Mechanobiology of Matrix Production by Corneal Fibroblasts

角膜成纤维细胞基质产生的力学生物学

• 批准号: 8539623
• 负责人: Jeffrey W Ruberti
• 金额: $ 36.93万
• 依托单位: NORTHEASTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-02-01 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8539623
• 关键词: Architecture; Ascorbic Acid; Basic Science; Behavior; Behavior Control; Belief; Bioreactors; Cells; Chemistry; Cicatrix; Clinical Trials; Collagen; Collagen Fibril; Collagen Type III; Connective Tissue; Cornea; Corneal Injury; Corneal Stroma; Cues; Custom; Deposition; Development; Epigenetic Process; Extracellular Matrix; Fibroblasts; Goals; Growth; Human; Image; Implant; In Situ; Invaded; Investigation; Knowledge; Lead; Life; Mechanics; Mesenchymal; Natural regeneration; Neural Crest; Output; Paper; Pattern; Physiologic Intraocular Pressure; Play; Process; Production; Refractory; Reporting; Research Personnel; Resolution; Role; Series; Signal Transduction; Stretching; System; Testing; Thick; Tissue Engineering; Tissues; Weight-Bearing state; Work; Wound Healing; base; cell motility; cellular imaging; connective tissue development; crosslink; design; follow-up; insight; migration; novel strategies; prospective; regenerative; repaired; response; scaffold; self organization; success; tissue regeneration; Architecture; Ascorbic Acid; Basic Science; Behavior; Behavior Control; Belief; Bioreactors; Cells; Chemistry; Cicatrix; Clinical Trials; Collagen; Collagen Fibril; Collagen Type III; Connective Tissue; Cornea; Corneal Injury; Corneal Stroma; Cues; Custom; Deposition; Development; Epigenetic Process; Extracellular Matrix; Fibroblasts; Goals; Growth; Human; Image; Implant; In Situ; Invaded; Investigation; Knowledge; Lead; Life; Mechanics; Mesenchymal; Natural regeneration; Neural Crest; Output; Paper; Pattern; Physiologic Intraocular Pressure; Play; Process; Production; Refractory; Reporting; Research Personnel; Resolution; Role; Series; Signal Transduction; Stretching; System; Testing; Thick; Tissue Engineering; Tissues; Weight-Bearing state; Work; Wound Healing; base; cell motility; cellular imaging; connective tissue development; crosslink; design; follow-up; insight; migration; novel strategies; prospective; regenerative; repaired; response; scaffold; self organization; success; tissue regeneration

DESCRIPTION (provided by applicant): Controlled fibroblast directional motility, self-organization and synthetic capacity are fundamental to connective tissue development, growth, remodeling and repair. However, the signals which drive these aspects of fibroblast behavior are poorly understood. It is becoming increasingly clear that fibroblasts in mesenchymal tissue possess intrinsic patterning and synthetic potential and, under appropriate conditions, could be harnessed to repair or regenerate highly-organized connective tissues such as the corneal stroma. In mammalian stromal development, neural crest-derived mesenchymal cells invade the prospective corneal space, organize themselves, then produce, extend and subsequently maintain stromal extracellular matrix (ECM) architecture under increasing intraocular pressure. Our understanding of this process is quite limited, particularly with respect to 1) drivers of inital corneal mesenchymal/fibroblast cell patterning, 2) mechanisms of local and global control of matrix deposition and 3) mechanisms which guide tissue extension (growth) under mechanical force. Our limited appreciation of factors which control matrix deposition, retention and resorption extends to tissue regeneration including stromal wound healing, scar resolution and implant remodeling. It has been a dogmatic belief that the highly-organized collagenous arrays found in the stroma are refractory to in situ regeneration (particularly in humans). Our long-term basic science goals are to determine how highly-organized connective tissue is constructed, extended during growth, maintained, repaired and remodeled. Our long-term translational goals are to utilize this knowledge to formulate new approaches to corneal wound repair and regeneration. To accomplish the long term goals, it is necessary to first determine what factors control fibroblast organizational behavior during matrix production and to determine by what mechanism they control the orientation and retention of deposited collagen. The specific objective of this proposal is to investigate the effect of mechanical signaling on the organization synthesis and retention of collagenous matrix. To accomplish this objective, we will utilize our expertise in live, dynamic cell imaging, bioreactor design, mechanobiology and mechanochemistry. Our central hypothesis for this proposal is that mechanical signaling provides a robust and persistent guidance cue to corneal fibroblasts which is capable of 1) controlling the global organization of the cells, 2) guiding the deposition and preferential retention of collagen and 3) controlling tissue growth. To examine these hypotheses we will utilize our recently developed mechanobioreactor which permits live, long-term imaging of fibroblasts during the production of tissue. Loads of varying magnitude will be placed on a culture system and the output of the cells will be recorded on a minute-to-minute basis. The results of the proposed investigations should not only fully test the central hypothesis, but provide quantitative details about the levels of force necessary to stimulate and control the behavior of PCFs during matrix production.

描述（由申请人提供）：受控的成纤维细胞方向运动，自组织和合成能力是结缔组织发展，生长，重塑和修复的基础。但是，驱动成纤维细胞行为这些方面的信号知之甚少。越来越清楚的是，间充质组织中的成纤维细胞具有内在的图案和合成潜力，并且在适当的条件下，可以利用修复或再生高度组织的结缔组织，例如角膜基质。在哺乳动物的基质发育中，神经rest衍生的间充质细胞会侵入前瞻性角膜空间，组织自身，然后产生，延伸并随后维持基质外细胞外基质（ECM）在增加的眼内压力下。我们对这一过程的理解非常有限，尤其是关于1）Inital角膜间充质/成纤维细胞模式的驱动因素，2）基质沉积的局部和全局控制的机制以及3）在机械力下指导组织扩展（生长）的机制。我们对控制基质沉积，保留和吸收的因素的良好程度扩展到组织再生，包括基质伤口愈合，疤痕分辨率和植入物重塑。这是一种教条信念，即在基质中发现的高度组织的胶原阵列对原位再生（尤其是在人类中）是难治性的。我们的长期基础科学目标是确定如何在生长，维护，修复和重塑过程中建造，扩展高度组织结缔组织。我们的长期翻译目标是利用这些知识来制定角膜伤口修复和再生的新方法。为了实现长期目标，有必要首先确定哪些因素控制矩阵生产过程中成纤维细胞的组织行为，并通过控制胶原蛋白的方向和保留的方式来确定哪种机制。该提案的具体目的是研究机械信号传导对胶原基质组织合成和保留的影响。为了实现这一目标，我们将利用我们的专业知识，用于实时，动态细胞成像，生物反应器设计，机械生物学和机械化学。我们对该提案的中心假设是，机械信号传导为角膜成纤维细胞提供了坚固且持续的引导提示，该提示能够1）控制细胞的全球组织，2）指导胶原蛋白的沉积和优先保留和3）控制组织生长。为了审查这些假设，我们将利用我们最近开发的机械反应器，该机械反应器允许在组织生产过程中对成纤维细胞的长期成像。各种大小的负载将放在培养系统上，并将以分钟至分钟的基础记录细胞的输出。拟议的研究的结果不仅应充分检验中心假设，而且还应提供有关刺激和控制矩阵生产过程中PCF行为所需的力量水平的定量细节。