Regeneration and control of human fibroblast cell density by intermittently delivered pulsed electric fields

Abstract

Proliferative scarring is a human disease with neither available effective treatment nor relevant animal model. One of the hypotheses for scar formation involves deregulation of fibroblast signaling and delayed apoptosis. Here, we introduce a new chemical-free method for fibroblast density control in culture by intermittently delivered pulsed electric fields (IDPEF), which cause irreversible damage to cell membranes. Using 5-100 pulses with electric field strength of 150 V/mm, pulse duration 70 µs, and frequency of 1 Hz, we investigated the effects of PEF application on growth, death, and regeneration of normal human dermal fibroblasts in culture. We found that the fraction of fibroblasts that survive depends on the number of pulses applied and follows a Weibull distribution. We have successfully developed an IDPEF protocol that controls fibroblasts density in culture. Specifically, through application of IDPEF every 72 h for 12 days, we maintain a normal human dermal fibroblast density in the 3.1 ± 0.2 × 10(5) -1.4 ± 0.2 × 10(5) cell/mL range. Our results suggest that IDPEFs may prove useful as a non-chemical method for fibroblast density control in human wound healing.

Rejuvenation of aged rat skin with pulsed electric fields

“…These electric properties changes, which can be attributed to water content (Peyman et al, ) and structural changes (Farage, Miller, Elsner, & Maibach, ), affect the distribution of the electric fields inside the skin, and, therefore, could change the local electric field strength to which specific cells are exposed to (Aström, Lemaire, & Wardell, ; Corovic et al, ; Golberg, Bruinsma, Uygun, & Yarmush, ). Exposure of cells to the different local electric field could lead to different responses (Golberg, Bei, Sheridan, & Yarmush, ).…”

Non-thermal, pulsed electric field cell ablation: A novel tool for regenerative medicine and scarless skin regeneration

“…These studies, however, were focused exclusively on PEF application for tumor ablation, rather than the phenomenon of scarless tissue regeneration, and thus reduced fibrosis was reported only as a favorable side-effect of PEF in comparison to other tissue ablation methods. Recently, we have successfully developed an intermittently delivered pulsed electric fields protocol that controls human normal skin fibroblasts density in culture 14 . We showed that in culture, the fraction of fibroblasts that survive the PEF regenerates to the 100% confluency 14 .…”

“…Recently, we have successfully developed an intermittently delivered pulsed electric fields protocol that controls human normal skin fibroblasts density in culture 14 . We showed that in culture, the fraction of fibroblasts that survive the PEF regenerates to the 100% confluency 14 . Based on these prior observations, we set out to evaluate PEF as a novel tool for use in regenerative medicine.…”

Preventing Scars after Injury with Partial Irreversible Electroporation

“…In that work we showed that a single application of pIRE to normal skin in rats triggers the secretion of new collagen, the formation of additional capillaries, the proliferation of the epidermis, and increases the total metabolic activity in the treated area (Golberg et al 2015b). In addition, we also have developed methods to control fibroblast cell populations using intermittently delivered pIRE in cell cultures (Golberg et al 2013a), and to disinfect third degree burns with complete IRE of the infecting bacteria (Golberg et al 2014). Here, we present a protocol for a therapeutic procedure based on intermittently delivering pIRE to burn wounds to prevent scarring.…”