In Vitro Model of Bacterial Biofilm Formation on Polyvinyl Chloride Biomaterial
In Vitro Model of Bacterial Biofilm Formation on Polyvinyl Chloride Biomaterial
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聚氯乙烯生物材料上细菌生物膜形成的体外模型
DOI：
10.1007/s12013-011-9220-6
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发表时间：
2011-11-01
影响因子：
2.6
通讯作者：
Lei, Yu-jie
中科院分区：
文献类型：
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作者：
Zhao, Guang-qiang;Ye, Lian-hua;Lei, Yu-jie
本研究旨在建立表皮葡萄球菌在聚氯乙烯（PVC）材料上形成生物被膜的体外模型，并运用共聚焦激光扫描显微镜（CLSM）和扫描电子显微镜（SEM）相结合的方法，探究细菌生物被膜的形成过程及其结构。将表皮葡萄球菌（菌株RP62A）与PVC片在 Tris 缓冲盐水中共同孵育以形成生物被膜。分别在6小时、12小时、18小时、24小时、30小时和48小时检测生物被膜的形成情况。测量这些生物被膜的厚度、生物被膜内活菌的数量及百分比。利用CLSM和环境扫描电镜获取生物被膜的CT扫描图像。研究结果表明，表皮葡萄球菌生物被膜是一种高度有序的多细胞结构。该生物被膜由大量活菌和死菌组成。研究发现，PVC材料表面细菌生物被膜的形成是一个动态过程，在12 - 18小时达到最大厚度，24小时时生物被膜成熟。生物被膜内层、中层和外层的活菌百分比存在显著差异（P < 0.05）。表皮葡萄球菌生物被膜结构复杂，CLSM和SEM相结合的方法是研究PVC材料上生物被膜的理想方法。
The aim of the study was to establish an in vitro model ofStaphylococcus epidermidisbiofilms on polyvinyl chloride (PVC) material, and to investigate bacterial biofilm formation and its structure using the combined approach of confocal laser scanning microscope (CLSM) and scanning electron microscope (SEM).Staphylococcus epidermidisbacteria (stain RP62A) were incubated with PVC pieces in Tris buffered saline to form biofilms. Biofilm formation was examined at 6, 12, 18, 24, 30, and 48 h. Thicknesses of these biofilms and the number, and percentage of viable cells in biofilms were measured. CT scan images of biofilms were obtained using CLSM and environmental SEM. The results of this study showed thatStaphylococcus epidermidisbiofilm is a highly organized multi-cellular structure. The biofilm is constituted of large number of viable and dead bacterial cells. Bacterial biofilm formation on the surface of PVC material was found to be a dynamic process with maximal thickness being attained at 12–18 h. These biofilms became mature by 24 h. There was significant difference in the percentage of viable cells along with interior, middle, and outer layers of biofilms (P< 0.05).Staphylococcus epidermidisbiofilm is sophisticated in structure and the combination method involving CLSM and SEM was ideal for investigation of biofilms on PVC material.