Engineering biomimetic knee menisci with zonal and anisotropic variations

具有分区和各向异性变化的仿生膝关节半月板工程

• 批准号: 9755363
• 负责人: Kyriacos A Athanasiou
• 金额: $ 33.28万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9755363
• 关键词: Address; Adhesions; Adhesives; Allogenic; American; Animals; Anisotropy; Biochemical; Biochemistry; Biological Assay; Biomechanics; Biomimetics; Cattle; Cell Culture Techniques; Cells; Characteristics; Chondroitin ABC Lyase; Clinical; Collagen; Complex; Cytology; Data; Disadvantaged; Engineering; Future; Generations; Goals; Growth; Histology; Human; Hydrostatic Pressure; Immunohistochemistry; Implant; Knee; LOXL2 gene; Laboratories; Magnetic Resonance Imaging; Mechanical Stimulation; Mechanics; Medial; Meniscus structure of joint; Methodology; Methods; Modeling; Molds; Molecular; Morphology; Natural regeneration; Operative Surgical Procedures; Oryctolagus cuniculus; Pathologic; Performance; Phase; Phenotype; Population; Positioning Attribute; Procedures; Production; Property; Reaction; Scanning Electron Microscopy; Serologic tests; Sheep; Source; Stimulus; Stress; Structure-Activity Relationship; Surgical Models; Surgical sutures; System; Techniques; Technology; Testing; Time; Tissue Donors; Tissue Engineering; Tissues; Traction; Translations; Variant; cellular engineering; design; improved; in vitro testing; in vivo; innovation; lysophosphatidic acid; mechanical properties; mimetics; novel; preservation; repaired; sample fixation; scaffold; success; synergism; trait

PROJECT SUMMARY This proposal aims to tissue engineer an anisotropic neo-meniscus that also captures the regional variations present in the native tissue. Subsequently, allogeneic neo-meniscal constructs will be implanted in a leporine model to achieve both meniscus repair and replacement. It is hypothesized that: 1) regionally variant, anisotropic, meniscus-shaped constructs can be engineered by optimizing cell culture and scaffold-free culture conditions; 2) the strategic temporal application of multi-level stimuli (at cellular-, molecular-, and construct- levels) will allow for synergisms across the different levels of action to enhance the functional properties of the maturing neo-menisci; and 3) allogeneic constructs can be successfully implanted in a leporine model. These hypotheses will be tested via the following three specific aims: 1) To create an anisotropic neo-meniscus with regional variations mimicking native tissue; 2) to enhance functional and organizational properties of the neo- meniscus via multi-level exogenous stimulation synergized by temporal coordination; and 3) to develop surgical fixation techniques and implant the neo-menisci in the rabbit. Previously, the native meniscus was found to be highly anisotropic and regionally variant both morphologically and biomechanically, motivating our current tissue engineering approach to mimic these characteristics. Allogeneic leporine cells will be used to form organizationally and regionally mimetic neo-meniscal constructs in Aim 1; this goal will be accomplished via the use of novel spatial and temporally variant seeding techniques. The anisotropic and organizational properties of the engineered neo-meniscus will then be enhanced by manipulating molecular-, cellular-, and construct-level targets in Aim 2. Specifically, TGF-β1 and hydrostatic pressure will act on the cellular level to increase matrix production; lysophosphatidic acid and chondroitinase-ABC will be used to align and compact the matrix at the molecular level; and meniscus-specific mechanical stimulation will direct anisotropy at the construct level. In this proposal, to avoid the use of primary cells, we will investigate the use of passaged allogeneic cells toward in vivo repair and replacement of the meniscus (Aim 3). Upon successful demonstration of repair/replacement in the leporine model, we will determine methods to likewise expand sheep and human cells for future studies. This approach seeks to address the issue of tissue scarcity and aims to provide a solution to the complex problem of meniscus repair and replacement.

项目概要 该提案旨在组织工程各向异性新半月板，该新半月板也能捕获区域差异 存在于天然组织中。随后，同种异体新半月板结构将被植入兔体内 模型实现半月板修复和置换。假设：1）区域差异， 可以通过优化细胞培养和无支架培养来设计各向异性的弯月形结构 状况; 2）多层次刺激的策略性时间应用（在细胞、分子和结构上） 水平）将允许不同水平的行动之间的协同作用，以增强 成熟的新半月板； 3)同种异体构建体可以成功植入兔类模型。这些 假设将通过以下三个具体目标进行测试：1）创建各向异性新半月板 模仿天然组织的区域差异； 2）增强新组织的功能和组织特性 通过时间协调协同的多层次外源刺激来调节半月板； 3）开发 手术固定技术并将新半月板植入兔子体内。以前，原生半月板是 发现在形态和生物力学上都具有高度各向异性和区域变异性，这激发了我们 当前的组织工程方法可以模仿这些特征。同种异体兔细胞将用于 在目标 1 中形成组织和区域模仿的新半月板结构；这个目标将会实现 通过使用新颖的空间和时间变异播种技术。各向异性和组织性 工程化的新半月板的特性将通过操纵分子、细胞和 目标 2 中的构建水平目标。具体而言，TGF-β1 和静水压将作用于细胞水平 增加基质产量；溶血磷脂酸和软骨素酶-ABC将用于对齐和压实 分子水平的基质；半月板特异性机械刺激将引导各向异性 构建水平。在本提案中，为了避免使用原代细胞，我们将研究传代细胞的使用 同种异体细胞用于半月板的体内修复和替换（目标 3）。演示成功后 在兔模型的修复/替换中，我们将确定同样扩展绵羊和人类的方法 用于未来研究的细胞。这种方法旨在解决组织稀缺的问题，旨在提供一种 解决复杂的半月板修复和更换问题。