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：
10468156
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 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10468156
关键词：
3-Dimensional 3D Print Adrenergic Agents Adult Advisory Committees Affect American Anisotropy Arteries Binding Biochemical Biology Biomechanics Biosensor 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 Persons 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 bioink biomechanical test bioprinting blood pressure elevation blood pressure reduction career career development cell dedifferentiation design fluid flow fluorescence imaging hypertensive 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.

项目概要/摘要高血压血管疾病是全球发病率和死亡率的主要危险因素，影响超过 116 万人。其特点是细胞外基质（ECM）成分、生物力学的改变特性和异常的分子信号传导导致血压升高和血管硬化。广泛的工作重点是开发改进的药物治疗、组织工程血管和使工程体积组织血管化，但常常忽视细胞信号传导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以及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。在这里我建议结合空间定义的 ECM 将成分和细胞排列成受血管启发的 3D 生物打印组织支架将产生设计了小动脉，可以在生理上控制血管张力，并有助于研究 ECM 如何成分和受体运输的改变会损害血管反应性并促进高血压表型。我将利用两个新颖的平台： FRESH 3D 生物打印直接从 ECM 制造可灌注脉管系统蛋白质，以及基于荧光的纳米机械生物传感器（NMBS），用于绘制体内组织应变和血管平滑肌收缩力目标 1：开发基于胶原蛋白的 3D 生物打印血管微流体集成受控流体流动和内皮化的平台，以复制血管 ECM 生物力学和内皮屏障功能；目标 2：使用 FRESH 直接构建 ECM 结构和细胞组织模式逐层打印再现阻力动脉血管平滑肌细胞和内皮细胞细胞功能；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墨水复制健康和高血压血管 ECM 成分的血管反应性材料特性。该提案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强制生化信号传导。这创建的微流体（K99）和工程血管组织（R00）将在药物筛选和疾病建模。在共同导师Drs.的指导下制定职业发展计划。范伯格和克莱曼，和我的顾问委员会将提供微流体、生物力学分析和血管方面的高级培训生物学/疾病建模。指导阶段利用跨学科的指导团队和卡内基梅隆大学、卡内基梅隆大学等拥有丰富的研究和专业发展资源匹兹堡和血管医学研究所。这款 K99/R00 结合了我之前在 3D 方面的专业知识生物打印、先进的荧光显微镜、定量图像分析和细胞/分子生物学，将帮助我过渡到独立职业，专注于 ECM 成分和结构如何改变受体驱动组织成熟和疾病进展的信号传导。

项目成果

期刊论文数量（8）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

3D Bioprinted Patient-Specific Extracellular Matrix Scaffolds for Soft Tissue Defects.

DOI：
10.1002/adhm.202200866
发表时间：
2022-12
期刊：
ADVANCED HEALTHCARE MATERIALS
影响因子：
10
作者：
Behre, Anne;Tashman, Joshua W.;Dikyol, Caner;Shiwarski, Daniel J.;Crum, Raphael J.;Johnson, Scott A.;Kommeri, Remya;Hussey, George S.;Badylak, Stephen F.;Feinberg, Adam W.
通讯作者：
Feinberg, Adam W.

Development of a high-performance open-source 3D bioprinter.

DOI：
10.1038/s41598-022-26809-4
发表时间：
2022-12-31
期刊：
Scientific reports
影响因子：
4.6
作者：
通讯作者：

Rapid model-guided design of organ-scale synthetic vasculature for biomanufacturing.

用于生物制造的器官级合成脉管系统的快速模型引导设计。

DOI：
发表时间：
2023
期刊：
ArXiv
影响因子：
0
作者：
Sexton,ZacharyA;Hudson,AndrewR;Herrmann,JessicaE;Shiwarski,DanJ;Pham,Jonathan;Szafron,JasonM;Wu,SeanM;Skylar-Scott,Mark;Feinberg,AdamW;Marsden,Alison
通讯作者：
Marsden,Alison

FRESH 3D Bioprinted Collagen-based Resistance Vessels and Multiscale Vascular Microfluidics.

FRESH 3D 生物打印的基于胶原蛋白的阻力血管和多尺度血管微流体。

DOI：
发表时间：
2022
期刊：
FASEB journal : official publication of the Federation of American Societies for Experimental Biology
影响因子：
0
作者：
Shiwarski,DanielJ;Hudson,Andrew;Tashman,Joshua;Straub,Adam;Feinberg,Adam
通讯作者：
Feinberg,Adam

Embedded 3D Printing of Thermally-Cured Thermoset Elastomers and the Interdependence of Rheology and Machine Pathing.

热固化热固性弹性体的嵌入式 3D 打印以及流变学和机器路径的相互依赖性。

DOI：
10.1002/admt.202200984
发表时间：
2023
期刊：
Advanced materials technologies
影响因子：
6.8
作者：
Stang,Maria;Tashman,Joshua;Shiwarski,Daniel;Yang,Humphrey;Yao,Lining;Feinberg,Adam
通讯作者：
Feinberg,Adam