Al-Ahmad Ali, Wunder Axel, Auschill Thorsten Mathias, Follo Marie, Braun Gabriele, Hellwig Elmar, Arweiler Nicole Birgit
Department of Operative Dentistry and Periodontology, Albert-Ludwigs-University, Freiburg, Germany.
Department of Hematology and Oncology, Core Facility, Albert-Ludwigs-University, Freiburg, Germany.
J Med Microbiol. 2007 May;56(Pt 5):681-687. doi: 10.1099/jmm.0.47094-0.
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 microm after 1 day to 49.3+/-11.6 microm 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.
牙菌斑生物膜在体内的形成和组成是影响牙龈炎、龋齿和牙周炎发展的重要因素。在体外模型中研究牙菌斑生物膜可能会使口腔实际情况过于简化。在本研究中，将牛牙釉质片原位固定在单个丙烯酸装置中，使用多重荧光原位杂交（FISH）和共聚焦激光扫描显微镜来量化牙菌斑的形成和组成。用于FISH的五个寡核苷酸探针中的每一个都对真细菌或属于牙齿表面早期和晚期定植菌的四种常见分离细菌成分之一具有特异性。形成的生物膜厚度从1天后的14.9±5.0微米增加到7天后的49.3±11.6微米。链球菌在1日龄牙菌斑中占主导地位，7天后显著减少（P = 0.0061）。与第一天相比，具核梭杆菌在2天后减少，7天后显著增加（P = 0.0006）。内氏放线菌含量在第2天和第7天的下降具有显著性（P = 0.0028）。韦荣氏菌属在研究期间的变化不显著（P>0.05）。结果表明，需要7天的体内观察期来检测链球菌和具核梭杆菌的显著变化。所使用的多重FISH适用于在流行病学研究中分析口腔生物膜中四种重要细菌成分的动态变化。
