Development of a Collagen-based 3D Bioprinted Microfluidic Platform for Vascular Tissue Engineering and Disease Modeling

开发基于胶原蛋白的 3D 生物打印微流体平台，用于血管组织工程和疾病建模

基本信息

批准号：
10301622
负责人：
Daniel J Shiwarski
金额：
$ 11.1万
依托单位：
CARNEGIE-MELLON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-15 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10301622
关键词：
3-Dimensional 3D Print Adrenergic Agents Adult Advisory Committees Affect American Anisotropy Arteries Binding Biochemical Biology Biomechanics Biosensor Blood Pressure Blood Vessels Calcium Signaling Caliber Cell Differentiation process Cell physiology Cells Cessation of life Collagen Collagen Type IV Complex Cues Custom Deposition Development Development Plans Disease Disease Progression Disease model Drug Screening Endothelial Cells Endothelium Engineering Extracellular Matrix Extracellular Matrix Proteins Fibronectins Fluorescence Fluorescence Microscopy Foundations G-Protein-Coupled Receptors Geometry Goals Growth Factor Health Health Care Costs Hydrogels Hypertension Image Analysis Impairment Institute of Medicine (U.S.)Investigation Knowledge Laminin Measures Mechanics Mediating Mentors Microfluidic Microchips Microfluidics Modeling Molecular Molecular and Cellular Biology Morbidity - disease rate Mus Optical Coherence Tomography Pathologic Pattern Perfusion Pharmacological Treatment Pharmacology Phase Phenotype Physiological Population Printing Production Property Proteomics Pulsatile Flow Receptor Signaling Research Resistance Resource Development Risk Factors Signal Transduction Silicones Smooth Muscle Myocytes Stimulus Structure System Technology Testing Time Tissue Engineering Tissues Training Tunica Adventitia Tunica Intima Universities Vascular Diseases Vascular Endothelial Growth Factors Vascular Smooth Muscle Work base bioprinting blood pressure reduction career career development cell dedifferentiation design fluid flow fluorescence imaging improved in vivo mechanical properties molecular pathology monolayer mortality nanomechanics new technology novel organ on a chip pressure quantitative imaging receptor response sensor three dimensional structure tissue support frame tool trafficking vascular smooth muscle cell proliferation vascular tissue engineering vasoconstriction

项目摘要

Project Summary/Abstract Hypertensive vascular disease is a leading global risk factor for morbidity and mortality, affecting over 116 million people. It is characterized by alterations to extracellular matrix (ECM) composition, biomechanical properties, and aberrant molecular signaling leading to increased blood pressure and vessel stiffening. Extensive work has focused on developing improved pharmacological treatments, tissue engineered blood vessels, and vascularizing engineered volumetric tissue, but often overlook the interdependence between cellular signaling and biomechanical forces leading to disease. Understanding the relationship between ECM biomechanics and receptor signaling, and developing tools to manipulate it, are essential for creating a healthy, mature vascular tissue while avoiding pathological changes. Here I propose that incorporation of spatially defined ECM composition and cellular alignment into a vascular-inspired 3D bioprinted tissue scaffold will produce an engineered small artery that physiologically controls vascular tone and facilitates investigation into how ECM composition and altered receptor trafficking impair vascular reactivity and promote a hypertensive phenotype. I will utilize two novel platforms: FRESH 3D bioprinting to directly fabricate perfusable vasculature from ECM proteins, and a fluorescence-based nanomechanical biosensor (NMBS) for mapping in vivo tissue strain and vascular smooth muscle contractility to Aim 1: Develop a collagen-based 3D bioprinted vascular microfluidic platform integrating controlled fluid flow and endothelialization to replicate vascular ECM biomechanics and endothelial barrier function; Aim 2: Directly pattern ECM structure and cellular organization using FRESH printing in a layer-by-layer manner to recapitulate resistance artery vascular smooth muscle cell and endothelial cell function; and Aim 3:Investigate how pathologic changes in the ECM alters receptor trafficking and impairs vascular reactivity using novel bioinks to replicate a healthy and hypertensive vessel ECM composition and material properties. This proposal seeks to enhance our knowledge of the interplay between biomechanical forces biochemical signaling during vascular development and hypertensive disease progression. The microfluidics (K99) and engineered vascular tissues (R00) created will have wide utility for drug screening and disease modeling. The career development plan, under the guidance of co-mentors Drs. Feinberg and Kleyman, and my advisory committee, will provide advanced training in microfluidics, biomechanical analysis, and vascular biology/disease modeling. The mentored phase capitalizes on an inter-disciplinary mentoring team and substantial research and professional development resources at Carnegie Mellon University, the University of Pittsburgh, and the Vascular Medicine Institute. This K99/R00, combined with my prior expertise in 3D bioprinting, advanced fluorescence microscopy, quantitative image analysis, and cellular/molecular biology, will facilitate my transition to an independent career focused on how ECM composition and structure alter receptor signaling to drive tissue maturation and disease progression.

项目摘要/摘要高血压血管疾病是发病率和死亡率的全球领先危险因素，影响了116多个百万人。它的特征是对细胞外基质（ECM）组成的改变，生物力学性质，以及异常的分子信号传导，导致血压和血管僵硬。广泛的工作重点是开发改进的药理治疗，组织工程的血管和血管化工程的容量组织，但通常忽略细胞信号之间的相互依赖性和导致疾病的生物力学力。了解ECM生物力学与受体信号传导和开发操纵它的工具对于创建健康，成熟的血管至关重要组织同时避免病理变化。在这里，我建议将空间定义的ECM合并成分和细胞对齐为血管启发的3D生物打印组织支架，将产生设计的小动脉，可以在生理上控制血管张力并促进研究ECM的研究组成和受体运输改变会损害血管反应性，并促进高血压表型。我将利用两个新型平台：新鲜的3D生物打印来直接制造ECM的蒸发脉管系统蛋白质和基于荧光的纳米力学生物传感器（NMB），用于在体内组织菌株和目标1：开发基于胶原蛋白的3D生物打印血管微流体的血管平滑肌收缩力平台整合受控的流体流和内皮化以复制血管ECM生物力学和内皮屏障功能； AIM 2：直接使用新鲜的ECM结构和蜂窝组织进行模式以逐层的方式打印，以概括抗性动脉血管平滑肌细胞和内皮细胞功能；目标3：研究ECM的病理变化如何改变受体运输并损害使用新型生物互联来复制健康且高血压的ECM组成和血管反应性材料特性。该建议旨在增强我们对生物力学之间相互作用的了解在血管发育和高血压疾病进展过程中的生化信号传导。这产生的微流体（K99）和工程性血管组织（R00）将具有广泛的药物筛查和疾病建模。职业发展计划，在联合委托人的指导下。 Feinberg和Kleyman，我的咨询委员会将提供微流体学，生物力学分析和血管的高级培训生物学/疾病建模。受过指导的阶段大利用跨学科的指导团队，卡内基梅隆大学的大量研究和专业发展资源，大学匹兹堡和血管医学研究所。这个K99/R00，加上我先前在3D方面的专业知识生物打印，晚期荧光显微镜，定量图像分析和细胞/分子生物学，将会促进我向独立职业的过渡，专注于ECM组成和结构如何改变受体信号传导以驱动组织成熟和疾病进展。