Formation of Pseudomonas aeruginosa Biofilms in Full-thickness Scald Burn Wounds in Rats
Formation of Pseudomonas aeruginosa Biofilms in Full-thickness Scald Burn Wounds in Rats
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DOI：
10.1038/s41598-019-50003-8
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发表时间：
2019-09-20
影响因子：
4.6
通讯作者：
Leung, Kai P.
中科院分区：
文献类型：
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作者：
Brandenburg, Kenneth S.;Weaver, Alan J., Jr.;Leung, Kai P.
使用 Sprague-Dawley 大鼠（350-450 g；n = 61）和最近更新的 Walker-Mason 大鼠烫伤模型，我们证明铜绿假单胞菌很容易在全层烧伤创面内形成生物膜。烧伤后，用磷酸盐缓冲盐水 (PBS) 中的铜绿假单胞菌对伤口进行表面接种，同时使用无菌 PBS 进行对照。在烧伤后第 1、3、7 和 11 天，对动物实施安乐死，并收集样本进行定量细菌学、细菌基因表达、全血细胞计数、组织学和髓过氧化物酶活性。在接种 1 x 10(4) CFU 铜绿假单胞菌的全层烧伤创面中形成了强大的生物膜感染。组织学和扫描电子显微镜均显示病原体遍布烧伤皮肤的组织学横截面。定量基因分析显示烧伤焦痂内藻酸盐和铜绿假单胞菌的薄膜生物膜基质基因显着上调。此外，烧伤创面环境中铜绿假单胞菌蛋白酶和铁载体的表达显着增加。有趣的是，与对照烧伤相比，焦痂或循环血液中宿主对病原体的中性粒细胞反应并未升高。这种新的全层烧伤生物膜感染模型将用于测试新的抗生物膜疗法，这些疗法可以与战斗中的士兵一起部署，以便在战场上的烧伤部位立即使用。
Using Sprague-Dawley rats (350-450 g; n = 61) and the recently updated Walker-Mason rat scald burn model, we demonstrated that Pseudomonas aeruginosa readily formed biofilms within full-thickness burn wounds. Following the burn, wounds were surface-inoculated with P. aeruginosa in phosphate-buffered saline (PBS), while sterile PBS was used for controls. On post-burn days 1, 3, 7, and 11, animals were euthanized and samples collected for quantitative bacteriology, bacterial gene expression, complete blood cell counts, histology, and myeloperoxidase activity. Robust biofilm infections developed in the full-thickness burn wounds inoculated with 1 x 10(4) CFU of P. aeruginosa. Both histology and scanning electron microscopy showed the pathogen throughout the histologic cross-sections of burned skin. Quantigene analysis revealed significant upregulation of alginate and pellicle biofilm matrix genes of P. aeruginosa within the burn eschar. Additionally, expression of P. aeruginosa proteases and siderophores increased significantly in the burn wound environment. Interestingly, the host's neutrophil response to the pathogen was not elevated in either the eschar or circulating blood when compared to the control burn. This new full-thickness burn biofilm infection model will be used to test new anti-biofilm therapies that may be deployed with soldiers in combat for immediate use at the site of burn injury on the battlefield.