Inactivation of Candida biofilms by non-thermal plasma and its enhancement for fungistatic effect of antifungal drugs.
We investigated the antifungal effect of non-thermal plasma, as well as its combination with common antifungal drugs, against Candida biofilms. A direct current atmospheric pressure He/O(2) (2%) plasma microjet (PMJ) was used to treat Candida biofilms in a 96-well plate. Inactivation efficacies of the biofilms were evaluated by XTT assay and counting colony forming units (CFUs). Morphological properties of the biofilms were evaluated by Scanning Electron Microscope (SEM). The sessile minimal inhibitory concentrations (SMICs) of fluconazole, amphotericin B, and caspofungin for the biofilms were also tested. Electron Spin Resonance (ESR) spectroscopy was used to detect the reactive oxygen species (ROS) generated directly and indirectly by PMJ. The Candida biofilms were completely inactivated after 1 min PMJ treatment, where severely deformed fungal elements were observed in SEM images. The SMICs of the tested antifungal drugs for the plasma-treated biofilms were decreased by 2-6 folds of dilution, compared to those of the untreated controls. ROS such as hydroxyl radical ((•)OH), superoxide anion radical ((•)O(2) (-)) and singlet molecular oxygen ((1)O(2)) were detected by ESR. We hence conclude that He/O(2) (2%) plasma alone, as well as in combination with common antifungal drugs, is able to inactivate Candida biofilms rapidly. The generation of ROS is believed to be one of the underlying mechanisms for the fungicidal activity of plasma.
Department of Dermatology and Venereolgy, Peking University First Hospital, Research Center for Medical Mycology, Peking University, Beijing, China.
Yi
Y
Sun
Shuang
S
Yu
Peng
P
Sun
Haiyan
H
Wu
Weidong
W
Zhu
Wei
W
Liu
Jue
J
Zhang
Jing
J
Fang
Ruoyu
R
Li
D000935:Antifungal Agents / Q000494:pharmacology*; D018441:Biofilms / Q000187:drug effects*; D002175:Candida / Q000187:drug effects* / Q000254:growth & development / Q000502:physiology* / Q000648:ultrastructure; D015169:Colony Count, Microbial; D003497:Cyclic N-Oxides / Q000378:metabolism; D005609:Free Radicals / Q000378:metabolism; D006371:Helium / Q000737:chemistry; D008826:Microbial Sensitivity Tests; D010100:Oxygen / Q000737:chemistry; D058626:Plasma Gases / Q000494:pharmacology*; D013482:Superoxide Dismutase / Q000378:metabolism
D000935:Antifungal Agents; D003497:Cyclic N-Oxides; D005609:Free Radicals; D058626:Plasma Gases; D006371:Helium; C069977:2,2-dimethyl-5-hydroxy-1-pyrrolidinyloxy; D013482:Superoxide Dismutase; D010100:Oxygen; C003959:TEMPO