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Antibiofilm Properties of Temporin-L on Pseudomonas fluorescens in Static and In-Flow Conditions

Biofilms consist of a complex microbial community adhering to biotic or abiotic surfaces and enclosed within a protein/polysaccharide self-produced matrix. The formation of this structure represents the most important adaptive mechanism that leads to antibacterial resistance, and therefore, closely...

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Autores principales: Di Somma, Angela, Recupido, Federica, Cirillo, Arianna, Romano, Alessia, Romanelli, Alessandra, Caserta, Sergio, Guido, Stefano, Duilio, Angela
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7696879/
https://www.ncbi.nlm.nih.gov/pubmed/33198325
http://dx.doi.org/10.3390/ijms21228526
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author Di Somma, Angela
Recupido, Federica
Cirillo, Arianna
Romano, Alessia
Romanelli, Alessandra
Caserta, Sergio
Guido, Stefano
Duilio, Angela
author_facet Di Somma, Angela
Recupido, Federica
Cirillo, Arianna
Romano, Alessia
Romanelli, Alessandra
Caserta, Sergio
Guido, Stefano
Duilio, Angela
author_sort Di Somma, Angela
collection PubMed
description Biofilms consist of a complex microbial community adhering to biotic or abiotic surfaces and enclosed within a protein/polysaccharide self-produced matrix. The formation of this structure represents the most important adaptive mechanism that leads to antibacterial resistance, and therefore, closely connected to pathogenicity. Antimicrobial peptides (AMPs) could represent attractive candidates for the design of new antibiotics because of their specific characteristics. AMPs show a broad activity spectrum, a relative selectivity towards their targets (microbial membranes), the ability to act on both proliferative and quiescent cells, a rapid mechanism of action, and above all, a low propensity for developing resistance. This article investigates the effect at subMIC concentrations of Temporin-L (TL) on biofilm formation in Pseudomonas fluorescens (P. fluorescens) both in static and dynamic conditions, showing that TL displays antibiofilm properties. Biofilm formation in static conditions was analyzed by the Crystal Violet assay. Investigation of biofilms in dynamic conditions was performed in a commercial microfluidic device consisting of a microflow chamber to simulate real flow conditions in the human body. Biofilm morphology was examined using Confocal Laser Scanning Microscopy and quantified via image analysis. The investigation of TL effects on P. fluorescens showed that when subMIC concentrations of this peptide were added during bacterial growth, TL exerted antibiofilm activity, impairing biofilm formation both in static and dynamic conditions. Moreover, TL also affects mature biofilm as confocal microscopy analyses showed that a large portion of preformed biofilm architecture was clearly perturbed by the peptide addition with a significative decrease of all the biofilm surface properties and the overall biomass. Finally, in these conditions, TL did not affect bacterial cells as the live/dead cell ratio remained unchanged without any increase in damaged cells, confirming an actual antibiofilm activity of the peptide.
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spelling pubmed-76968792020-11-29 Antibiofilm Properties of Temporin-L on Pseudomonas fluorescens in Static and In-Flow Conditions Di Somma, Angela Recupido, Federica Cirillo, Arianna Romano, Alessia Romanelli, Alessandra Caserta, Sergio Guido, Stefano Duilio, Angela Int J Mol Sci Article Biofilms consist of a complex microbial community adhering to biotic or abiotic surfaces and enclosed within a protein/polysaccharide self-produced matrix. The formation of this structure represents the most important adaptive mechanism that leads to antibacterial resistance, and therefore, closely connected to pathogenicity. Antimicrobial peptides (AMPs) could represent attractive candidates for the design of new antibiotics because of their specific characteristics. AMPs show a broad activity spectrum, a relative selectivity towards their targets (microbial membranes), the ability to act on both proliferative and quiescent cells, a rapid mechanism of action, and above all, a low propensity for developing resistance. This article investigates the effect at subMIC concentrations of Temporin-L (TL) on biofilm formation in Pseudomonas fluorescens (P. fluorescens) both in static and dynamic conditions, showing that TL displays antibiofilm properties. Biofilm formation in static conditions was analyzed by the Crystal Violet assay. Investigation of biofilms in dynamic conditions was performed in a commercial microfluidic device consisting of a microflow chamber to simulate real flow conditions in the human body. Biofilm morphology was examined using Confocal Laser Scanning Microscopy and quantified via image analysis. The investigation of TL effects on P. fluorescens showed that when subMIC concentrations of this peptide were added during bacterial growth, TL exerted antibiofilm activity, impairing biofilm formation both in static and dynamic conditions. Moreover, TL also affects mature biofilm as confocal microscopy analyses showed that a large portion of preformed biofilm architecture was clearly perturbed by the peptide addition with a significative decrease of all the biofilm surface properties and the overall biomass. Finally, in these conditions, TL did not affect bacterial cells as the live/dead cell ratio remained unchanged without any increase in damaged cells, confirming an actual antibiofilm activity of the peptide. MDPI 2020-11-12 /pmc/articles/PMC7696879/ /pubmed/33198325 http://dx.doi.org/10.3390/ijms21228526 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Di Somma, Angela
Recupido, Federica
Cirillo, Arianna
Romano, Alessia
Romanelli, Alessandra
Caserta, Sergio
Guido, Stefano
Duilio, Angela
Antibiofilm Properties of Temporin-L on Pseudomonas fluorescens in Static and In-Flow Conditions
title Antibiofilm Properties of Temporin-L on Pseudomonas fluorescens in Static and In-Flow Conditions
title_full Antibiofilm Properties of Temporin-L on Pseudomonas fluorescens in Static and In-Flow Conditions
title_fullStr Antibiofilm Properties of Temporin-L on Pseudomonas fluorescens in Static and In-Flow Conditions
title_full_unstemmed Antibiofilm Properties of Temporin-L on Pseudomonas fluorescens in Static and In-Flow Conditions
title_short Antibiofilm Properties of Temporin-L on Pseudomonas fluorescens in Static and In-Flow Conditions
title_sort antibiofilm properties of temporin-l on pseudomonas fluorescens in static and in-flow conditions
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7696879/
https://www.ncbi.nlm.nih.gov/pubmed/33198325
http://dx.doi.org/10.3390/ijms21228526
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