Optimization of nitric oxide donors for investigating biofilm dispersal response in Pseudomonas aeruginosa clinical isol
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APPLIED MICROBIAL AND CELL PHYSIOLOGY
Optimization of nitric oxide donors for investigating biofilm dispersal response in Pseudomonas aeruginosa clinical isolates Yu-ming Cai 1
&
Jeremy S. Webb 1
Received: 2 July 2020 / Revised: 13 August 2020 / Accepted: 23 August 2020 # The Author(s) 2020
Abstract Pseudomonas aeruginosa biofilms contribute heavily to chronic lung infection in cystic fibrosis patients, leading to morbidity and mortality. Nitric oxide (NO) has been shown to disperse P. aeruginosa biofilms in vitro, ex vivo and in clinical trials as a promising anti-biofilm agent. Traditional NO donors such as sodium nitroprusside (SNP) have been extensively employed in different studies. However, the dosage of SNP in different studies was not consistent, ranging from 500 nM to 500 μM. SNP is light sensitive and produces cyanide, which may lead to data misinterpretation and inaccurate predictions of dispersal responses in clinical settings. New NO donors and NO delivery methods have therefore been explored. Here we assessed 7 NO donors using P. aeruginosa PAO1 and determined that SNP and Spermine NONOate (S150) successfully reduced > 60% biomass within 24 and 2 h, respectively. While neither dosage posed toxicity towards bacterial cells, chemiluminescence assays showed that SNP only released NO upon light exposure in M9 media and S150 delivered much higher performance spontaneously. S150 was then tested on 13 different cystic fibrosis P. aeruginosa (CF-PA) isolates; most CF-PA biofilms were significantly dispersed by 250 μM S150. Our work therefore discovered a commercially available NO donor S150, which disperses CF-PA biofilms efficiently within a short period of time and without releasing cyanide, as an alternative of SNP in clinical trials in the future. Key points • S150 performs the best in dispersing P. aeruginosa biofilms among 7 NO donors. • SNP only releases NO in the presence of light, while S150 releases NO spontaneously. • S150 successfully disperses biofilms formed by P. aeruginosa cystic fibrosis clinical isolates. Keywords Nitric oxide . Pseudomonas aeruginosa . Biofilm . Cystic fibrosis . Chemiluminescence
Introduction The major cause of morbidity and mortality in cystic fibrosis (CF) patients is the chronic bacterial colonization of patients’ lungs and airways leading to pulmonary dysfunction and infection (Gilligan 1991; Govan and Deretic 1996). When Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00253-020-10859-7) contains supplementary material, which is available to authorized users. * Yu-ming Cai [email protected] 1
Institute for Life Sciences, University of Southampton, Southampton SO17 1BJ, UK
bacteria invade healthy individuals, opportunistic pathogens that overcome mucociliary clearance can be targeted by phagocytic cells and specific opsonizing antibodies (Govan and Deretic 1996). However, in CF patients, the dehydrated surface liquid on respiratory epithelium results in defective mucociliary clearance and frustrated phagocytosis du
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