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Mechanism of Action of Surface Immobilized Antimicrobial Peptides Against Pseudomonas aeruginosa

Bacterial colonization and biofilm development on medical devices can lead to infection. Antimicrobial peptide-coated surfaces may prevent such infections. Melimine and Mel4 are chimeric cationic peptides showing broad-spectrum antimicrobial activity once attached to biomaterials and are highly bioc...

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Autores principales: Yasir, Muhammad, Dutta, Debarun, Hossain, Khondker R., Chen, Renxun, Ho, Kitty K. K., Kuppusamy, Rajesh, Clarke, Ronald J., Kumar, Naresh, Willcox, Mark D. P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6987417/
https://www.ncbi.nlm.nih.gov/pubmed/32038530
http://dx.doi.org/10.3389/fmicb.2019.03053
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author Yasir, Muhammad
Dutta, Debarun
Hossain, Khondker R.
Chen, Renxun
Ho, Kitty K. K.
Kuppusamy, Rajesh
Clarke, Ronald J.
Kumar, Naresh
Willcox, Mark D. P.
author_facet Yasir, Muhammad
Dutta, Debarun
Hossain, Khondker R.
Chen, Renxun
Ho, Kitty K. K.
Kuppusamy, Rajesh
Clarke, Ronald J.
Kumar, Naresh
Willcox, Mark D. P.
author_sort Yasir, Muhammad
collection PubMed
description Bacterial colonization and biofilm development on medical devices can lead to infection. Antimicrobial peptide-coated surfaces may prevent such infections. Melimine and Mel4 are chimeric cationic peptides showing broad-spectrum antimicrobial activity once attached to biomaterials and are highly biocompatible in animal models and have been tested in Phase I and II/III human clinical trials. These peptides were covalently attached to glass using an azidobenzoic acid linker. Peptide attachment was confirmed using X-ray photoelectron spectroscopy and amino acid analysis. Mel4 when bound to glass was able to adopt a more ordered structure in the presence of bacterial membrane mimetic lipids. The ability of surface bound peptides to neutralize endotoxin was measured along with their interactions with the bacterial cytoplasmic membrane which were analyzed using DiSC(3)-5 and Sytox green, Syto-9, and PI dyes with fluorescence microscopy. Leakage of ATP and nucleic acids from cells were determined by analyzing the surrounding fluid. Attachment of the peptides resulted in increases in the percentage of nitrogen by 3.0% and 2.4%, and amino acid concentrations to 0.237 nmole and 0.298 nmole per coverslip on melimine and Mel4 coated surfaces, respectively. The immobilized peptides bound lipopolysaccharide and disrupted the cytoplasmic membrane potential of Pseudomonas aeruginosa within 15 min. Membrane depolarization was associated with a reduction in bacterial viability by 82% and 63% for coatings melimine and Mel4, respectively (p < 0.001). Disruption of membrane potential was followed by leakage of ATP from melimine (1.5 ± 0.4 nM) or Mel4 (1.3 ± 0.2 nM) coated surfaces compared to uncoated glass after 2 h (p < 0.001). Sytox green influx started after 3 h incubation with either peptide. Melimine coatings yielded 59% and Mel4 gave 36% PI stained cells after 4 h. Release of the larger molecules (DNA/RNA) commenced after 4 h for melimine (1.8 ± 0.9 times more than control; p = 0.008) and after 6 h with Mel4 (2.1 ± 0.2 times more than control; p < 0.001). The mechanism of action of surface bound melimine and Mel4 was similar to that of the peptides in solution, however, their immobilization resulted in much slower (approximately 30 times) kinetics.
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spelling pubmed-69874172020-02-07 Mechanism of Action of Surface Immobilized Antimicrobial Peptides Against Pseudomonas aeruginosa Yasir, Muhammad Dutta, Debarun Hossain, Khondker R. Chen, Renxun Ho, Kitty K. K. Kuppusamy, Rajesh Clarke, Ronald J. Kumar, Naresh Willcox, Mark D. P. Front Microbiol Microbiology Bacterial colonization and biofilm development on medical devices can lead to infection. Antimicrobial peptide-coated surfaces may prevent such infections. Melimine and Mel4 are chimeric cationic peptides showing broad-spectrum antimicrobial activity once attached to biomaterials and are highly biocompatible in animal models and have been tested in Phase I and II/III human clinical trials. These peptides were covalently attached to glass using an azidobenzoic acid linker. Peptide attachment was confirmed using X-ray photoelectron spectroscopy and amino acid analysis. Mel4 when bound to glass was able to adopt a more ordered structure in the presence of bacterial membrane mimetic lipids. The ability of surface bound peptides to neutralize endotoxin was measured along with their interactions with the bacterial cytoplasmic membrane which were analyzed using DiSC(3)-5 and Sytox green, Syto-9, and PI dyes with fluorescence microscopy. Leakage of ATP and nucleic acids from cells were determined by analyzing the surrounding fluid. Attachment of the peptides resulted in increases in the percentage of nitrogen by 3.0% and 2.4%, and amino acid concentrations to 0.237 nmole and 0.298 nmole per coverslip on melimine and Mel4 coated surfaces, respectively. The immobilized peptides bound lipopolysaccharide and disrupted the cytoplasmic membrane potential of Pseudomonas aeruginosa within 15 min. Membrane depolarization was associated with a reduction in bacterial viability by 82% and 63% for coatings melimine and Mel4, respectively (p < 0.001). Disruption of membrane potential was followed by leakage of ATP from melimine (1.5 ± 0.4 nM) or Mel4 (1.3 ± 0.2 nM) coated surfaces compared to uncoated glass after 2 h (p < 0.001). Sytox green influx started after 3 h incubation with either peptide. Melimine coatings yielded 59% and Mel4 gave 36% PI stained cells after 4 h. Release of the larger molecules (DNA/RNA) commenced after 4 h for melimine (1.8 ± 0.9 times more than control; p = 0.008) and after 6 h with Mel4 (2.1 ± 0.2 times more than control; p < 0.001). The mechanism of action of surface bound melimine and Mel4 was similar to that of the peptides in solution, however, their immobilization resulted in much slower (approximately 30 times) kinetics. Frontiers Media S.A. 2020-01-22 /pmc/articles/PMC6987417/ /pubmed/32038530 http://dx.doi.org/10.3389/fmicb.2019.03053 Text en Copyright © 2020 Yasir, Dutta, Hossain, Chen, Ho, Kuppusamy, Clarke, Kumar and Willcox. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Yasir, Muhammad
Dutta, Debarun
Hossain, Khondker R.
Chen, Renxun
Ho, Kitty K. K.
Kuppusamy, Rajesh
Clarke, Ronald J.
Kumar, Naresh
Willcox, Mark D. P.
Mechanism of Action of Surface Immobilized Antimicrobial Peptides Against Pseudomonas aeruginosa
title Mechanism of Action of Surface Immobilized Antimicrobial Peptides Against Pseudomonas aeruginosa
title_full Mechanism of Action of Surface Immobilized Antimicrobial Peptides Against Pseudomonas aeruginosa
title_fullStr Mechanism of Action of Surface Immobilized Antimicrobial Peptides Against Pseudomonas aeruginosa
title_full_unstemmed Mechanism of Action of Surface Immobilized Antimicrobial Peptides Against Pseudomonas aeruginosa
title_short Mechanism of Action of Surface Immobilized Antimicrobial Peptides Against Pseudomonas aeruginosa
title_sort mechanism of action of surface immobilized antimicrobial peptides against pseudomonas aeruginosa
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6987417/
https://www.ncbi.nlm.nih.gov/pubmed/32038530
http://dx.doi.org/10.3389/fmicb.2019.03053
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