Cell and matrix interactions in diabetic vascular tissue engineering models

糖尿病是血管疾病的主要危险因素，几乎影响所有血管类型和 口径。在糖尿病中，血糖和血脂水平的升高与长寿之间存在不可逆转的相互作用。 蛋白质，如血管壁上的胶原蛋白和弹性蛋白，通过氧化和交联 过程，导致形成晚期糖基化终末产物(AGEs)；其后果是 血管僵硬是糖尿病的标志。此外，血管细胞对糖尿病做出反应- 与激活相关的环境改变并导致病理性重塑和发病 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、愈合受阻、纤维化和异位钙化。 AGEs与其受体RAGE的相互作用刺激活性氧的产生 导致血管壁功能重建(硬化、纤维化和钙化)的物种。 该项目的目标是表征糖尿病对血管外膜成纤维细胞及其功能的影响。 与血管病理有关。通过使用基于组织工程学原理的3D模型，我们 可以控制种植在基于血管基质的支架上的细胞类型，同时提供必要的 生理学生物反应器中的生化和机械刺激。组织工程化构建物可以 还可植入糖尿病动物模型，探讨细胞外基质氧化和增龄的影响 外膜成纤维细胞命运的积聚。抗氧化剂和抗炎作用 还可以监控代理。 我们的假设是成纤维细胞被ROS激活，并在功能障碍中起作用 糖尿病所致损伤时血管壁的重塑。这一假设将是 在以下两个目标上进行了测试。 在特定的目标1中，我们将研究糖尿病外膜成纤维细胞在糖尿病血管病变中的作用。 病理性血管壁重塑。基于血管ECM的支架(无细胞动脉)将 接种于人内皮细胞、平滑肌细胞和成纤维细胞，以及a)在 生理性血管生物反应器在糖尿病介质中2个月，b)作为移植物植入 取正常大鼠和糖尿病大鼠的腹主动脉，连续观察3个月和6个月。移植物将被监测 氧化应激和炎症。 在特定的目标2中，我们将探讨糖尿病外膜成纤维细胞在存在 抗氧化剂和消炎剂。人种植血管ECM支架材料的研究 血管细胞将在生理性血管生物反应器中在糖尿病培养液中培养2个月 和b)作为移植物植入正常和糖尿病裸鼠的腹主动脉内，共3次 6个月后，在抗氧化剂多酚化合物、二甲双胍、胰岛素- 致敏药物和免疫调节间充质细胞(分成两组)。移植物将会是 监测氧化应激和炎症。 期望：在这项研究的结论中，我们将获得有关专业的重要信息 可能由外膜成纤维细胞引发的糖尿病相关的血管壁改变 提供针对这些活动的途径。

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.