Inflammatory potential of monospecies biofilm matrix components

Abstract

Aim To assess the inflammatory potential of biofilm matrix constituents of Enterococcus faecalis and Pseudomonas aeruginosa monospecies biofilms on macrophages. Methodology In vitro biofilms of E. faecalis and P. aeruginosa were grown (7 days) in aerobic and anaerobic conditions. The biofilm matrix components: exopolysaccharides (EPS) and extracellular DNA (eDNA) were extracted and quantified. The inflammatory potential of EPS and eDNA was assessed on macrophage cell lines (RAW 267.4) using nitric oxide (NO), and enzyme‐linked immunosorbent assay for tumour necrosis factor (TNF‐α) and interleukin‐6 (IL‐6) expressions. LPS from P. aeruginosa and planktonic bacteria were positive controls. One‐way analysis of variance and the Tukey post hoc test were used for statistical analysis. Results Extracted EPS from both biofilm strains was associated with higher levels than eDNA in both growth conditions (P < 0.05). The biofilm components had less inflammatory potential compared to planktonic bacteria and LPS. EPS produced higher levels of inflammatory response compared to eDNA for both strains (P < 0.05). IL‐6 and TNF‐α, and NO expression showed no difference for E. faecalis EPS (P ≥ 0.05). In contrast, P. aeruginosa EPS and eDNA had significant levels of IL‐6 compared to TNF‐α and NO (P < 0.05). Conclusions Monospecies biofilm matrix EPS and eDNA from the bacterial strains tested had the ability to induce a low‐grade inflammatory response when compared to planktonic bacteria and LPS. This study highlights the potential of biofilm matrix/components, devoid of bacteria to induce low‐grade chronic inflammation.

Biofilm-Forming Ability and Effect of Sanitation Agents on Biofilm-Control of Thermophile Geobacillus sp. D413 and Geobacillus toebii E134

“…Many microorganisms release eDNA within their biofilm matrix. Moreover, eDNA was reported as a component of the EPS matrix of numerous Gram-negative and Gram-positive bacteria ( Ibáñez de Aldecoa et al 2017 ; Ramirez et al 2019 ). However, as far as it is known, there is no information about the eDNA of Geobacillus strain.…”

Type III Secretion Protein, PcrV, Impairs Pseudomonas aeruginosa Biofilm Formation by Increasing M1 Macrophage-Mediated Anti-bacterial Activities

“…In vitro co-culture of P. aeruginosa biofilm matrix exopolysaccharides (EPS) and extracellular DNA (eDNA) with RAW264.7 cells induced a lower-grade inflammatory response than that induced by planktonic bacteria-or LPS-treated cells (Ramirez et al, 2019). In accordance with these reports, we have demonstrated that compared to planktonic bacteria, P. aeruginosa biofilms impaired the production of proinflammatory cytokines, as well as iNOS, while promoting the expression of the anti-inflammatory enzyme, Arg1, both in vitro and in vivo.…”

“…Chronic P. aeruginosa biofilm infections in cystic fibrosis patients are dominated by a Th2 response with increased and decreased levels of IL4 and IFNγ, respectively ( Moser et al, 2000 ; Hartl et al, 2006 ), suggesting that P. aeruginosa biofilms persist by reducing host proinflammatory responses to infection. In vitro co-culture of P. aeruginosa biofilm matrix exopolysaccharides (EPS) and extracellular DNA (eDNA) with RAW264.7 cells induced a lower-grade inflammatory response than that induced by planktonic bacteria‐ or LPS-treated cells ( Ramirez et al, 2019 ). In accordance with these reports, we have demonstrated that compared to planktonic bacteria, P. aeruginosa biofilms impaired the production of proinflammatory cytokines, as well as iNOS, while promoting the expression of the anti-inflammatory enzyme, Arg1, both in vitro and in vivo .…”

Trans-cinnamaldehyde attenuatesEnterococcus faecalisvirulence and inhibits biofilm formation

“…CC-BY 4.0 International license made available under a (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is The copyright holder for this preprint this version posted March 15, 2021. ; https://doi.org/10.1101/2021.03.15.435450 doi: bioRxiv preprint 7 Deionized water (20 µl), 5% phenol solution (20 µl) (Sigma Aldrich, St. Louis, MO, USA) and 98% sulfuric acid (100 µl) were added to the wells and incubated at 90°C for 30 min [41]. The absorbance was measured at 492 nm using a microplate reader, and the concentration of biofilm exopolysaccharides was determined using a standard curve generated using glucose standards.…”