Developing a Novel Vascularized Bone Microdevice for Investigating the Post-Stroke Bone Microenvironment

开发新型血管化骨微装置用于研究中风后骨微环境

• 批准号: 10664856
• 负责人: Sandra Jeanne Stangeland-Molo
• 金额: $ 3.88万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10664856
• 关键词: 3-Dimensional; Acceleration; Address; Adult; Affect; Aging; Alkaline Phosphatase; Apoptosis; Arthritis; Automobile Driving; Bed rest; Behavior; Biological Assay; Biomimetics; Blood Vessels; Bone Matrix; Bone remodeling; Cardiovascular Diseases; Cell Communication; Cell Culture Techniques; Cell Maintenance; Cells; Cellular Morphology; Cessation of life; Chitosan; Chronic; Clinical Research; Collagen; Complex; Computer Models; Deposition; Devices; Dextrans; Electron Microscopy; Endothelial Cells; Endothelium; Environment; Enzyme-Linked Immunosorbent Assay; Event; Exposure to; Extracellular Matrix; Fracture; Gait; Gene Expression; Genetic Diseases; Geometry; Goals; Height; Hematopoietic; Human; Impairment; In Vitro; Individual; Inflammatory; Inflammatory Response; Interferon Type II; Interferons; Interleukin-1 beta; Interleukin-6; Interleukins; Investigation; Ischemia; Ischemic Stroke; Label; Liquid substance; Maintenance; Mechanics; Minerals; Modeling; Mus; Nitric Oxide; Nutrient; Osteoblasts; Paracrine Communication; Pathology; Patients; Permeability; Persons; Pharmacologic Substance; Physiologic calcification; Porosity; Production; Property; Protein Secretion; Public Health; Qualifying; Radiation; Regulation; Research; Reverse Transcription; Sensorimotor functions; Serum; Signal Transduction; Spectroscopy, Fourier Transform Infrared; Stroke; TdT-Mediated dUTP Nick End Labeling Assay; Techniques; Technology; Tight Junctions; Tissues; Training; United States; Vascularization; Work; X ray spectroscopy; biomaterial compatibility; bone; bone cell; bone fragility; bone health; bone loss; bone mass; bone scaffold; cell behavior; clinical decision-making; comorbidity; crosslink; cytokine; cytotoxicity; density; disability; experience; extracellular; fabrication; fluorescence imaging; fracture risk; improved; in vivo; inflammatory milieu; insight; manufacture; mechanical load; mechanical properties; microdevice; mineralization; mortality; nanoindentation; novel; organ on a chip; osteoclastogenesis; paracrine; post stroke; programs; response; scaffold; stroke patient; stroke survivor; treatment strategy

PROJECT SUMMARY Ischemic stroke is a serious condition affecting nearly 800,000 people in the United States annually and is a leading cause of long-term disability. Although stroke survivors commonly experience accelerated bone loss and higher fracture risk compared to typically aging adults, the underlying causes remain poorly understood and cannot be explained solely by bedrest. Inflammatory cytokines are present in serum post-stroke, but whether cytokine dysregulation is a driving factor for altered bone remodeling following stroke – as seen in inflammatory- related bone loss in other conditions like arthritis – is unknown. Our primary hypothesis is that the inflammatory environment seen post-stroke stimulates a pro-inflammatory response in bone vasculature that acts to suppress osteoblast activity and drive an increase in secreted proteins to activate bone resorptive programs. Microdevices and “organ-on-chip” constructs are effective 3D in vitro platforms for mechanistically probing different cell-cell interactions in controlled microenvironments. Such microdevices have been used successfully to examine the response of different niches within bone to pharmaceuticals, radiation, and genetic disorders, but they have not yet been used to examine mineralized bone and vascular interactions. The overall goal of this project is to expand understanding of underlying factors contributing to stroke-related bone loss, specifically inflammatory factors, by developing and implementing a bone-vascular microdevice platform that mimics the mineralized bone microenvironment. Aim 1 will determine optimal manufacturing conditions for producing a mineralized extracellular matrix (ECM) scaffold for osteoblast support in the microdevice. Aim 2 will develop the novel bone- vascular microdevice platform and investigate osteoblast-endothelial cell interactions under homeostatic conditions. Aim 3 will determine the effects of inflammatory cytokines interleukin-6, interleukin-1β, and interferon- γ on osteoblast-endothelial paracrine signaling using the microdevice platform. We will accomplish these aims by leveraging traditional ECM scaffold fabrication techniques with passive mineral deposition, computational fluid dynamics modeling of fluidic shear and nutrient transport, and biological assays of osteoblast activity and vascular barrier function. This work will create a new in vitro platform that enables mechanistic probing of complex bone-vascular interactions and advance understanding of inflammatory regulation of bone loss post- stroke, thereby providing a framework that may inform better treatment strategies to mitigate stroke-related bone fragility.

项目总结 缺血性中风是一种严重的疾病，每年在美国影响近80万人，是一种 长期残疾的主要原因。尽管中风幸存者通常会经历加速的骨丢失和 与典型的老年人相比，骨折风险更高，其根本原因仍然鲜为人知，而且 不能仅仅用卧床休息来解释。中风后血清中存在炎性细胞因子，但是否 细胞因子失调是中风后骨重塑改变的驱动因素-如炎症- 其他情况下的相关骨质丢失--如关节炎--尚不清楚。我们的主要假设是炎症性疾病 中风后环境刺激骨血管中的促炎反应，从而抑制 成骨细胞活性，并推动分泌蛋白的增加，以激活骨吸收计划。微型设备 芯片上的器官结构是机械探测不同细胞-细胞的有效3D体外平台 受控微环境中的相互作用。这种微型设备已被成功地用于检测 骨中不同的生态位对药物、辐射和遗传疾病的反应，但它们没有 还被用来研究矿化骨和血管的相互作用。这个项目的总体目标是扩大 通过以下方式了解导致中风相关骨丢失的潜在因素，特别是炎症因素 模拟矿化骨的骨血管微装置平台的开发与实现 微环境。目标1将确定生产矿化矿的最佳制造条件 微设备中用于成骨细胞支持的细胞外基质(ECM)支架。目标2将开发新的骨骼- 血管微装置平台和研究动态平衡条件下成骨细胞-内皮细胞的相互作用 条件。目的3将确定炎症细胞因子白介素6、白介素1β和干扰素的作用。 利用微设备平台对成骨细胞-内皮旁分泌信号进行γ。我们将实现这些目标 通过利用传统的ECM支架制造技术和被动矿物沉积，计算流体 流体剪切和营养物质运输的动力学模拟，以及成骨细胞活性和营养物质的生物分析 血管屏障功能。这项工作将创造一种新的体外平台，使机械探测成为可能 复杂的骨-血管相互作用与骨丢失后炎症调节的研究进展 中风，从而提供了一个框架，可以为更好的治疗策略提供信息，以减轻中风相关的骨骼 脆弱。