Critical nitric oxide concentration for Pseudomonas aeruginosa biofilm reduction on polyurethane substrates
Critical nitric oxide concentration for Pseudomonas aeruginosa biofilm reduction on polyurethane substrates
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DOI：
10.1116/1.4962266
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发表时间：
2016-09-01
期刊：
影响因子：
2.1
通讯作者：
Reynolds, Melissa M.
中科院分区：
文献类型：
--
作者：
Neufeld, Bella H.;Reynolds, Melissa M.
细菌菌落存在于被称为生物膜的表面上，传统抗生素在本质上是无法穿透的，这最终推动了对替代治疗方法的研究。一氧化氮（NO）因其生物学上的有益特性而受到关注，特别是其抗菌能力。在生理条件下（如s -亚硝基硫醇）可以释放分子的NO供体可在临床环境中用于对抗细菌生物膜感染。在这里，作者描述了确定必要的临界浓度的一氧化氮，以使在医用级聚氨酯薄膜上生长的铜绿假单胞菌生物膜减少约90%。生物膜在最佳培养条件下[在37℃的营养肉汤培养基（NBM）中]生长24小时，然后在NBM中添加NO供体s -亚硝基谷胱甘肽（GSNO）再生长24小时。在挑战期后，使用不同浓度的NO测试生物膜的细胞活力，以确定导致细菌生物膜活力至少降低90%所需的临界量。GSNO的临界浓度为10mM，对应于2.73mM NO。在4、8、12和16 h分别使用临界量NO对24 h生物膜进行时间杀伤实验，结果表明，12 h生物膜活力降低90%，并持续整个24 h。随后通过一氧化氮分析仪测试该临界浓度的总NO释放。12 h激发期内释放的NO总量为5.97+/-0.66 × 10(-6) mol NO，对应于1.49+/-0.17 μ mol NO/ml NBM。这是第一次确定在医学相关聚合物上引发这种生物反应所需的临界NO浓度。(C) 2016年美国真空学会。
Bacterial colonies that reside on a surface, known as biofilms, are intrinsically impenetrable to traditional antibiotics, ultimately driving research toward an alternative therapeutic approach. Nitric oxide ( NO) has gained attention for its biologically beneficial properties, particularly centered around its antibacterial capabilities. NO donors that can release the molecule under physiological conditions ( such as S-nitrosothiols) can be utilized in clinical settings to combat bacterial biofilm infections. Herein the authors describe determining a critical concentration of NO necessary to cause > 90% reduction of a Pseudomonas aeruginosa biofilm grown on medical grade polyurethane films. The biofilm was grown under optimal culture conditions [ in nutrient broth media ( NBM) at 37 degrees C] for 24 h before the addition of the NO donor S-nitrosoglutathione ( GSNO) in NBM for an additional 24 h. The cellular viability of the biofilm after the challenge period was tested using varying concentrations of NO to determine the critical amount necessary to cause at least a 90% reduction in bacterial biofilm viability. The critical GSNO concentration was found to be 10mM, which corresponds to 2.73mM NO. Time kill experiments were performed on the 24 h biofilm using the critical amount of NO at 4, 8, 12, and 16 h and it was determined that the 90% biofilm viability reduction occurred at 12 h and was sustained for the entire 24 h challenge period. This critical concentration was subsequently tested for total NO release via a nitric oxide analyzer. The total amount of NO released over the 12 h challenge period was found to be 5.97+/-0.66 x 10(-6) mol NO, which corresponds to 1.49+/-0.17 mu mol NO/ml NBM. This is the first identification of the critical NO concentration needed to elicit this biological response on a medically relevant polymer. (C) 2016 American Vacuum Society.