Improved Wound Healing of Airway Epithelial Cells Is Mediated by Cold Atmospheric Plasma: A Time Course-Related Proteome Analysis.

Christian Scharf,Christine Eymann,Katrin Darm,Thomas Von Woedtke,Achim Beule,Leif Steil,Jörg Bernhardt,Martin Wilhelm,Georg Daeschlein,Fabian Görries,Werner Hosemann,Philipp Emicke,Jörn Winter

doi:10.1155/2019/7071536

Abstract

The promising potential of cold atmospheric plasma (CAP) treatment as a new therapeutic option in the field of medicine, particularly in Otorhinolaryngology and Respiratory medicine, demands primarily the assessment of potential risks and the prevention of any direct and future cell damages. Consequently, the application of a special intensity of CAP that is well tolerated by cells and tissues is of particular interest. Although improvement of wound healing by CAP treatment has been described, the underlying mechanisms and the molecular influences on human tissues are so far only partially characterized. In this study, human S9 bronchial epithelial cells were treated with cold plasma of atmospheric pressure plasma jet that was previously proven to accelerate the wound healing in a clinically relevant extent. We studied the detailed cellular adaptation reactions for a specified plasma intensity by time-resolved comparative proteome analyses of plasma treated vs. nontreated cells to elucidate the mechanisms of the observed improved wound healing and to define potential biomarkers and networks for the evaluation of plasma effects on human epithelial cells. K-means cluster analysis and time-related analysis of fold-change factors indicated concordantly clear differences between the short-term (up to 1 h) and long-term (24-72 h) adaptation reactions. Thus, the induction of Nrf2-mediated oxidative and endoplasmic reticulum stress response, PPAR-alpha/RXR activation as well as production of peroxisomes, and prevention of apoptosis already during the first hour after CAP treatment are important cell strategies to overcome oxidative stress and to protect and maintain cell integrity and especially microtubule dynamics. After resolving of stress, when stress adaptation was accomplished, the cells seem to start again with proliferation and cellular assembly and organization. The observed strategies and identification of marker proteins might explain the accelerated wound healing induced by CAP, and these indicators might be subsequently used for risk assessment and quality management of application of nonthermal plasma sources in clinical settings.

Highlights

The prospective application of cold atmospheric plasma (CAP) on the human body for the therapy of several diseases and nonhealing, chronic infected wounds of several locations requires an exact evaluation of possible risks and long-term effects and a deep but comprehensive analysis and description of molecular effects on human cells and tissues in response to cold plasma treatment.The term plasma in physics is defined as the fourth state of matter and is generated almost completely or partly by ionization of gas
As the mucosa of the mouth and upper airway are accessible, we evaluated the specific requirements for otorhinolaryngologic applications: the mucosa of the upper airways and the oral cavity is different from skin surfaces elsewhere on the human body, less resistant to mechanic trauma, and might be much more sensitive to external plasma applications
As recently shown by Lendeckel et al [17], S9 airway epithelial cells were clearly affected by the 120 s intensity of nonthermal plasma as demonstrated by (I) an increased wound healing rate in an in vitro wound healing model comprising plasma intensities in the range of 30 s to 360 s and (II) the highest number of regulated protein spots revealed by a plasma intensity-dependent 2D-DIGE approach

Summary

Introduction

The prospective application of cold atmospheric plasma (CAP) on the human body for the therapy of several diseases (e.g., cancer) and nonhealing, chronic infected wounds of several locations requires an exact evaluation of possible risks and long-term effects and a deep but comprehensive analysis and description of molecular effects on human cells and tissues in response to cold plasma treatment.The term plasma in physics is defined as the fourth state of matter and is generated almost completely or partly by ionization of gas. The prospective application of cold atmospheric plasma (CAP) on the human body for the therapy of several diseases (e.g., cancer) and nonhealing, chronic infected wounds of several locations requires an exact evaluation of possible risks and long-term effects and a deep but comprehensive analysis and description of molecular effects on human cells and tissues in response to cold plasma treatment. According to the thermal equilibrium state of electrons and heavy particles (e.g., atoms and ions), two temperature plasma forms can be classified: (I) thermal plasmas can reach several thousand degrees Kelvin and were used for instance for the ablation and cauterisation of biological tissue [1,2,3,4]. (II) Nonequilibrium plasmas can be created artificially under low and atmospheric pressure conditions and are characterized by a strong imbalance of electron and heavy particle temperature. Further promising are applications in cancer treatment [19,20,21,22,23]

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Conclusion