Targeted disruption of the extracellular polymeric network of Pseudomonas aeruginosa biofilms by alginate oligosaccharides

被引:131
|
作者
Powell, Lydia C. [1 ]
Pritchard, Manon F. [1 ]
Ferguson, Elaine L. [1 ]
Powell, Kate A. [1 ]
Patel, Shree U. [1 ]
Rye, Phil D. [2 ]
Sakellakou, Stavroula-Melina [3 ]
Buurma, Niklaas J. [3 ]
Brilliant, Charles D. [4 ]
Copping, Jack M. [4 ]
Menzies, Georgina E. [4 ]
Lewis, Paul D. [4 ]
Hill, Katja E. [1 ]
Thomas, David W. [1 ]
机构
[1] Cardiff Univ, Sch Dent, Adv Therapies Grp, Heath Pk, Cardiff CF14 4XY, S Glam, Wales
[2] AlgiPharma AS, Sandvika, Norway
[3] Cardiff Univ, Sch Chem, Phys Organ Chem Ctr, Cardiff, S Glam, Wales
[4] Swansea Univ, Resp Diagnost Grp, Swansea, W Glam, Wales
关键词
CYSTIC-FIBROSIS; LUNG INFECTION; IN-VITRO; OLIGOG CF-5/20; MECHANICAL-PROPERTIES; DNA; POLYSACCHARIDE; CALCIUM; MATRIX; PSL;
D O I
10.1038/s41522-018-0056-3
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Acquisition of a mucoid phenotype by Pseudomonas sp. in the lungs of cystic fibrosis (CF) patients, with subsequent overproduction of extracellular polymeric substance (EPS), plays an important role in mediating the persistence of multi-drug resistant (MDR) infections. The ability of a low molecular weight (Mn = 3200 g mol(-1)) alginate oligomer (OligoG CF-5/20) to modify biofilm structure of mucoid Pseudomonas aeruginosa (NH57388A) was studied in vitro using scanning electron microscopy (SEM), confocal laser scanning microscopy (CLSM) with Texas Red (TxRd (R))-labelled OligoG and EPS histochemical staining. Structural changes in treated biofilms were quantified using COMSTAT image-analysis software of CLSM z-stack images, and nanoparticle diffusion. Interactions between the oligomers, Ca2+ and DNA were studied using molecular dynamics (MD) simulations, Fourier transform infrared spectroscopy (FTIR) and isothermal titration calorimetry (ITC). Imaging demonstrated that OligoG treatment (>= 0.5%) inhibited biofilm formation, revealing a significant reduction in both biomass and biofilm height (P < 0.05). TxRd (R)-labelled oligomers readily diffused into established (24 h) biofilms. OligoG treatment (>= 2%) induced alterations in the EPS of established biofilms; significantly reducing the structural quantities of EPS polysaccharides, and extracellular (e)DNA (P < 0.05) with a corresponding increase in nanoparticle diffusion (P< 0.05) and antibiotic efficacy against established biofilms. ITC demonstrated an absence of rapid complex formation between DNA and OligoG and confirmed the interactions of OligoG with Ca2+ evident in FTIR and MD modelling. The ability of OligoG to diffuse into biofilms, potentiate antibiotic activity, disrupt DNA-Ca2+-DNA bridges and biofilm EPS matrix highlights its potential for the treatment of biofilm-related infections.
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页数:10
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