POC Biosensor for Periodontitis

用于牙周炎的 POC 生物传感器

• 批准号: 10244668
• 负责人: TOSHIHISA KAWAI
• 金额: $ 2.91万
• 依托单位: NOVA SOUTHEASTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10244668
• 关键词: ADORA2A gene; Adenosine; Affect; Anti-Inflammatory Agents; Aspartate Transaminase; BMP2 gene; Binding; Biological; Biological Markers; Biosensor; CD100 antigen; Calculi; Caring; Cells; Clinical; Collection; Consumption; Detection; Development; Devices; Diagnosis; Disease; Elderly; Endothelial Cells; Enzyme-Linked Immunosorbent Assay; Enzymes; Evaluation; Future; Gingiva; Gingival Indexes; Gingivitis; Health; Hemorrhage; Immune; Inflammation; Inflammation Mediators; Inflammatory; Inflammatory Response; Interleukin-1 beta; Lab-On-A-Chips; Lesion; Ligation; Liquid substance; MMP8 gene; Measurement; Measures; Mental Retardation; Monitor; Oral cavity; Osteogenesis; Outcome; P2X-receptor; Paper; Pathogenicity; Patients; Periodontal Pocket; Periodontitis; Periodontium; Plant Roots; Population; Prediction of Response to Therapy; Predictive Value; Procedures; Production; Regenerative response; Regulatory T-Lymphocyte; Reporting; Risk Factors; Role; Sampling; Series; Severities; Site; Standardization; Study Subject; Sum; TNF gene; Temporomandibular Joint; Testing; Time; Tissues; Tooth structure; aged; base; biomarker panel; bone; cell injury; cohort; collagenase; cytokine; extracellular; healing; neutrophil; next generation; novel; novel strategies; operation; osteogenic; outcome forecast; periapical; receptor; recruit; regenerative; response; tissue regeneration; tool; tooth crowding; treatment response

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.

近一个世纪以来，医生通常依靠牙周探查来诊断牙周炎。基于 牙周袋深度与牙周炎严重程度之间存在很强的正相关性，其主要目的是 牙周探诊是测量牙齿周围的牙周袋深度，以确定牙齿的健康状况 牙周组织。然而，牙周探诊是一项耗时的椅旁手术，并且几乎没有预测性。 在确定当前疾病活动、未来进展的预后或治疗后的愈合方面的价值， 主要是因为通过测量口袋深度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 中的细胞外腺苷 (eADO)。更具体地说，使用纸点从患者收集的 GCF 将 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® 可以量化 GCF 中的 eATP 和 eADO。 最近的研究表明，受损细胞或活化的中性粒细胞释放的 eATP 是导致 通过与一系列嘌呤能 P2X 和 P2Y 受体结合来调节炎症反应。该 eATP 诱导 来自先天免疫细胞的促炎因子，例如 TNF-α 和 IL-1β，表明 eATP 是一种早期 反应炎症介质。同时，由 Treg 和 Breg 产生的核酸外切酶 CD39 和 CD73 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 BMP2 的产生。这些证据表明 eATP 与 与炎症有关，但 eADO 可能作为抗炎和促骨生成因子 牙周炎。我们的假设认为，GCF 中 eATP/eADO 的 SMARTChip® 测量将有助于 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 为了检验这一假设，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骨再生活动的预测。三个队列将 为这项研究招募的； 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 将进行牙龈指数）和标准化根尖周X光检查。然后，GCF将被收集 PerioPaper（GCF 收集条）位于每个象限最深的牙周袋或牙龈缝隙，以及 eATP 和 eADO 的浓度将在椅边使用 SMARTChip® 进行监测。一个面板 将使用 Luminex® Multiplex 在 GCF 中监测生物标志物，并将其与 eATP/eADO 进行比较 与袋深度和临床附着丧失以及治疗的临床测量的相关性 研究对象的相关骨再生活动。