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

Pattawika Lertpatipanpong; Chanin Sillapachaiyaporn; Garam Oh; Yeong-Hun Kang; Cheol-Yong Hwang; Seung Joon Baek

doi:10.3389/fcell.2023.1105692

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

Front Cell Dev Biol. 2023 Jan 25:11:1105692. doi: 10.3389/fcell.2023.1105692. eCollection 2023.

Authors

Pattawika Lertpatipanpong¹, Chanin Sillapachaiyaporn^{1

2}, Garam Oh¹, Yeong-Hun Kang³, Cheol-Yong Hwang³, Seung Joon Baek¹

Affiliations

¹ Laboratory of Signal Transduction, Research Institute for Veterinary Science, College of Veterinary Medicine, Seoul National University, Seoul, South Korea.
² Program in Clinical Biochemistry and Molecular Medicine, Department of Clinical Chemistry, Faculty of Allied Health Sciences, Chulalongkorn University, Bangkok, Thailand.
³ Laboratory of Veterinary Dermatology, Research Institute for Veterinary Science, College of Veterinary Medicine, Seoul National University, Seoul, South Korea.

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.

Keywords: RNA-Seq; cold plasma (CP); keratinocyte; microwave plasma; wound healing.

Grants and funding

This work was supported by the Project for Cooperative R&D between Industry, Academy, and Research Institute (S3057012 and S3247061) funded by the Korea Ministry of SMEs and Startups in 2022.