In Situ Hardening Cell-Laden Constructs for Osteochondral Tissue Engineering

用于骨软骨组织工程的原位硬化细胞负载结构

• 批准号: 9144318
• 负责人: ANTONIOS G. MIKOS
• 金额: $ 33.36万
• 依托单位: RICE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-15 至 2020-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9144318
• 关键词: Acrylates; Address; Aldehydes; Amines; Behavior; Bone Regeneration; Cartilage; Cell Communication; Cell Transplantation; Cell-Matrix Junction; Cells; Characteristics; Chemicals; Chondrocytes; Chondrogenesis; Chondroitin Sulfates; Coculture Techniques; Coupled; Crosslinker; Custom; Defect; Development; Diamines; Dose; Encapsulated; Engineering; Formulation; Gel; Glycolates; Goals; Hydrogels; In Situ; In Vitro; Injectable; Joints; Kinetics; Laboratories; Lactones; Lysine; Mechanics; Mesenchymal Stem Cells; Modeling; Modification; N-isopropylacrylamide; Natural regeneration; Oryctolagus cuniculus; Osteogenesis; Patients; Physical condensation; Population; Property; Signal Pathway; Signal Transduction; Site; Societies; Stem cells; Structure; Swelling; System; Technology; Time; Tissue Adhesives; Tissue Engineering; Tissues; Transplantation; Vertebral column; Work; abstracting; acrylic acid; base; bone; butyrolactone; cartilage regeneration; cartilage repair; chemical addition; crosslink; design; dosage; implantation; improved; in vivo; innovation; mechanical behavior; novel; osteochondral repair; osteochondral tissue; osteogenic; poly-N-isopropylacrylamide; repaired; tissue regeneration; tissue repair

Project Summary/Abstract The ultimate goal of this proposal is to develop an innovative and modular technology for osteochondral tissue repair comprising injectable, thermally responsive, in situ forming, and biodegradable hydrogel constructs capable of sustaining the delivery of encapsulated chondrogenic and osteogenic cell populations in a spatially directed fashion to promote native tissue regeneration. We hypothesize that a cytocompatible hydrogel system consisting of non-shrinking, injectable hydrogels with fully soluble degradation products will be formed through the combination of custom poly(N-isopropylacrylamide)-based thermogelling macromers and lysine-based crosslinking macromers that also contain sites for covalent attachment of chondroitin sulfate (CS) to enhance the integration of resultant constructs. Additionally, we hypothesize that the incorporation of poly(L-lysine) (PLL) within the thermogelling hydrogel will enhance the chondrogenic capacity of co-encapsulated articular chondrocyte and mesenchymal stem cell (AC-MSC) cocultures via the induction of developmentally relevant condensation signals. Finally, we hypothesize that a bilayered construct combining the CS-modified chondrogenic hydrogel layer with an osteogenic hydrogel layer of designer mineralizing capability will be leveraged to promote effective osteochondral tissue repair. Three Specific Aims are proposed to address these hypotheses. First, a lysine-based polyesterurethane macromer comprising a biodegradable poly(DL-lactic-co- glycolic acid) mid-block and chemically crosslinkable diamine functionalities will be developed, covalently modified with CS, combined with the thermogelling macromer and thoroughly assessed to establish structure- property relationships. Second, PLL will be incorporated into the hydrogels and its effects on the chondrogenesis of encapsulated AC-MSC cocultures will be evaluated. Further, the combined effects of PLL presentation, AC-MSC coculture, and CS-modification of hydrogel constructs on cartilage tissue integration will be also evaluated ex vivo. Third, the hydrogels developed in Specific Aim 1 and optimized for chondrogenic potential in Specific Aim 2 will be merged with hydrogel formulations with high mineralizing capability to yield bilayered hydrogel constructs comprising chondrogenic and osteogenic layers for the effective repair of osteochondral defects. The potential synergistic effects of encapsulated cells in the osteogenic and chondrogenic layers with PLL delivery will be evaluated in vitro and in vivo to determine the most effective configuration for osteochondral tissue repair in a well-established rabbit osteochondral defect model. The proposed system will address persisting significant challenges associated with osteochondral defect repair by enabling stable integration of the construct with the surrounding native cartilage tissue through a highly modular two-component design, while promoting the chondrogenic and osteogenic differentiation of respective cell populations delivered to effect both cartilage and bone regeneration, respectively.

项目摘要/摘要 该提案的最终目标是开发骨软骨组织的创新和模块化技术 维修包括可注射的，热响应的原位形成和可生物降解的水凝胶构建体 能够维持在空间中递送包裹的软骨和成骨细胞种群 指示促进天然组织再生的时尚。我们假设是细胞增强的水凝胶系统 由非碎裂，可注射的水凝胶与完全溶解的降解产物组成 定制聚（N-异丙基丙烯酰胺）的基于赖氨酸基于赖氨酸的定制聚（N-异丙基酰胺）的组合 交联的宏观分子也包含用于共价硫酸软骨素（CS）的位点以增强 合成结构的整合。此外，我们假设掺入聚（L-赖氨酸） （PLL）在热凝胶水凝胶中将增强共囊关节的软骨天性能力 软骨细胞和间质干细胞（AC-MSC）共培养通过发育相关 冷凝信号。最后，我们假设结合了CS修饰的双层结构 软骨水凝胶层具有矿化能力的成骨水凝胶层，将是 杠杆以促进有效的骨软骨组织修复。提出了三个特定目标来解决这些问题 假设。首先，一种赖氨酸的聚酯素雄烷宏，包括可生物降解的聚（DL乳酸化） 乙醇酸）将开发中间块和化学交联的二胺功能，共价 用CS修饰，与热凝胶宏组合结合并进行彻底评估以建立结构 - 财产关系。其次，PLL将纳入水凝胶及其对 将评估封装的AC-MSC共培养的软骨形成。此外，PLL的综合作用 在软骨组织整合中的水凝胶构建体的表现，AC-MSC共培养和CS修饰将会 也可以在体内进行评估。第三，水凝胶在特定的目标1中开发并针对软骨的优化 特定目标2的潜力将与具有高矿化能力的水凝胶配方合并 双层水凝胶构建体，包括软骨和成骨层，可有效修复 骨软骨缺陷。成骨和封装细胞在成骨和 将在体外和体内评估具有PLL递送的软骨层，以确定最有效的 在良好的兔骨软骨缺损模型中骨软骨组织修复的构型。这 拟议的系统将解决与骨软骨缺陷维修相关的持续重大挑战 通过高度将构建体与周围天然软骨组织的稳定整合 模块化两组分的设计，同时促进了各自的软骨质和成骨分化 分别输送的细胞群体分别实现软骨和骨再生。