Cell and matrix interactions in diabetic vascular tissue engineering models

糖尿病血管组织工程模型中细胞和基质的相互作用

基本信息

批准号：
9383944
负责人：
Agneta Simionescu
金额：
$ 32万
依托单位：
CLEMSON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-01 至 2022-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9383944
关键词：
Advanced Glycosylation End Products Affect American Animal Model Animals Anti-Inflammatory Agents Anti-inflammatory Antioxidants Arterial Fatty Streak Arteries Artificial Implants Atherosclerosis Binding Biochemical Biocompatible Materials Bioreactors Blood Glucose Blood Vessels Caliber Cardiovascular system Cells Chemicals Clinical Research Collagen Cytoskeleton Deposition Development Devices Diabetes Mellitus Disease Dyslipidemias Elastin Endothelial Cells Endothelium Environment Event Extracellular Matrix Fibroblasts Fibrosis Future Glucose Goals Growth Heart Valves Human Hypertrophy Impaired wound healing Implant Incubated Inflammation Inflammatory Response Injury Kidney Light Link Literature Mechanics Mesenchymal Metformin Modeling Modification Monitor Morphology Myocardium NADPH Oxidase Nerve Nude Rats Operative Surgical Procedures Oxidative Stress Pathologic Pathology Patients Physiological Problem Solving Process Production Proliferating Proteins Rattus Reactive Oxygen Species Resistance Risk Factors Role Smooth Muscle Myocytes Stimulus TLR4 gene Testing Tissue Engineering Tissues Tunica Adventitia Vascular Diseases Vascular remodeling abdominal aorta base blood lipid calcification cardiovascular risk factor cell type crosslink diabetic diabetic patient emergency service responder expectation high risk human tissue immunoregulation implantation injured insulin sensitizing drugs monocyte non-diabetic oxidation receptor repaired response scaffold three-dimensional modeling tissue support frame vasa vasorum vascular tissue engineering

项目摘要

Diabetes is a major risk factor for vascular diseases that affects nearly all blood vessel types and calibers. In diabetes, elevated levels of blood glucose and lipids interact irreversibly with long-lived proteins, such as collagen and elastin from the blood vessel wall, via oxidation and crosslinking processes, resulting in formation of advanced glycation end products (AGEs); the consequence is vascular stiffening, the hallmark of diabetes. Furthermore, vascular cells respond to diabetes- related altered environment by activation and leading to pathological remodeling and to the onset and progression of vascular disease. Together, these severe cell and extracellular matrix (ECM) changes result in activation of inflammation, impaired healing, fibrosis, and ectopic calcification. The interaction of AGEs with their receptor, RAGE, stimulates the production of reactive oxygen species, leading to dysfunctional remodeling of the vascular wall (stiffening, fibrosis, and calcification). The goal of this project is to characterize the effect of diabetes on the adventitial fibroblasts and their involvement in vascular pathology. By using 3D models based on tissue engineering principles, we can control the type of cells seeded on a vascular matrix-based scaffold while providing the necessary biochemical and mechanical stimuli in a physiologic bioreactor. The tissue engineered construct can also be implanted in diabetic animal models, to explore the effect of ECM oxidation and AGE accumulation on the fate of adventitial fibroblasts. The effect of antioxidant and anti-inflammatory agents can also be monitored. Our hypothesis is that fibroblasts are activated by ROS and contribute to the dysfunctional remodeling of the vascular wall in response to diabetes-induced injuries. This hypothesis will be tested in the following two aims. In specific Aim 1 we will investigate the contribution of diabetic adventitial fibroblasts to the pathological vascular wall remodeling. Vascular ECM-based scaffolds (acellular arteries) will be seeded with human endothelial cells, smooth muscle cells, and fibroblasts and a) incubated in a physiologic vascular bioreactor for 2 months in diabetic media, b) implanted as transposition grafts in the abdominal aorta of normal and diabetic nude rats for 3 and 6 months. Grafts will be monitored for oxidative stress and inflammation. In specific Aim 2 we will explore the fate of diabetic adventitial fibroblasts in the presence of antioxidant and anti-inflammatory agents. Vascular ECM-based scaffolds seeded with human vascular cells will be a) incubated in a physiologic vascular bioreactor for 2 months in diabetic media and b) implanted as transposition grafts in the abdominal aorta of normal and diabetic nude rats for 3 and 6 months, in the presence of antioxidant polyphenolic compounds, metformin, an insulin- sensitizer drug, and immunomodulatory mesenchymal cells (in separate groups). Grafts will be monitored for oxidative stress and inflammation. Expectations: at the conclusion of this study, we would gain important information about the major diabetes-related alterations in the vascular wall initiated by the adventitial fibroblasts, potentially offering avenues for targeting these events.

糖尿病是血管疾病的主要危险因素，影响几乎所有血管类型和口径。在糖尿病中，血糖和血脂水平升高与长寿不可逆地相互作用通过氧化和交联产生蛋白质，例如血管壁中的胶原蛋白和弹性蛋白过程，导致高级糖基化终产物（AGE）的形成；结果是血管硬化，糖尿病的标志。此外，血管细胞对糖尿病有反应- 通过激活和导致病理重塑以及发病和发生相关改变的环境血管疾病的进展。总之，这些严重的细胞和细胞外基质 (ECM) 变化导致炎症激活、愈合受损、纤维化和异位钙化。 AGE 与其受体 RAGE 相互作用，刺激活性氧的产生种，导致血管壁重塑功能失调（硬化、纤维化和钙化）。该项目的目标是表征糖尿病对外膜成纤维细胞及其功能的影响参与血管病理学。通过使用基于组织工程原理的 3D 模型，我们可以控制接种在基于血管基质的支架上的细胞类型，同时提供必要的生理生物反应器中的生化和机械刺激。组织工程构建体可以也可植入糖尿病动物模型，探讨ECM氧化和AGE的影响积累对外膜成纤维细胞命运的影响。抗氧化和抗炎作用代理也可以被监控。我们的假设是成纤维细胞被 ROS 激活并导致功能失调血管壁重塑以应对糖尿病引起的损伤。这个假设将是在以下两个目标中进行测试。在具体目标 1 中，我们将研究糖尿病外膜成纤维细胞对糖尿病的贡献病理性血管壁重塑。基于血管 ECM 的支架（无细胞动脉）将接种人内皮细胞、平滑肌细胞和成纤维细胞，并且 a) 在生理血管生物反应器在糖尿病培养基中培养 2 个月，b) 作为转位移植物植入正常和糖尿病裸鼠的腹主动脉3个月和6个月。将监测移植物氧化应激和炎症。在具体目标 2 中，我们将探讨糖尿病外膜成纤维细胞在存在以下因素的情况下的命运：抗氧化剂和抗炎剂。基于血管 ECM 的支架植入了人类血管细胞将 a) 在生理血管生物反应器中的糖尿病培养基中孵育 2 个月 b) 将转位移植物植入正常和糖尿病裸鼠的腹主动脉 3 次 6 个月，在抗氧化剂多酚化合物二甲双胍（一种胰岛素）存在下敏化剂药物和免疫调节间充质细胞（分在不同的组中）。移植物将是监测氧化应激和炎症。期望：在本研究结束时，我们将获得有关该专业的重要信息由外膜成纤维细胞引发的与糖尿病相关的血管壁改变，可能提供针对这些事件的途径。