Brun, Paola

Castagliuolo, Ignazio

Leonardi, Andrea

Martines, Emilio

Vono, Maria

Zuin, Matteo

Firenze University Press: E-Journals

IJAE Vol. 115, n. 1/2 (Supplement), 2010
25
Effects of cold plasma on pathogenic microorganisms, 
cells and corneal tissues
Paola Brun1, Maria Vono1, Ignazio Castagliuolo1, Matteo Zuin2, Emilio Martines2, Andrea Leonardi3
1 Department of Histology, Microbiology and Medical Biotechnology, Histology Unit, University of Padua, Italy
2 Consorzio RFX, Associazione Euratom-ENEA sulla Fusione, Padua, Italy
3 Department of Neuroscience, Ophthalmology Unit, University of Padua, Italy
background Low temperature plasma is a partially ionized gas that has long been 
considered an important tool for material processing and sterilization.
Aim The objective of the present study was to evaluate the effects of plasma on dif-
ferent microorganisms, cells and corneal tissues.
Methods A prototype of a plasma source has been developed and its effects on dif-
ferent microorganisms and on conjunctival fibroblasts and corneal keratocytes were as-
sessed at different exposure times. Microbial viability was expressed in CFU, the antiviral 
effect by analyzing the plaque forming units developed after infection of Vero cells and 
viability of human cells evaluated with MTT test. ROS production was analysed using 
the 2’,7’-dichlorodihydrofluorescein test. The micronucleus test was used to ascertain 
induction of DNA damage in human cells. Human corneas were also infected ex-vivo 
with bacteria (P. aerugionosa or S. aureus) or virus (HSV type 1) and treated with plasma. 
Bacterial CFU or viral plaques were then counted and histology observed. Apoptosis 
and UV damage of tissues exposed to plasma were also analyzed with the Tunel test and 
immunohistochemical analysis, respectively.
Results Application of plasma for several minutes caused a significant reduction 
of microbial viability. Conversely, treatment of human cells did not cause any signifi-
cant reduction of viability. No effect on HSV vitality was observed. High levels of ROS 
were found in both microorganisms and human cell cultures. Plasma application to in-
fected corneas caused a significant reduction of microbial viability with no evidence of 
morphological damage. No induction of thymine dimer formation nor apoptosis was 
observed in treated tissues. Conversely, a significant increase in the frequency of micro-
nucleated cells was found at all treatment times.
Conclusion Cold plasma reduced the viability of microorganisms but not that of 
human cells. The measurable effects caused by plasma consisted in high ROS produc-
tion and increases of micronucleated cells. Further study is necessary to investigate the 
evolution of DNA damage in treated cells.


Effects of cold plasma on pathogenic microorganisms, cells and corneal tissues

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Effects of cold plasma on pathogenic microorganisms, cells and corneal tissues

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