GROWTH FACTOR MEDIATION OF HEALING OF VASCULAR GRAFTS

生长因子对血管移植物愈合的调节

• 批准号: 6330034
• 负责人: HOWARD P GREISLER
• 金额: $ 27.55万
• 依托单位: LOYOLA UNIVERSITY CHICAGO
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-07-01 至 2002-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6330034
• 关键词: biomaterial interface interaction; biomechanics; blood vessel prosthesis; cardiovascular transplantation; cell migration; cell proliferation; chimeric proteins; dogs; endarterectomy; extracellular matrix; fibrin; fibroblast growth factor; heparin; histology; immunocytochemistry; insulinlike growth factor; mutant; site directed mutagenesis; vascular endothelial growth factors; vascular endothelium; vascular smooth muscle; wound healing

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.

描述(改编自申请者摘要)：所有当前治疗 动脉粥样硬化的介入治疗结果令人失望。 主要是由于血液：动脉壁和血液：材料的界面。 主要问题是早期血栓形成和晚期血栓形成的比率都很高。 肌内膜增生。这两种故障模式可能都会减少 募集正常功能的内皮细胞(ECM)单层。这个 调查员和他的同事们开发了一部小说并对其进行了表征 生物活性生长因子的受控局部输送方法 和/或植入血管移植物或血管壁的细胞。vbl.使用 研究人员已经证明了该系统的诱导能力 跨壁毛细血管生成和融合血管生成 内皮化。这些研究是在移植物上进行的，只要 30厘米的狗，这些研究表明显著抑制 犬球囊内皮剥脱术后肌内膜增生 颈动脉。已经确定了该系统的局限性，并 提出了解决方案。本提案的假设是 先前描述的添加成纤维细胞生长因子-1的纤维蛋白胶的性能 促进血管治疗干预模式中的内皮化 可以在不同时促进肌内膜增生的情况下维持 使用成纤维细胞生长因子-1的EC特异性定点突变体和/或通过包含 增加有丝分裂信号的EC特异性的其他因素。 本提案是围绕三个具体目标提出的。特定目标 #1旨在表征现有的成纤维细胞生长因子-1突变体和嵌合蛋白 并生成和表征逻辑派生的第二代 基于现有的体外检测的突变。具体目标#2旨在 评估优化的成纤维细胞生长因子-1突变体的潜在改善性能 嵌合体通过添加可能的因素补充纤维蛋白胶 使用EC和EC提高EC有丝分裂潜能的特异性 SMC文化和共文化。在具体目标#3中，调查人员提出 对移植组织的生物力学完整性进行量化并评价 体外优化处方的细胞特异性反应 体外循环动物模型体内添加纤维蛋白原的实验研究 移植和动脉内膜切除术。申请者将系统地评估 血管内皮细胞和平滑肌细胞生长动力学及功能特点 利用生化、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 手术后信使核糖核酸的产生与时间的关系。终极目标 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，以优化愈合与减少 动脉内膜切除术的再狭窄，并在未来的研究中修改 含有这些生长因子的导管输送技术 经腔内手术后动脉的悬浮物，包括 血管成形术和动脉粥样硬化切除术。此外，希望这项技术能够 将为转基因技术的应用提供一个最佳的系统 以基因产品输送为目的的转基因ECs。