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 据估计，2700万年龄≥25岁的美国人患有骨关节炎（OA），预计这一数字将 到2029年将增加到6 500多万人，直接费用估计为286亿美元。降低发病率， 通过有效治疗软骨缺损来改善骨关节炎的效果将是一个显著的社会经济效益。作为 合适的软骨移植物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具有显著的临床影响。 在多尺度水平上对工程化软骨组织的检查表明局部可变ECM含量 在单细胞水平，其中例如对ECM表现出强烈异染性染色的细胞是 与其他的并置，具有相对较少的异染性染色。我们推测这种内在的细胞间的 ECM生产能力的可变性破坏或限制了整个组织可获得的峰值组织特性， 细胞群，也可能影响工程化软骨整合修复潜力。 H1）细胞周期引发导致协调的细胞组织 精细加工能力，从而通过以下方式加速功能组织特性的发育和峰值 降低源自临床相关软骨细胞的工程化软骨中的局部空间不均匀性。 H2）细胞周期引发部分由同步化后发生率增加的初级纤毛介导。H3） 细胞工程软骨修复全层骨软骨缺损的实验研究 周期）同步的软骨细胞将上级非同步的（对照）软骨细胞， 与细胞周期引发相关的功能组织发育的前所未有的加速， 与临床骨软骨同种异体移植物相关的软骨移植物更接近。 相应的目标将在体外研究人和犬软骨细胞（具体目标1）， 工程犬软骨结构在体内与全层骨软骨局灶性缺损修复模型， 2.特殊目的（Special Aim 2） 这项NIH R21应用将探索细胞周期引发作为一种新平台技术的潜力 用于功能性组织工程，并在6周内产生具有天然机械性能的组织，或 少我们将确定细胞同步对3D软骨组织形成的功能益处是否 来源于细胞间变异性的降低和细胞ECM输出的均质化。虽然这个概念 细胞同步化在细胞生物学中已经建立，其在工程软骨中的应用， 我们的初步数据表明，这是一种创新。