Artery-on-a-chip with perivascular adipose tissue for pressure myography

在健康的心血管功能中，血管平滑肌细胞围绕在血管周围，通过收缩或放松来改变血管直径。最近的研究表明，大多数血管周围的脂肪层，称为血管周围脂肪组织（PVAT），可能调节血管的收缩性。在健康受试者中，PVAT使邻近的血管松弛，使血液更容易流动。然而，在高血压和动脉粥样硬化等疾病中，PVAT可能导致血管收缩。虽然人类和动物都可以测量血管的收缩和扩张，但在这些模型中很难研究PVAT对血管的影响。研究者的长期目标是了解PVAT如何影响健康和疾病中的动脉功能。该项目的目标是创建一个动脉芯片模型，可以测量血管收缩性，并在未来与PVAT集成。具体来说，该项目将利用微带和纳米带制造一条动脉，使可收缩的血管平滑肌细胞沿空心通道圆周排列。然后，通道内部将内衬内皮细胞。这种芯片上的动脉装置将通过生化分析和与小鼠血管测量和人体研究的比较来验证。芯片上的动脉可以用于药物测试，并在未来提高对PVAT变化如何影响血管功能的理解。此外，将扩大本科生参与研究经验的机会。具体而言，该研究项目将被纳入基于课程的本科研究经验（CURE），在该课程中，整个班级将解决科学界感兴趣的研究问题。然后，课程的组成部分将被翻译成细胞和器官水平的模块，供参加生物医学暑期学院的高中生使用。该项目的重点是创造一种芯片动脉，能够在机械和生化刺激下测量血管收缩力。血管收缩是由血管平滑肌细胞（vSMC）和血管周围脂肪组织（PVAT）介导的，对血压调节、血液分布和损伤反应至关重要。虽然已知PVAT在健康状态下对邻近的血管系统发挥抗收缩作用，而在高血压、动脉粥样硬化、动脉瘤和肥胖等疾病中失去其抗收缩作用，但大多数将PVAT与血管疾病联系起来的证据仍然是相关的，因为很少有方法可以在不影响其他脂肪储存点或血管本身的情况下特异性地操纵PVAT。目前的体外动脉系统不包括PVAT和少数能够血管收缩性测量。该项目开发的芯片动脉平台将通过实现两个目标来满足这一需求。第一个目标是制造一个内皮化的周向排列、可收缩的vSMCs管。研究旨在探索排列的微带和纳米带如何在圆柱形通道中创建排列的内皮化vSMC合胞体，并确定微带和纳米带的组成和机械性能对vSMC血管收缩和血管舒张的影响。该芯片上的动脉将通过小鼠动脉压肌图和人类血管反应性数据进行验证。第二个目标是确定如何利用微带和纳米带的独特特性来测量vSMC产生的三维牵引力。vSMC收缩测量将与芯片上动脉收缩进行比较，并将使用小鼠动脉压力肌图和人类血管反应性数据进行验证。这项研究将首次创建人体动脉芯片，以测试血管收缩性，并在未来有可能纳入PVAT。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

Vascular smooth muscle cells surround blood vessels and contract or relax to change the vessel diameter during healthy cardiovascular function. Recent studies have shown that the layer of fat surrounding most blood vessels, called perivascular adipose tissue (PVAT), may modulate vascular contractility. In healthy subjects, PVAT relaxes the adjacent blood vessel, making it easier for blood to flow. However, in diseases such hypertension and atherosclerosis, PVAT may cause the blood vessels to constrict. While blood vessel constriction and dilation can be measured in humans and animals, it is difficult to study the effects of PVAT on the blood vessels in these models. The investigator's long-term goal is to understand how PVAT affects arterial function in health and disease. The goal of this project is to create an artery-on-a-chip model that enables measurement of vascular contractility and could in the future be integrated with PVAT. Specifically, the project will create an artery using micro- and nano-ribbons to align contractile vascular smooth muscle cells circumferentially around a hollow channel. The inside of the channel will then be lined with endothelial cells. The artery-on-a-chip device will be validated using biochemical assays and comparisons to mouse blood vessel measurements and to human studies. The artery-on-a-chip can then be used for drug testing and in the future to improve understanding of how changes in PVAT affect vascular function. In addition, opportunities for undergraduate students to participate in research experiences will be expanded. Specifically, the research project will be incorporated into a Course-based Undergraduate Research Experience (CURE), in which the entire class addresses a research question of interest to the scientific community. Components of the course will then be translated into cell and organ level modules for high school students attending a Biomed Summer Academy.The project is focused on creating an artery-on-a-chip that enables vascular contractility measurements in response to mechanical and biochemical stimuli. Vascular contractility, which is essential to blood pressure regulation, blood distribution and injury response, is mediated by vascular smooth muscle cells (vSMC) and perivascular adipose tissue(PVAT). While PVAT is known to exert an anti-contractile effect on the adjacent vasculature in healthy conditions and to lose its anti-contractile effect in diseases such hypertension, atherosclerosis, aneurysm and obesity, most of the evidence linking PVAT to vascular disease remains correlative because there are few ways to specifically manipulate PVAT without affecting the other adipose depots or the vessel itself. Current in vitro arterial systems do not incorporate PVAT and few enable vascular contractility measurements. The artery-on-a chip platform developed in this project will address this need by achieving two objectives. The FIRST Objective is to create an endothelialized tube of circumferentially aligned, contractile vSMCs. Studies are designed to explore how aligned micro- and nano-ribbons can create an aligned endothelialized vSMC syncytium in a cylindrical channel and to determine the effect of micro- and nano-ribbons composition and mechanical properties on vSMC vasoconstriction and vasodilation. The artery-on-a-chip will be validated using mouse artery pressure myography and human vasoreactivity data. The SECOND Objective is to determine how to measure 3D traction forces generated by vSMC using the unique properties of the micro and nano-ribbons. The vSMC contractile measurements will be compared to artery-on-a-chip constriction and will be validated using mouse artery pressure myography and human vasoreactivity data. This research will be the first to create a human artery-on-a-chip to test vascular contractility with future potential to incorporate PVAT.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.