纳米银离子对细菌生物膜作用的探讨 - 第三军医大学学报.docVIP

纳米银离子对铜绿假单胞菌生物膜的细菌死亡率的影响 邹志慧，余加林(，刘官信，李芳，林丽华，林雅茵 (重庆医科大学附属儿童医院新生儿科，重庆400014) 摘要：目的 探讨纳米银离子对细菌生物膜（Biofilm，BF）的细菌死亡率的影响。方法 采用摇床法，以纳米银离子含量不同的乙烯-醋酸乙烯酯thylene-Vinyl acetate， EVA）塑料为细菌粘附载体，模拟体内铜绿假单胞菌(P.aeruginosa,PA)BF形成的微环境，建立体外BF模型；将培养3d的空白标本分别在扫描电子显微镜（scanning electron microscopy, SEM）下及用FITC-ConA染色后荧光显微镜下观察不含纳米银EVA中BF的形成情况；将生长0.5、1、2、3、5d的BF模型行SYTO9∕PI染色，激光共聚焦扫描电镜（confocal laser scanning microscopy, CLSM）下摄取不同层面的图像，然后采用Image Pro Plus 6.0分析软件分析不同干预条件下BF的细菌的死亡率。结果 运用SEM及荧光显微镜的方法，在以不含纳米银EVA塑料为细菌粘附载体上培养3d的标本中均观察到流线状的BF形成；纳米银离子含量、作用时间对BF的细菌死亡率均有明显影响（F值分别为84.62, 85.67，P 0.01）；不同时间点含有纳米银离子材料的BF细菌死亡率均显著高于空白对照组（P0.05），但纳米银离子含量不同的2种材料上BF的细菌死亡率无明显影响（P0.05）；纳米银离子含量不同的（0、0.05%、0.1%）材料上的BF，其第1、2、3天细菌死亡率均分别显著高于第0.5、5d的细菌死亡率（P0.05）；含纳米银离子0.1%的材料上BF在作用的第2天其细菌死亡率最高（88.53±1.88）％。结论：运用摇床法成功建立了体外BF模型；纳米银离子对BF内的细菌有明显杀灭作用。 关键词：细菌生物膜；纳米银离子；模型 Silver nanoparticle on the role of bacterial mortality of P.aeruginosa biofilms ZOU－Zhihui,YU－Jialin,LIU－Guanxin,LI－Fang,LIN－Lihua, LIN－Yayin (Department of Neonatology，Children’s Hospital，Chongqing Medical University，Chongqing 400014，China) Abstract： Objective Analysis of the bacteria mortality of P.aeruginosa biofilms in order to explore the effect of silver nanoparticle to the biofilm. Method The ethylene-vinyl acetate(EVA) plastics in different concentration of silver nanoparticle were used as carriers for the bacteria adhesion, on which bacterial biofilm models in vitro were built by using shaking method to simulate the micro-environment in vivo.The samples of not contain silver nanoparticle Cultured 3 day were scanned by Scanning Electron Microscopy (SEM) and were observed by fluorescence microscope after stained with FITC-ConA to observe biofilm formation .In 0.5 day, 1 day, 2 day, 3 day, 5 day , The specimens were be taken out the vitro model of the shaking method, stained by the fluorescent antibody SYTO9/PI were detected under Confocal Laser Scanning Microscopy(CLSM). The images of different levels were intaked by CLSM .The images of CLSM w