Gene delivery approaches to vascular graft elastogenesis

描述(申请人提供)：组织工程血管(TEBV)生成的主要障碍之一是种子细胞无法产生足够数量的交联性弹性蛋白来实现正常动脉的机械特性。本研究的目的是优化成人平滑肌细胞的增殖、排列和弹力形成的条件，以实现TEBV在培养中的快速发展。一种基于蛋白多糖V3剪接变异体过表达的基因工程策略，在体外和体内诱导成年哺乳动物细胞弹性形成，但抑制细胞增殖。这项建议的目的是为V3和其他弹性基因的表达生成可调控的载体，以便可以控制弹性蛋白合成的大小和时间。我们假设，该系统将使细胞覆盖率和弹性蛋白沉积在生物材料支架上达到最优化。为了提供机械支撑和促进排列，细胞将被种植到有凹槽的胶原膜上。排列好的单元片将形成管状结构。这些构建物将在生物反应器中受到径向应变，并将评估细胞预排列和弹性形成的力学贡献。

DESCRIPTION (provided by applicant): Among the primary obstacles to generating tissue engineered blood vessels (TEBV) is the inability of seeded cells to produce sufficient amounts of cross-linked elastin to achieve the mechanical characteristics of normal arteries. The objective of this proposal is to optimize conditions for adult smooth muscle cell proliferation, alignment and elastogenesis to achieve rapid development of TEBV in culture. A genetic engineering strategy based on overexpression of a splice variant of the proteoglycan versican, V3, induces elastogenesis in adult mammalian cells both in vitro and in vivo, but attenuates cell proliferation. The objectives of this proposal are to generate regulatable vectors for expression of V3 and other elastogenic genes so that the magnitude and timing of elastin synthesis can be controlled. We hypothesize that this system will enable optimization of cell coverage and elastin deposition on biomaterial scaffolds. To provide mechanical support and promote alignment, cells will be seeded onto grooved collagen membranes. Sheets of aligned cells will be formed into tubular constructs. The constructs will be subjected to radial strain in a bioreactor and the mechanical contribution of cell pre-alignment and elastogenesis will be assessed.