A microphysiological system of tendon inflammation and fibrosis for drug screening and efficacy testing: MPS Database Engagement

摘要 微生理系统(MPS)作为临床前研究工具的持续发展对 克服人类疾病动物模型中固有的低吞吐量和不准确性。它的局限性 临床前动物模型，最常见的是小鼠，在炎症性疾病中尤其明显，这些疾病是 已知对炎症有不同的遗传和细胞因子反应。建立MPS替代方案， 然而，需要就最适合于解决特定问题的协议和系统达成科学共识 疾病。由于目前的MPS时代的特点是方法的扩散，微生理学 匹兹堡大学创建的系统数据库(MPS-db)是一个有价值的工具，可以加速 制定MPS标准。因为mps-db的成功需要mps的积极参与。 开发人员和用户，我们寻求补充资金，以贡献设计、协议和结果 MPS系统，模拟肌腱愈合过程中炎症和纤维化之间的相互作用(UG3TR00287)。 重要的是，结缔组织损伤的损伤和修复在目前的mps-db中没有描述，但是 每年超过850万次临床手术，其中包括200万次大手术。我们的人类 芯片上肌腱(HToC)模型关注肌腱修复的早期炎症阶段，其中及时 干预措施可能会促进无疤痕愈合。HToC的特点是血管和胶原蛋白隔间 在建立的新生血管微环境模拟中的交换可溶性和细胞因子 在血液凝结后不久。单核细胞的渗透被认为在肿瘤的发生中起着重要作用。 收缩肌成纤维细胞进入衰老并释放单核细胞激活因子 会导致疤痕组织的反馈环。该模型使用来自原代人类肌腱细胞的IPSCs来创造 血管内皮细胞和单核细胞在以患者为中心的同源三种培养中。有了补充资金，我们 将分享：1)肌腱损伤和纤维炎症修复过程的机制描述；2) HToC的设计细节包括用于流动和集成光子的器件组件和模块 基于传感；3)细胞培养和设备协议，包括表型特征和操作 参数，如内皮细胞启动的流速和免疫细胞的引入；4)设计和 在基线和炎症/修复条件下进行研究的基本原理；以及5)结果，包括对 研究内重复性。

Abstract The continued advancement of microphysiological systems (MPS) as pre-clinical research tools is vital to overcome the low throughput and inaccuracies inherent in animal models of human disease. The limitations of pre-clinical animal models, most commonly mice, are particularly apparent in inflammatory diseases which are known to have distinct genetic and cytokine responses to inflammation. The establishment of MPS alternatives, however, will require scientific consensus on the protocols and systems best suited to address particular diseases. As the current MPS era is characterized by a proliferation of approaches, the Microphysiological System Data Base (MPS-db) created by the University of Pittsburgh is a valuable tool to hasten the development of MPS standards. Because the success of the MPS-db requires the active participation by MPS developers and users, we seek supplemental funding to contribute the designs, protocols and results for an MPS system that models the interplay between inflammation and fibrosis in tendon healing (UG3TR00287). Importantly, the injury and repair of connective tissue injury is not represented in the current MPS-db but accounts for more than 8.5 million clinical procedures annually, including 2 million major surgeries. Our human tendon-on-a-chip (hToC) model focuses on the early inflammatory stages of tendon repair, where timely interventions may promote scarless healing. The hToC features vascular and collagen compartments which exchange soluble and cellular factors in a simulation of the neovascularized microenvironment established shortly after blood clotting. Monocyte infiltration is hypothesized to play an essential role in the generation of contractile myofibroblasts which progress to senescence and release monocyte activating factors in a positive feedback loop that causes scar tissue. The model uses iPSCs derived from primary human tenocytes to create vascular endothelial cells and monocytes in an isogenic, patient-centric triculture. With supplemental funding we will share: 1) descriptions of the mechanisms of the tendon injury and fibroinflammatory repair process; 2) design details for the hToC including device components and modules for both flow and integrated photonic- based sensing; 3) cell culture and device protocols including phenotypic characteristics and operational parameters such as flow rates for priming of ECs and the introduction of immune cells; 4) The design and rationale for studies under baseline and inflammation/repair conditions; and 5) Results including an analysis of intra-study reproducibility.