A novel physiologically realistic microfluidic in-vitro blood-brain barrier model

一种新颖的生理真实微流控体外血脑屏障模型

基本信息

批准号：
8469865
负责人：
BALABHASKAR PRABHAKARPANDIAN
金额：
$ 61.9万
依托单位：
CFD RESEARCH CORPORATION
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-04-01 至 2015-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8469865
关键词：
Animal Model Apical Astrocytes Basic Science Biochemical Biological Biological Assay Biology Blood - brain barrier anatomy Blood flow Brain Cell Line Cells Characteristics Chemicals Coculture Techniques Collaborations Conditioned Culture Media Connexins Data Devices Drug Delivery Systems Drug usage Electrical Resistance Electrodes Endothelial Cells Engineering Evaluation Failure Generations Imagery Immigration In Vitro Inflammatory Internet Journals Knowledge Leukocytes Libraries Measurement Measures Microcirculation Microfluidic Microchips Microfluidics Modeling Monitor Neuraxis Neuroglia Neurologic Neurons Optics Pediatric Hospitals Penetration Permeability Pharmaceutical Preparations Pharmacologic Substance Phase Physiological Plastics Process Publications Rattus Reagent Research Research Personnel Saint Jude Children&apos s Research Hospital Scientist Side Simulate Staging Techniques Technology Testing Therapeutic Effect Therapeutic Human Experimentation Tight Junctions Time Toxic effect Universities Up-Regulation Visual Work base cost data modeling design drug candidate drug development drug discovery electrical measurement immortalized cell improved in vitro Model in vivo instrumentation interest lithography meetings migration novel novel strategies particle protein expression receptor expression screening shear stress symposium therapeutic development

项目摘要

DESCRIPTION (provided by applicant): The overall objective of this study is to develop a novel in vitro microfluidic platform to test a drug or delivery vehicle's ability to permeate the Blood-Brain Barrier (BBB). In contrast to current in-vitro models, our proposed device, SIM-BBB, comprises of a microfluidic two-compartment chamber. The chamber is designed in such a way as to permit visualization-friendly evaluation of transport/permeation under appropriate microcirculatory size and flow conditions, while simultaneously simplifying device fabrication. The apical side is seeded with endothelial cells and the basolateral side supports glial cell co-cultures. The increased physiological realism substantially improves BBB characteristics including formation of tight junctions and expression of relevant transporters. The new platform offers greater throughput, increased library coverage, lower cost, rapid turnaround times and increased mechanistic knowledge benefiting drug discovery efforts. In Phase I, the first generation microfluidic SIM-BBB device was designed and fabricated using soft lithography. Brain endothelial cells were cultured in the microfluidic constructs with a perfusate of astrocyte conditioned media. Biochemical analysis showed upregulation of tight junction molecules while optical analysis showed intactness of the BBB in the microfluidic device. Finally, transporters assay was successfully demonstrated in the device. Phase II efforts will focus on optimization of the microfluidic device for enhanced physiological fidelity. Electrodes will be integrated for non-visual monitoring of the endothelial cell layers and tight junction formation via trans-endothelial electrical resistance (TEER) measurements. Finally, the developed technology will be demonstrated for diverse applications including drug penetration studies and leukocyte migration under inflammatory conditions. A multi-disciplinary partnership with expertise in engineering and biology has been assembled for successful completion of the project.

描述（由申请人提供）：这项研究的总体目的是开发一种新型的体外微流体平台，以测试药物或递送工具渗透血脑屏障（BBB）的能力。与当前的体外模型相反，我们提出的设备SIM-BBB包括一个微流体两室室。该腔室的设计是允许在适当的微循环尺寸和流动条件下对传输/渗透的可视化评估，同时简化设备的制造。顶端用内皮细胞接种，基底外侧侧支持神经胶质细胞共培养。生理现实主义的增加大大改善了BBB的特征，包括形成紧密的连接和相关转运蛋白的表达。新平台提供了更大的吞吐量，增加的图书馆覆盖范围，较低的成本，快速的周转时间以及增加的机械知识，从而使药物发现工作受益。在第一阶段，使用软光刻设计和制造了第一代微流体SIM-BBB设备。将脑内皮细胞培养在具有星形胶质细胞调节培养基的微流体构建体中。生化分析表明，紧密连接分子的上调，而光学分析显示微流体装置中BBB的完整性。最后，在设备中成功证明了转运蛋白测定法。第二阶段的工作将集中于优化微流体设备，以增强生理保真度。电极将集成，以通过跨内皮电阻（TEER）测量值进行非视觉监测内皮细胞层和紧密的连接形成。最后，将在炎症条件下进行包括药物渗透研究和白细胞迁移在内的各种应用，包括开发的技术。与工程和生物学专业知识的多学科合作伙伴关系已被组装，以成功完成该项目。