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Prevention of ESKAPE pathogen biofilm formation by antimicrobial peptides WLBU2 and LL37.

ABSTRACT: OBJECTIVES:Bacterial biofilm-dependent infections (e.g. cystic fibrosis, surgical sites, and medical implants) are associated with enhanced drug-resistance and are thus difficult to eradicate. The goal of this study was to systematically compare three distinct classes of antimicrobial peptides (AMPs) that include the clinically used antibiotic colistin, the natural AMP LL37, the engineered cationic-AMP WLBU2, and four commonly used antibiotics with different bactericidal mechanisms (tobramycin, ciprofloxacin, ceftazidime, and vancomycin) for biofilm prevention properties. METHODS:Using biofilm-prevention assays, we detected bacterial biomass post-attachment in subinhibitory concentrations (1/3 of the minimum inhibitory concentration [MIC]) for each AMP by the crystal violet method, to distinguish the commonly known bactericidal activity from potentially distinct mechanisms of biofilm prevention. Biofilm regulatory gene expression was assessed using RT-qPCR for correlation with biofilm growth inhibition. RESULTS:Commonly used antibiotics at 1x MIC showed modest ESKAPE biofilm prevention while 1/3 MIC of AMPs demonstrated up to 90% biofilm prevention. WLBU2 was generally more effective in preventing bacterial attachment than colistin and LL37. Changes in bacterial biofilm regulatory gene expression were consistent with biofilm prevention. CONCLUSION:The data warrant further exploration of AMPs with optimized structures to fill a knowledge gap on the potential application of AMPs for difficult-to-cure bacterial biofilm-related infections.

SUBMITTER: Lin Q

PROVIDER: S-EPMC6230315 | biostudies-literature | 2018 Nov

REPOSITORIES: biostudies-literature

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Prevention of ESKAPE pathogen biofilm formation by antimicrobial peptides WLBU2 and LL37.

Lin Qiao Q Deslouches Berthony B Montelaro Ronald C RC Di Y Peter YP

International journal of antimicrobial agents 20180510 5

<h4>Objectives</h4>Bacterial biofilm-dependent infections (e.g. cystic fibrosis, surgical sites, and medical implants) are associated with enhanced drug-resistance and are thus difficult to eradicate. The goal of this study was to systematically compare three distinct classes of antimicrobial peptides (AMPs) that include the clinically used antibiotic colistin, the natural AMP LL37, the engineered cationic-AMP WLBU2, and four commonly used antibiotics with different bactericidal mechanisms (tobr ...[more]

PMID: 29753132

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Project description:Staphylococcus aureus infections represent the major cause of titanium based-orthopaedic implant failure. Current treatments for S. aureus infections involve the systemic delivery of antibiotics and additional surgeries, increasing health-care costs and affecting patient's quality of life. As a step toward the development of new strategies that can prevent these infections, we build upon previous work demonstrating that the colonization of catheters by the fungal pathogen Candida albicans can be prevented by coating them with thin polymer multilayers composed of chitosan (CH) and hyaluronic acid (HA) designed to release a ?-amino acid-based peptidomimetic of antimicrobial peptides (AMPs). We demonstrate here that this ?-peptide is also potent against S. aureus (MBPC?=?4??g/mL) and characterize its selectivity toward S. aureus biofilms. We demonstrate further that ?-peptide-containing CH/HA thin-films can be fabricated on the surfaces of rough planar titanium substrates in ways that allow mammalian cell attachment and permit the long-term release of ?-peptide. ?-Peptide loading on CH/HA thin-films was then adjusted to achieve release of ?-peptide quantities that selectively prevent S. aureus biofilms on titanium substrates in vitro for up to 24?days and remained antimicrobial after being challenged sequentially five times with S. aureus inocula, while causing no significant MC3T3-E1 preosteoblast cytotoxicity compared to uncoated and film-coated controls lacking ?-peptide. We conclude that these ?-peptide-containing films offer a novel and promising localized delivery approach for preventing orthopaedic implant infections. The facile fabrication and loading of ?-peptide-containing films reported here provides opportunities for coating other medical devices prone to biofilm-associated infections. STATEMENT OF SIGNIFICANCE: Titanium (Ti) and its alloys are used widely in orthopaedic devices due to their mechanical strength and long-term biocompatibility. However, these devices are susceptible to bacterial colonization and the subsequent formation of biofilms. Here we report a chitosan and hyaluronic acid polyelectrolyte multilayer-based approach for the localized delivery of helical, cationic, globally amphiphilic ?-peptide mimetics of antimicrobial peptides to inhibit S. aureus colonization and biofilm formation. Our results reveal that controlled release of this ?-peptide can selectively kill S. aureus cells without exhibiting toxicity toward MC3T3-E1 preosteoblast cells. Further development of this polymer-based coating could result in new strategies for preventing orthopaedic implant-related infections, improving outcomes of these titanium implants.

Dataset Information

Prevention of ESKAPE pathogen biofilm formation by antimicrobial peptides WLBU2 and LL37.

Publications

Prevention of ESKAPE pathogen biofilm formation by antimicrobial peptides WLBU2 and LL37.

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