Enamel biomineralization; the role of pH cycling

牙釉质生物矿化；

基本信息

批准号：
10737481
负责人：
Pamela K Den Besten
金额：
$ 55.43万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-03-07 至 2028-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10737481
关键词：
AKT Signaling Pathway Actins Adhesions Affect Ameloblasts Amelogenesis Imperfecta Apical Biocompatible Materials Calcium Cell Adhesion Cell membrane Cell physiology Cells Cellular Morphology Code Cytoskeleton Defect Dental Enamel Dental caries Developmental Process Enamel Formation Enamel Organ Endocytosis Environmental Risk Factor Enzymes Epigenetic Process Epithelium Esthetics Etiology Extracellular Matrix FRAP1 gene Fluorides Focal Adhesions Fracture Gene Expression Genes Genetic Goals Human body Hydrolysis Hydroxyapatites In Vitro Integrins Knowledge Maintenance Maturation-Stage Ameloblast Mediating Morphogenesis PIK3CG gene Pathway Analysis Polymers Predisposition Process Protein Secretion Proteins Proto-Oncogene Proteins c-akt Recombinants Regulation Resistance Risk Reduction Role Signal Transduction Structure System Testing Thinness Time Tissue Engineering Tissues Tooth structure amelogenin biomineralization conditional knockout crosslink deamidation enamel matrix proteins extracellular fluorosis improved in vivo Model mTOR Signaling Pathway mineralization mouse model mutant novel polymerization protein structure repaired therapeutic protein three dimensional cell culture transcriptome transglutaminase 2

项目摘要

PROJECT SUMMARY/ABSTRACT Mineralized enamel is the hardest tissue in the human body, and its proper formation is critical for the protection and maintenance of healthy teeth for a lifetime. Enamel biomineralization is directed by the epithelial-derived ameloblasts. Ameloblasts secrete enamel matrix proteins first, which are then hydrolyzed and replaced by hydroxyapatite crystals in the maturation process. During the maturation process, the pH of the enamel matrix modulates periodically to be acidic in approximately 80% of the maturation stage, with the remaining approximately 20% of the time remaining neutral. Maturation stage ameloblasts also modulate periodically along with the matrix pH cycling, changing cell morphology and functions. When pH cycling is disrupted by gene mutants or environmental factors that dysregulate ameloblast modulation, enamel maturation is dysregulated, resulting in hypomineralized, poorly formed enamel. However, the mechanisms that drive both matrix pH cycling and ameloblast modulation interactions are poorly understood. In preliminary studies, we found amelogenins can differentially affect cell morphology and adhesion in a pH-dependent manner; and ameloblast modulation is associated with changes in focal adhesion-P13k-AKT signaling pathways. Furthermore, we identified possible contributions of matrix pH to the function of the pH-dependent multifunctional enzyme, transglutaminase 2, in regulating amelogenin hydrolysis in the maturation of enamel matrix. In these proposed studies we will address the central hypothesis that enamel matrix pH regulates ameloblast function, protein structure, and matrix mineralization. We will use both in vitro and in vivo models with alterations in endocytosis, calcium transport, matrix hydrolysis and pH, to test our central hypothesis with the following specific aims. 1) To determine the effect of extracellular pH on adhesion and modulation of maturation stage ameloblasts; 2) To determine the effect of pH and calcium on integrin-mediated focal adhesion - PI3K-AKT-mTOR signaling pathways of ameloblasts; 3) To determine the significance of matrix pH on Transglutaminase 2 activity in the maturation stage enamel matrix. These studies will allow us to better understand the etiology of enamel hypomineralization, and they will allow us to apply this knowledge to reduce the risk of enamel defects. Furthermore, the findings from these studies will contribute to our long-term goal to identify strategies for enamel tissue bioengineering, repair, and the use of enamel proteins therapeutic purposes.

项目概要/摘要矿化牙釉质是人体最坚硬的组织，其正确形成对于保护牙釉质至关重要并终生维护健康的牙齿。牙釉质生物矿化是由上皮衍生的成釉细胞。成釉细胞首先分泌牙釉质基质蛋白，然后水解并被羟基磷灰石晶体在成熟过程中。在成熟过程中，牙釉质基质的pH值在大约 80% 的成熟阶段周期性地调节成酸性，其余的大约 20% 的时间保持中立。成熟阶段成釉细胞也周期性调节随着基质pH值循环，改变细胞形态和功能。当 pH 循环被基因破坏时突变体或环境因素导致成釉细胞调节失调，牙釉质成熟失调，导致牙釉质矿化不足、形成不良。然而，驱动基质 pH 循环的机制和成釉细胞调节相互作用知之甚少。在初步研究中，我们发现牙釉蛋白可以以 pH 依赖性方式对细胞形态和粘附产生不同的影响；成釉细胞调节是与粘着斑-P13k-AKT 信号通路的变化相关。此外，我们还确定了可能的基质 pH 对 pH 依赖性多功能酶转谷氨酰胺酶 2 功能的贡献调节牙釉质基质成熟过程中的釉原蛋白水解。在这些拟议的研究中，我们将解决核心假设是牙釉质基质 pH 调节成釉细胞功能、蛋白质结构和基质矿化。我们将使用体外和体内模型来改变内吞作用、钙转运、基质水解和 pH 值，以测试我们的中心假设，其具体目标如下。 1) 确定细胞外pH值对成熟阶段成釉细胞粘附和调节的影响； 2）确定 pH 和钙对整合素介导的粘着斑的影响 - PI3K-AKT-mTOR 信号通路成釉细胞； 3) 确定基质 pH 对转谷氨酰胺酶 2 活性的显着性成熟阶段牙釉质基质。这些研究将使我们更好地了解牙釉质的病因矿化不足，它们将使我们能够应用这些知识来降低牙釉质缺陷的风险。此外，这些研究的结果将有助于我们确定牙釉质策略的长期目标组织生物工程、修复以及使用釉质蛋白的治疗目的。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Three-Dimensional Culture of Ameloblast-Originated HAT-7 Cells for Functional Modeling of Defective Tooth Enamel Formation.

成釉细胞来源的 HAT-7 细胞的三维培养，用于缺陷牙釉质形成的功能建模。

DOI：
发表时间：
2021
期刊：
影响因子：
0
作者：
Földes, Anna;Sang;Kádár, Kristóf;Rácz, Róbert;Zsembery, Ákos;DenBesten, Pamela;Steward, Martin C;Varga, Gábor
通讯作者：
Varga, Gábor

Fluoride Alters Signaling Pathways Associated with the Initiation of Dentin Mineralization in Enamel Fluorosis Susceptible Mice.

氟化物改变与牙釉质氟中毒易感小鼠牙本质矿化启动相关的信号通路。

DOI：
发表时间：
2021-08
期刊：
影响因子：
3.9
作者：
Kao, Yu;Igarashi, Nanase;Abduweli Uyghurturk, Dawud;Li, Zhu;Zhang, Yan;Ohshima, Hayato;MacDougall, Mary;Takano, Yoshiro;Den Besten, Pamela;Nakano, Yukiko
通讯作者：
Nakano, Yukiko

An extracellular matrix culture system for induced pluripotent stem cells derived from human dental pulp cells.

用于源自人牙髓细胞的诱导多能干细胞的细胞外基质培养系统。

DOI：
10.1016/j.freeradbiomed.2022.06.223
发表时间：
2022-08-01
期刊：
European review for medical and pharmacological sciences
影响因子：
0
作者：
Hines, Madeline R.;Goetz, Jessica E.;Gomez-Contreras, Piedad C.;Rodman, Samuel N., III;Liman, Suryamin;Femino, Elise L.;Kluz, Paige N.;Wagner, Brett A.;Buettner, Garry R.;Kelley, Eric E.;Coleman, Mitchell C.
通讯作者：
Coleman, Mitchell C.