Cell Response to 3D Engineered Gradients of FGF-2

细胞对 FGF-2 3D 工程梯度的反应

• 批准号: 7060419
• 负责人: PHIL GORDON CAMPBELL
• 金额: $ 30.41万
• 依托单位: CARNEGIE-MELLON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-05-01 至 2009-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7060419
• 关键词: angiogenesis; bioengineering /biomedical engineering; biomimetics; biotechnology; cell adhesion; cell migration; cell proliferation; chorioallantoic membrane; extracellular matrix; fibrin; fibroblast growth factor; fluorescence microscopy; fluorescent dye /probe; intravital microscopy; osteoblasts; scanning electron microscopy; stem cells; therapy design /development; tissue engineering; vascular endothelium

DESCRIPTION (provided by applicant): Spatial concentration gradients of endogenous soluble growth factors play critical roles directing cell migration, proliferation, and differentiation during development, homeostasis, and wound healing. This grant application addresses the question: Could an exogenous solid-phase growth factor immobilized in specified directional gradients within an engineered biomimetic extracellular matrix (bECM) be an effective approach to recruit and guide surrounding host cells into a bECM and promote a desired biological outcome? The goal is to determine if this growth factor delivery paradigm would be relevant for future therapy development in tissue engineering applications. The paradigm in vitro models will include human microvascular endothelial cell, MG-63 human preosteoblastic cell, and human adult mesenchymal stem cell responses to solid-phase human fibroblast growth factor-2 (FGF-2) in fibrin bECMs. The in vivo model will be a chicken embryo chorioallantoic membrane (CAM) assay to assess directed angiogenic response within fibrin bECM/FGF-2 structures placed on the CAM. The research will systematically progress from two-dimensional patterns of FGF-2 on fibrin films to three-dimensional (3D) patterns of FGF-2 within fibrin bECMs, and from isolated cell responses in vitro to angiogenic responses in vivo. The overall hypotheses are spatial gradients of solid phase FGF-2 in a fibrin bECM will direct cell migration in vitro in register with gradient directionality, and cell migrational flux will be greater in response to gradient patterns than patterns with uniform distributions. And in vivo, a directed angiogenic response will be greater in designs with gradients in comparison with uniform distributions. The bECM/FGF-2 structures will be fabricated with a solid freeform fabrication printing process that produces specified 3D spatial patterns of growth factors within 3D fibrin bECMs. Fluorescent based quantum dot imaging technology and fluorescence optical sectioning microscopy will be used to validate printed structures and assess cell and CAM responses. The research will be carried out by multidisciplinary bioengineering with expertise in endocrinology, advanced manufacturing, and biophysical imaging.

描述（由申请人提供）： 内源性可溶性生长因子的空间浓度梯度在发育，稳态和伤口愈合过程中指导细胞迁​​移，增殖和分化的关键作用。该赠款申请解决了以下问题：在工程仿生的细胞外基质（BECM）中，固定在指定方向性梯度中的外源固相生长因子是否可以成为一种有效的方法，可以募集和引导周围的宿主细胞进入BECM并促进所需的生物学结果？目的是确定这种生长因子递送范式是否与组织工程应用中的未来治疗开发有关。体外模型的范式将包括人类微血管内皮细胞，MG-63人类前细胞细胞和人类成人间充质干细胞对纤维蛋白BECM中对固相的人成纤维细胞生长因子2（FGF-2）的反应。体内模型将是一种鸡肉胚胎绒毛膜膜（CAM）测定，以评估放置在CAM上的纤维蛋白BECM/FGF-2结构中的有针对性的血管生成反应。该研究将系统地从纤维蛋白膜上的FGF-2的二维模式到纤维蛋白BECM中FGF-2的三维（3D）模式，从体外体外对血管生成反应的分离细胞反应。总体假设是纤维蛋白becm中固相FGF-2的空间梯度，将在带有梯度方向性的注册中引导细胞迁移，并且与具有统一分布的模式相比，细胞迁移通量对梯度模式的响应将更大。在体内，与均匀分布相比，在具有梯度的设计中，定向的血管生成反应将更大。 BECM/FGF-2结构将使用固体自由形式制造工艺制造，该过程在3D纤维蛋白BECM中产生指定的3D空间生长因子的空间模式。基于荧光的量子点成像技术和荧光光学切片显微镜将用于验证印刷结构并评估细胞和CAM响应。这项研究将通过多学科生物工程具有内分泌，高级制造和生物物理成像方面的专业知识进行。