The Structure and Function of Dental Lymphatics (R21)

牙齿淋巴管的结构和功能（R21）

• 批准号: 10388309
• 负责人: PHILIP D KING
• 金额: $ 23.4万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-08 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10388309
• 关键词: 3-Dimensional; Address; Animal Model; Antigen-Presenting Cells; Antigens; Apical; Biological Assay; Biology; Blood; Blood Circulation; Blood Vessels; Blood capillaries; Caliber; Cardiovascular system; Cell Death; Cells; Cessation of life; Clinical; Consensus; Data; Dental; Dental Pulp; Dental Research; Drainage procedure; Endodontics; Floor; Fluid Balance; Fusobacterium nucleatum; Future; Host Defense; Hour; Image; Immune; Immune response; Immunity; Immunology; In Vitro; Infection; Inflammatory; Intercellular Fluid; Knowledge; Lead; Life; Link; Liquid substance; Lymphatic; Lymphatic Endothelial Cells; Lymphatic System; Lymphatic function; Mediating; Methods; Minerals; Mission; Modeling; Mus; Organ; Outcome; Peripheral; Plant Roots; Play; Polymer Chemistry; Positioning Attribute; Prevention therapy; Public Health; Pulp Canals; Pulp Chambers; Regulation; Research; Role; Science; Site; Structure; System; Testing; Time; Tissues; Tooth structure; Travel; United States National Institutes of Health; adaptive immune response; adaptive immunity; body system; burden of illness; dental infection; dental structure; disability; draining lymph node; follow-up; improved; innovation; lymph nodes; lymphatic vessel; macrophage; man; mouse model; multidisciplinary; novel; novel strategies; novel therapeutics; particle; pathogen; promoter; response; three dimensional structure; tool; two photon microscopy; two-photon; uptake; 3-Dimensional; Address; Animal Model; Antigen-Presenting Cells; Antigens; Apical; Biological Assay; Biology; Blood; Blood Circulation; Blood Vessels; Blood capillaries; Caliber; Cardiovascular system; Cell Death; Cells; Cessation of life; Clinical; Consensus; Data; Dental; Dental Pulp; Dental Research; Drainage procedure; Endodontics; Floor; Fluid Balance; Fusobacterium nucleatum; Future; Host Defense; Hour; Image; Immune; Immune response; Immunity; Immunology; In Vitro; Infection; Inflammatory; Intercellular Fluid; Knowledge; Lead; Life; Link; Liquid substance; Lymphatic; Lymphatic Endothelial Cells; Lymphatic System; Lymphatic function; Mediating; Methods; Minerals; Mission; Modeling; Mus; Organ; Outcome; Peripheral; Plant Roots; Play; Polymer Chemistry; Positioning Attribute; Prevention therapy; Public Health; Pulp Canals; Pulp Chambers; Regulation; Research; Role; Science; Site; Structure; System; Testing; Time; Tissues; Tooth structure; Travel; United States National Institutes of Health; adaptive immune response; adaptive immunity; body system; burden of illness; dental infection; dental structure; disability; draining lymph node; follow-up; improved; innovation; lymph nodes; lymphatic vessel; macrophage; man; mouse model; multidisciplinary; novel; novel strategies; novel therapeutics; particle; pathogen; promoter; response; three dimensional structure; tool; two photon microscopy; two-photon; uptake

The lymphatic vessel (LV) system is a parallel circulatory system to the blood circulatory system present in almost all tissues. One major function of the LV system is the return of extravasated interstitial fluid to the blood vessel (BV) circulation. In addition, LV act as conduits for the traffic of antigens and antigen-presenting cells from peripheral tissue sites to draining lymph nodes, thereby permitting the induction of adaptive immunity. The existence of LV in dental pulp is still a controversial issue; no consensus as to whether LV are present in the dental pulp has hitherto been reached. In addition, even in those studies that support the existence of dental LV, the overall organization of the dental LV system and its three-dimensional (3D) structure has not been demonstrated. Furthermore, whether a putative dental LV system is involved in immune-inflammatory regulation, similar to the role of LV in other organ systems, is an important question that has not been addressed. We have recently identified a 3D network of LV in dental pulp using a Prox1-eGFP mouse model in which the Prox1 promoter drives expression of eGFP specifically in lymphatic endothelial cells (LEC). As imaged by Two-photon microscopy, the network comprises of microcapillaries (≃3-5 um in diameter) that form dense plex- uses mainly in the pulp chamber and which connect to larger vessels (≃10-15 um in diameter) that travel as a bundle from the pulp chamber floor through the root canals. In addition, we found that infection of dental pulp with Fusobacterium nucleatum resulted in disappearance of coronal microcapillary LV plexuses within 48 hours, suggesting that pulpal infection induces rapid death of LEC in the dental pulp. The structural and functional relationship of components of the identified dental LV network to each other and to the different types of LV that have been identified in other organs is unknown. In addition, the mechanism of infection-induced LV loss has not been determined. Answers to these questions are expected to inform upon the role of dental LV in host immunity to dental pathogens. In this proposal, the assembled multidisciplinary team will further investigate the structure, function, and pathophysiological changes of the dental pulp LV network using an innovative animal model, two-photon micros- copy, a dental infection model, and a fluorescent particle-tracking assay to confirm that the identified LV network conveys pulpal material to draining lymph nodes. There are two specific aims as follows: Aim 1. To characterize normal lymphatics in mouse dental pulp Aim 2. To examine the response of dental pulp lymphatics to pulpal infection Successful completion of the proposed aims will significantly advance our understanding of LV biology, pulp biology, and the immune response to dental infection and is expected to lead to follow-up projects that focus upon host pathogen relationships in the dental pulp.

淋巴管（LV）系统是存在血液回路系统的平行电路系统 在几乎所有组织中。 LV系统的一个主要功能是将间质流体返回到 血管（BV）循环。此外，LV充当抗原和抗原交通流量的导管 从周围组织部位到排水淋巴结的细胞，从而诱导适应性免疫史。 牙纸浆中LV的存在仍然是一个有争议的问题。关于LV是否存在，尚无共识 在牙纸浆中已经达到了牙膏。此外，即使在支持存在的研究中 牙科LV，牙科液压系统的整体组织及其三维（3D）结构尚未 证明。此外，假定的牙齿液伏系统是否参与免疫炎症调节， 与LV在其他器官系统中的作用相似，这是尚未解决的重要问题。 我们最近使用Prox1-EGFP鼠标模型确定了牙浆中LV的3D网络，其中 Prox1启动子驱动EGFP在淋巴内皮细胞（LEC）中的表达。如成像 两光子显微镜，该网络由微毛细血管（直径3-5 um）组成，形成密集的plex- 主要在纸浆室中使用，并连接到较大的容器（直径为≃10-15UM） 从纸浆腔室的地板束穿过根管。此外，我们发现牙髓的感染 核杆菌核核核杆菌在48小时内导致冠状微毛细管LV丛消失， 表明牙髓感染会诱导LEC在牙髓中的快速死亡。 已确定的牙科LV网络组件的结构和功能关系彼此之间 对于在其他器官中鉴定出的不同类型的LV是未知的。另外，机制 尚未确定感染引起的LV损失。这些问题的答案应告知 牙齿LV在宿主对牙科病原体免疫中的作用。 在此提案中，组装的多学科团队将进一步研究结构，功能和 使用创新的动物模型，两光子微型 - 复制，牙科感染模型和荧光颗粒跟踪测定法，以确认已鉴定的LV网络 将牙髓材料传达为排水淋巴结。有两个具体的目的如下： 目的1。表征小鼠牙髓中的正常淋巴样 目的2。检查牙髓淋巴细胞对牙髓感染的反应 成功完成拟议的目标将大大提高我们对LV生物学的理解， 纸浆生物学以及对牙齿感染的免疫反应，并有望导致关注的后续项目 在牙髓中的宿主病原体关系上。