POC Biosensor for Periodontitis

近世纪来，从业者通常依靠牙周探测来诊断牙周炎。基于 牙周袋深度与牙周炎严重程度之间存在强正相关性， 牙周探测是测量牙齿周围的牙周袋深度，以确定牙齿的健康状态。 牙周组织然而，牙周探测是一个耗时的椅旁程序，几乎没有预测 在确定当前疾病活动性、未来进展的预后或治疗后的愈合方面的价值， 主要是因为从测量囊袋深度获得的定量生物信息很少。在本研究中， 我们将使用“芯片实验室”设备SMARTChip®生物传感器来测量细胞外ATP（eATP）， GCF中的细胞外腺苷（eADO）。更具体地说，使用纸尖从患者收集的GCF将 将其稀释到PBS中并立即应用于椅旁的SMARTChip。与其他主观POC设备不同， 基于酶的电流分析SMARTChip®可以定量GCF中的eATP和eADO。 最近的研究表明，从受损细胞或激活的中性粒细胞释放的eATP是因果关系 通过与一系列嘌呤能P2 X和P2 Y受体结合来进行炎症反应。这种eATP诱导 促炎因子，如TNF-α和IL-1β，从先天免疫细胞，这表明eATP是一个早期- 炎症反应介质。同时，由Treg和布雷格产生的胞外核苷酸酶CD 39和CD 73也可被抑制。 细胞以及内皮细胞将ATP转化为抗炎eADO。我们了解到，eADO通过 与其特异性ADO受体连接，通过下调TNF-α和IL-1β抑制炎症 表达，并促进BMP 2的产生。这些证据表明eATP与 但eADO可能作为一种抗炎和促骨生成因子， 牙周炎我们的假设认为，SMARTChip®测量GCF中的eATP/eADO将有助于 确定牙周炎病变的炎症状态，并作为骨再生反应的预测因子。在 为了验证这一假设，我们将验证GCF中eATP和eADO的POC SMARTChip®测量结果， 牙周炎的诊断和治疗相关的骨再生活性的预测。三支队伍将在 本研究招募的受试者：A）对照健康受试者，B）牙龈炎患者和C）牙周炎患者。 常规牙周测量（牙周袋深度、临床附着丧失、探诊出血和 牙龈指数）和标准化根尖X线片。然后，GCF将由一个 在每个象限的最深牙周袋或牙龈缝隙处的PerioPaper（GCF收集条），以及 eATP和eADO的浓度将在椅旁使用SMARTChip®进行监测。一组 将使用Luminex® Multiplex在GCF中监测生物标志物，将其与eATP/eADO进行比较， 与囊袋深度和临床附着丧失的临床测量值以及治疗的相关性- 相关的骨再生活动的研究对象。

For nearly a century, practitioners have typically relied on periodontal probing to diagnose periodontitis. Based on a strong positive correlation between pocket depth and severity of periodontitis, the primary purpose of periodontal probing is to measure pocket depths around a tooth in order to establish the state of health of the periodontium. However, periodontal probing is a time-consuming chairside procedure and has little predictive value in determining current disease activity, prognosis of future progression, or healing in response to treatment, essentially because scant quantitative biological information gained from measuring pocket depth. In this study, we will use “lab-on-a-chip” device, SMARTChip® Biosensor, to measure extracellular ATP (eATP) and extracellular Adenosine (eADO) in GCF. More specifically, GCF collected from patients using a paper point will be diluted into PBS and immediately applied to SMARTChip at chairside. Unlike other subjective POC devices, enzyme-based amperometric SMARTChip® can quantify eATP and eADO in GCF. Recent studies have demonstrated that eATP released from damaged cells or activated neutrophils is causal for inflammatory responses by binding with a series of purinergic P2X and P2Y receptors. This eATP induces proinflammatory factors, such as TNF-α and IL-1β, from innate immune cells, suggesting that eATP is an early- response inflammatory mediator. Meanwhile, ectonucleotidases, CD39 and CD73, produced by Treg and Breg cells, as well as endothelial cells convert ATP to anti-inflammatory eADO. We have learned that eADO, by ligation with its specific ADO receptor, suppresses inflammation by down-modulating TNF-α and IL-1β expressions, and promotes the production of BMP2. These lines of evidence suggest that eATP is associated with inflammation, but that eADO may function as an anti-inflammatory and pro-osteogenesis factor in periodontitis. Our hypothesis holds that SMARTChip® measurements of eATP/eADO in GCF will serve to determine inflammatory status in periodontitis lesion and act as a predictor of bone regenerative responses. In order to test this hypothesis we will validate POC SMARTChip® measurements of eATP and eADO in GCF for the diagnosis periodontitis and prediction of treatment-related bone regenerative activities. Three cohorts will be recruited for this study; A) control healthy subjects, B) gingivitis patients and C) periodontitis patients. Conventional periodontal measurements (pocket depth, clinical attachment loss, bleeding on probing and gingival index) and standardized periapical radiograph will be performed. Then, GCF will be collected by a PerioPaper (GCF Collection Strip) at the deepest periodontal pocket or gingival crevice from each quadrant, and the concentrations of eATP and eADO will be monitored at chairside using a SMARTChip®. A panel of biomarkers will be monitored in GCF using Luminex® Multiplex which will be compared to eATP/eADO for correlation with the clinical measurements of pocket depth and clinical attachment loss as well as treatment- related bone regenerative activities in the study subjects.