摘要
目的：研究5种口腔细菌在体外形成生物膜的能力、形成过程及不同培养条件对生物膜形成的影响，并通过激光共聚焦扫描显微镜（CISM）观察细菌生物膜的形成及结构特征。方法：选择与龋病发生关系密切的5种口腔细菌，分别接种在含牛心脑浸液培养基（BHI）、人工唾液（BM5）以及人唾液培养基的标准96孔板中，在培养6h、8h、12h、24h、36h、48h和60h后取出相应的微孔板，11％Crystal Violet染色生物膜，95％乙醇复吸Crystal Violet，HTS7000PLUS多孔板高效分析仪测定各时段细菌生物膜的生长情况，并绘制生长曲线。CLSM观察细菌生物膜形成的结构变化。结果：在BHI及BM5中5种口腔细菌均能形成生物膜，在人唾液中没有形成生物膜。细菌生物膜形成表现为缓慢的非线性生长，出现了一个相对的生长停滞期，这个时期出现在36～48h之间，不同细菌略有差别。激光共聚焦扫描显微镜（asM）观察发现在培养6h后仅有少量细菌粘附形成散在微菌落，24h后出现生物膜的基本结构，48h形成成熟的生物膜。结论：研究发现在体外细菌形成生物膜的能力与培养基条件密切相关，细菌生物膜形成缓慢，其间有一个相对生长停滞期。细菌定植，粘附，共集聚是生物膜形成的重要步骤。
Objective：This research was to standardize a polyvinyl chloride （PVC） micro-titer plate assay to compare the ability of 5 cariogenic bacteria to form biofilms and to explore its characteristic, ultra structure, and influences by different medium. Method： A total of 5 cariogenic bacteria strains were grown in three different medium included Bill, BM5 and saliva medium in PVC micro-titer plate wells. Biofilm formation was indirectly assessed by staining with 1% crystal violet and measuring crystal violet absorbance, using distaining solution. The biofilm growth curves of 5 laboratory strains were made. The ultrastructures of bioilm were observed under Co focal Laser Scanning Microscope （CLSM）. Result：5 cariogenic bacteria strains formed biofilm structure in Bill and BM5 medium, but in saliva medium no biofilm were observed. The developmental step in biofilm formation include initial attachment to a surface, the formation of micro colonies and an mature biofilm structure. Conclusion：The ability of cariogenic bacteria to form biofilm was influenced by different strains and culture medium. The necessary steps of biofilm formation include initial attachment, the formation of micro colonies and co-aggregation.
出处
《临床口腔医学杂志》
2006年第6期323-325,共3页
Journal of Clinical Stomatology
基金
国家自然科学基金资助项目(30430800)
