Bioengineered Substrata to Probe Cellular Behavior

用于探测细胞行为的生物工程基质

• 批准号: 6941688
• 负责人: JOYCE Y WONG
• 金额: $ 37.15万
• 依托单位: BOSTON UNIVERSITY (CHARLES RIVER CAMPUS)
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-22 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6941688
• 关键词: biomechanics; biotechnology; cell adhesion; cell components; cell growth regulation; cell morphology; cell motility; cell proliferation; cytoskeletal proteins; extracellular matrix; immunocytochemistry; integrins; phenotype; phosphorylation; restenosis; tissue engineering; vascular smooth muscle; video microscopy

DESCRIPTION (provided by applicant): Restenosis due to intimal hyperplasia and vasoconstriction remains a major problem in treatments of arterial occlusive disease. Smooth muscle cells play a major role in vascular remodeling, and local control of their cellular phenotype would greatly enhance efforts to reduce the occurrence of restenosis. A common result of vascular injury is excessive remodeling of the extracellular matrix, which becomes rich in collagens and proteoglycans. While this leads to changes in biochemical properties, it also significantly alters biomechanical properties. Based on recent work that cells respond to substrata with varying mechanical properties, our central hypothesis is that the biomechanical properties of the substratum will modulate vascular smooth muscle cell cellular phenotype that is relevant for restenosis. Current therapies for restenosis largely involve soluble factors, but little attention has been focused on understanding the effects of substratum biomechanical properties on cellular phenotype. The objective of the proposed research is to create model systems that will recapitulate the biomechanical environment during vascular remodeling and to identify key relationships between substrate compliance and cellular phenotype associated with restenosis. This objective will be achieved by investigating the effect of substrate compliance on the cellular phenotype of smooth muscle cells on model bioengineered substrata that are designed to exhibit a systematic variation in their compliance ranging from the microscopic to macroscopic length scales. The outcome of this research will be a novel in vitro model system that will more closely mimic the biomechanical environment of the remodeled matrix in which one can test the effects of agents on vascular smooth muscle cell phenotype. This model system can also be applied to other pathophysiologic systems. Synthetic hydrogels will be used as model substrata in order to control the local mechanical compliance. Aim 1 is to develop bioengineered substrata with well-defined mechanical compliance at the macro- and microscales. Results from studies in Aims 2 and 3 will be used in the refinement of Aim 1. Aim 2 will establish and quantify relationships between substrate compliance and vascular smooth muscle cell phenotypes associated with restenosis. Aim 3 will test the effects of substrate compliance on the expression, localization, and activity of putative mechanosensing cellular components (integrins, cytoskeleton, FAK, paxillin, Rho GTPases). These studies will advance new insights on the physical factors that control phenotypic modulation of vascular smooth muscle cells with the aim of developing therapies to block restenosis.

描述（由申请人提供）： 由于内膜增生和血管收缩引起的再狭窄仍然是动脉闭塞性疾病治疗的主要问题。平滑肌细胞在血管重塑中起着主要作用，对其细胞表型的局部控制将大大加强减少再狭窄的努力。血管损伤的常见结果是对细胞外基质的重塑过多，该基质富含胶原蛋白和蛋白聚糖。尽管这导致生化特性的变化，但它也显着改变了生物力学特性。基于最近的工作，即细胞对基质具有不同机械性能的反应，我们的中心假设是底层的生物力学特性将调节与再狭窄相关的血管平滑肌细胞细胞表型。当前对再狭窄的疗法在很大程度上涉及可溶性因子，但是很少关注理解基质生物力学特性对细胞表型的影响。拟议的研究的目的是创建模型系统，该系统将在血管重塑过程中概括生物力学环境，并确定与再狭窄相关的底物合规性和细胞表型之间的关键关系。通过研究底物依从性对平滑肌细胞细胞表型对模型生物工程底层的细胞表型的影响，该目标将实现，这些底层旨在表现出从微观到宏观长度量表的依从性中的系统变化。这项研究的结果将是一种新型的体外模型系统，它将更加模仿重塑基质的生物力学环境，其中人们可以测试药物对血管平滑肌细胞表型的影响。该模型系统也可以应用于其他病理生理系统。合成水凝胶将用作模型基质，以控制局部机械依从性。 AIM 1是在宏观和显微镜下开发具有定义明确的机械合规性的生物工程底层。 AIM 2和3中研究的结果将用于AIM 1的完善。AIM2将建立和量化与再狭窄相关的底物依从性与血管平滑肌细胞表型之间的关系。 AIM 3将测试底物依从性对推定的机械感应细胞成分的表达，定位和活性的影响（整联蛋白，细胞骨架，FAK，Paxillin，Rho GTPase）。这些研究将提高对控制血管平滑肌细胞表型调节的物理因素的新见解，目的是开发疗法阻止再狭窄。