Multi-Scale In Vitro 3D Tissue Model of Vascularized Bone-Cartilage Interactions

摘要 目前的血管化骨组织体外模型不能模拟体内微环境 由通过基质屏障相互通信的不同类型的细胞组成。在……里面 此外，由于缺乏血管系统的实时可视化和量化，它们受到阻碍。 骨和骨-软骨的相互作用。相比之下，动物模型虽然提供了有用的 信息既耗时又昂贵，近年来越来越多地提高了道德标准 担忧。此外，动物研究对机械性行为的理解有限。 与控制良好的体外研究相比。因此，对体外平台的需求尚未得到满足 用于改进血管化骨-软骨相互作用的监测和分析。 在第一阶段，我们成功地开发和演示了一个多尺度的体外模型，包括 微型微流控装置和中观生物反应器来模拟体内的条件。 我们成功地在平台患者体内分化出了人骨髓间充质干细胞 (HMSCs)向成骨和成软骨方向发展，突出与血管的相互作用 内皮细胞。在详细的功能描述之后，我们演示了该功能 以评估消炎疗法的功能。在第二阶段，我们将 测试其他模拟天然骨软骨的促炎成分 微环境。我们还将使用我们的多尺度系统来(A)机械地理解 以及(B)对炎性侮辱后的候选治疗方法进行治疗性筛选。最后，我们 将开发基础设施，通过多路复用平台来提高吞吐量 以实现自动化。 多学科产业-学术合作伙伴关系，具有基于细胞的微流控技术专业知识 化验和肌肉骨骼生物学和组织再生已经组装成功 完成这一项目。通过提供准确、定量和可预测的模型 生理互动，开发的多尺度平台有望建立新的 治疗药物生理反应的体外评估范例。

Abstract Current in vitro models of vascularized bone tissues do not mimic the in vivo microenvironment comprising of diverse cell types in communication with each other through stromal barriers. In addition, they are hampered by lack of real-time visualization and quantitation of vasculature- bone as well as bone-cartilage interactions. In contrast, animal models while providing useful information are time consuming, expensive and in recent years, have increasingly raised ethical concerns. Furthermore, animal studies provide limited understanding of mechanistic behavior compared to well-controlled in vitro studies. Thus, there is an unmet need for an in vitro platform for improved monitoring and analysis of vascularized bone-cartilage interactions. In Phase I we successfully developed and demonstrated a multi-scale in vitro model comprising of a micro scale microfluidic device and a meso scale bioreactor to mimic the in vivo conditions. We successfully differentiated in the platform patient derived human mesenchymal stem cells (hMSCs) towards osteogenic and chondrogenic lineages highlighting interactions with vascular endothelial cells. Following detailed functional characterizations, we demonstrated the capability of the platform to evaluate functionality for an anti-inflammatory therapeutic. In Phase II we will test additional pro-inflammatory components that mimic the native osteochondral microenvironment. We will also use our multi-scale system for (a) mechanistic understanding and (b) therapeutic screening of candidate treatments following inflammatory insults. Finally, we will develop the infrastructure to increase the throughput capability by multiplexing the platform for automation. A multi-disciplinary industry-academic partnership with expertise in microfluidics cell-based assays and musculoskeletal biology and tissue regeneration has been assembled for successful completion of this project. By providing an accurate, quantitative and predictive model of physiological interactions, the developed multi-scale platform promises to establish a new paradigm for in vitro assessment of the physiological response to therapeutics.