Cell Cycle-Mediated Optimization of Cartilage Tissue Development

细胞周期介导的软骨组织发育优化

• 批准号: 9896522
• 负责人: Clark T. Hung
• 金额: $ 13.71万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-03-19 至 2022-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9896522
• 关键词: 3-Dimensional; Acceleration; Address; Adult; Age; Allografting; American; Applications Grants; Arthroscopy; Autologous; Biologic Development; Bioreactors; Canis familiaris; Cartilage; Cell Cycle; Cells; Cellular biology; Chondrocytes; Cilia; Clinical; Data; Defect; Degenerative polyarthritis; Development; Diagnostic Imaging; Direct Costs; Engineering; Exhibits; Extracellular Matrix; Financial cost; Generations; Histology; Histopathologic Grade; Human; Hydrogels; Immunohistochemistry; In Vitro; Incidence; Individual; Joint repair; Joints; Knee joint; Length; Lesion; Marrow; Measures; Mechanics; Mediating; Microscopy; Modeling; Monitor; Output; Pain; Phenotype; Population; Prevention; Procedures; Production; Property; Prosthesis; Protocols documentation; Quality of life; Reporter; Research; Role; Stains; Synovial Fluid; Technology; Testing; Thick; Time; Tissue Engineering; Tissues; Tolonium chloride; United States National Institutes of Health; base; cartilage development; clinically relevant; cost estimate; effective therapy; gait examination; implantation; in vivo; innovation; mechanical properties; novel; osteochondral tissue; promoter; real time monitoring; repaired; scaffold; socioeconomics

PROJECT SUMMARY An estimated 27 million Americans age ≥25 have osteoarthritis (OA) and this number is projected to escalate to more than 65 million by 2029 at a direct cost estimated at $28.6 billion. Reducing the incidence and effects of OA through effective treatment of cartilage defects would be a significant socioeconomic benefit. As the supply of suitable cartilage grafts is unable to meet clinical demand, the development of tissue engineered osteochondral grafts with mechanically functional properties would have a significant clinical impact. Examination of engineered cartilage tissues at a multi-scale level suggests local variable ECM content at the single cell level, where cells, for example, exhibiting intense metachromatic staining for ECM are juxtaposed to others with relatively little metachromatic staining. We speculate that this intrinsic cell-to-cell variability in ECM production capacity undermines or limits the peak tissue properties attainable by the whole cell population, and may also impact engineered cartilage integrative repair potential. This proposal will test the following hypotheses: H1) Cell cycle priming leads to coordinated cell tissue elaboration capacity, thereby expediting development and peak magnitude of functional tissue properties by decreasing local spatial inhomogeneity in engineered cartilage derived from clinically-relevant chondrocytes. H2) Cell cycle priming is mediated in part by primary cilia that increase in incidence post synchronization. H3) Repair of full thickness osteochondral defects with engineered cartilage constructs derived from initially (cell cycle) synchronized chondrocytes will be superior to non-synchronized (control) chondrocytes due to the unprecedented acceleration of functional tissue development associated with cell cycle priming that leads to cartilage grafts that better approximate the cartilage associated with clinical osteochondral allografts. The corresponding aims will study human and canine chondrocytes in vitro (Specific Aim 1) and engineered canine cartilage constructs in vivo with a full-thickness ostechondral focal defect repair model in the dog (Specific Aim 2). This NIH R21 application will explore the potential for cell cycle priming as a novel platform technology for functional tissue engineering and generation of tissues with native mechanical properties in 6 weeks or less. We will determine if the functional benefits of cell synchronization on 3D cartilage tissue formation are derived from the reduction of cell-to-cell variability and homogenization of cell ECM output. While the concept of cell synchronization is well-established in cell biology, its application for engineering cartilage, as demonstrated by our preliminary data, represents an innovation.

项目摘要 估计有2700万美国人≥25岁患有骨关节炎（OA），这一数字预计为 到2029年，直接成本估计为286亿美元。减少事件和 OA通过有效治疗软骨缺陷的影响将是一个重大的社会经济益处。作为 合适的软骨移植物的供应无法满足临床需求，组织的发展 具有机械功能特性的骨软骨移植物将具有显着的临床影响。 在多尺度上对工程软骨组织的检查表明局部可变ECM含量 在单细胞水平上，例如，细胞在ECM中表现出强烈的正常染色为 与其他相对较少的实质性染色并置。我们推测这个内在的细胞到细胞 ECM生产能力的可变性破坏或限制了总体可实现的峰组织特性 细胞种群，还可能影响工程软骨的整合维修潜力。 该建议将检验以下假设：H1）细胞周期启动导致协调的细胞组织 详细说明能力，从而加快了功能组织特性的发展和峰值幅度 降低了源自临床上与软骨细胞的工程软骨的局部空间不均匀性。 H2）细胞周期启动部分是由原发性纤毛介导的，后者同步后炎症增加。 H3） 用最初衍生的工程软骨构建体修复全厚度骨软骨缺陷（细胞 循环）同步软骨细胞将优于非同步（对照）软骨细胞 与细胞周期启动相关的功能组织发育的空前加速，导致 软骨移植物可以更好地近似于与临床骨软骨外形相关的软骨。 相应的目标将在体外研究人类和犬软骨细胞（特定目标1）和 工程犬软骨在体内具有全厚度的Osthondral局灶性缺陷修复模型 狗（特定目标2）。 该NIH R21应用程序将探索作为一种新型平台技术的细胞周期启动的潜力 在6周内或 较少的。我们将确定细胞同步在3D软骨组织形成上的功能益处是否是 来自细胞间变异性的降低和细胞ECM输出均匀化的衍生得出。而这个概念 细胞同步在细胞生物学（其工程软骨的应用）中已经建立了良好的 通过我们的初步数据证明，代表了一种创新。