Atmospheric-Pressure Cold Plasma Induces Transcriptional Changes in Ex Vivo Human Corneas

PLoS One. 2015 Jul 23;10(7):e0133173. doi: 10.1371/journal.pone.0133173. eCollection 2015.

Abstract

Background: Atmospheric pressure cold plasma (APCP) might be considered a novel tool for tissue disinfection in medicine since the active chemical species produced by low plasma doses, generated by ionizing helium gas in air, induces reactive oxygen species (ROS) that kill microorganisms without substantially affecting human cells.

Objectives: In this study, we evaluated morphological and functional changes in human corneas exposed for 2 minutes (min) to APCP and tested if the antioxidant n-acetyl l-cysteine (NAC) was able to inhibit or prevent damage and cell death.

Results: Immunohistochemistry and western blotting analyses of corneal tissues collected at 6 hours (h) post-APCP treatment demonstrated no morphological tissue changes, but a transient increased expression of OGG1 glycosylase that returned to control levels in 24 h. Transcriptome sequencing and quantitative real time PCR performed on different corneas revealed in the treated corneas many differentially expressed genes: namely, 256 and 304 genes showing expression changes greater than ± 2 folds in the absence and presence of NAC, respectively. At 6 h post-treatment, the most over-expressed gene categories suggested an active or enhanced cell functioning, with only a minority of genes specifically concerning oxidative DNA damage and repair showing slight over-expression values (<2 folds). Moreover, time-related expression analysis of eight genes up-regulated in the APCP-treated corneas overall demonstrated the return to control expression levels after 24 h.

Conclusions: These findings of transient oxidative stress accompanied by wide-range transcriptome adjustments support the further development of APCP as an ocular disinfectant.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylcysteine / pharmacology
  • Aged
  • Air
  • Antioxidants / pharmacology
  • Atmospheric Pressure
  • Cold Temperature
  • Cornea / drug effects*
  • Cornea / metabolism
  • DNA Damage
  • DNA Glycosylases / biosynthesis
  • DNA Glycosylases / genetics
  • Disinfection / methods*
  • Enzyme Induction / drug effects
  • Equipment Design
  • Eye Proteins / genetics
  • Gene Expression Profiling
  • Helium
  • Humans
  • In Vitro Techniques
  • Middle Aged
  • Oxidative Stress / genetics
  • Plasma Gases / pharmacology*
  • RNA, Messenger / biosynthesis
  • RNA, Messenger / genetics
  • Time Factors
  • Transcription, Genetic / drug effects*
  • Transcriptome / drug effects*

Substances

  • Antioxidants
  • Eye Proteins
  • Plasma Gases
  • RNA, Messenger
  • Helium
  • DNA Glycosylases
  • oxoguanine glycosylase 1, human
  • Acetylcysteine

Grants and funding

This study was supported by the Italian Research and University Ministry (MIUR) and by the University of Padova, Italy.