Effect of cold atmospheric microwave plasma (CAMP) on wound healing in canine keratinocytes

Abstract

Cutaneous wound healing is a biological process that occurs upon skin injury and involves different mechanisms to repair tissue damage. Improper healing or prolonged curation period of wound lesions may induce unpleasant complications. Cold atmospheric microwave plasma (CAMP) is an upcoming medical therapeutic option for skin infection and wound treatment. However, the molecular mechanisms of CAMP-mediated canine wound healing are not well characterized. Wound-healing activity was examined to elucidate the biological effects and molecular mechanisms of CAMP. Canine keratinocytes (CPEKs) were treated using CAMP, and their wound-healing activities were evaluated. The molecular mechanisms of that effect were examined, based on RNA-Seq analysis data, and verified using immunoblotting and polymerase chain reaction. It was found that the CAMP-treated cells exhibited a significant increase in cell migration evaluated by scratch assay in human keratinocytes (HaCaT) and canine keratinocytes (CPEK). Additionally, CAMP-treated CPEK cells showed a significant positive effect on cell invasion. The RNA-Seq data revealed that CAMP alters different genes and pathways in CPEK cells. Gene expression involved in the cell cycle, cell proliferation, angiogenesis, cell adhesion, and wound healing was upregulated in CAMP-treated cells compared with gas-activated media used as a control. The Hippo pathway was also analyzed, and the protein and mRNA levels of YAP were significantly increased in CAMP-treated cells. CAMP-treated CPEK cells indicated the downregulation of E-cadherin and upregulation of vimentin, Snail, and Slug at transcription and translation levels, contributing to a favorable effect on cell migration. Our findings suggested that CAMP treatment provided beneficial effects on the curative wound process through the induction of genes involved in wound healing, promotion of EMT, and increase in the molecular targets in the Hippo signaling pathway.

A Multiparametric Analysis Reveals Differential Behavior of Spheroid Cultures on Distinct Ultra-Low Attachment Plates Types

“…Previous work showed that YAP1 nuclear localization in adherent HaCaT cultures is dependent on cell density [24] , that YAP1 activity is primarily confined to the basal, proliferating keratinocyte layer of the epidermis [43] , and that YAP1 activity is increased in activated keratinocytes during wound healing [44] . This prompted us to investigate a potential correlation between the reduced amount of Ki-67+ HaCaT cells in spheroids from plate type A.…”

Clinical application of cold atmospheric-pressure plasma: mechanisms and irradiation conditions

“…Studies on wound healing and cancer cell treatment with CAP describe the characteristics of the plasma generator and treatment conditions, and report the effects of different plasma treatment conditions. In wound healing treatments using animal models, the operating voltage for plasma generation ranges from 7 to 32 kV, and the SLM ranges from 0.5 to 5.0 l min −1 (figure 1(G)) [121][122][123][124][125]. Ar is predominantly used as the CAP gas, but He, O 2 , N 2 or a mixture of these has also been utilized [38,94,[126][127][128] He-CAP effectively coagulates blood vessels in wound tissues, and new blood vessels form better in plasma-irradiated tissues than in non-plasma-treated tissues [37].…”

Advances in Gas Therapeutics for Wound Healing: Mechanisms, Delivery Materials, and Prospects

“…[42] Cold atmospheric microwave plasma-treated cells indicate the downregulation of E-cadherin and upregulation of vimentin, Snail, and Slug at transcription and translation levels, thereby promoting cell migration. [63] These cells promote the cytoskeletal transformation of keratinocytes and migration due to changes in the expression of integrin-dependent focal adhesion molecules and MMPs. [64]…”