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: 6851399
• 负责人: PHIL GORDON CAMPBELL
• 金额: $ 31.36万
• 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/6851399
• 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在指导细胞迁​​移、增殖和分化方面发挥着关键作用。该拨款申请解决了以下问题：在工程仿生细胞外基质 (bECM) 内以指定方向梯度固定的外源固相生长因子能否成为招募和引导周围宿主细胞进入 bECM 并促进所需生物学结果的有效方法？目标是确定这种生长因子递送范例是否与组织工程应用中的未来治疗开发相关。体外模型范例将包括人微血管内皮细胞、MG-63 人前成骨细胞和人成人间充质干细胞对纤维蛋白 bECM 中固相人成纤维细胞生长因子 2 (FGF-2) 的反应。体内模型将是鸡胚绒毛尿囊膜 (CAM) 测定，以评估置于 CAM 上的纤维蛋白 bECM/FGF-2 结构内的定向血管生成反应。该研究将从纤维蛋白膜上 FGF-2 的二维模式系统地进展到纤维蛋白 bECM 内 FGF-2 的三维 (3D) 模式，以及从体外分离的细胞反应到体内血管生成反应。总体假设是纤维蛋白 bECM 中固相 FGF-2 的空间梯度将指导体外细胞迁移，与梯度方向性一致，并且响应梯度模式的细胞迁移通量将大于响应均匀分布模式的细胞迁移通量。在体内，与均匀分布相比，梯度设计的定向血管生成反应会更大。 bECM/FGF-2 结构将采用固体自由成型制造印刷工艺制造，该工艺可在 3D 纤维蛋白 bECM 内产生生长因子的指定 3D 空间图案。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打印结构并评估细胞和 CAM 响应。该研究将由具有内分泌学、先进制造和生物物理成像专业知识的多学科生物工程人员进行。