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危险因素的认识有所提高，但其患病率没有下降的迹象。事实上，更长的牙齿保留，加上人口老龄化，可能会导致未来受牙周破坏影响的人数增加。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进展的受试者和部位。因此，患者往往得不到适当的治疗。GCF样本中牙周病的生化标志物的分析可以克服这些局限性，并为临床医生提供疾病的诊断和预后生物标志物。本研究的目的是为临床医生提供一个有区别的和可靠的分析，以帮助诊断疾病的状态，在个别科目和网站。Specific Aim 1将优化和验证棋盘免疫印迹（CBIB）技术，以在大量样本中定量GCF的多种组分。在大量样本中对几种生物标志物进行平行检查可能会导致具有更显著诊断价值的测试。此外，一次研究几种分子的能力将允许推断这些介质调节免疫应答和组织破坏的机制。一旦灵敏的检测方法得到优化，将进行横断面初步研究，作为选择疾病进展可能生物标志物的第一步。在具体目标2中，将从30名牙周健康受试者和30名牙周炎受试者中获得GCF样本，以评估不同GCF生物标志物在区分受试者和部位水平上的健康和疾病方面的诊断特性。一旦确定了最佳候选标志物，将评价使用标志物组合来提高诊断能力。这项研究的最终目标是对GCF生物标志物的水平进行纵向研究，以确定它们作为疾病活动预测因子的价值，并更好地了解它们的交叉调节。这一目标与NIDCR的使命是雅阁的，即识别疾病标志物并阐明与炎症相关的生物学事件，希望这些知识将导致牙周病治疗的新疗法。此外，这种方法是在蛋白质组学的新方法的最前沿，并在不久的将来进行翻译应用的巨大承诺。

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.