Ameloblast Differentiation and Amelogenesis: Next-Generation Models to Define Key Mechanisms and Factors Involved in Biological Enamel Formation

成釉细胞分化和成釉细胞：定义生物牙釉质形成涉及的关键机制和因素的下一代模型

基本信息

批准号：
10460290
负责人：
Tom Diekwisch
金额：
$ 18.87万
依托单位：
TEXAS A&M UNIVERSITY HEALTH SCIENCE CTR
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-01 至 2023-07-31
项目状态：
已结题

项目摘要

Abstract Amelogenesis is a biological process by which highly specialized enamel organ epithelial cells called ameloblasts transport substantial amounts of calcium ions and enamel proteins into the secretory enamel matrix and manufacture a biomaterial of exceptional structural and mechanical properties: tooth enamel (Pandya and Diekwisch 2018). Recent studies have identified some of the calcium channels in dental enamel cells at the basal ameloblast aspect (Nurbaeva et al. 2015, Lacruz 2017). However, there is remarkably little agreement on the mechanisms of ion and protein trafficking at the secretory ameloblast pole and throughout the ameloblast cell body. In support of the present application we have developed three highly innovative models that will address knowledge gaps and advance our understanding of physiological and pathological amelogenesis, including (i) a conditional clathrin deletion mouse model, (ii) an ameloblast 3D bioreactor cell culture model, and (iii) a new liquid cell atomic resolution imaging technology for life in situ imaging of vesicular and extracellular enamel matrices. Establishment of a clathrin knockout model represents significant progress in the area of vesicular trafficking research and a powerful tool to study the function of coated vesicles during amelogenesis. Clathrin-coated vesicles are among the most abundant cellular vesicles, and loss of clathrin has been associated with severe and usually lethal phenotypes (Robinson 2015). Here we present exciting preliminary data demonstrating that clathrin depletion during amelogenesis resulted in altered enamel prism structure and crystal density. Our 3D bioreactor amelogenesis model marks another milestone in enamel research as it promoted the propagation of elongated amelogenin secreting cells, overcoming shortcomings of traditional 2D ameloblast cell culture technology. Third, our atomic resolution liquid chamber model facilitates unprecedented in situ imaging of vesicular contents and native enamel matrix, allowing for the identification of matrix/mineral clusters at the earliest stages of amelogenesis. In response to RFA-DE- 19-004 we have now designed a research plan to develop and optimize these model systems (UG3 phase) and to validate their physiological relevance and usefulness for understanding mechanisms of enamel development and disease during the UH3 phase.

抽象的醒目是一种生物学过程，高度专业化的搪瓷器官上皮细胞称为成熟的细胞将大量钙离子和搪瓷蛋白转运到分泌搪瓷矩阵并制造出特殊结构和机械性能的生物材料：牙齿搪瓷（Pandya and Diekwisch 2018）。最近的研究已经确定了牙齿中的一些钙通道基础成成木细胞的搪瓷细胞（Nurbaeva等，2015，Lacruz，2017）。但是，有关于分泌成分细胞的离子和蛋白质运输机制的一致性很小杆和整个成成熟细胞体。为了支持本应用程序，我们已经开发了三种高度创新的模型将解决知识差距并提高我们对生理和病理学症，包括（i）有条件的网格蛋白缺失小鼠模型，（ii）成熟的3D生物反应器细胞培养模型和（iii）新的液体细胞原子分辨率成像囊泡和细胞外搪瓷矩阵的原位成像的生命技术。建立网格蛋白敲除模型代表了囊泡贩运研究领域的重大进展和在没有变性过程中研究涂层囊泡功能的强大工具。网格蛋白涂层的囊泡是在最丰富的细胞囊泡中，网格蛋白的丧失与严重和通常是致命的表型（Robinson 2015）。在这里，我们提供令人兴奋的初步数据在没有变性过程中的网状蛋白消耗导致牙釉质棱镜结构和晶体密度改变。我们的3D生物反应器的反应症模型标志着搪瓷研究中的另一个里程碑，因为它促进了该模型伸长蛋白蛋白分泌细胞的传播，克服传统2D的缺点成熟的细胞培养技术。第三，我们的原子分辨率液体室模型促进囊泡内容物和天然搪瓷基质的原位成像，允许在最早的弱点阶段鉴定基质/矿物簇。响应rfa-de- 19-004我们现在已经设计了一个研究计划来开发和优化这些模型系统（UG3 阶段）并验证其生理相关性和有用性，以理解 UH3期间的搪瓷发育和疾病。