Effects of exogenous glucose on <i>Pseudomonas aeruginosa</i> biofilm formation and antibiotic resistance

She, Pengfei; Wang, Yanle; Liu, Yiqing; Tan, Fang; Chen, Lihua; Luo, Zhen; Wu, Yong

doi:10.1002/mbo3.933

Cited by 48 publications

(36 citation statements)

References 45 publications

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“…As previously reported across several studies [20,71], after 24 hours of growth in LB medium, P. aeruginosa and S. aureus formed biofilms that displayed the presence of metabolic activity (Figure 3). This is also observed in the presence of host components, as in FBS and IVWM, where both P. aeruginosa and S. aureus formed metabolicallyactive biofilms (Figure 2).…”

Section: Biofilm Formation Metabolic Activity and Interspecies Intersupporting

confidence: 80%

“…To determine the effects of the IVWM on the planktonic growth of P. aeruginosa and S. aureus (single-species), we performed growth curves in the IVWM, and in LB medium and FBS for comparison (Figure 1). Luria-Bertani (LB) broth is widely-used to study biofilms under in vitro conditions [20,38,[71][72][73], however, its composition (refined yeast extract and tryptone) poorly mimics the infection state. To recapitulate factors relevant to the wound milieu, several biofilm models incorporate fetal bovine serum (FBS), in varying concentrations, into growth conditions [22,26,74,75].…”

Section: Development Of An In Vitro Wound Milieu (Ivwm) That Mimics Hmentioning

confidence: 99%

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Milieu matters: Anin vitrowound milieu to recapitulate key features of, and probe new insights into, polymicrobial biofilms

Kadam

Vandana

Bhide

et al. 2021

Preprint

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Bacterial biofilms are a major cause of delayed wound healing. Consequently, the study of wound biofilms, particularly in host-relevant conditions, has gained importance. Most in vitro biofilm studies employ refined laboratory media to study biofilms, conditions that are not relevant to the infection state. To mimic the wound milieu, in vitro biofilm studies often incorporate serum or plasma in growth conditions, or employ clot or matrix-based biofilm models. While incorporating serum or plasma alone is a minimalistic approach, the more complex in vitro wound models are technically demanding, and poorly compatible with standard biofilm assays. Based on previous reports of clinical wound fluid composition, we have developed an in vitro wound milieu (IVWM) that includes, in addition to serum (to recapitulate wound fluid), matrix elements and biochemical factors. In comparison with Luria-Bertani broth and Fetal Bovine Serum (FBS), the IVWM was used to study planktonic growth and biofilm features, including interspecies interactions, of common wound pathogens, Staphylococcus aureus and Pseudomonas aeruginosa. We demonstrate that the IVWM recapitulates widely reported in vivo biofilm features such as metabolic activity, increased antibiotic tolerance, 3D structure, and interspecies interactions for single- and co-species biofilms. Further, the IVWM is simple to formulate, uses laboratory-grade components, and is compatible with standard biofilm assays. Given this, it holds potential as a tractable approach to study wound biofilms under host-relevant conditions.

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Section: Biofilm Formation Metabolic Activity and Interspecies Intersupporting

confidence: 80%

Section: Development Of An In Vitro Wound Milieu (Ivwm) That Mimics Hmentioning

confidence: 99%

Milieu matters: Anin vitrowound milieu to recapitulate key features of, and probe new insights into, polymicrobial biofilms

Kadam

Vandana

Bhide

et al. 2021

Preprint

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“…These authors attributed this difference to an enrichment of the biofilm matrix in polysaccharides in ASM(Ϫ). This explanation may, however, not apply to our study because TGN is supplemented by glucose, which upregulates the expression of the extracellular polysaccharide-related gene pslA, required for biofilm formation (24). Second, the addition of frozen mucus from patients with CF to preformed biofilms was shown to decrease the activity of tobramycin, which was related to a reduced transport of this polycationic drug in the biofilm (25) due to its strong ionic interactions with components of the biofilm matrix and/or mucus (26).…”

Section: Discussionmentioning

confidence: 80%

Activity of Antibiotics against Pseudomonas aeruginosa in an In Vitro Model of Biofilms in the Context of Cystic Fibrosis: Influence of the Culture Medium

Iglesias

Bambeke

2020

Antimicrob Agents Chemother

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Pseudomonas aeruginosa is a major cause of respiratory biofilm-related infections in patients with cystic fibrosis. We developed an in vitro pharmacodynamic model to study the activity of antipseudomonal antibiotics against PAO1 biofilms grown in artificial sputum medium with agar [ASM(+)] versus that against biofilms grown in Trypticase soy broth supplemented with glucose and NaCl (TGN). We measured bacterial counts, metabolic activity (fluorescein diacetate [FDA] hydrolysis), and biomass (crystal violet absorbance). Biofilms grew slower in ASM(+) than in TGN but reached the same CFU counts and metabolic activity in both media and a slightly higher biomass after 48 h in ASM(+) than in TGN. The concentration-response curves of the antibiotics after 24 h of incubation with mature biofilms showed maximal effects ranging from a 3 (ciprofloxacin)- to a 1.5 (ceftazidime, meropenem)-log10-CFU decrease, with tobramycin and colistin showing intermediate values. These maximal reductions in the numbers of CFU were similar in both media for ciprofloxacin and β-lactams but lower in ASM(+) than in TGN for tobramycin and colistin; they were reached at concentrations lower than the human maximum concentration in plasma for ciprofloxacin and β-lactams only. The reductions in metabolic activity and in biomass were low in both media. Small-colony variants were selected by tobramycin in ASM(+) and by ciprofloxacin in both media. The model was then successfully applied to 4 isolates from patients with cystic fibrosis. These biofilms showed CFU counts similar to those of PAO1 biofilms in ASM(+) but a higher biomass than PAO1 biofilms in ASM(+) and moderate differences in their susceptibility to antibiotics from that of PAO1 biofilms grown in this medium. This model proved useful to establish the pharmacodynamic profile of drugs against P. aeruginosa biofilms in the context of cystic fibrosis.

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“…Hyperglycemia and high glycation products are the most striking features of diabetes ( Huijberts et al, 2008 ). In many in vitro studies, glucose was found to induce biofilm formation of bacteria ( Rode et al, 2007 ; Waldrop et al, 2014 ; Kyoui et al, 2016 ; She et al, 2019 ), it is not enough to explain the difference in severity of infection and antibiotic therapy effects between patients with similar blood sugar levels. As this study showed, we did not find differences in blood glucose levels between DB+ and DB- patients.…”

Section: Discussionmentioning

confidence: 99%

Advanced Glycation End Products Enhance Biofilm Formation by Promoting Extracellular DNA Release Through sigB Upregulation in Staphylococcus aureus

Xie

Liu

et al. 2020

Front. Microbiol.

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Bacterial biofilms do serious harm to the diabetic foot ulcer (DFU) because they play a crucial role in infection invasion and spread. Staphylococcus aureus , the predominant Gram-positive bacteria in diabetic foot infection (DFI), is often associated with colonization and biofilm formation. Through biofilm formation tests in vitro , we observed that S. aureus bacteria isolated from DFU wounds were more prone to form biofilms than those from non-diabetic patients, while there was no difference in blood sugar between the biofilm (+) diabetics (DB+) and biofilm (-) diabetics (DB-). Furthermore, we found that advanced glycation end products (AGEs) promoted the biofilm formation of S. aureus in clinical isolates and laboratory strains in vitro , including a methicillin-resistant strain. Analysis of biofilm components demonstrated that the biofilms formed mainly by increasing extracellular DNA (eDNA) release; remarkably, the S. aureus global regulator sigB was upregulated, and its downstream factor lrgA was downregulated after AGE treatments. Mechanism studies using a sigB -deleted mutant (Newman-Δ sigB ) confirmed that AGEs decreased expression of lrgA via induction of sigB , which is responsible for eDNA release and is a required component for S. aureus biofilm development. In conclusion, the present study suggests that AGEs promote S. aureus biofilm formation via an eDNA-dependent pathway by regulating sigB . The data generated by this study will provide experimental proof and theoretical support to improve DFU infection healing.

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Effects of exogenous glucose on Pseudomonas aeruginosa biofilm formation and antibiotic resistance

Cited by 48 publications

References 45 publications

Milieu matters: Anin vitrowound milieu to recapitulate key features of, and probe new insights into, polymicrobial biofilms

Milieu matters: Anin vitrowound milieu to recapitulate key features of, and probe new insights into, polymicrobial biofilms

Activity of Antibiotics against Pseudomonas aeruginosa in an In Vitro Model of Biofilms in the Context of Cystic Fibrosis: Influence of the Culture Medium

Advanced Glycation End Products Enhance Biofilm Formation by Promoting Extracellular DNA Release Through sigB Upregulation in Staphylococcus aureus

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