Inactivation of Candida Biofilms by Non-Thermal Plasma and Its Enhancement for Fungistatic Effect of Antifungal Drugs

Yi Sun,Jue Zhang,Weidong Zhu,Haiyan Wu,Shuang Yu,Peng Sun,Wei Liu,Ruoyu Li,Jing Fang,Tianzi Jiang

doi:10.1371/journal.pone.0040629

Yi Sun, Jue Zhang + Show 8 more

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https://doi.org/10.1371/journal.pone.0040629

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Abstract

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/O2 (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 (•O2 -) and singlet molecular oxygen (1O2) were detected by ESR. We hence conclude that He/O2 (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.

Highlights

Candidiasis, caused by Candida species, is the most common fungal infection in humans [1,2]
CFUtreated/CFUcontrol) 6100%) showed that 80% of the Candida biofilms was inactivated after 10 s plasma microjet (PMJ) treatment
The increased using of implanted devices has facilitated the formation of Candida biofilms, which further accelerates infections caused by Candida species

Summary

Introduction

Candidiasis, caused by Candida species, is the most common fungal infection in humans [1,2]. Candida species are the microorganism exhibiting planktonic unicellular form, they commonly show filamentous growth or complex multicellular structure in the infected tissues [5]. These structured microbial communities, known as biofilms, can attach to surfaces and encase within a matrix of exopolymeric materials [5,6], and can form on various implanted medical devices such as vascular and urinary catheters, joint prostheses, cardiac valves, artificial vascular bypass devices, and those being topically used including contact lens and dentures [6,7,8,9]. To develop novel approaches to inactivate candidal biofilm has great clinical practicability in treating candidiasis, especially those associated with biofilms

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