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于 克服了人类疾病动物模型中固有的低通量和不准确性。的局限性 临床前动物模型，最常见的是小鼠，在炎性疾病中特别明显， 已知对炎症有不同的遗传和细胞因子反应。制定MPS替代方案， 然而，需要就最适合于解决特定问题的议定书和系统达成科学共识， 疾病由于当前MPS时代的特征是方法的扩散，微生理学 由匹兹堡大学创建的系统数据库（MPS-db）是一个有价值的工具， 制定MPS标准。由于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，以促进设计，协议和结果， 对肌腱愈合中炎症和纤维化之间的相互作用进行建模的MPS系统（UG 3 TR 00287）。 重要的是，结缔组织损伤的损伤和修复在目前的MPS-db中没有表现， 每年有超过850万例临床手术，包括200万例大手术。我们人类 tendon-on-a-chip（hToC）模型关注肌腱修复的早期炎症阶段， 干预可以促进无疤痕愈合。hToC具有血管和胶原蛋白隔室， 在模拟建立的新生血管化微环境中交换可溶性和细胞因子， 血液凝固后不久。单核细胞浸润被假设为在肿瘤的产生中起重要作用。 收缩性肌成纤维细胞逐渐衰老并释放单核细胞活化因子， 导致疤痕组织的反馈回路。该模型使用来源于原代人类肌腱细胞的iPSC来创建 血管内皮细胞和单核细胞在一个同基因的，以病人为中心的triculture。通过补充资金，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，包括用于流和集成光子的器件组件和模块， 3）细胞培养和装置方案，包括表型特征和操作特性; 参数，例如用于引发EC和引入免疫细胞的流速; 4）设计和 基线和炎症/修复条件下研究的基本原理;和5）结果，包括 研究内重现性。

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.