Quantification of Host Markers in GCF Using CBIB

描述(由申请人提供)：尽管我们对牙周炎的病因和风险因素的了解有所进步，但其患病率并没有下降的迹象。事实上，牙齿保留的时间更长，加上人口老龄化，可能会导致未来受牙周破坏影响的受试者数量增加。目前临床诊断方法的严重局限性，导致无法区分牙周病发生和发展的高危受试者和部位。因此，患者经常受到不适当的治疗。分析牙周病的生化标志物可以克服这些限制，为临床医生提供疾病的诊断和预后生物标志物。本调查的目的是为临床医生提供一种歧视性和可靠的分析，以帮助诊断个别受试者和地点的疾病状况。具体目标1将优化和验证棋盘免疫印迹(CBIB)技术，以定量大量样本中GCF的多种成分。大量样本中几个生物标志物的平行检测可能会导致一项具有更重要诊断价值的测试。此外，同时研究几个分子的能力将允许对这些介体调节免疫反应和组织破坏的机制进行推断。一旦一种灵敏的分析被优化，将进行一项横断面先导研究，作为选择疾病进展的可能生物标记物的第一步。在具体目标2中，将从30名牙周健康受试者和30名牙周炎受试者身上获取牙周循环液样本，以评估不同牙周病生物标志物在受试者和地点水平上区分健康和疾病的诊断特性。一旦确定了最佳候选标记物，将对使用标记物组合以提高诊断能力进行评估。这一系列研究的最终目标是对GCF生物标志物水平进行纵向研究，以确定它们作为疾病活动预测指标的价值，并更好地了解它们的交叉调节。这一目标与NIDCR确定疾病标志和阐明与炎症相关的生物学事件的使命是一致的，希望这一知识将导致牙周病治疗的新疗法。此外，这种方法处于蛋白质组学新方法的前沿，并在不久的将来为翻译应用带来了巨大的希望。

DESCRIPTION (provided by applicant): Despite advances in our knowledge of the causes and risk factors associated with periodontitis, there are no signs of a decline in its prevalence. Indeed, longer retention of teeth, coupled with an aging population might lead to future increases in the number of subjects affected by periodontal destruction. The severe limitations of the present clinical methods of diagnosis, result in an inability to distinguish subjects and sites at risk for periodontal disease initiation and progression. Thus, patients are often inappropriately treated. Analyses of biochemical markers of periodontal disease in GCF samples may overcome such limitations and provide clinicians with diagnostic and prognostic biomarkers of disease. The goal of the present investigation is to provide clinicians with a discriminatory and reliable analysis to help to diagnose disease status in individual subjects and sites. Specific Aim 1 will optimize and validate the checkerboard immunoblotting (CBIB) technique to quantify multiple components of the GCF, in a large number of samples. The parallel examination of several biomarkers in a large number of samples could result in a test with more significant diagnostic value. Moreover, the ability to investigate several molecules at once would allow inferences on the mechanisms of modulation of the immune response and tissue destruction by these mediators. Once a sensitive assay has been optimized, a cross-sectional pilot study will be performed as a first step in selecting possible biomarkers of disease progression. In Specific Aim 2, GCF samples will be obtained from 30 periodontally healthy and 30 periodontitis subjects in order to assess the diagnostic properties of the different GCF biomarkers in distinguishing health and disease on a subject and site level. Once the best candidate markers have been determined, the use of combinations of markers for improving diagnostic ability will be evaluated. The ultimate goal of this line of research is to conduct longitudinal studies on levels of GCF biomarkers, in order to determine their value as predictors of disease activity, and to better understand their cross-regulation. This goal is in accord with NIDCR's mission to identify markers of disease and elucidate biological events associated with inflammation, with the hope that this knowledge will lead to new therapies in the treatment of periodontal diseases. Moreover, this methodology is at the forefront of new approaches in proteomics, and carries enormous promise for translational applications in the near future.