Collagen Assembly in Tissue-Engineered Cartilage

组织工程软骨中的胶原蛋白组装

• 批准号: 8053852
• 负责人: RUSSELL J FERNANDES
• 金额: $ 30.33万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8053852
• 关键词: Absorbable Implants; Address; Adult; Antibodies; Appearance; Architecture; Award; Benchmarking; Bio-Base; Biological Markers; Biomechanics; Biomedical Engineering; Caliber; Cartilage; Cartilage Matrix; Chondrocytes; Chondrogenesis; Collagen; Collagen Arthritis; Collagen Fibril; Collagen Type II; Collagen Type IX; Collagen Type XI; Complex; Defect; Dorsal; Electron Microscopy; Engineering; Enzyme-Linked Immunosorbent Assay; Event; Extracellular Matrix; Fibrillar Collagen; Fingerprint; Foundations; Future; Goals; Growth; Healed; Human; Hyaline Cartilage; Knowledge; Mass Spectrum Analysis; Measures; Mesenchymal Stem Cells; Methodology; Methods; Minor; Molecular; Molecular Profiling; Monitor; National Institute of Arthritis and Musculoskeletal and Skin Diseases; Nude Mice; Outcome Measure; Pain; Pathway interactions; Pattern; Peptides; Phenotype; Polymers; Property; Protein Chemistry; Proteomics; Shock; Stem cells; Structure; Techniques; Time; Tissue Engineering; Tissues; Transplantation; Western Blotting; articular cartilage; base; biodegradable polymer; crosslink; density; fibrillogenesis; healing; in vivo; indexing; joint function; molecular assembly/self assembly; public health relevance; response; sound; subcutaneous; tool

DESCRIPTION (provided by applicant): The ability of cartilage to act as a shock absorber depends on the quality of the collagen fibrillar network that frames cartilage. This network is made up of three different types of collagen (types II, IX and XI) assembled and precisely cross-linked together into heteropolymers. Despite many advances in the field of cartilage tissue engineering, a continuous challenge has been to increase the collagen content of bioengineered cartilage to levels observed in native articular cartilage. Since the collagen heteropolymer is a crucial template of the mature fibrillar architecture and for the ongoing stability of the mature tissue, we hypothesize that a cross-linked template of type II/IX/XI collagen heterofibrils must sequentially assemble in the matrix of newly forming cartilage to allow type II collagen fibril growth, an increased collagen content and so achieve the biomechanical properties of functional articular cartilage. Whether correct cross-linked assemblies of the minor collagen template (types IX and XI) with type II collagen form in neo-cartilage, is not well characterized. Our goal is to determine if a collagen network typical of hyaline cartilage is assembled in chondrocyte based and mesenchymal stem cell based bio-engineered tissues. Since the minor collagens regulate the organization of the network, (e.g., collagen fibril diameter modulated by type XI/V collagen), fingerprinting the pattern of collagen inter-type cross-linking can provide a screen for normal matrix assembly and a valuable tool in understanding the sequence of events in the fibrillogenesis of the type II/IX/XI collagen fibril in adult mesenchymal stem cells undergoing chondrogenesis. The aim is to use Western blot, ELISA, LC-mass spectrometry, and other advanced methods in protein chemistry to establish a molecular fingerprint of the collagen fibril assembly and a set of parameters to use as a biomarker to better evaluate the cross-linked collagen network in normal and engineered cartilage. The ultimate goal is to validate proteomic methods to use as an outcome measure of tissue engineered cartilage that has the collagen content, quality and structural properties of articular cartilage needed to transplant into cartilage defects. The goals also include laying the foundation for evaluating the changing quality of cartilage produced as a healing response. PUBLIC HEALTH RELEVANCE: The ability of cartilage to act as a shock absorber depends on the quality of the collagen heterofibrillar network that frames cartilage. In arthritis, this collagen network is broken down leading to a loss of normal joint function and severe pain. The goal of this study is to develop and authenticate a biomarker to monitor if this collagen heterofibrillar network assembled in bioengineered cartilage is typical of normal cartilage.

描述（由申请人提供）：软骨作为减震器的能力取决于构成软骨的胶原纤维网络的质量。该网络由三种不同类型的胶原蛋白（II 型、IX 型和 XI 型）组成，它们组装并精确交联在一起形成杂聚物。 尽管软骨组织工程领域取得了许多进展，但将生物工程软骨的胶原蛋白含量增加到天然关节软骨中观察到的水平一直是一个持续的挑战。由于胶原异聚物是成熟原纤维结构的重要模板，对于成熟组织的持续稳定性来说，我们假设 II/IX/XI 型胶原异原纤维的交联模板必须顺序组装在新形成的软骨基质中使 II 型胶原原纤维生长，增加胶原蛋白含量，从而实现功能性关节软骨的生物力学特性。新软骨中次要胶原蛋白模板（IX 型和 XI 型）与 II 型胶原蛋白形式的正确交联组装是否正确尚不清楚。 我们的目标是确定透明软骨典型的胶原蛋白网络是否组装在基于软骨细胞和基于间充质干细胞的生物工程组织中。由于次要胶原蛋白调节网络的组织（例如，由 XI/V 型胶原蛋白调节的胶原原纤维直径），因此对胶原蛋白类型间交联模式进行指纹识别可以为正常基质组装提供筛选，并为研究提供有价值的工具。了解正在经历软骨形成的成体间充质干细胞中 II/IX/XI 型胶原原纤维的原纤维形成的事件顺序。目的是利用蛋白质印迹、ELISA、LC-质谱和蛋白质化学中的其他先进方法来建立胶原原纤维组装的分子指纹和一组参数，以用作生物标志物，以更好地评估交联胶原蛋白正常和工程软骨中的网络。 最终目标是验证蛋白质组学方法，以用作组织工程软骨的结果测量，该软骨具有移植软骨缺损所需的关节软骨的胶原蛋白含量、质量和结构特性。这些目标还包括为评估作为愈合反应而产生的软骨质量变化奠定基础。 公众健康相关性：软骨作为减震器的能力取决于构成软骨的胶原异原纤维网络的质量。在关节炎中，这种胶原蛋白网络被破坏，导致正常关节功能丧失和剧烈疼痛。本研究的目的是开发和验证一种生物标记物，以监测生物工程软骨中组装的胶原异原纤维网络是否是正常软骨的典型特征。