Engineering Developmental Microenvironments: Cartilage Formation and Maturation

工程发育微环境：软骨的形成和成熟

• 批准号: 9250130
• 负责人: Jason A Burdick
• 金额: $ 34.91万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-13 至 2019-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9250130
• 关键词: Acceleration; Address; Adult; Autologous; Binding; Biomedical Engineering; Biophysics; Bioreactors; Book Chapters; Cadherin Domain; Cartilage; Cartilage Matrix; Cell Communication; Cell Density; Cell Maturation; Cells; Chemicals; Chondrogenesis; Cues; Data; Defect; Degenerative polyarthritis; Development; Disease; Doctor of Philosophy; Dose; Encapsulated; Engineering; Environment; Event; Evolution; Extracellular Domain; Extracellular Matrix; Family suidae; Fostering; Funding; Gel; Generations; Grant; Growth; Growth Factor; Hand; Hyaluronic Acid; Hydrogels; Implant; In Vitro; Incidence; Instruction; Joints; Limb Bud; Long-Term Effects; Manuscripts; Measures; Mechanics; Mediating; Mesenchymal Differentiation; Mesenchymal Stem Cells; Modeling; N-Cadherin; Nature; Orthopedic Surgery procedures; Orthopedics; Patients; Peptides; Phenotype; Population Heterogeneity; Porosity; Process; Progress Reports; Property; Proteolysis; Research; Role; Signal Transduction; Skeletal Development; Slide; Sports Medicine; Stimulus; Surface; Surgeon; System; Techniques; Technology; Testing; Time; Tissue Engineering; Tissues; Transforming Growth Factors; Translations; Trauma; Variant; Weight-Bearing state; Work; articular cartilage; base; cartilage development; cartilage regeneration; cartilage repair; clinical translation; clinically relevant; crosslink; density; design; healing; improved; in vivo; innovation; intercellular communication; interdisciplinary approach; mechanical load; mechanical properties; novel; peptidomimetics; permissiveness; public health relevance; receptor; repaired; response; student mentoring; three dimensional cell culture

DESCRIPTION (provided by applicant): Articular cartilage lines the surfaces of joints and transmits the forces generated with loading. Due to limitations in the natural healing capacity of cartilage, and given the increasing incidence of osteoarthritis, there exists a growing demand for cell-based strategies for repair. Tissue engineering, and particularly those approaches based on autologous mesenchymal stem cells (MSCs), is evolving as a clinically relevant technique to promote cartilage regeneration. Yet, the formed tissue properties as well as the stability of phenotype and heterogeneous cellular response within constructs are concerns that currently limit translation of this technology. Our general approach to MSC-based cartilage repair addresses the differences observed between early rapid stages of cartilage formation and the gradual remodeling (maturation) that results in a tissue capable of adult function. This transformative process is driven by a multitude of temporal factors (chemical, mechanical, and soluble). Our progress during the ongoing grant has shown a role for matrix and cellular density, the timing of material degradation, introduction of soluble inductive factors, and mechanical loading (both compression and sliding contact) in guiding cartilage formation and maturation. Here, we build from these studies by introducing a developmentally relevant signal, namely cell-cell interactions through N-cadherin that are found during limb bud development, into our engineered hydrogel systems. In the first Aim, MSCs will be encapsulated in HA hydrogels modified with peptides that mimic the extracellular domain of N-cadherin, and the influence of peptide density on chondrogenesis and cartilage maturation will be investigated, in addition to the influence of the peptide on population heterogeneity and phenotypic stability. In the second Aim, the temporal presentation of the peptides will be investigated by introducing linkers that undergo cell-mediated proteolysis of the peptides from the HA hydrogels. The influence of the temporal peptide presentation on chondrogenesis, cartilage maturation, population heterogeneity, and phenotypic stability will be assessed as in the first Aim, in addition to the responsiveness to mechanical loading based on the acceleration of chondrogenesis. In the third Aim, N-cadherin peptide modified hydrogels, including both stable and transient presentation, will be investigated in a clinically-relevant load-bearing porcine defect model to assess the role of these interactions in an implanted hydrogel in cartilage defect repair. These Aims were designed to allow the testing of our hypotheses that control over the MSC microenvironment, and inclusion of signals present during normal development that are both permissive and instructive for cartilage formation and maturation, will lead to the generation of constructs with properties akin to native tissue and improved repair.

描述（由申请人提供）：关节软骨线的关节表面并传输带负荷产生的力。由于软骨的自然愈合能力的局限性以及骨关节炎发生率的增加，人们对基于细胞的修复策略的需求不断增长。组织工程，尤其是基于自体间充质干细胞（MSC）的方法，正在发展为促进软骨再生的临床相关技术。然而，构造中形成的组织特性以及表型和异质细胞反应的稳定性是当前限制该技术翻译的关注点。我们对基于MSC的软骨修复的一般方法解决了软骨形成的早期快速阶段与逐渐重塑（成熟）之间观察到的差异，从而导致具有成人功能的组织。这种变革过程是由多种时间因素（化学，机械和可溶性）驱动的。我们在正在进行的赠款期间的进展显示了矩阵和细胞密度，材料降解的时机，引入可溶性电感因子以及机械负载（压缩和滑动接触）在指导软骨形成和成熟中的作用。在这里，我们通过引入与开发的信号，即通过N-钙粘着蛋白的细胞 - 细胞相互作用，在肢体芽发育过程中发现的细胞 - 细胞相互作用，并将其源于我们的工程水凝胶系统。在第一个目的中，MSC将封装在模仿N-钙粘着蛋白的细胞外域的肽修饰的HA水凝胶中，除了肽密度对软骨生成和软骨成熟的影响，还将研究肽对种群异质性和现象型稳定性的影响。在第二个目标中，将通过引入细胞介导的蛋白水解的肽的肽的肽从HA水凝胶中进行蛋白水解来研究肽的时间表现。除了基于软骨发生的加速度对机械负荷的反应外，颞肽表现对软骨发生，软骨成熟，种群异质性和表型稳定性的影响还将像第一个目标一样评估。在第三个目的中，将在临床上与临床相关的承重猪缺陷模型中研究N-钙粘蛋白修饰的水凝胶，包括稳定和瞬态表现，以评估这些相互作用在软骨缺陷修复中的植入水凝胶中这些相互作用的作用。这些目标旨在允许控制控制MSC微环境的假设，并在正常发育过程中纳入信号，这些信号既允许又具有启发性的软骨形成和成熟，这将导致具有类似于天然组织和改善修复的特性的结构。