Development and pyrosequencing analysis of an in-vitro oral biofilm model.
Development and pyrosequencing analysis of an in-vitro oral biofilm model.
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DOI：
10.1186/s12866-015-0364-1
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发表时间：
2015-02-10
期刊：
影响因子：
4.2
通讯作者：
Wade WG
中科院分区：
文献类型：
--
作者：
Kistler JO;Pesaro M;Wade WG
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。主要的预防方法是机械去除牙菌斑，并通过牙膏和漱口水中掺入的活性剂来增强。需要包括复杂口腔细菌生物膜的体外测定来准确预测新型活性剂的体内功效。本研究的目的是使用卡尔加里生物膜装置（CBD）开发口腔生物膜模型，该装置接种了天然唾液接种物并通过下一代测序进行分析。具体目标是通过比较来自（i）不同时间点的相同志愿者和（ii）不同志愿者组的生物膜随时间的组成，以确定模型的再现性和稳定性。过滤后，焦磷酸测序产生了 280,093 个序列，平均长度为 432 个碱基。在汇集的唾液和生物膜样本中分别检测到平均 320 个和 250 个 OTU。基于群落成员和结构的主坐标分析 (PCoA) 图表明，重复的生物膜样本高度相似并聚集在一起。此外，使用分子方差分析 (AMOVA)，在不同时间源自同一组的生物膜之间没有显着差异。不同组的生物膜之间存在显着差异（AMOVA，P < 0.002）。 PCoA 显示生物膜组成在 7 至 14 天之间发生了变化（AMOVA，P<0.001）。 Veillonella parvula、Veillonella atypica/dispar/parvula 和消化链球菌是 7 天生物膜中检测到的主要 OTU，而口腔普雷沃菌、V. parvula 和星座链球菌是 14 天生物膜中检测到的主要 OTU。使用 CBD 成功生长和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有抗菌剂的口腔护理产品，也可用于筛选旨在改变牙菌斑成分的新方法，例如益生菌或益生菌。本文的在线版本 (doi:10.1186/s12866-015-0364-1) 包含补充材料，可供授权用户使用。
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. The online version of this article (doi:10.1186/s12866-015-0364-1) contains supplementary material, which is available to authorized users.
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DOI：
10.1111/j.1365-2672.1996.tb03199.x
发表时间：
1996-02-01
期刊：
JOURNAL OF APPLIED BACTERIOLOGY
影响因子：
--
作者：
Bradshaw, DJ;Marsh, PD;Cummins, D
通讯作者：
Cummins, D
影响因子：
15.5
作者：
Edlund A;Yang Y;Hall AP;Guo L;Lux R;He X;Nelson KE;Nealson KH;Yooseph S;Shi W;McLean JS
通讯作者：
McLean JS
DOI：
10.1099/13500872-140-12-3407
发表时间：
1994-12-01
期刊：
MICROBIOLOGY-UK
影响因子：
--
作者：
BRADSHAW, DJ;HOMER, KA;BEIGHTON, D
通讯作者：
BEIGHTON, D
影响因子：
4
作者：
Johnson, SA;Goddard, PA;Handley, PS
通讯作者：
Handley, PS
影响因子：
7
作者：
Nasidze, Ivan;Li, Jing;Stoneking, Mark
通讯作者：
Stoneking, Mark