Dige I, Raarup M K, Nyengaard J R, Kilian M, Nyvad B
Department of Dental Pathology, Operative Dentistry and Endodontics, School of Dentistry, Aarhus University, Vennelyst Boulevard 9, 8000 Aarhus C, Denmark.
Stereology and Electron Microscopy Research Laboratory and MIND Center, Aarhus University, Ole Worms Allé 8, 8000 Aarhus C, Denmark.
Microbiology (Reading). 2009 Jul;155(Pt 7):2116-2126. doi: 10.1099/mic.0.027706-0. Epub 2009 Apr 30.
The combined use of confocal laser scanning microscopy (CLSM) and fluorescent in situ hybridization (FISH) offers new opportunities for analysis of the spatial relationships and temporal changes of specific members of the microbiota of intact dental biofilms. The purpose of this study was to analyse the patterns of colonization and population dynamics of Actinomyces naeslundii compared to streptococci and other bacteria during the initial 48 h of biofilm formation in the oral cavity. Biofilms developed on standardized glass slabs mounted in intra-oral appliances worn by ten individuals for 6, 12, 24 and 48 h. The biofilms were subsequently labelled with probes against A. naeslundii (ACT476), streptococci (STR405) or all bacteria (EUB338), and were analysed by CLSM. Labelled bacteria were quantified by stereological tools. The results showed a notable increase in the number of streptococci and A. naeslundii over time, with a tendency towards a slower growth rate for A. naeslundii compared with streptococci. A. naeslundii was located mainly in the inner part of the multilayered biofilm, indicating that it is one of the species that attaches directly to the acquired pellicle. The participation of A. naeslundii in the initial stages of dental biofilm formation may have important ecological consequences.
共聚焦激光扫描显微镜（CLSM）与荧光原位杂交（FISH）的联合应用为分析完整牙菌斑微生物群特定成员的空间关系和时间变化提供了新机会。本研究的目的是分析在口腔生物膜形成的最初48小时内，与链球菌和其他细菌相比，内氏放线菌的定殖模式和种群动态。生物膜在安装于10名个体佩戴的口腔矫治器中的标准化玻璃板上形成，持续6、12、24和48小时。随后用针对内氏放线菌（ACT476）、链球菌（STR405）或所有细菌（EUB338）的探针标记生物膜，并通过CLSM进行分析。用体视学工具对标记细菌进行定量。结果显示，随着时间的推移，链球菌和内氏放线菌的数量显著增加，与链球菌相比，内氏放线菌的生长速度有减缓的趋势。内氏放线菌主要位于多层生物膜的内部，表明它是直接附着于获得性薄膜的物种之一。内氏放线菌参与牙菌斑形成的初始阶段可能具有重要的生态意义。
