Mechanical Causation of Corneal Stromal Matrix Synthesis and Fibrosis

角膜基质基质合成和纤维化的机械原因

• 批准号: 10659976
• 负责人: Jeffrey W Ruberti
• 金额: $ 55.23万
• 依托单位: NORTHEASTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10659976
• 关键词: Acceleration; Access to Information; Address; Animals; Biopolymers; Cell Culture Techniques; Cells; Cicatrix; Collagen; Collagen Fibril; Communication; Connective Tissue; Cornea; Corneal Stroma; Coupled; Couples; Custom; Deposition; Development; Diffusion; Engineering; Environment; Etiology; Event; Exclusion; Experimental Designs; Extracellular Matrix; Eye; Fibroblasts; Fibronectins; Fibrosis; Genetic; Glaucoma; Goals; Growth; Homeostasis; Human; Image; In Vitro; Integrins; Intervertebral disc structure; Investigation; Keratoconus; Knowledge; Laser In Situ Keratomileusis; Length; Ligaments; Measurement; Measures; Mechanics; Medicine; Mesenchymal; Methods; Microscopy; Minor; Modeling; Molecular; Morphogenesis; Myofibroblast; Myopia; Natural regeneration; Nature; Pathologic; Pathological Dilatation; Pattern; Phase; Positioning Attribute; Production; Recording of previous events; Retinal Detachment; Role; Sclera; Shapes; Speed; Stress; Structure; Tendon structure; Testing; Tissue Engineering; Tissues; Traction; Tunic; Vertebrates; Vision; Weight-Bearing state; Work; blebbistatin; corneal scar; fibrillogenesis; improved; light transmission; live cell microscopy; mechanical force; mechanical signal; member; molecular assembly/self assembly; monomer; organizational structure; pressure; prospective; regenerative; response; single molecule; temporal measurement; theories; tool; wound healing

Project Summary The cornea and sclera are the principal load bearing members in the tough fibrous ocular tunic which we consider to be an integrated mechano-biological structure. During development, the shape of the eye has been tuned to conform to a specific mechanical environment through a long time-scale integration of its loading history with its initial genetic patterning. Although mechanics are known to contribute to the development of many connective tissues, the ocular globe is particularly sensitive to pressure (tensile wall stress) during the expansive phase of growth. Even in the mature ocular tunic, mechanical instabilities often manifest as conditions which disrupt vision (e.g. myopia, keratoconus, post-LASIK ectasia, tractional retinal detachments and glaucoma). While the underlying causes of tissue structural instabilities are poorly understood, we suspect that they are mechanobiological in nature and potentially reflect an imbalance in the tensional homeostasis that exists between mesenchymal cells, their local ECM and the global mechanical environment. We know that during development, disruption of the mechanical connection between fibroblastic cells and their ECM severely retards ocular growth in a manner analogous to pressure loss, suggesting that mechanical communication is critical to proper ocular morphogenesis. However, the effect of mechanical forces on the mechanisms which drive tissue formation and growth are not well characterized. It is remarkable that we still do not fully understand how the most important structural molecule in vertebrates, collagen, is efficiently assembled into highly-organized, functional, load-bearing tissues which are massively expanded into macroscale structures during growth. However, if we are able to uncover new mechanisms which control tissue formation and growth, we will have access to information which can inform therapies for a variety of pathological conditions including fibrosis, myopia, keratoconus and potentially, glaucoma. Additionally, if we understand how tissue is produced, then there will be implications for engineering corneas de novo and for improving approaches to regenerative corneal medicine. In the proposed work, we plan combine our human cell culture model of corneal stromal tissue elaboration with live-cell mechanodynamics imaging to directly observe single collagen molecules during their transition from solution to fibrils. We will thus directly test a new hypothesis which directly couples local and globally applied forces directly to molecular assembly of collagen during fibrillogenesis and growth. The working hypothesis for this proposal is that force causes corneal stromal ECM elaboration to regulate fibril assembly, remodeling and growth. If the hypothesis is correct, there are myriad mechanotherapeutic opportunities and more critically, our basic understanding of collagenous tissue formation and growth, will be fundamentally altered.

项目摘要 角膜和巩膜是坚韧的纤维眼底上衣中的主要负载轴承成员 成为综合的机械生物结构。在开发过程中，眼睛的形状已调整为 通过长期整合其加载历史记录，符合特定的机械环境 它的最初遗传模式。尽管已知力学有助于许多人的发展 结缔组织，眼球在膨胀期间对压力（拉伸壁应力）特别敏感 增长阶段。即使在成熟的眼中衣中，机械不稳定性也常常表现为 破坏视力（例如，近视，角膜肌肉，lasik ectasia，牵引性视网膜脱离和青光眼）。 虽然对组织结构不稳定性的根本原因了解较少，但我们怀疑它们是 机械生物学本质上，并有可能反映出存在的紧张稳态的失衡 在间充质细胞，其本地ECM和全球机械环境之间。我们知道在 发展，成纤维细胞与其ECM之间机械连接的破坏严重贴 眼科生长的一种类似于压力损失的方式，表明机械通信对于 正确的眼形形态发生。但是，机械力对驱动组织的机制的影响 形成和生长没有很好地表征。值得注意的是，我们仍然不完全了解 脊椎动物中最重要的结构分子有效地组装成高度组织的， 功能性，承载的组织在生长过程中大规模扩展为宏观结构。 但是，如果我们能够发现控制组织形成和生长的新机制，我们将拥有 获取可以为疗法提供有关多种病理状况的疗法的信息，包括纤维化， 近视，圆锥角膜以及青光眼。另外，如果我们了解组织的产生方式，那么 对从头开始工程和改进再生角膜的方法将有影响 药品。在拟议的工作中，我们计划结合角膜基质组织的人类细胞培养模型 用活细胞机械动力学成像阐述，直接观察单个胶原蛋白分子。 从溶液到原纤维的过渡。因此，我们将直接检验一个新的假设，该假设直接构造本地和 在原纤维发生和生长过程中，全球施用的力直接应用于胶原蛋白的分子组装。工作 该提议的假设是，力会导致角膜基质ECM阐述调节原纤维组件， 重塑和增长。如果该假设是正确的，则有无数的机械性机会和 更重要的是，我们对胶原组织形成和生长的基本理解将是从根本上是 改变。