Disinfection of ocular cells and tissues by atmospheric-pressure cold plasma

Paola Brun, Paola Brun, Maria Vono, Paola Venier, Elena Tarricone, Velika Deligianni, Emilio Martines, Matteo Zuin, Silvia Spagnolo, Roberto Cavazzana, Romilda Cardin, Ignazio Castagliuolo, Alvise la Gloria Valerio, Andrea Leonardi

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Abstract

Background: Low temperature plasmas have been proposed in medicine as agents for tissue disinfection and have received increasing attention due to the frequency of bacterial resistance to antibiotics. This study explored whether atmospheric-pressure cold plasma (APCP) generated by a new portable device that ionizes a flow of helium gas can inactivate ocular pathogens without causing significant tissue damage. Methodology/Principal Findings: We tested the APCP effects on cultured Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Candida albicans, Aspergillus fumigatus and Herpes simplex virus-1, ocular cells (conjunctival fibroblasts and keratocytes) and ex-vivo corneas. Exposure to APCP for 0.5 to 5 minutes significantly reduced microbial viability (colony-forming units) but not human cell viability (MTT assay, FACS and Tunel analysis) or the number of HSV-1 plaque-forming units. Increased levels of intracellular reactive oxygen species (ROS) in exposed microorganisms and cells were found using a FACS-activated 2′,7′-dichlorofluorescein diacetate probe. Immunoassays demonstrated no induction of thymine dimers in cell cultures and corneal tissues. A transient increased expression of 8-OHdG, genes and proteins related to oxidative stress (OGG1, GPX, NFE2L2), was determined in ocular cells and corneas by HPLC, qRT-PCR and Western blot analysis. Conclusions: A short application of APCP appears to be an efficient and rapid ocular disinfectant for bacteria and fungi without significant damage on ocular cells and tissues, although the treatment of conjunctival fibroblasts and keratocytes caused a time-restricted generation of intracellular ROS and oxidative stress-related responses. © 2012 Brun et al.
Original languageEnglish
Article numbere33245
Pages (from-to)-
JournalPLoS One
Volume7
Issue number3
DOIs
Publication statusPublished - 14 Mar 2012
Externally publishedYes

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All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Brun, P., Brun, P., Vono, M., Venier, P., Tarricone, E., Deligianni, V., Martines, E., Zuin, M., Spagnolo, S., Cavazzana, R., Cardin, R., Castagliuolo, I., la Gloria Valerio, A., & Leonardi, A. (2012). Disinfection of ocular cells and tissues by atmospheric-pressure cold plasma. PLoS One, 7(3), -. [e33245]. https://doi.org/10.1371/journal.pone.0033245