GOALI: Engineering an In Vitro Assembled Corneal Stroma

摘要：研究人员试图开发一种体外组装的角膜基质，最终用作替代组织来帮助视力受损的人。这项工作的动机是将从黄瓜中提取的胶原纤维组装成模仿角膜基质的能力，这是该团队已经存在的一项技术。将这种能力扩展到哺乳动物材料是一个首要目标。拟议的工作中心预期的研究成果围绕定义细胞外基质和结缔组织的胶原支架组装应用于组织工程的原则。尽管有大量关于胶原蛋白形成的宏观结构的机械性能的数据，但在胶原蛋白分子或纤维水平上的机制知之甚少。这些特性将被测量，以及相关ECM的蛋白多糖成分之间的相互作用，提供合理设计组织体外等效物所需的信息。用于构建基质的材料与自然系统的材料将在大分子和宏观尺度上进行机械匹配。通过与MiMedx，Inc.的这一大学-工业合作研究项目，该团队获得了胶原蛋白，并在胶原蛋白组装和组织工程应用方面获得了丰富的经验。此外，行业合作提供了一种简化的机制，通过这种机制，研究成果将转化为以患者为导向的产品。智力价值：智力价值有三个方面。1.将首次在多个长度尺度上系统和全面地探索胶原组织成分之间的力学和相互作用，提供丝状自组织系统，特别是I型胶原的力学性质的完整图景。2.研究结果将解决如何在胶原组织中支持负荷的重要问题。3.研究结果还将解决如何建立和维持胶原蛋白的层次结构。更广泛的影响：1.结果本身将有利于那些努力a.开发结缔组织的仿生材料，b.开发用作组织工程支架的材料，以及c.合理地设计由纳米级构建块构建的自组织三维系统。2.这项工作是高度跨学科的，因此吸引了对生物医学工程、材料科学、物理、生物学和生物化学感兴趣的学生。这种广度增强了从代表性不足的群体中招收学生的能力。实验室由研究生、本科生和高中生组成。这些学生将有机会在学术和工业环境中共同工作。4.已经建立了一个外联项目，在该项目中，首席调查员向美国和国外的年轻学生(K-12)提供机会，通过在他们自己的教室远程访问仪器来参加研究实验。5.原子力显微镜的新界面正在开发中，它将允许视力受损的人进行力光谱实验。6.最后，正在研究的材料在作为角膜替代物的应用方面具有巨大的社会效益潜力。

0854023MatthewsSummary: The investigators seek to develop an in vitro assembled corneal stroma for eventual use as a replacement tissue to aid the visually impaired. The work is motivated by the ability to assemble collagen fibrils derived from Cucumaria frondosa into mimics of corneal stroma, a technique which already exists within the team. Extension of this ability to mammalian materials is an overarching goal. The research outcomes expected from the proposed work center around defining the principles by which the collagen scaffolds of extracellular matrices (ECMs) and connective tissues may be assembled for application to tissue engineering. Despite the vast amounts of data available on the mechanical properties of the macroscopic constructs formed by collagen, little is known of the mechanics at the level of the collagen molecule or fibril. These properties will be measured, as well as the interactions between the proteoglycan components of the relevant ECM, providing the information needed to rationally design in vitro equivalents of the tissue. Mechanically matching the materials used in the construction of the stroma to those of the native system will be done at the macromolecular and macroscopic scales. Through this university-industrial collaborative research project with MiMedx, Inc., the team has access to collagens and to a wealth of experience in collagen assembly and applications to tissue engineering. Additionally, the industrial collaboration provides a streamlined mechanism by which the research outcomes will be translated into patient oriented products. Intellectual Merit: The intellectual merit is threefold. 1. For the first time, the mechanics of and interactions between the constituents of collagenous tissues will be systematically and comprehensively explored on multiple length scales, providing a complete picture of how the mechanical properties of filamentous self-organizing systems, in particular type I collagen, are derived. 2. The results will resolve the important question of how load is supported within collagenous tissues. 3. The results also will resolve how the collagen hierarchal forms are established and maintained. Broader Impacts: 1. The results themselves will benefit those endeavoring to a. develop biomimetic materials for connective tissues, b. develop materials for use as scaffolds in tissue engineering, and c. rationally design self-organizing, three-dimensional systems constructed from nanoscale building blocks. 2. The work is highly interdisciplinary, and as such draws students with interest in biomedical engineering, material science, physics, biology, and biochemistry. The ability to recruit students from underrepresented groups is enhanced by this breadth. 3. The laboratories consist of graduate, undergraduate, and high school students. These students will have the opportunity to work jointly in an academic and an industrial setting. 4. An outreach project has been established in which the principal investigator provides young students (K-12) within the US and abroad the opportunity to participate in research experiments by remotely accessing the instrumentation from their own classroom. 5. A new interface for the atomic force microscope is being developed which will allow the visually impaired to perform force spectroscopy experiments. 6. Finally, the material under study has tremendous potential for societal benefit in its applications as a cornea replacement.