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.

项目总结 据估计，年龄在≥25岁的美国人有2,700万人患有骨关节炎(OA)，这一数字预计将 到2029年上升到6500万以上，直接成本估计为286亿美元。减少发病率和 通过有效治疗软骨缺损，骨性关节炎的效果将是显著的社会经济效益。AS 合适的软骨移植物的供应无法满足临床需求，组织工程的发展 具有机械功能特性的骨软骨移植物将会对临床产生重大影响。 多尺度水平的工程化软骨组织检查提示局部ECM含量变化 在单细胞水平，例如，对ECM显示强烈异染染色的细胞是 与其他细胞并列，异色染色相对较少。我们推测这种固有的细胞对细胞 ECM生产能力的可变性破坏或限制了整体可达到的最高组织特性 细胞群，也可能影响工程软骨的综合修复潜力。 这一提议将检验以下假设：1)细胞周期启动导致细胞组织协调 精加工能力，从而通过以下方式加速功能组织特性的发展和峰值 减少来自临床相关软骨细胞的工程化软骨的局部空间不均一性。 H2)细胞周期启动在一定程度上是由同步化后发病率增加的初级纤毛介导的。H3) 原代(细胞)来源的工程化软骨构建修复全层骨软骨缺损 周期)同步的软骨细胞将优于非同步(对照)的软骨细胞，因为 与细胞周期启动相关的功能组织发育前所未有的加速，导致 软骨移植更接近临床同种异体骨软骨移植的软骨。 相应的目标将在体外研究人和狗的软骨细胞(特定目标1)和 犬工程化软骨体内构建全层骨软骨缺损区修复模型 狗(特定目标2)。 NIH R21的应用将探索细胞周期启动作为一种新的平台技术的潜力 用于功能性组织工程和在6周内生成具有天然机械性能的组织 较少。我们将确定细胞同步化对3D软骨组织形成的功能益处是否 源于减少细胞间的变异性和细胞间ECM输出的均质化。虽然这个概念 细胞同步化的概念在细胞生物学中已经确立，它在工程软骨中的应用，如 我们的初步数据表明，这是一项创新。