Time-dependent effects of low-temperature atmospheric-pressure argon plasma on epithelial cell attachment, viability and tight junction formation in vitro

Abstract

The application of physical plasma to living tissues is expected to promote wound healing by plasma disinfection and stimulation of tissue regeneration. However, the effects of plasma on healthy cells must be studied and understood. In our experiments we used an argon plasma jet (kINPen®09) to gain insights into time-dependent plasma effects on cell attachment, viability and tight junction formation in vitro. Murine epithelial cells mHepR1 were suspended in complete cell culture medium and were irradiated with argon plasma (direct approach) for 30, 60 and 120 s. Suspecting that physical plasma may exert its effect via the medium, cell culture medium alone was first treated with argon plasma (indirect approach) and immediately afterwards, cells were added and also cultured for 24 h. Cell morphology and vitality were verified using light microscopy and an enzyme-linked immunosorbent assay. Already after 30 s of treatment the mHepR1 cells lost their capability to adhere and the cell vitality decreased with increasing treatment time. Interestingly, the same inhibitory effect was observed in the indirect approach. Furthermore, the argon plasma-treated culture medium-induced large openings of the cell's tight junctions, were verified by the zonula occludens protein ZO-1, which we observed for the first time in confluently grown epithelial cells.

Introduction to DIN-specification 91315 based on the characterization of the plasma jet kINPen® MED

“…One explanation for that is that different cell lines were used whereas GM00637 seems to be more sensitive to plasma application than mHepR1. Also, the weaker gas pressure used in the study of Hoentsch et al (2012) with 1.9 slm compared to 5 slm in the present study can be an explanation [50]. Still, our results reveal a strong and fast cytotoxic effect of the tested plasma jet.…”

“…Treatments for 10 s had no effects on the keratinocytes, whereas 30 s of argon plasma treatment led to a decreased number of cells which could re-attach and furthermore the expression levels of some adhesion molecules changed. However, a time-dependent decrease of cell vitality was also shown 2012 by Hoentsch et al [50]. They also treated suspended murine epithelial cells (mHepR1) in complete cell culture medium for 30, 60 and 120 s with the kINPen ® 09.…”

Plasma-activated medium induces A549 cell injury via a spiral apoptotic cascade involving the mitochondrial–nuclear network

“…Recent studies reported that plasma affected cancer cells not only directly, but also by the indirect treatment of cells with previously prepared medium or water irradiated by plasma, termed plasma-activated medium (PAM) or plasma-activated water (PAW), respectively [8,[13][14][15]. The effectiveness of PAM promoted the potential of clinical applications of plasma medicine because PAM can be prepared in advanced and stored until use.…”

“…In these studies on clinical applications, a low dose of plasma was reported to be able to induce the proliferation of cells, while a high dose induced apoptosis in cells [4,9]. Several studies in which applications for cancer therapy were designed demonstrated that plasma treatments killed cancer cells more effectively than normal cells [10][11][12].Recent studies reported that plasma affected cancer cells not only directly, but also by the indirect treatment of cells with previously prepared medium or water irradiated by plasma, termed plasma-activated medium (PAM) or plasma-activated water (PAW), respectively [8,[13][14][15]. The effectiveness of PAM promoted the potential of clinical applications of plasma medicine because PAM can be prepared in advanced and stored until use.…”

Cold plasma on full-thickness cutaneous wound accelerates healing through promoting inflammation, re-epithelialization and wound contraction

“…Although the mechanism of the interaction between cold plasma and cells or living tissue is still unclear 21 , several studies have reported the effects of cold plasma on key wound-related cells or sub-cells, included promoting theproliferationoffibroblasts 22 and endothelial cells 23 , as well as the growth of epithelial cells 24 , inhibiting the migration of fibroblasts 25 and their surface expression 26 , and activating integrin of fibroblasts and epithelial cells 27 . Interestingly, some of these effects are likely to be similar to the activities of natural ROS and/or NO during wound healing, particularly cold plasma's effects on the proliferation of both fibroblasts and endothelial cells 19 .…”