the Special Issue Applications of Pulsed Electric Field (PEF) Interactions with Biological Cells)

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18 pages, 1385 KB

Open AccessArticle

The Impact of Pulsed Electric Field Treatment of Beetroots on the Physicochemical Properties of Juice, Dried Juice, and Dried Pomace

by Aleksandra Jedlińska, Alicja Barańska-Dołomisiewicz, Katarzyna Samborska, Katarzyna Rybak, Artur Wiktor, Dorota Witrowa-Rajchert and Małgorzata Nowacka

Appl. Sci. 2025, 15(7), 3834; https://doi.org/10.3390/app15073834 - 31 Mar 2025

Cited by 4 | Viewed by 1801

Abstract

The aim of this work was to determine the impact of pulsed electric field treatment of beetroots on select physicochemical properties of the obtained juice, spray-dried juice, and freeze-dried pomace. Pulsed electric field pretreatment of beetroots was conducted with a different number of pulses (3 or 44 pulses) at a constant electric field strength of E = 1.07 kV/cm, which resulted in the total energy levels of 0.5 and 4 kJ/kg. The juice was dehumidified and air spray-dried with NUTRIOSE^® as a drying carrier at 90/60 °C, while pomace was freeze-dried at 0.63 Pa and a shelf temperature of 20 °C. Pulsed electric field (PEF) treatment caused an increase in pigment content in beetroots and the juice. In the case of powders, the following effects were noted: a decrease in drying efficiency, changed powder color, decreased powder particle diameter. Dried juice and pomace treated with a higher pulsed electric field (PEF) treatment were characterized by the highest hygroscopicity. Full article

(This article belongs to the Special Issue Applications of Pulsed Electric Field (PEF) Interactions with Biological Cells)

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15 pages, 3029 KB

Open AccessArticle

Pulsed Electric Fields as an Infrared–Convective Drying Pretreatment: Effect on Drying Course, Color, and Chemical Properties of Apple Tissue

by Aleksandra Skarżyńska, Ewa Gondek, Małgorzata Nowacka and Artur Wiktor

Appl. Sci. 2025, 15(5), 2348; https://doi.org/10.3390/app15052348 - 22 Feb 2025

Cited by 5 | Viewed by 1483

Abstract

In this research, the impact of the pulsed electric fields (PEF) pretreatment on the infrared–convective drying (IR-CD) of apples and selected quality indicators (antioxidant capacity, color, and total phenolic content) of dried apples was evaluated. PEF pretreatment was carried out at the following parameters: electric field strength of 1 kV/cm, specific energy inputs of 1, 3.5, and 6 kJ/kg. Moreover, variable IR-CD process parameters were also assessed (peak wavelength of 1.2 µm, distance between the infrared lamps and the apple slices of 10, 20, and 30 cm). PEF pretreatment, by disintegrating apple cells, reduced the drying time of that fruit by 11–20%. The IR-CD of apples was the most effective at the shortest distance (10 cm) between the infrared source and the apple slices; it was associated with the intense heating of its surface. PEF-pretreated samples exhibited lower retention of antioxidants and were darker and redder than untreated ones. PEF could increase the activity of enzymes responsible for oxidizing phenolic compounds to brown pigments. The use of a medium distance (20 cm) during IR-CD promoted the highest retention of antioxidant compounds (relatively prompt drying with moderate heating of the sample surface). Full article

(This article belongs to the Special Issue Applications of Pulsed Electric Field (PEF) Interactions with Biological Cells)

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15 pages, 1711 KB

Open AccessArticle

Exploration of Machine Learning Models for Prediction of Gene Electrotransfer Treatment Outcomes

by Alex Otten, Michael Francis and Anna Bulysheva

Appl. Sci. 2024, 14(24), 11601; https://doi.org/10.3390/app142411601 - 12 Dec 2024

Viewed by 1536

Abstract

Gene electrotransfer (GET) is a physical method of gene delivery to various tissues utilizing pulsed electric fields to transiently permeabilize cell membranes to allow for genetic material transfer and expression. Optimal pulsing parameters dictate gene transfer efficiency and cell survival, which are critical for the wide adaptation of GET as a gene therapy technique. Tissue heterogeneity complicates the delivery process, requiring the extensive optimization of pulsing protocols currently empirically optimized. These experiments are time-consuming and resource-intensive, requiring large numbers of animals for in vivo optimization. Advances in machine learning (ML) and computing power, data analysis, and model generation using ML techniques, such as neural networks, enable predictive modeling for GET. ML models have been used previously to predict ablation performance in irreversible electroporation procedures and single-cell electroporation platforms. In this work, we present ML predictive models that could be used to optimize pulsing parameters based on already completed experiments. The models were trained on 132 data points from 19 papers with the Matlab Statistics and Machine Learning Toolbox. An artificial neural network (ANN) was generated that could predict binary treatment outcomes with an accuracy of 71.8%. Support vector machines (SVMs) using selected features based on

χ^{2}

tests were also explored. All models used a maximum of 24 features as input, spread across target species, needle configuration, pulsing parameters, and plasmid parameters. Pulse voltage and pulse width dominated as the critical parameters, followed by field strength, dose, and electrode with the greatest impact on GET efficiency. This study elucidates areas where predictive ML algorithms may ideally inform GET study design to accelerate optimization and improve efficiencies upon the further training of these models. Full article

(This article belongs to the Special Issue Applications of Pulsed Electric Field (PEF) Interactions with Biological Cells)

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12 pages, 8031 KB

Open AccessArticle

Localized In Vivo Electro Gene Therapy (LiveGT)-Mediated Skeletal Muscle Protein Factory Reprogramming

by Jacob Hensley, Michael Francis, Alex Otten, Nadezhda Korostyleva, Tina Gagliardo and Anna Bulysheva

Appl. Sci. 2024, 14(23), 11298; https://doi.org/10.3390/app142311298 - 4 Dec 2024

Viewed by 2541

Abstract

Gene electrotransfer (GET) has gained significant momentum as a non-viral gene delivery method for various clinical applications, primarily in the cancer immunotherapy and vaccine development space. Preclinical studies have demonstrated exogenous gene delivery and expression in various tissues, including the liver, skin, cardiac muscle, and skeletal muscle. However, protein replacement applications of this technology have yet to be fully actuated. Plasmid DNA skeletal muscle delivery has been shown to maintain expression for up to 18 months. In the current study, we evaluated localized skeletal muscle delivery for protein replacement applications. We developed localized in vivo electro gene therapy (liveGT) protocols utilizing mono- and biphasic pulse sequences for localized pulse delivery directly to skeletal muscle with a custom monopolar platinum electrode. Plasmid DNA encoding human insulin and human glucokinase were chosen for this study to evaluate the liveGT platform for protein replacement potential. Initial in vitro GET was performed in mouse myoblasts to evaluate human insulin and glucokinase co-delivery. This was followed by liveGT-mediated reporter gene delivery in the skeletal muscle of Sprague–Dawley rats for pulse sequence selection. Protein replacement potential was evaluated in healthy (non-diabetic) rats with liveGT-mediated human insulin and glucokinase co-delivery to skeletal muscle. Human and rat insulin levels were measured via ELISA over the course of 3 months. Fed-state blood glucose measurements were monitored in correlation with serum human insulin levels. LiveGT-mediated skeletal muscle reprogramming successfully produced physiological levels of human insulin in serum over the course of 3 months. Hypo- and hyperglycemic events were not observed. Therefore, liveGT is a safe and viable platform for potential protein replacement therapies. Full article

(This article belongs to the Special Issue Applications of Pulsed Electric Field (PEF) Interactions with Biological Cells)

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12 pages, 1643 KB

Open AccessArticle

Challenges and Opportunities for Pilot Scaling-Up Extraction of Olive Oil Assisted by Pulsed Electric Fields: Process, Product, and Economic Evaluation

by Sara Dias, Enrique Pino-Hernández, Diogo Gonçalves, Duarte Rego, Luís Redondo and Marco Alves

Appl. Sci. 2024, 14(9), 3638; https://doi.org/10.3390/app14093638 - 25 Apr 2024

Cited by 11 | Viewed by 3421

Abstract

This study aimed to investigate the impact of Pulsed Electric Fields (PEF) technology in the extraction of olive oil on a pilot scale, using the “Galega Vulgar” olive variety as raw material. The extraction assisted by PEF had a malaxation time of 30 min and was compared with the traditional process of 45 min of malaxation. The main quality parameters of olive oil and the PEF’s cost-benefit assessment were performed. The incorporation of PEF in olive oil production reduced the malaxation stage by 33% without compromising the yield or extra-virgin classification. This efficiency leads to a potential 12.3% increase in annual olive oil production, with a 12.3% and 36.8% rise in revenue and gross profit, respectively. For small-scale production, the considerable upfront investment required for PEF equipment may be a challenge in terms of return on investment. In this scenario, opting for a renting scheme is the best economic solution, especially given the seasonal nature of olive oil production. In medium- to large-scale production, the investment in PEF is a sound investment since it is possible to achieve, with an equipment cost of EUR 450,000 and a production output of 5 tons per hour, an annual ROI of 20%. Full article

(This article belongs to the Special Issue Applications of Pulsed Electric Field (PEF) Interactions with Biological Cells)

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23 pages, 3247 KB

Open AccessArticle

Comparative Analysis of Pulsed Electric Fields (PEF) and Traditional Pasteurization Techniques: Comparative Effects on Nutritional Attributes and Bacterial Viability in Milk and Whey Products

by Aivaras Šalaševičius, Dovilė Uždavinytė, Mindaugas Visockis, Paulius Ruzgys and Saulius Šatkauskas

Appl. Sci. 2023, 13(22), 12127; https://doi.org/10.3390/app132212127 - 8 Nov 2023

Cited by 11 | Viewed by 4980

Abstract

With the current upsurge in the desire to foster healthy lifestyles and consume nutritious food products, the food industry has been propelled to develop novel food processing technologies. In our study, we critically evaluated the influence of pulsed electric field (PEF) processing by comparing it to conventional thermal pasteurization protocols—low temperature, long time (LTLT), high temperature, short time (HTST), and microfiltration (MF)—and its ramifications on the nutritional properties inherent in raw milk, which comprises vitamins, whey protein, amino acids, cholesterol, and fatty acids. A significant difference in β-lactoglobulin content was observed in PEF-treated liquid whey samples compared to those treated with high-temperature (HT) pasteurization, where 4.8-fold reduction with a concentration of 0.80 mg/mL was observed. Liquid whey samples treated with PEF, LTLT, HTST and MF retained β-lactoglobulin content, PEF-treated samples yielded 3.85 mg/mL, while HTST, LTLT, and MF-treated samples had β-lactoglobulin content of 3.62 mg/mL, 3.63 mg/mL, and 3.62 mg/mL compared to raw whey control (RWC) at 3.81 mg/mL. The concentrations of nutritional properties, like vitamins (A, D, E), amino acids, cholesterol, and fatty acids, remained approximately consistent across all the pasteurization methodologies. Moreover, the bacterial viability in the context of various pasteurization methodologies was scrutinized, with an absence of colonies observed in whey specimens subjected to thermal pasteurization. PEF-treated samples exhibited a substantial 1.6-log reduction in coliform colony count to less than 4 CFU/mL after curd reduction, in contrast to raw milk samples. Full article

(This article belongs to the Special Issue Applications of Pulsed Electric Field (PEF) Interactions with Biological Cells)

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23 pages, 2937 KB

Open AccessArticle

Pulsed Electric Fields vs. Pectolytic Enzymes in Arinto Vinification: Effects on Yield and Oenological Parameters

by Mafalda Aguiar-Macedo, Luis M. Redondo, Marcos Teotónio Pereira and Carlos Silva

Appl. Sci. 2023, 13(14), 8343; https://doi.org/10.3390/app13148343 - 19 Jul 2023

Cited by 5 | Viewed by 2271

Abstract

The increase in awareness of consumers and producers regarding the sustainable production and consumption of food commodities is motivating the emergence of new technologies to improve the efficiency of pre-established methods and reduce or supplant the usage of production factors. Thus, innovative technologies, such as the nonthermal application of pulsed electric fields (PEFs), may play a crucial role in the optimization of processes, both economically and environmentally (shrinkage of wastage, energy efficiency and decrease in the use of food additives), without compromising the quality of the final product. Thus, a comparison was made between the application of commercial-grade enzymes and PEF treatment to assess the impact on cuvée white grape must and on wine yield and quality. Oenological parameters were evaluated during alcoholic fermentation and after 3 months, with tartaric stability measured after 6 months. For this, assays resorting to 1.5 tons of Arinto grapes were separated into nine similar batches: three control, three treated with enzymes (1.5 g/100 kg) and three subjected to PEFs (1 kV/cm; 2 kJ/kg) at a rate of 4 ton/h. PEFs presented the highest increase in cuvée wine yield of 5.47%; a reduction of 19% of wine lees production was also determined. The effect of PEFs on pH, total acidity, turbidity, total phenols, color intensity, %Ye, total dry extract, volatile acidity and tartaric stability was studied and compared with control and enzymatic treatment. PEF and enzyme usage direct costs were determined; the employment of PEFs represented a direct cost of 0.12 EUR/ton, while enzyme usage was 1.80 EUR/ton. Full article

(This article belongs to the Special Issue Applications of Pulsed Electric Field (PEF) Interactions with Biological Cells)

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