Antibiofilm Properties of Temporin-L on Pseudomonas fluorescens in Static and In-Flow Conditions.
Antibiofilm Properties of Temporin-L on Pseudomonas fluorescens in Static and In-Flow Conditions.
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DOI：
10.3390/ijms21228526
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发表时间：
2020-11-12
影响因子：
5.6
通讯作者：
Duilio A
中科院分区：
文献类型：
--
作者：
Di Somma A;Recupido F;Cirillo A;Romano A;Romanelli A;Caserta S;Guido S;Duilio A
生物膜由粘附于生物或非生物表面并封闭在蛋白质/多糖自生基质内的复杂微生物群落组成。这种结构的形成代表了导致抗菌抗性的最重要的适应机制，因此与致病性密切相关。抗菌肽（Antimicrobial Peptides，AMP）因其独特的生物学特性而成为设计新型抗生素的候选药物。AMP显示出宽的活性谱、对它们的靶标（微生物膜）的相对选择性、作用于增殖和静止细胞的能力、快速的作用机制，以及最重要的是，产生抗性的低倾向。本文研究了亚MIC浓度的Temporin-L（TL）在静态和动态条件下对荧光假单胞菌（P. fluorescens）生物膜形成的影响，结果表明TL显示出生物膜特性。通过结晶紫测定法分析静态条件下的生物膜形成。在由微流室组成的商业微流体装置中进行动态条件下的生物膜的研究，以模拟人体中的真实的流动条件。使用共聚焦激光扫描显微镜检查生物膜形态，并通过图像分析进行定量。TL对荧光假单胞菌影响的研究表明，当在细菌生长过程中加入低于MIC浓度的该肽时，TL发挥生物膜活性，在静态和动态条件下均损害生物膜形成。此外，TL还影响成熟的生物膜，因为共聚焦显微镜分析显示，大部分预先形成的生物膜结构明显受到肽添加的干扰，所有生物膜表面性质和总体生物量显著降低。最后，在这些条件下，TL不影响细菌细胞，因为活/死细胞比率保持不变，而损伤细胞没有任何增加，证实了肽的实际生物膜活性。
Biofilms consist of a complex microbial community adhering to biotic or abiotic surfaces and enclosed within a protein/polysaccharide self-produced matrix. The formation of this structure represents the most important adaptive mechanism that leads to antibacterial resistance, and therefore, closely connected to pathogenicity. Antimicrobial peptides (AMPs) could represent attractive candidates for the design of new antibiotics because of their specific characteristics. AMPs show a broad activity spectrum, a relative selectivity towards their targets (microbial membranes), the ability to act on both proliferative and quiescent cells, a rapid mechanism of action, and above all, a low propensity for developing resistance. This article investigates the effect at subMIC concentrations of Temporin-L (TL) on biofilm formation in Pseudomonas fluorescens (P. fluorescens) both in static and dynamic conditions, showing that TL displays antibiofilm properties. Biofilm formation in static conditions was analyzed by the Crystal Violet assay. Investigation of biofilms in dynamic conditions was performed in a commercial microfluidic device consisting of a microflow chamber to simulate real flow conditions in the human body. Biofilm morphology was examined using Confocal Laser Scanning Microscopy and quantified via image analysis. The investigation of TL effects on P. fluorescens showed that when subMIC concentrations of this peptide were added during bacterial growth, TL exerted antibiofilm activity, impairing biofilm formation both in static and dynamic conditions. Moreover, TL also affects mature biofilm as confocal microscopy analyses showed that a large portion of preformed biofilm architecture was clearly perturbed by the peptide addition with a significative decrease of all the biofilm surface properties and the overall biomass. Finally, in these conditions, TL did not affect bacterial cells as the live/dead cell ratio remained unchanged without any increase in damaged cells, confirming an actual antibiofilm activity of the peptide.
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影响因子：
28.3
作者：
Kim, Minyoung Kevin;Ingremeau, Francois;Zhao, Aishan;Bassler, Bonnie L.;Stone, Howard A.
通讯作者：
Stone, Howard A.
影响因子：
4.3
作者：
Patel, Saurabh Kumar;Pratap, Chandra Bhan;Nath, Gopal
通讯作者：
Nath, Gopal
影响因子：
3.9
作者：
Rusconi, Roberto;Lecuyer, Sigolene;Stone, Howard A.
通讯作者：
Stone, Howard A.
影响因子：
64.5
作者：
Persat A;Nadell CD;Kim MK;Ingremeau F;Siryaporn A;Drescher K;Wingreen NS;Bassler BL;Gitai Z;Stone HA
通讯作者：
Stone HA
影响因子：
3.1
作者：
Overhage, Joerg;Campisano, Andrea;Hancock, Robert E. W.
通讯作者：
Hancock, Robert E. W.