Collagen Assembly in Tissue-Engineered Cartilage

组织工程软骨中的胶原蛋白组装

• 批准号: 8234165
• 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/8234165
• 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(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。这个网络由三种不同类型的胶原蛋白(II型、II型和XI型)组装而成，并精确地交叉连接在一起形成杂多聚合物。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含量提高到天然关节软骨的水平一直是一个持续的挑战。由于胶原异质聚合物是成熟的纤维结构和成熟组织的持续稳定性的重要模板，我们假设II/IX/XI型胶原异质纤维的交联型模板必须顺序地组装在新形成的软骨基质中，以允许II型胶原纤维生长，增加胶原含量，从而获得功能性关节软骨的生物力学性能。在新生软骨中，次要胶原模板(IX和XI型)是否与II型胶原形成正确的交联组装，目前还没有很好的描述。我们的目标是确定在以软骨细胞和间充质干细胞为基础的生物工程组织中是否组装了典型的透明软骨的胶原网络。由于次胶原调节网络的组织(例如，由XI/V型胶原调节的胶原纤维直径)，指纹图谱可以为正常的基质组装提供筛查，也是了解正在进行软骨形成的成人间充质干细胞II/IX/XI型胶原纤维形成过程中的一系列事件的有价值的工具。本研究的目的是利用蛋白质化学中的蛋白质印迹、酶联免疫吸附试验、液质联用等先进方法，建立胶原纤维组装的分子指纹图谱和一系列参数作为生物标志物，以更好地评价正常和工程化软骨中的交联型胶原网络。最终目标是验证蛋白质组学方法作为组织工程软骨的结果衡量标准，该组织工程软骨具有移植到软骨缺损处所需的关节软骨的胶原含量、质量和结构特性。目标还包括为评估作为愈合反应产生的软骨质量的变化奠定基础。 与公共健康相关：软骨作为减震器的能力取决于构成软骨的胶原蛋白异质纤维网络的质量。在关节炎中，这种胶原蛋白网络被破坏，导致失去正常的关节功能和剧烈的疼痛。这项研究的目的是开发和鉴定一种生物标志物，以监测在生物工程软骨中组装的胶原异质纤维网络是否为正常软骨的典型。