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Silver Nanoparticle on the Roleof Bacterial Mortality of P.Aeruginosa Biofilms

Author: ZouZhiHui
Tutor: YuJiaLin
School: Chongqing Medical University
Course: Pediatrics
Keywords: Bacterial biofilms Model Nano silver ions Mortality rate Quantitative Analysis
CLC: R378.991
Type: Master's thesis
Year: 2009
Downloads: 118
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Abstract

Part I: Objective】 mimic in vivo environment in vitro bacterial biofilm model for further in-depth study of the biological characteristics of bacterial biofilm foundation. 【Methods】 adhesion to the carrier in the microenvironment aerosol method and shaker law mimic in vivo bacterial biofilm formation in Pseudomonas aeruginosa strains cultured for 3 days, remove the specimens respectively conducted by FITC-ConA staining and SYTO9 of? PI staining, and morphological characteristics of the fluorescence microscope and confocal laser detection, observation of the formation of the bacterial biofilm; scanning electron microscope observation of the formation of the bacterial biofilm. 【Results】 aerosol micro-environment, a fluorescence microscope after staining of FITC-of ConA observed bright into a sheet of bacterial biofilms; SYTO9? PI staining in the laser confocal detection, observed flaky, layered as cumulus like, batt kind of bacterial biofilms; to a large number of bacteria into a group gathered in the scanning electron microscope observation, slug profusion prominent surface, with a three-dimensional structure of the bacterial biofilm. Three detection methods are observed in the micro-environment of the shaker method, linear bacterial biofilm into a stream. [Conclusion] The use of aerosol method, shaking method can be successfully established were simulated in vivo respiratory and circulation, the micro-environment of the urinary system, biofilm formation model. Part II: [Objective] To investigate the bacterial mortality and the impact of the spatial structure of the nano-silver ions on bacterial biofilm (Biofilm, BF). 【Method】 shaker method, nano silver ion content of ethylene - vinyl acetate (Ethylene-Vinyl acetate, EVA) plastic for bacterial adhesion carrier mimic in vivo Pseudomonas aeruginosa (P.aeruginosa, PA) BF formation of micro-environments, and to establish in vitro the BF models; the culture 3d blank for each specimen in a scanning electron microscope (scanning electron microscopy, SEM) under excluding nano silver, the formation of the EVA in the BF with the observed under a fluorescence microscope after staining of FITC-ConA ; the the BF model line SYTO9 growth 0.5,1,2,3,5 d? PI staining, laser confocal scanning electron microscopy (confocal laser scanning microscopy, CLSM) intake of different levels of image, and then apply the Image Pro Plus 6.0 analysis software the PAO1 strains BF bacteria under different intervention conditions mortality and ISA analysis software PAO1 strain BF spatial structure parameters quantitative data. [Results] ① using SEM and fluorescence microscopy, culture the 3d specimens were observed to flow linear BF not contain nano-silver EVA plastic carrier for bacterial adhesion formation. ② Image Pro Plus 6.0 analysis software quantitative analysis shows that nano silver ion content, the role of of PAO1 strain BF bacterial mortality have significant impact (F = 84.62,85.67, p lt; 0.0001); different time groups containing nano silver ions The PAO1 strain BF material of bacteria mortality are significantly higher than the control group (P lt; 0.05); the PAO1 strain BF bacterial nano silver ion content material on mortality did not significantly affect (P gt; 0.05); nano silver ion content (0, 0.05%, 0.1%) PAO1 strain on the material BF, 1,2,3 days bacterial mortality were significantly higher than the first 0.5,5 d bacterial mortality (P lt; 0.05); 0.1% of the material containing nano-silver ions BF in the first two days of the role of bacteria the highest mortality (88.53 ± 1.88)%. The ③ ISA software Quantitative analysis showed that the silver ion content, duration of action of BF spatial structure significantly affected different time group contains the regional porosity of nano silver ion material on BF (Areal Porosity AP) average diffusion distance (Average Diffusion Distance , ADD) and structural entropy (Textural Entropy, TE) are significantly higher than the control group (P lt; 0.05), but no significant difference in the impact of two kinds of nano-silver ion materials BF AP, ADD and TE ( P gt; 0.05). With prolonged incubation time, each containing nano silver ion materials group the PAO1 strains of BF AP value ADD value no obvious trend, the same time the group containing nano silver ion materials group the PAO1 strains of BF AP values ??are higher than the blank control AP value of the group; same time containing nano silver ion materials group the PAO1 strains of BF ADD value lower than the value of the blank control group ADD. Each containing nano silver ion materials group the PAO1 strains of BF TE values ??with time, increased first increased and then decreased trend, 2d are the highest value; same time group TE values ??with increasing containing nano silver ions were tested to reduce the trend. (4) with time, the average thickness of each containing nano silver ion materials group the PAO1 strains of BF were tested first increased and then decreased the 3d peak value; nano silver ion content of BF thickness was not statistically differences (F = 2.11, p gt; 0.1), the duration of action of BF thickness statistically significant difference (F = 985.81, p lt; 0.001). [Conclusion] ① use of shaking method successfully established in vitro the PAO1 strains of BF model; significant killing effect the ② nano silver ions on the bacteria PAO1 strain BF, and significantly affect the spatial structure of the the PAO1 strains of BF.

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