Antibiofilm Properties of Temporin-L on <i>Pseudomonas fluorescens</i> 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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MDPI AG
2020-11-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/21/22/8526 |
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| author | Angela Di Somma Federica Recupido Arianna Cirillo Alessia Romano Alessandra Romanelli Sergio Caserta Stefano Guido Angela Duilio |
| author_facet | Angela Di Somma Federica Recupido Arianna Cirillo Alessia Romano Alessandra Romanelli Sergio Caserta Stefano Guido Angela Duilio |
| author_sort | Angela Di Somma |
| collection | DOAJ |
| 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 <i>Pseudomonas fluorescens</i> (<i>P. fluorescens</i>) 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 <i>P. fluorescens</i> 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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| language | English |
| last_indexed | 2024-03-10T14:54:14Z |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-bf13effb1a6f43499e17d4fe6ff08b122023-11-20T20:45:18ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672020-11-012122852610.3390/ijms21228526Antibiofilm Properties of Temporin-L on <i>Pseudomonas fluorescens</i> in Static and In-Flow ConditionsAngela Di Somma0Federica Recupido1Arianna Cirillo2Alessia Romano3Alessandra Romanelli4Sergio Caserta5Stefano Guido6Angela Duilio7Department of Chemical Science, University of Naples “Federico II”, Via Cinthia, 80126 Naples, ItalyDivision of Chemical Technology, School of Chemistry, Aristotle University of Thessaloniki, University Box 116, 54 124 Thessaloniki, GreeceCEINGE—Biotecnologie Avanzate, 80145 Naples, ItalyCEINGE—Biotecnologie Avanzate, 80145 Naples, ItalyDepartment of Pharmaceutical Sciences, University of Milan, Via Venezian 21, 20133 Milan, ItalyDepartment of Chemical, Materials and Industrial Production Engineering, University of Naples “Federico II”, Piazzale V. Tecchio 80, 80125 Naples, ItalyDepartment of Chemical, Materials and Industrial Production Engineering, University of Naples “Federico II”, Piazzale V. Tecchio 80, 80125 Naples, ItalyDepartment of Chemical Science, University of Naples “Federico II”, Via Cinthia, 80126 Naples, ItalyBiofilms 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 <i>Pseudomonas fluorescens</i> (<i>P. fluorescens</i>) 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 <i>P. fluorescens</i> 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.https://www.mdpi.com/1422-0067/21/22/8526biofilmantimicrobial peptidebiofilm formation inhibitiondynamic and static condition |
| spellingShingle | Angela Di Somma Federica Recupido Arianna Cirillo Alessia Romano Alessandra Romanelli Sergio Caserta Stefano Guido Angela Duilio Antibiofilm Properties of Temporin-L on <i>Pseudomonas fluorescens</i> in Static and In-Flow Conditions International Journal of Molecular Sciences biofilm antimicrobial peptide biofilm formation inhibition dynamic and static condition |
| title | Antibiofilm Properties of Temporin-L on <i>Pseudomonas fluorescens</i> in Static and In-Flow Conditions |
| title_full | Antibiofilm Properties of Temporin-L on <i>Pseudomonas fluorescens</i> in Static and In-Flow Conditions |
| title_fullStr | Antibiofilm Properties of Temporin-L on <i>Pseudomonas fluorescens</i> in Static and In-Flow Conditions |
| title_full_unstemmed | Antibiofilm Properties of Temporin-L on <i>Pseudomonas fluorescens</i> in Static and In-Flow Conditions |
| title_short | Antibiofilm Properties of Temporin-L on <i>Pseudomonas fluorescens</i> in Static and In-Flow Conditions |
| title_sort | antibiofilm properties of temporin l on i pseudomonas fluorescens i in static and in flow conditions |
| topic | biofilm antimicrobial peptide biofilm formation inhibition dynamic and static condition |
| url | https://www.mdpi.com/1422-0067/21/22/8526 |
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