Engineered Artery Growth in Vitro, Cell-Remodeled Fibrin

体外工程动脉生长、细胞重塑纤维蛋白

• 批准号: 7069323
• 负责人: ROBERT T TRANQUILLO
• 金额: $ 55.49万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-06-01 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7069323
• 关键词: angiogenesis; artery; biomaterial compatibility; biomaterial development /preparation; biomaterial interface interaction; bioreactors; blood vessel prosthesis; fibrin; fibrinolysis; hemodynamics; human tissue; laboratory rat; medical implant science; swine; tissue /cell culture; tissue engineering; tissue support frame; ultrasound blood flow measurement; vascular smooth muscle

DESCRIPTION (provided by applicant): The ultimate goal of this project is a completely biological artificial artery (bioartificial artery) that is suitable as a small diameter vascular graft, such as for use in coronary bypass. We will build upon our two major discoveries: (1) when neonatal smooth muscle cells (SMCs) are entrapped in fibrin gel formed as a tube around a nonadhesive rod, the SMCs contract the gel around the rod, causing the fibrin fibrils and SMCs to become circumferentially aligned. As the SMCs subsequently degrade the fibrin, they produce extensive cross-linked collagen and elastic fibers, which also are circumferentially aligned. This remodeling provides the construct with tensile mechanical properties approaching values of arterial tissue; (2) endothelial cells cultured from blood can be expanded to 10A8 cells in less than six weeks, the expansion occurring from a putative circulating marrow-derived angioblast. In Specific Aim 1, we will compare the properties of remodeled fibrin gel tubes prepared with neonatal SMCs (rat, pig, and human) under the static culture condition used to date with tubes subject to cyclic distension, imparting a mechanical signal expected to increase tissue growth as we found in collagen gel-based tubes, and controlled transmural flow of culture medium, minimizing gradients of soluble factors and potentially leading to increased and near-uniform tissue growth. In Aim 2, we will study the properties of bioartificial arteries, the remodeled fibrin/SMC constructs described above plus an endothelium generated using traditional late-stage seeding of the tubular construct with the blood outgrowth endothelial cells (BOECs) noted above, including their adhesion strength in pulsatile flow at physiological shear stress and their state of activation. Once such constructs have been validated in vitro we will proceed with testing the viability, patency, and hemocompatibility properties in animal models. The bioartificial artery will be implanted into the aorta of the rat, first in the syngeneic setting and subsequently in the "allogeneic" setting (with respect to the SMCs; autologous BOECs will be used). A similar allogeneic study will subsequently be performed in the pig. We will also fabricate bioartificial arteries from human SMC and BOECs, so that these results should be directly relevant for a future clinical study.

描述（由申请人提供）： 该项目的最终目标是完全生物人造动脉（生物人造动脉），适合作为小直径的血管移植物，例如用于冠状动脉旁路。我们将建立在两个主要发现的基础上：（1）当新生儿平滑​​肌细胞（SMC）被捕获在纤维蛋白凝胶中，形成了围绕非粘附杆的管，SMCS在杆周围凝胶收缩，从而导致纤维蛋白原纤维和SMC，以使其圆周平整。随后SMC降解纤维蛋白时，它们会产生广泛的交联胶原蛋白和弹性纤维，它们也会圆周排列。这种重塑为构建体提供了接近动脉组织值的拉伸机械性能。 （2）从血液培养的内皮细胞可以在不到六周的时间内将其扩展到10A8细胞，这是由推定的循环骨髓衍生的血管细胞进行的。在特定目标1中，我们将比较在静态培养条件下用新生儿SMC（大鼠，猪和人）制备的重塑纤维蛋白凝胶管的性能，该条件与静态培养物条件相比，与管道延伸的试管相关，这会散发出一种机械信号，从而在我们在基于胶原型的胶原型和控制型的跨度型培养基范围内发现的机械信号，并会增加组织的生长。几乎均匀的组织生长。在AIM 2中，我们将研究生物人工动脉的特性，上述重塑的纤维蛋白/SMC构建体以及使用管状结构的传统后期播种产生的内皮，并具有上述血液生长内皮细胞（BOEC），包括上述引起的血液外壳内皮细胞（BOEC），包括它们在生理学的脉动流动及其状态的脉动流动强度和他们的态度。一旦在体外验证了此类构建体，我们将继续测试动物模型中的生存​​力，通畅性和血流相容性。生物人工动脉将植入大鼠的主动脉，首先在合成环境中，然后在“同种异体”设置中（相对于SMC；将使用自体BOEC）。随后将在猪中进行类似的同种异体研究。我们还将从人类SMC和BOEC中捏造生物人工动脉，以便这些结果应与未来的临床研究直接相关。