Kistler James O, Pesaro Manuel, Wade William G
Centre for Immunology and Infectious Disease, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London, UK.
Symrise AG, Holzminden, Germany.
BMC Microbiol. 2015 Feb 10;15:24. doi: 10.1186/s12866-015-0364-1.
Dental caries and periodontal disease are the commonest bacterial diseases of man and can result in tooth loss. The principal method of prevention is the mechanical removal of dental plaque augmented by active agents incorporated into toothpastes and mouthrinses. In-vitro assays that include complex oral bacterial biofilms are required to accurately predict the efficacy of novel active agents in vivo. The aim of this study was to develop an oral biofilm model using the Calgary biofilm device (CBD) seeded with a natural saliva inoculum and analysed by next generation sequencing. The specific objectives were to determine the reproducibility and stability of the model by comparing the composition of the biofilms over time derived from (i) the same volunteers at different time points, and (ii) different panels of volunteers.
Pyrosequencing yielded 280,093 sequences with a mean length of 432 bases after filtering. A mean of 320 and 250 OTUs were detected in pooled saliva and biofilm samples, respectively. Principal coordinates analysis (PCoA) plots based on community membership and structure showed that replicate biofilm samples were highly similar and clustered together. In addition, there were no significant differences between biofilms derived from the same panel at different times using analysis of molecular variance (AMOVA). There were significant differences between biofilms from different panels (AMOVA, P < 0.002). PCoA revealed that there was a shift in biofilm composition between seven and 14 days (AMOVA, P < 0.001). Veillonella parvula, Veillonella atypica/dispar/parvula and Peptostreptococcus stomatis were the predominant OTUs detected in seven-day biofilms, whilst Prevotella oralis, V. parvula and Streptococcus constellatus were predominant in 14-day biofilms.
Diverse oral biofilms were successfully grown and maintained using the CBD. Biofilms derived from the same panel of volunteers were highly reproducible. This model could be used to screen both antimicrobial-containing oral care products and also novel approaches aiming to modify plaque composition, such as pre- or probiotics.
龋齿和牙周病是人类最常见的细菌性疾病，可导致牙齿脱落。主要的预防方法是通过刷牙和使用漱口水等主动措施辅助机械清除牙菌斑。需要包含复杂口腔细菌生物膜的体外试验来准确预测新型活性剂在体内的功效。本研究的目的是使用接种天然唾液接种物的卡尔加里生物膜装置（CBD）开发一种口腔生物膜模型，并通过下一代测序进行分析。具体目标是通过比较不同时间点来自（i）同一志愿者和（ii）不同志愿者组的生物膜组成，确定模型的可重复性和稳定性。
过滤后焦磷酸测序产生280,093条序列，平均长度为432个碱基。在混合唾液和生物膜样品中分别平均检测到320个和250个操作分类单元（OTU）。基于群落成员和结构的主坐标分析（PCoA）图显示，重复的生物膜样品高度相似并聚集在一起。此外，使用分子方差分析（AMOVA），同一组在不同时间产生的生物膜之间没有显著差异。不同组的生物膜之间存在显著差异（AMOVA，P < 0.002）。PCoA显示，7天至14天之间生物膜组成发生了变化（AMOVA，P < 0.001）。细小韦荣球菌、非典型/差异/细小韦荣球菌和口腔消化链球菌是在7天生物膜中检测到的主要OTU，而口腔普雷沃菌、细小韦荣球菌和星座链球菌在14天生物膜中占主导地位。
使用CBD成功培养并维持了多种口腔生物膜。来自同一组志愿者的生物膜具有高度可重复性。该模型可用于筛选含抗菌剂的口腔护理产品以及旨在改变菌斑组成的新方法，如益生元或益生菌。
