GROWTH FACTOR MEDIATION OF HEALING OF VASCULAR GRAFTS

描述（改编自申请人摘要）：所有当前治疗 动脉粥样硬化的介入治疗方法的结果令人失望 这主要是由于血液：动脉壁和血液：材料界面。 主要问题是早期血栓形成和晚期血栓形成的发生率都很高。 肌内膜增生 这两种失效模式可能会减少， 募集正常功能的内皮细胞（ECM）单层。 的 研究人员和他的同事们开发并描述了一种小说 生物活性生长因子的受控局部递送方法 和/或浸渍到血管移植物或血管壁中的细胞。 使用 FGF-1，研究人员已经证明了该系统诱导 通过透壁毛细血管化和融合的血管生成 内皮化 这些研究已经在移植物中进行， 30 cm，这些研究表明， 犬球囊血管内皮剥脱术后肌内膜增生 颈动脉 该系统的局限性已被确定， 提出的解决方案。 本提案的假设是， 先前描述的补充FGF-1的纤维蛋白胶在 促进血管治疗干预模型中的内皮化 可以维持而不同时促进肌内膜增生， 使用FGF-1的位点定向EC特异性突变体和/或通过包含 增加促有丝分裂信号的EC特异性的额外因子。 本提案围绕三个具体目标组织。 具体目标 #1设计用于表征现有FGF-1突变体和嵌合蛋白 并生成和表征逻辑导出的第二代 基于现有的体外测定的突变。 具体目标#2旨在 评估优化的FGF-1突变体的潜在改进性能 嵌合体通过添加可能导致 使用EC增加EC的促有丝分裂潜能的特异性， SMC培养和共培养。 在具体目标#3中，研究人员建议 对移植组织的生物力学完整性进行定量， 细胞对体外优化制剂的特异性反应 补充FG在良好表征的旁路动物模型中的体内试验 移植和动脉内膜切除术。 申请人将系统地评估 EC和SMC向内生长的动力学和 使用生物化学、组织学、免疫组织化学、 以及细胞增殖和蛋白质的分子生物学分析， 手术后mRNA产生随时间的变化。 最终目标 这项工作的目的是应用这项技术来提高临床疗效， 小直径血管移植物，以优化愈合， 再狭窄的动脉内膜切除术，并在未来的研究，以修改 用于导管递送这些含有生长因子的技术 在经腔手术后将混悬液注入动脉，包括 血管成形术和斑块切除术。 此外，希望这项技术 将为转染的细胞的应用提供最佳系统， 用于基因产物递送目的的遗传修饰的EC。

DESCRIPTION (Adapted from applicant's abstract): All current therapeutic interventional modalities in atherosclerosis have had disappointing results due largely to the blood:arterial wall and the blood:material interface. The main problems have been high rates of both early thrombosis and late myointimal hyperplasia. Both failure modes would likely be reduced by recruitment of a normal functioning endothelial cell (ECM) monolayer. The investigator and his colleagues have developed and characterized a novel method for controlled local delivery of biologically active growth factors and/or cells impregnated into a vascular graft or a vessel wall. Using FGF-1, the investigators have shown the system's capability of inducing angiogenesis via transmural capillarization and confluent endothelialization. These studies have been conducted in grafts as long as 30 cm in dogs and these studies have shown significant inhibition of myointimal hyperplasia following balloon de-endothelialization of canine carotid arteries. Limitations of this system have been identified and solutions proposed. The hypothesis of the present proposal is that the previously described performance of the FGF-1 supplemented fibrin glue in promoting endothelialization in models of vascular therapeutic interventions can be maintained without simultaneously promoting myointimal hyperplasia by the use of site directed EC specific mutants of FGF-1 and/or by inclusion of additional factors that increase the EC specificity of the mitogenic signal. The present proposal was organized about three specific aims. Specific Aim #1 is designed to characterize existing FGF-1 mutants and chimeric proteins and to generate and characterize logically derived second generation mutations based on existing in vitro assays. Specific Aim #2 is designed to evaluate the potential improved performance of the optimized FGF-1 mutant chimera supplemented fibrin glue through the addition of factors likely to increase the specificity of the mitogenic potential for the EC using EC and SMC cultures and co-cultures. In Specific Aim #3, the investigators propose to quantitate the biomechanical integrity of the graft tissue and evaluate cell specific responses to the in vitro based optimized formulation of supplemented FG in vivo in well characterized animal models of bypass grafting and of endarterectomy. The applicants will systematically evaluate the kinetics of EC and SMC ingrowth and functional characteristics of ingrowing ECs and SMCs using biochemical, histologic, immunohistochemical, and molecular biologic analysis of cellular proliferation and protein and mRNA production as a function of time following surgery. The ultimate goals of this work are to apply the technology to improve the clinical efficacy of small diameter vascular grafts, to optimize healing with diminished restenosis of endarterectomy procedures, and in future studies to modify the technology for catheter delivery of these growth factor containing suspensions to arteries following transluminal procedures including angioplasty and atherectomy. Furthermore, it is hoped that this technology will provide an optimal system for the application of transfected genetically modified ECs for purposes of gene product delivery.