Differential Cell Death and Regrowth of Dermal Fibroblasts and Keratinocytes After Application of Pulsed Electric Fields

Abstract

Background: High-powered pulsed electric fields (PEF) may be used for tissue debridement and disinfection, while lower PEF intensities may stimulate beneficial cellular responses for wound healing. We investigated the dual effects of nonuniform PEF on cellular death and stimulation. Methods: Dermal fibroblast or keratinocyte monolayers were exposed to PEF induced by two needle electrodes (2 mm apart). Voltages (100-600 V; 1 Hz; 70 micros pulse width; 90 pulses/cycle) were applied between the two electrodes. Controls consisted of similar monolayers subjected to a scratch mechanical injury. Results: Cell growth and closure of the cell-free gap was faster in PEF-treated cell monolayers versus scratched ones. Media conditioned from cells pre-exposed to PEF, when applied to responder cells, stimulated greater proliferation than media from scratched monolayers. Conclusions: PEF treatment causes the release of soluble factors that promote cell growth, and thus may play a role in the accelerated healing of wounds post PEF.

Application of pulsed electric field technology to skin engineering

“…Enhanced proliferative and migratory responses of fibroblasts in 2D culture following PEF treatment was observed in several studies ( Table 2 ) ( Das et al, 2020 ; Gouarderes et al, 2020 ). When embedded in 3D collagen lattices, PEF-treated fibroblasts had enhanced alignment and contractility ( Liu et al, 2018 ; Gouarderes et al, 2022 ).…”

“…Control of cell fate is a critical part of producing successful tissue constructs ( Demidova-Rice et al, 2012b ; Lanza et al, 2020 ; Battafarano et al, 2021 ) and application of an exogenous electric field to cells using PEF has the capacity to influence their migration, proliferation, and functionality ( Ren et al, 2019 ; Gouarderes et al, 2020 ; Gouarderes et al, 2022 ). PEF treatment of skin cells has focused largely on epidermal and dermal cell types including fibroblasts ( Liu et al, 2018 ; Nguyen et al, 2018 ; Das et al, 2020 ; Gouarderes et al, 2020 ) and keratinocytes ( Ren et al, 2019 ; Das et al, 2020 ) ( Table 2 ).…”

“…PEF treatment of cell suspensions or monolayers has been successfully executed using modified electrochambers ( Hess et al, 2012 ; Liu et al, 2018 ; Ren et al, 2019 ) or electroporation cuvettes comprised of two opposing electrode plates on a cuvette ( Hargrave and Li, 2012 ; Hargrave and Li, 2015 ) ( Figure 2 ). Needle electrodes composed of two opposing needle electrodes or multi-needle arrays have also been arranged to deliver electric fields to multi-well plates ( Figure 2 ) ( Basu et al, 2014 ; Chang et al, 2017 ; Das et al, 2020 ).…”

“…When embedded in 3D collagen lattices, PEF-treated fibroblasts had enhanced alignment and contractility ( Liu et al, 2018 ; Gouarderes et al, 2022 ). Studies investigating the effect of PEF on keratinocytes have been limited to 2D culture and showed variable effects on their migration ( Ren et al, 2019 ; Das et al, 2020 ).…”

The Emerging Role of Ionic Liquid-Based Approaches for Enhanced Skin Permeation of Bioactive Molecules: A Snapshot of the Past Couple of Years

“…Latest examples on skin applications of electroporation techniques include the permeation of low concentration formic and acetic acids for wound disinfection [ 16 , 17 ]. This type of electroporation applications was found to promote differential regrowth of dermal fibroblasts and keratinocytes [ 18 ].…”