The in vivo dynamics of Streptococcus spp., Actinomyces naeslundii, Fusobacterium nucleatum and Veillonella spp. in dental plaque biofilm as analysed by five-colour multiplex fluorescence in situ hybridization
The in vivo dynamics of Streptococcus spp., Actinomyces naeslundii, Fusobacterium nucleatum and Veillonella spp. in dental plaque biofilm as analysed by five-colour multiplex fluorescence in situ hybridization
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DOI：
10.1099/jmm.0.47094-0
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发表时间：
2007-05-01
影响因子：
3
通讯作者：
Arweiler, Nicole Birgit
中科院分区：
文献类型：
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作者：
Al-Ahmad, Ali;Wunder, Axel;Arweiler, Nicole Birgit
菌斑生物膜的形成和组成是影响牙龈炎、龋病和牙周炎发生发展的重要因素。在体外模型中研究牙菌斑生物膜可能会导致对口腔中真实的条件的过度简化。在这项研究中，牛牙釉质板固定在一个单独的丙烯酸装置原位定量牙菌斑的形成和组合物，使用多重荧光原位杂交（FISH）和共聚焦激光扫描显微镜。用于FISH的五种寡核苷酸探针中的每一种都对真细菌或属于牙齿表面早期和晚期定植者的四种经常分离的细菌成分之一具有特异性。形成的生物膜的厚度从1天后的14.9 +/- 5.0 μ m增加到7天后的49.3 +/- 11.6 μ m。链球菌属在1 d龄菌斑中占主导地位，7 d后明显下降（P=0.0061）。与第1天相比，具核梭杆菌在第2天后减少，在第7天后显著增加（P=0.0006）。内氏放线菌含量在第2天和第7天下降显著（P=0.0028）。韦荣氏球菌属的变化在研究期内，两组间无显著性差异（P > 0.05）。结果表明，需要7天的体内观察期才能检测到链球菌属的显著变化。和F.核质多重荧光原位杂交技术适用于流行病学研究中口腔生物膜中四种重要细菌成分的动态分析。
The formation and composition of dental plaque biofilm in vivo are important factors which influence the development of gingivitis, caries and periodontitis. Studying dental plaque biofilm in in vitro models can cause an oversimplification of the real conditions in the oral cavity. In this study, bovine enamel slabs were fixed in an individual acrylic appliance in situ to quantify dental plaque formation and composition using multiplex fluorescence in situ hybridization (FISH) and confocal laser scanning microscopy. Each of the five oligonucleotide probes used for FISH was specific for either eubacteria or one of four frequently isolated bacterial constituents belonging to early and late colonizers of tooth surfaces. The thickness of formed biofilm increased from 14.9 +/- 5.0 mu m after 1 day to 49.3 +/- 11.6 mu m after 7 days. Streptococcus spp. were predominant in 1-day-old dental plaque and decreased significantly after 7 days (P=0.0061). Compared to the first day, Fusobacterium nucleatum decreased after 2 days and increased significantly after 7 days (P=0.0006). The decreases of Actinomyces naeslundii content on day 2 and day 7 were significant (P=0.0028). Changes in Veillonella spp. were not significant during the study period (P > 0.05). The results showed that an in vivo observation period of 7 days was required to detect significant changes in Streptococcus spp. and F. nucleatum. The multiplex FISH used is suitable for analysing the dynamics of four important bacterial constituents in the oral biofilm in epidemiological studies.