DENTIN BIOMINERALIZATION

牙本质生物矿化

基本信息

批准号：
7231485
负责人：
CHARLES SFEIR
金额：
$ 28.31万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-08-01 至 2010-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7231485
关键词：
Address Adopted Affect Apatites Applications Grants Atomic Force Microscopy Biological C-terminal Calcium Binding Cell Line Cells Characteristics Charge Chemicals Chimeric Proteins Classification Code Collagen Collagen Type I Condition Core Facility Crystal Formation Data Dentin Dentin Formation Deposition Disease Electron Microscopy Electron energy loss spectroscopy Environment Escherichia Event Extracellular Matrix Proteins Fibroblasts Future Genes Goals Growth Habits Heart Valve Prosthesis In Vitro Invertebrates Kidney Calculi Kinetics Knowledge Learning Localized Mammalian Cell Mediating Minerals Modeling Modification Nature Odontoblasts Osteoblasts Pattern Phosphoproteins Phosphorylation Phosphorylation Site Physiologic Ossification Physiological Play Post-Translational Protein Processing Process Property Protein Sequence Analysis Proteins Proteomics Reaction Recombinants Regulation Relative (related person)Research Proposals Resolution Rest Role Scanning Scanning Transmission Electron Microscopy Procedures Serine Shapes Site Specific qualifier value Spectroscopy, Fourier Transform Infrared Spectrum Analysis Structure Surface System Testing Time Tissues Universities Vertebrates Work biomineralization bone calcification casein kinase II concept dentinal phosphophoryn extracellular in vitro Model in vivo in vivo Model inorganic phosphate interfacial macromolecule mass spectrometer mineralization mutant novel phosphophoryn research study secretory protein size therapy design

项目摘要

DESCRIPTION (provided by applicant): Biomineralization is one of the most widespread and important processes in both vertebrates and invertebrates. We have directed our efforts to study the mineralization of bone and dentin in which apatite crystals are specifically grown within type I collagen matrices. Although it is likely that the overall mechanisms involved in mineralization of the two tissues are similar, we have adopted the dentin system since it represents a relatively simpler system than bone. The basic working hypothesis has been that certain acidic, phosphorylated, non-collagenous extracellular matrix protein macromolecules (NCP) are first localized by interaction with the collagen I fibril surfaces, which then direct the nucleation of crystal deposits and orientation of certain specific crystallographic habit directions relative to the fibril axes. Further interactions of the NCP with the growing crystals are postulated to specifically regulate crystal growth in terms of habit, shape and size. Although there are many NCP present in dentin, we have focused our efforts on the primary phosphoprotein of dentin: Phosphophoryn ([PP]). PP is very acidic in nature and characterized by a very high content of serine residues, at about an 85 percent level of phosphorylation. In this proposal we plan to introduce the gene encoding for PP to over-express it in fibroblast, osteoblast and odontoblast cells, respectively. This will enable us to establish an in vivo system to study biomineralization. We will characterize the mineralization of the crystals in these three separate cells to ascertain the nucleation and crystal growth directions. This approach will allow us to address many important questions: Is the post-translationnal modification of PP similar in fibroblast in contrast to osteoblast and/or odontoblast cells? Will the mineralized crystal have the same crystallographic properties when the nucleating protein, such as PP, is expressed in a fibroblast without the rest of the NCP, which is usually expressed in an osteoblast or odontoblast? We will also study the effect of phosphorylation on the Mineralization process in vitro by expressing the recombinant PP and studying its in vitro phosphorylation on the mineral. These studies should yield new information related to the specific role of phosphoproteins in biomineralization. This knowledge will be extremely valuable in determining the mechanisms involved in the biomineralization process particularly since the studies are conducted in an in vivo model in comparison to the traditional approaches of in vitro studies of biomineralization. This data will also provide basic knowledge for designing treatment of ectopic mineralization specifically in diseases as such kidney stones or failing heart valve prosthesis.

描述（由申请人提供）：生物矿化是脊椎动物和无脊椎动物中最广泛和最重要的过程之一。我们致力于研究骨和牙本质的矿化，其中磷灰石晶体专门生长在 I 型胶原蛋白基质中。尽管两种组织矿化所涉及的总体机制可能相似，但我们采用了牙本质系统，因为它代表了比骨骼相对简单的系统。基本的工作假设是，某些酸性、磷酸化、非胶原细胞外基质蛋白大分子 (NCP) 首先通过与 I 型胶原原纤维表面相互作用而定位，然后指导晶体沉积物的成核和某些特定晶体习惯方向的定向相对于原纤维轴。假设 NCP 与生长晶体的进一步相互作用可以在习性、形状和尺寸方面专门调节晶体生长。尽管牙本质中存在许多NCP，但我们将重点放在牙本质的主要磷蛋白：磷酸蛋白（[PP]）上。 PP 本质上呈强酸性，其特点是丝氨酸残基含量非常高，磷酸化水平约为 85%。在本提案中，我们计划引入编码 PP 的基因，使其分别在成纤维细胞、成骨细胞和成牙本质细胞中过度表达。这将使我们能够建立一个体内系统来研究生物矿化。我们将表征这三个独立单元中晶体的矿化，以确定成核和晶体生长方向。这种方法将使我们能够解决许多重要问题：与成骨细胞和/或成牙本质细胞相比，成纤维细胞中的 PP 翻译后修饰是否相似？当成核蛋白（例如 PP）在成纤维细胞中表达而没有其余 NCP（通常在成骨细胞或成牙本质细胞中表达）时，矿化晶体是否具有相同的晶体学特性？我们还将通过表达重组PP并研究其对矿物质的体外磷酸化来研究磷酸化对体外矿化过程的影响。这些研究应该会产生与磷蛋白在生物矿化中的具体作用相关的新信息。这些知识对于确定生物矿化过程中涉及的机制非常有价值，特别是因为与传统的生物矿化体外研究方法相比，这些研究是在体内模型中进行的。这些数据还将提供设计异位矿化治疗的基础知识，特别是针对肾结石或人工心脏瓣膜衰竭等疾病的治疗。

项目成果

期刊论文数量（6）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Primary structure and phosphorylation of dentin matrix protein 1 (DMP1) and dentin phosphophoryn (DPP) uniquely determine their role in biomineralization.

DOI：
10.1021/bm2005214
发表时间：
2011-08-08
期刊：
BIOMACROMOLECULES
影响因子：
6.2
作者：
Deshpande, Atul Suresh;Fang, Ping-An;Zhang, Xiaoyuan;Jayaraman, Thottala;Sfeir, Charles;Beniash, Elia
通讯作者：
Beniash, Elia

Possible role of DMP1 in dentin mineralization.

DOI：
10.1016/j.jsb.2010.11.013
发表时间：
2011-04
期刊：
JOURNAL OF STRUCTURAL BIOLOGY
影响因子：
3
作者：
Beniash, Elia;Deshpande, Atul S.;Fang, Ping An;Lieb, Nicholas S.;Zhang, Xiaoyuan;Sfeir, Charles S.
通讯作者：
Sfeir, Charles S.

Synthesis of bone-like nanocomposites using multiphosphorylated peptides.

DOI：
10.1016/j.actbio.2014.01.007
发表时间：
2014-05
期刊：
ACTA BIOMATERIALIA
影响因子：
9.7
作者：
Sfeir, Charles;Fang, Ping-An;Jayaraman, Thottala;Raman, Aparna;Zhang Xiaoyuan;Beniash, Elia
通讯作者：
Beniash, Elia

Cell derived hierarchical assembly of a novel phosphophoryn-based biomaterial.

新型基于磷酸基的生物材料的细胞衍生分层组装。