Skip to main content

Research Repository

Advanced Search

A Simple Polymicrobial Biofilm Keratinocyte Colonization Model for Exploring Interactions Between Commensals, Pathogens and Antimicrobials

Jordana-Lluch, Elena; Garcia, Vanina; Kingdon, Alexander D. H.; Singh, Nishant; Alexander, Cameron; Williams, Paul; Hardie, Kim R.

Authors

Elena Jordana-Lluch

Alexander D. H. Kingdon

Nishant Singh



Abstract

© 2020 Jordana-Lluch, Garcia, Kingdon, Singh, Alexander, Williams and Hardie. Skin offers protection against external insults, with the skin microbiota playing a crucial defensive role against pathogens that gain access when the skin barrier is breached. Linkages between skin microbes, biofilms and disease have not been well established although single-species biofilm formation by skin microbiota in vitro has been extensively studied. Consequently, the purpose of this work was to optimize and validate a simple polymicrobial biofilm keratinocyte model for investigating commensal, pathogen and keratinocyte interactions and for evaluating therapeutic agents or health promoting interventions. The model incorporates the commensals (Staphylococcus epidermidis and Micrococcus luteus) and pathogens (Staphylococcus aureus and Pseudomonas aeruginosa) which form robust polymicrobial biofilms on immortalized keratinocytes (HaCat cells). We observed that the commensals reduce the damage caused to the keratinocyte monolayer by either pathogen. When the commensals were combined with P. aeruginosa and S. aureus, much thinner biofilms were observed than those formed by the pathogens alone. When P. aeruginosa was inoculated with S. epidermidis in the presence or absence of M. luteus, the commensals formed a layer between the keratinocytes and pathogen. Although S. aureus completely inhibited the growth of M. luteus in dual-species biofilms, inclusion of S. epidermidis in triple or quadruple species biofilms, enabled M. luteus to retain viability. Using this polymicrobial biofilm keratinocyte model, we demonstrate that a quorum sensing (QS) deficient S. aureus agr mutant, in contrast to the parent, failed to damage the keratinocyte monolayer unless supplied with the exogenous cognate autoinducing peptide. In addition, we show that treatment of the polymicrobial keratinocyte model with nanoparticles containing an inhibitor of the PQS QS system reduced biofilm thickness and P. aeruginosa localization in mono- and polymicrobial biofilms.

Citation

Jordana-Lluch, E., Garcia, V., Kingdon, A. D. H., Singh, N., Alexander, C., Williams, P., & Hardie, K. R. (2020). A Simple Polymicrobial Biofilm Keratinocyte Colonization Model for Exploring Interactions Between Commensals, Pathogens and Antimicrobials. Frontiers in Microbiology, 11, Article 291. https://doi.org/10.3389/fmicb.2020.00291

Journal Article Type Article
Acceptance Date Feb 10, 2020
Online Publication Date Feb 26, 2020
Publication Date Feb 26, 2020
Deposit Date Mar 16, 2020
Publicly Available Date Mar 16, 2020
Journal Frontiers in Microbiology
Electronic ISSN 1664-302X
Publisher Frontiers Media
Peer Reviewed Peer Reviewed
Volume 11
Article Number 291
DOI https://doi.org/10.3389/fmicb.2020.00291
Keywords Microbiology (medical); Microbiology
Public URL https://nottingham-repository.worktribe.com/output/4127391
Publisher URL https://www.frontiersin.org/articles/10.3389/fmicb.2020.00291/full