Bioengineered Substrata to Probe Cellular Behavior

用于探测细胞行为的生物工程基质

• 批准号: 7555766
• 负责人: JOYCE Y WONG
• 金额: $ 4.44万
• 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 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7555766
• 关键词: Adhesions; Affect; American; Angioplasty; Antibodies; Arterial Occlusive Diseases; Atomic Force Microscopy; Attention; Back; Behavior; Biochemical; Biological Models; Biomechanics; Biomedical Engineering; Blood Vessels; Cell Line; Cell Proliferation; Cells; Cellular biology; Classification; Collaborations; Collagen; Collagen Type I; Coupled; Crosslinker; Cytoskeletal Proteins; Cytoskeleton; Deposition; Dimensions; Dominant-Negative Mutation; Engineering; Environment; Exhibits; Extracellular Matrix; Extracellular Matrix Proteins; Family; Focal Adhesion Kinase 1; Gel; Goals; Guanosine Triphosphate Phosphohydrolases; Hydrogels; Hyperplasia; Image; Immunofluorescence Immunologic; Injury; Integrins; Lead; Length; Letters; Location; Maps; Matrix Metalloproteinases; Mechanics; Methods; Microfluidics; Microscopic; Modeling; Molecular; Morphology; Numbers; Outcomes Research; PTK2 gene; Phenotype; Phosphorylation; Phosphotyrosine; Play; Process; Production; Property; Proteoglycan; Range; Research; Research Personnel; Role; Signaling Molecule; Smooth Muscle Myocytes; Speed; Stents; Stress; Substrate Interaction; System; Techniques; Testing; Time; Traction; Variant; Vascular Graft; Vascular Smooth Muscle; Vascular remodeling; Video Microscopy; Western Blotting; Work; antibody inhibitor; base; cell behavior; cell motility; clinically significant; design; feeding; in vitro Model; insight; intracellular protein transport; monomer; mouse Smc1l1 protein; mouse Smc1l2 protein; novel; paxillin; photopolymerization; programs; protein expression; protein localization location; response; restenosis; rho; rho GTP-Binding Proteins; success; time use; vasoconstriction

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 micro- scales. 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.

由于内膜增生和血管收缩引起的再狭窄仍然是动脉治疗的主要问题 闭塞性疾病。平滑肌细胞在血管重塑中起主要作用，局部控制其 细胞表型将极大地加强减少再狭窄的努力。一个常见的结果 血管损伤是细胞外基质的过度重塑，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，但也显着改变 生物力学特性。基于最近的工作，细胞对基质有不同的机械响应 属性，我们的中心假设是底层的生物力学特性会调节 血管平滑肌细胞细胞表型与再狭窄相关。当前的再狭窄疗法 很大程度上涉及可溶性因素，但很少关注理解基质的影响 细胞表型上的生物力学特性。拟议研究的目的是创建模型 在血管重塑过程中会概括生物力学环境并识别钥匙的系统 与再狭窄相关的底物合规性与细胞表型之间的关系。这个目标 将通过研究底物符合对光滑细胞表型的影响来实现 模型生物工程基质上的肌肉细胞旨在在其系统上表现出系统变化 从显微镜到宏观长度尺度的合规性。这项研究的结果将是 一种新型的体外模型系统，将更加模仿重塑的生物力学环境 基质可以测试药物对血管平滑肌细胞表型的影响。这个模型 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。目的 1是在宏观和微观方面开发具有明确机械符合性的生物工程基质 秤。 AIM 2和3中研究的结果将用于AIM 1的完善。AIM 2将建立和 量化底物合规性与血管平滑肌细胞表型之间的关系 与再狭窄。 AIM 3将测试底物遵守对表达，本地化和 推定的机械感应细胞成分（整联蛋白，细胞骨架，FAK，Paxillin，Rho）的活性 GTPases）。这些研究将提高对控制表型调节的物理因素的新见解 血管平滑肌细胞的旨在开发疗法以阻断再狭窄。