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Applications of Pulsed Electric Field (PEF) Interactions with Biological Cells

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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: 20 June 2025 | Viewed by 6613

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Special Issue Editors

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

E-Mail Website
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 (5 papers)

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Research

15 pages, 1711 KiB

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 378

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 KiB

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 740

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 KiB

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 1 | Viewed by 952

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 KiB

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 3 | Viewed by 2172

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 KiB

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 1 | Viewed by 1440

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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