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单物种生物膜基质成分的炎症潜能。

Inflammatory potential of monospecies biofilm matrix components.

机构信息

Faculty of Dentistry, Universidad de Costa Rica, San José, Costa Rica.

Faculty of Dentistry, University of Toronto, Toronto, ON, Canada.

出版信息

Int Endod J. 2019 Jul;52(7):1020-1027. doi: 10.1111/iej.13093. Epub 2019 Mar 5.

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.

摘要

目的

评估粪肠球菌和铜绿假单胞菌单物种生物膜的生物膜基质成分对巨噬细胞的炎症潜能。

方法

在需氧和厌氧条件下培养粪肠球菌和铜绿假单胞菌的体外生物膜(7 天)。提取并定量生物膜基质成分:胞外多糖(EPS)和细胞外 DNA(eDNA)。使用一氧化氮(NO)以及酶联免疫吸附试验评估 EPS 和 eDNA 对巨噬细胞系(RAW 267.4)的炎症潜能,用于肿瘤坏死因子(TNF-α)和白细胞介素-6(IL-6)的表达。铜绿假单胞菌 LPS 和浮游菌为阳性对照。采用单因素方差分析和 Tukey 事后检验进行统计学分析。

结果

两种生物膜菌株提取的 EPS 在两种生长条件下的含量均高于 eDNA(P<0.05)。生物膜成分的炎症潜能低于浮游菌和 LPS。与 eDNA 相比,两种菌株的 EPS 产生的炎症反应更高(P<0.05)。粪肠球菌 EPS 的 IL-6 和 TNF-α以及 NO 表达无差异(P≥0.05)。相比之下,铜绿假单胞菌 EPS 和 eDNA 的 IL-6 水平显著高于 TNF-α和 NO(P<0.05)。

结论

与浮游菌和 LPS 相比,测试的细菌单物种生物膜基质 EPS 和 eDNA 具有诱导低度炎症反应的能力。本研究强调了缺乏细菌的生物膜基质/成分具有诱导低度慢性炎症的潜力。

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