POC Biosensor for Periodontitis

近一个世纪以来，医生通常依靠牙周探查来诊断牙周炎。基座 关于牙周袋深度与牙周炎严重程度之间的强正相关， 牙周探查是测量牙齿周围口袋的深度，以确定牙齿的健康状况 牙周组织。然而，牙周探查是一项耗时的主席旁程序，几乎没有预测性。 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， 本质上是因为从测量口袋深度中获得的定量生物信息很少。在这项研究中， 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)和 GCF中的胞外腺苷(EADO)。更具体地说，使用纸尖从患者那里收集的GCF将 被稀释成PBS，并立即应用于主席端的SmartChip。与其他主观PoC设备不同， 基于酶的安培SmartChip®可定量测定GCF中的eATP和EADO。 最近的研究表明，eATP从受损的细胞或激活的中性粒细胞中释放出来是导致 通过与一系列嘌呤能P2X和P2Y受体结合进行炎症反应。这是eATP诱导的 促炎症因子，如肿瘤坏死因子-α和IL-1β，来自先天性免疫细胞，表明eATP是一种早期的- 反应性炎症介质。与此同时，Treg和Breg产生的胞外核苷酸酶CD39和CD73 细胞和内皮细胞将三磷酸腺苷转化为抗炎EADO。我们已经了解到，伊多，通过 通过下调肿瘤坏死因子-α和白介素1β的表达抑制炎症 表达，促进BMP2的产生。这些证据表明eATP与 与炎症有关，但EADO可能作为一种抗炎和促成骨因子在 牙周炎。我们的假设认为，SmartChip®对GCF中eATP/EADO的测量将有助于 确定牙周炎病变中的炎症状态，并作为骨再生反应的预测因子。在……里面 为了验证这一假设，我们将在GCF中验证eATP和EADO的POC SmartChip®测量结果 牙周炎的诊断和与治疗相关的骨再生活动的预测。三个队列将是 受试者分为A)正常对照组、B)牙周炎患者和C)牙周炎患者。 常规牙周测量(牙槽深度、临床附着丧失、探诊出血和 牙龈指数)和标准化的根尖周X线片。然后，GCF将由一个 每一象限最深的牙周口袋或牙龈缝隙处的PerioPaper(GCF收集条)，以及 EATP和EADO的浓度将在主席端使用SmartChip®进行监测。一组 将使用Luminex®Multiplex在GCF中监测生物标记物，并将其与eATP/EADO进行比较 与眼袋深度和临床附着丧失的临床测量以及治疗的相关性 研究对象的相关骨再生活动。

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.