Transcriptional analysis and target genes discovery of Pseudomonas aeruginosa biofilm developed ex vivo chronic wound model

Abstract Bacterial biofilms formation is one of the major reasons for treatment failure in chronic wound infections. Therefore, diagnostic biomarkers remain the best option for prevention and treatment of chronic wound infections by biofilms. Herein, Pseudomonas aeruginosa PAO1 was used to mimic bio...

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Autores principales:	Xiaojuan Tan, Xi Cheng, Mei Hu, Yifan Zhang, Aiqun Jia, Jinwei Zhou, Guoping Zhu
Formato:	article
Lenguaje:	EN
Publicado:	SpringerOpen 2021
Materias:	Pseudomonas aeruginosa Biofilms Chronic wound infections Alkaline phosphatase Nitric oxide Biotechnology TP248.13-248.65 Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/ae1020211790460dbcca92b237da641c
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spelling	oai:doaj.org-article:ae1020211790460dbcca92b237da641c2021-11-28T12:10:48ZTranscriptional analysis and target genes discovery of Pseudomonas aeruginosa biofilm developed ex vivo chronic wound model10.1186/s13568-021-01317-22191-0855https://doaj.org/article/ae1020211790460dbcca92b237da641c2021-11-01T00:00:00Zhttps://doi.org/10.1186/s13568-021-01317-2https://doaj.org/toc/2191-0855Abstract Bacterial biofilms formation is one of the major reasons for treatment failure in chronic wound infections. Therefore, diagnostic biomarkers remain the best option for prevention and treatment of chronic wound infections by biofilms. Herein, Pseudomonas aeruginosa PAO1 was used to mimic biofilm development in porcine skin explants wells as ex vivo wound model. The microscopic imaging showed that PAO1 in porcine skin explants wells formed micro-colonies at 24 h, developed mushroom-like structure at 48 h, and at 72 h mushroom-like structure disappeared, remaining a thin bacterial lawn. RNA-seq data analysis revealed that the expression levels of genes involved in the type II hxc secretion system were significantly higher in biofilms than in planktonic cells, especially the expression of lapA encoding alkaline phosphatase. However, the expression levels of genes associated with denitrification pathway were markedly decreased in biofilms, especially the transcription of nirS encoding nitrite reductase to produce nitric oxide (NO). Therefore, their expressions and products were further detected using RT-qPCR and biochemical assays, respectively. The results found that the expression of lapA and alkaline phosphatase activity were induced, but the expression of nirS and intracellular NO were reduced at the whole biofilms cycle. The study indicates that LapA and NO would play an important role for P. aeruginosa biofilm formation in chronic wound infections. LapA would serve as potential target to monitor chronic wound infections by P. aeruginosa biofilms. Inducing NO would be used to treat chronic wound infections due to P. aeruginosa biofilms.Xiaojuan TanXi ChengMei HuYifan ZhangAiqun JiaJinwei ZhouGuoping ZhuSpringerOpenarticlePseudomonas aeruginosaBiofilmsChronic wound infectionsAlkaline phosphataseNitric oxideBiotechnologyTP248.13-248.65MicrobiologyQR1-502ENAMB Express, Vol 11, Iss 1, Pp 1-12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Pseudomonas aeruginosa Biofilms Chronic wound infections Alkaline phosphatase Nitric oxide Biotechnology TP248.13-248.65 Microbiology QR1-502
spellingShingle	Pseudomonas aeruginosa Biofilms Chronic wound infections Alkaline phosphatase Nitric oxide Biotechnology TP248.13-248.65 Microbiology QR1-502 Xiaojuan Tan Xi Cheng Mei Hu Yifan Zhang Aiqun Jia Jinwei Zhou Guoping Zhu Transcriptional analysis and target genes discovery of Pseudomonas aeruginosa biofilm developed ex vivo chronic wound model
description	Abstract Bacterial biofilms formation is one of the major reasons for treatment failure in chronic wound infections. Therefore, diagnostic biomarkers remain the best option for prevention and treatment of chronic wound infections by biofilms. Herein, Pseudomonas aeruginosa PAO1 was used to mimic biofilm development in porcine skin explants wells as ex vivo wound model. The microscopic imaging showed that PAO1 in porcine skin explants wells formed micro-colonies at 24 h, developed mushroom-like structure at 48 h, and at 72 h mushroom-like structure disappeared, remaining a thin bacterial lawn. RNA-seq data analysis revealed that the expression levels of genes involved in the type II hxc secretion system were significantly higher in biofilms than in planktonic cells, especially the expression of lapA encoding alkaline phosphatase. However, the expression levels of genes associated with denitrification pathway were markedly decreased in biofilms, especially the transcription of nirS encoding nitrite reductase to produce nitric oxide (NO). Therefore, their expressions and products were further detected using RT-qPCR and biochemical assays, respectively. The results found that the expression of lapA and alkaline phosphatase activity were induced, but the expression of nirS and intracellular NO were reduced at the whole biofilms cycle. The study indicates that LapA and NO would play an important role for P. aeruginosa biofilm formation in chronic wound infections. LapA would serve as potential target to monitor chronic wound infections by P. aeruginosa biofilms. Inducing NO would be used to treat chronic wound infections due to P. aeruginosa biofilms.
format	article
author	Xiaojuan Tan Xi Cheng Mei Hu Yifan Zhang Aiqun Jia Jinwei Zhou Guoping Zhu
author_facet	Xiaojuan Tan Xi Cheng Mei Hu Yifan Zhang Aiqun Jia Jinwei Zhou Guoping Zhu
author_sort	Xiaojuan Tan
title	Transcriptional analysis and target genes discovery of Pseudomonas aeruginosa biofilm developed ex vivo chronic wound model
title_short	Transcriptional analysis and target genes discovery of Pseudomonas aeruginosa biofilm developed ex vivo chronic wound model
title_full	Transcriptional analysis and target genes discovery of Pseudomonas aeruginosa biofilm developed ex vivo chronic wound model
title_fullStr	Transcriptional analysis and target genes discovery of Pseudomonas aeruginosa biofilm developed ex vivo chronic wound model
title_full_unstemmed	Transcriptional analysis and target genes discovery of Pseudomonas aeruginosa biofilm developed ex vivo chronic wound model
title_sort	transcriptional analysis and target genes discovery of pseudomonas aeruginosa biofilm developed ex vivo chronic wound model
publisher	SpringerOpen
publishDate	2021
url	https://doaj.org/article/ae1020211790460dbcca92b237da641c
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_version_	1718408160959528960

Transcriptional analysis and target genes discovery of Pseudomonas aeruginosa biofilm developed ex vivo chronic wound model

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