Use of Proteoglycan-Genes to Engineer Vascular Tissue

描述（由申请人提供）： 血管组织工程中的主要限制之一是工程化组织不能形成功能性和完整的弹性纤维。 我们实验室最近的工作已经发现，通过基因工程动脉平滑肌细胞过表达细胞外基质蛋白聚糖，多功能蛋白聚糖，弹性纤维的特定亚型在体外和体内形成时，转导的细胞接种到受伤的动脉。 我们假设这项技术可以用于制造具有完整和功能性弹性纤维的血管替代物和血管移植物。 为了解决这一假设，本建议有以下具体目标：1。 优化基因递送系统，以便以受控方式实现最佳表达水平，以最大限度地增加弹性纤维形成。 2. 在体外用完整的弹性纤维创建工程细胞的“片材”，以用于形成表现出最大弹性特性的小直径管。 3. 评估将工程细胞接种到围绕框架缠绕的基于微纤维的可生物降解聚合物支架上以产生散布有功能性弹性纤维的平滑肌细胞层的可行性; 4. 确定将这些弹性纤维生成细胞整合到基于胶原的管状支架中是否会导致形成具有适当拉伸强度和弹性特性的管。 本申请的总体目标是创建具有细胞外基质组分的适当混合物的血管中间层，所述细胞外基质组分模拟天然血管并含有完整和功能性弹性纤维。

DESCRIPTION (provided by applicant): One of the major limitations in vascular tissue engineering is the failure of the engineered tissue to form functional and intact elastic fibers. Recent work in our laboratory has found that by genetically engineering arterial smooth muscle cells to overexpress a specific isoform of the extracellular matrix proteoglycan, versican, elastic fibers form in vitro and in vivo when the transduced cells are seeded into injured arteries. We hypothesize that this technology can be used to create blood vessel substitutes and vascular grafts with intact and functional elastic fibers. To address this hypothesis, this proposal has the following specific aims: 1. Optimize the gene delivery system so that optimal levels of expression can be achieved in a controlled fashion to maximize elastic fiber formation. 2. Create "sheets" of engineered cells in vitro with intact elastic fibers to be used to form small diametric tubes which exhibit maximal elastogenic properties. 3. Assess the feasibility of seeding engineered cells onto micro-fiber based biodegradable polymer scaffolds wrapped around a frame to create layers of smooth muscle cells interspersed with functional elastic fibers and; 4. Determine if integrating these elastic fiber generating cells into collagen based tubular scaffolds will result in the formation of tubes with appropriate tensile strength and elastogenic properties. The overall goal of this application is to create a vascular medial layer that possesses an appropriate mixture of extracellular matrix components that mimic the natural vessel and contain intact and functional elastic fibers.