Clinical Validation of a Biofilm Signature for Early Caries Activity Assessment

V摘要 龋齿是一种多因素疾病，它是由于口腔生物膜、口腔和口腔之间的相互作用造成的。 个体的环境和牙齿结构。尽管横断面研究表明一些 物种通常与龋齿部位有关，而其他物种则与无龋齿表面有关，3，4 或缺乏特定的物种并不能预测未来的龋齿发展或龋齿 进步。新的技术进步使人们了解了许多 疾病，但尽管我们知道龋齿的基本概念，牙齿生物膜的相互作用 微生物区系可能作为一个重要的龋病风险指标，1-4还不是很清楚。我们建议 分析牙齿生物膜代谢产物与生物膜相关表面的龋齿状态的关系。我们的 目的是了解牙齿生物膜的代谢活性，最终确定其特征 龋齿活跃度。 我们目前的资助(NIH 5R01DE017890-05)，有300多名龋病活跃儿童的详细数据， 以国际龋病检测和评估系统(ICDAS)和定量光为特点 诱发荧光(QLF)在过去四年中对他们的龋病活动性和龋病风险的纵向影响 好几年了。这为我们提供了一个独特的机会窗口，以确定是否存在与 通过选择无龋齿(C-F)或龋齿活跃的儿童和特定牙面来患龋齿 (C-A)(有进展性病变)。这项研究的假设是龋损活动反映了 覆盖的牙齿生物膜的活性；因此，活动性病变可以与停滞性病变和 根据它们的新陈代谢活动，它们的表面是健全的。因此，我们建议，选择并纵向考察 并收集了450名5-19岁儿童(300名C-A和150名C-F)的生物膜，以确定：(1)是否有 用气相色谱-质谱法测定牙科生物膜代谢活动是否存在显著差异 现场特定分析的光谱(GC-MS)，可将C-F表面与 C-A儿童的C-A损害；(2)覆盖的生物膜中的代谢物信号是否有区别 与C-A儿童相比，C-F儿童的声音表面；以及(3)代谢物信号是否有变化 形成空化的覆盖表面的牙科生物膜。精确定位代谢物信号 与声音和C-A表面以及龋齿进展相关的将允许识别生物标记物(S) 对于可以在龋齿诊断测试中开发的具有前所未有的预测能力的龋齿活动。 这项工作是与生物分析化学专家密切合作的，特别是 代谢组学和有机挥发性化合物。我们的研究团队现在包括Soini博士和Novotny博士 伊利诺伊大学化学系和信息素研究所，以及目前在 波多黎各大学，我们的研究人群所在的地方。

v Abstract Dental caries is a multifactorial disease that results from the interaction between the dental biofilm, oral environment and tooth structure of an individual. Although cross-sectional studies have suggested some species are associated more commonly with carious sites and others with caries-free surfaces,3, 4 the presence or absence of a specific species has not been shown to be predictive of future caries development or caries progression. New technological advances have led to the understanding of the molecular mechanisms of many diseases, but despite our knowledge of the basic concepts of dental caries, the interaction of the dental biofilm microbiota, which may serve as an important caries risk indicator,1-4 is not well understood. We propose to analyze dental biofilm metabolites in relation to the caries status of the surface associated with the biofilm. Our goal is to understand the metabolic activity of the dental biofilm to ultimately identify a signature for caries activity. Our current grant (NIH 5R01DE017890-05), has detailed data on over 300 caries active children, well characterized by the International Caries Detection and Assessment System (ICDAS) and by Quantitative Light Induced Fluorescence (QLF) longitudinally in terms of their caries activity and caries risk for the past four years. This allows us a unique window of opportunity to identify if there is a potential signature associated with dental caries by selecting children and specific tooth surfaces that are either caries-free (C-F), or caries-active (C-A) (have progressing lesions). The hypothesis for this study is that caries lesion activity is a reflection of the activity of the overlaying dental biofilm; therefore, active lesions can be distinguished from arrested lesions and sound surfaces based upon their metabolic activity. Thus, we propose, to select and longitudinally examine and collect biofilm of 450 children (300 C-A and 150 C-F) between the ages of 5-19 to determine: (1) if there are marked differences between dental biofilm metabolic activities using gas chromatography - mass spectrometry (GC-MS) on a site-specific analysis that may categorize C-F surfaces compared to surfaces with C-A lesions in C-A children; (2) if there is a distinction between the metabolite signals in the biofilm overlaying sound surfaces from C-F children compared to C-A children; and (3) if there is a shift in the metabolite signals of the dental biofilm overlaying surfaces that progress to cavitation. Pinpointing the metabolite signals associated with sound and C-A surfaces and with caries progression will allow the identification of biomarker(s) for caries activity that can be developed in a caries diagnostic test with unprecedented prediction power. This work is being done is close collaboration with experts in bioanalytical chemistry, specifically metabolomics and organic volatile compounds. Our research team now includes Drs. Soini and Novotny from the IU Department of Chemistry, and Institute for Pheromone Research as well as current collaborators in the University of Puerto Rico, where our study population is located.