Disc-on-a-chip: microfluidic nutrition and biomechanical loading integrated mouse disc culture system

项目摘要 背痛是全世界最常见的残疾原因，终生患病率为80%。 腰间盘(IVD)退变是腰痛的主要原因。目前，对IVD退行性疾病的治疗 只是症状上的。我们有效治疗椎间盘疾病的能力在很大程度上受到1)不完整的 对IVD发生、功能和疾病的生物学过程的了解；2)缺乏有效的 由于研究平台有限，简化了新药发现的协议。 为了解决上述问题，我们建议开发一种新型的微流控芯片上的光盘 为小鼠IVD量身定制的器官培养平台。我们面向应用的微流控设备旨在 解决关键致病因素，包括生化因素(如营养素和炎性细胞因子)、机械因素 和遗传因素，导致椎间盘退行性疾病。我们的短期目标是提出证据- Of-Concept微流控设备，专为小鼠IVD多通量培养(目标1)和生理培养量身定制 模拟(目标2)方式，并演示各自面向应用的体外IVD桥接策略 培养方法包括体内IVD生物学/病理学和模拟药物评价。我们的长期目标是 为新的治疗和再生策略建立简化的药物发现协议，使用我们的 集成微流控小鼠IVD培养平台。 微流控器件已被证明在以下方面比传统的同类器件更具优势 许多方面，因为微型化、自动化和集成的流体控制系统允许更小的试剂 消耗、更低的成本、更短的周转时间和更高的吞吐量。我们提出的微流控小鼠IVD 培养装置不仅填补了芯片上光盘的方法学空白，而且还展示了一些 在解决光盘退化的道路上的革命性好处：a)文化媒体不断 提供/更新以模拟体内血液循环，提供一个独特的平台来研究各种 氧、营养素、细胞因子和生长因子等生化因素对IVD代谢的影响；b)与多个 吞吐量平台，我们可以轻松培养数十个IVD样本，有利于加快药物发现和 精确定义的微环境中的评估方案(目标1)；c)我们的生理模拟 小鼠IVD上的机械负荷使原本不可能实现的遗传因素之间的相互作用研究成为可能 和机械应力(目标2)；以及d)可以定制我们的微尺度的“片上光盘”装置， 商业化，大规模生产，并被所有研究实验室使用，将 传统光盘研究的范例，同时在战斗中加速药物发现过程 防止椎间盘退变。

Project Summary Back pain is the most common cause of disability worldwide with a lifetime prevalence of 80%. Intervertebral disc (IVD) degeneration is a major cause of low back pain. Currently, IVD degeneration is treated only symptomatically. Our ability to effectively treat disc disease is largely hampered by 1) an incomplete understanding of the biological processes of IVD development, function and disease; 2) The lack of an efficient and streamlined protocol for new drug discovery due to limited research platforms. To address the aforementioned problems, we propose to develop a novel microfluidic “disc-on-a-chip” organ culture platform tailored for mouse IVD. Our application-oriented microfluidic devices are designed to address key pathogenic factors, including biochemical (e.g. nutrients and inflammatory cytokines), mechanical and genetic aspects, contributing to disc degenerative diseases. Our short-term objective is to develop proof- of-concept microfluidic devices tailored for mouse IVD culture in multi-throughput (Aim 1) and physiologically simulated (Aim 2) manners and demonstrate respective application-oriented strategies for bridging in vitro IVD culture methodology with in vivo IVD biology/pathology and simulated drug evaluation. Our long-term goal is to establish a streamlined drug discovery protocol for new therapeutic and regenerative strategies using our integrated microfluidic mouse IVD culture platform. Microfluidic devices have been proven to be advantageous over their conventional counterparts in many aspects, since miniaturization, automation, and integration of fluidic control systems allow smaller reagent consumption, lower cost, shorter turnaround time, and higher throughput. Our proposed microfluidic mouse IVD culture devices will not only fill in the methodological blank of “disc-on-a-chip”, but also demonstrate a number of revolutionary benefits along the road to tackle disc degeneration: a) Culture media is continuously supplied/refreshed to mimick in vivo blood circulation, providing a unique platform to study the impacts of various biochemical factors such as oxygen, nutrients, cytokines, and growth factors on IVD metabolism; b) With multi- throughput platform, we can easily culture dozens of IVD samples, benefiting expedited drug discovery and evaluation protocols within a precisely defined microenvironment (Aim 1); c) Our physiologically simulated mechanical loading on mouse IVD empowers otherwise impossible cross-talking studies between genetic factor and mechanical stress (Aim 2); and d) Our micro-scale “disc-on-a-chip” device can be customized, commercialized, manufactured in mass production, and utilized by all research laboratories, shifting the paradigm of conventional disc research and meanwhile accelerating drug discovery processes in the battle against disc degeneration.