MATRIX BASED MINERAL ENAMEL-BIOMIMETICS

基于基质的矿物釉质仿生学

基本信息

批准号：
10399526
负责人：
Janet M. Oldak
金额：
$ 40.5万
依托单位：
UNIVERSITY OF SOUTHERN CALIFORNIA
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：
2019-05-01 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10399526
关键词：
Affect Ameloblasts Amelogenesis Animal Model Binding Biocompatible Materials Biological Biological Models Biomimetics Cell Culture Techniques Cell membrane Cell-Matrix Junction Cells Chemical Models Chemicals Complex Crystal Formation Crystallization Data Dental Dental Enamel Dental Materials Enamel Formation Event Exons Extracellular Matrix Future Generations Genetic Goals Growth Hydroxyapatites In Situ In Vitro Investigation Knowledge Liposomes Mediating Minerals Molecular Morphology Mutation NMR Spectroscopy Outcome Pathologic Pattern Point Mutation Procedures Process Property Proteins Publishing Resolution Structure Transmission Electron Microscopy ameloblastin amelogenin base biomineralization calcium phosphate cell motility design enamel matrix proteins gene product in vivo insight malformation migration mineralization mouse model mutant nanoscale novel overexpression polarized cell restorative material

项目摘要

Project Summary / Abstract To achieve the long-term goal of restoring dental enamel, it is necessary to understand the fundamental chemical and biological principles of extracellular matrix assembly and the manner they control mineral nucleation and growth. There is still a large gap in our understanding of the underlying molecular mechanisms by which enamel matrix proteins assemble and interact with cells to control nucleation and oriented growth of hydroxyapatite crystals, and possibly cell movement and polarization. This is particularly true of ameloblastin protein, which is the focus of our proposed study. The goal of this proposal is therefore to advance our understanding of ameloblastin’s structure and function through a systematic investigation of its interactions with different targets. We hypothesize that the highly organized carbonated hydroxyapatite crystals in enamel continuously grow and form prismatic structures by means of complex ameloblastin-cell, ameloblastin- amelogenin, and ameloblastin-mineral interactions. Two major aims are proposed to systematically examine the above hypothesis by applying in vitro chemical models, cell culture and animal models for amelogenesis. Aim I: To investigate ameloblastin-cell membrane interactions and identify the interacting domains using in vitro synthetic liposomes and ameloblast-like cell culture model systems. We will design several mouse models with point mutations in the interacting domains identified, and examine the consequence of these mutations on enamel prismatic structure, ameloblast morphology, and the attachment of the cells to the matrix. We hypothesize that ameloblastin interacts with ameloblast cells via a domain in the sequence encoded by exon 5 and functions to anchor the mineralizing extracellular matrix to the enamel-forming cells affecting cell polarization, migration, and the formation of Tomes’ processes. Aim II: To investigate ameloblastin-amelogenin interactions on the nanoscale in vivo and in vitro, and to identify their interacting domains using solution NMR spectroscopy. We will study the dynamics of calcium phosphate mineralization events on the nanoscale when ameloblastin is combined with amelogenin, using high resolution in situ atomic force (AFM) and Cryo- transmission electron microscopy (TEM). We hypothesize that amelogenin and ameloblastin form hetereomolecular entities that are functional during different stages of amelogenesis to control crystal formation. We anticipate to gain more insight into the structure, assembly properties and function of ameloblastin. We will define a novel cell- membrane- binding domain on ameloblastin protein. Novel protein- protein-interacting domains on the ameloblastin and amelogenin sequences will be identified. The effect of ameloblastin combined with amelogenin on mineralization will be elucidated and mineral-binding domains will be identified. These studies will advance understanding of the molecular function of ameloblastin in enamel biomineralization and will contribute to our efforts to fabricate synthetic enamel.

项目摘要 /摘要为了实现恢复牙齿搪瓷的长期目标，有必要了解基本细胞外基质组件的化学和生物学原理及其控制次要的方式成核和生长。我们对基本分子机制的理解仍然存在很大的差距通过此，牙釉质基质蛋白组装并与细胞相互作用，以控制成核和定向生长羟基磷灰石晶体以及可能的细胞运动和极化。 amelobolastin尤其如此蛋白质，这是我们提出的研究的重点。因此，该提议的目标是促进我们的通过对其相互作用进行系统研究，了解氨基蛋白细胞的结构和功能具有不同的目标。我们假设在搪瓷中高度有组织的碳酸羟基磷灰石晶体通过复杂的蛋白细胞，蛋白 - 蛋白 - 连续生长并形成棱柱结构氨基蛋白蛋白和杏仁蛋白矿物质相互作用。提出了两个主要目标来系统检查上述假设通过应用体外化学模型，细胞培养和动物模型进行休闲发生。目的I：研究Ameloblastin-Cell膜相互作用并使用IN识别相互作用域体外合成脂质体和成成细胞样细胞培养模型系统。我们将设计几种鼠标模型相互作用域中的点突变被鉴定出来，并检查这些突变对搪瓷棱柱结构，木质细胞形态和细胞附着在基质上。我们假设蛋白细胞蛋白通过外显子5编码的序列中的结构域与成成细胞相互作用和功能可将矿化细胞外基质锚定在影响细胞的牙釉质形成细胞上极化，迁移和TOMES过程的形成。 AIM II：研究蛋白蛋白 - 氨酰基蛋白在体内和体外的纳米级相互作用，并使用溶液NMR识别其相互作用域光谱法。我们将研究纳米级磷酸钙矿化事件的动力学使用高分辨率原位原子力（AFM）和冷冻 - 透射电子显微镜（TEM）。我们假设蛋白质蛋白和蛋白蛋白形式在不同阶段的杂型实体，可控制晶体的不同阶段格式。我们预计会对结构，组装属性和功能有更多的了解 ameloblastin。我们将定义一个新型的细胞膜结合结构蛋白上的细胞膜结合结构域。新蛋白质将鉴定在蛋白质细胞蛋白和蛋白质蛋白序列上的蛋白质相互作用结构域。效果氨基蛋白蛋白与氨基蛋白蛋白结合在矿化中的蛋白质将被阐明，矿物结合域将被识别。这些研究将提高人们对搪瓷中氨基细胞蛋白的分子功能的理解生物矿化，将有助于我们制造合成搪瓷的努力。

项目成果

期刊论文数量（59）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（2）

Controlled remineralization of enamel in the presence of amelogenin and fluoride.

DOI：
10.1016/j.biomaterials.2008.10.019
发表时间：
2009-02
期刊：
BIOMATERIALS
影响因子：
14
作者：
Fan, Yuwei;Sun, Zhi;Moradian-Oldak, Janet
通讯作者：
Moradian-Oldak, Janet

In situ AFM study of amelogenin assembly and disassembly dynamics on charged surfaces provides insights on matrix protein self-assembly.

DOI：
10.1021/ja206849c
发表时间：
2011-11-02
期刊：
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
影响因子：
15
作者：
Chen, Chun-Long;Bromley, Keith M.;Moradian-Oldak, Janet;DeYoreo, James J.
通讯作者：
DeYoreo, James J.

Protein-mediated enamel mineralization.

DOI：
10.2741/4034
发表时间：
2012-06-01
期刊：
Frontiers in bioscience (Landmark edition)
影响因子：
0
作者：
Moradian-Oldak J
通讯作者：
Moradian-Oldak J

Amelogenin-assisted ex vivo remineralization of human enamel: Effects of supersaturation degree and fluoride concentration.

DOI：
10.1016/j.actbio.2011.01.028
发表时间：
2011-05
期刊：
ACTA BIOMATERIALIA
影响因子：
9.7
作者：
Fan, Yuwei;Nelson, James R.;Alvarez, Jason R.;Hagan, Joseph;Berrier, Allison;Xu, Xiaoming
通讯作者：
Xu, Xiaoming

How amelogenin orchestrates the organization of hierarchical elongated microstructures of apatite.