Tissue Chip Modeling of Synovial Joint Pathologies: Effects of Inflammation and Adipose-Mediated Diabetic Complications

创伤、炎症、感染和衰老会对关节组织造成损害，最终导致关节炎。 疾病，如骨关节炎(OA)、感染性关节炎和炎症性关节炎，导致肢体残疾 这影响了生活质量；然而，目前还没有有效的治疗方法。在这方面的有限进展 疾病修饰药物的发展主要是因为：(1)机制不足 了解疾病的发生/发展；(2)无法涵盖三维(3D)和多组织 滑膜关节的性质在体外药物发现的早期阶段；以及(3)临床前动物的应用有限 早期临床疗效和毒性预测研究(缺乏“早期失败/快速失败”能力)，导致 出乎意料且代价高昂的临床试验失败。此外，患者特定的病因、进展和药物敏感性 个人资料强调了个性化治疗开发的必要性。为了满足这些需求，我们建议 设计一种3D人体微关节芯片(MJoint)，在生理上类似于天然关节，并能够 为DMM筛查/发展建立关节疾病发病机制模型。UG3-目标1：工程连接 骨软骨复合体、滑膜和脂肪的成分将使用原代细胞进行工程设计， 人骨髓间充质干细胞(MSCs)或诱导多能干细胞(IPSC)来源的MSCs被包裹在 光交联水凝胶支架，包括巨噬细胞，以评估其关键功能 调节/调节炎症，并用分子、生化和组织学表征表型 分析。目标2：产生正常和患病的小关节一个生物反应器将被设计成容纳所有 关节元素(MJoint)，模拟各自在体内的组织条件，并暴露于各种致病物质 模拟骨性关节炎、炎症性关节炎和脂肪介导的糖尿病关节并发症的试剂和条件， 将根据每个关节组件的组织学和结构变化以及 生物标志物。UH3-Aim 3：研究组织相互作用并开发特定的生物标记物 我们将评估正常情况下关节组织成分的贡献和相互作用 以及病态的情况。不同病因的关节疾病(见目标2)将被模拟，组织相互作用 分析，并开发了预测关节健康的潜在生物标记物。目标4：测试已知药物和 筛选候选DMM我们将使用mJoint评估已知和候选DMM的有效性 疾病模型，包括白细胞介素4、核因子-κB诱骗寡核苷酸、他汀类药物、金属蛋白酶抑制剂和 还侧重于目标3中确定的生物标记物的适用性。目标5：测试细胞潜力-- 基于治疗的人骨髓间充质干细胞及其产物，如外切体和条件化的疗效 将对媒体和其他生物制品进行检查，以探索被广泛感知的效用的科学基础 以干细胞为基础的肌肉骨骼疾病治疗。综上所述，mJoint在体外代表了一种高实用性 建立滑膜关节病理模型和筛选治疗关节疾病的治疗方法的平台。

Trauma, inflammation, infection, and aging can cause damages to joint tissues, ultimately leading to arthritic disorders, such as osteoarthritis (OA), septic arthritis, and inflammatory arthritis, resulting in physical disabilities that compromise quality of life; however, no efficacious therapies are currently available. The limited progress in the development of disease-modifying medications (DMMs) is principally because of: (1) insufficient mechanistic understanding of disease onset/progression; (2) inability to encompass the 3-dimensional (3D) and multi-tissue nature of the synovial joint in early phase in vitro drug discovery; and (3) limited utility of pre-clinical animal studies for early stage clinical efficacy and toxicity prediction (lacking “fail early/fail fast” capabilities), resulting in unanticipated and costly clinical trial failures. Also, patient-specific etiology, progression, and drug sensitivity profiles underscore the need for personalizable therapy development. To address these needs, we propose engineering a 3D human micro-joint chip (mJoint), physiologically analogous to the native joint and capable of modeling pathogenesis of joint diseases for DMM screening/development. UG3 - Aim 1: Engineering joint components The osteochondral complex, synovium, and adipose, will be engineered using primary cells, human mesenchymal stem cells (MSCs) or induced pluripotent stem cell (iPSC) derived MSCs encapsulated in a photocrosslinked hydrogel scaffold, with macrophages included to evaluate their critical function in mediating/regulating inflammation, and phenotype-characterized using molecular, biochemical and histological analyses. Aim 2: Generating normal and diseased mJoint A bioreactor will be designed to house all of the joint elements (mJoint), simulating the respective in vivo tissue conditions, and exposed to various pathogenic agents and conditions to model OA, inflammatory arthritis, and adipose-mediated diabetic joint complications, which will be assessed based on changes in histology and structure in each individual joint component as well as biomarkers. UH3 - Aim 3: Investigating tissue interactions and developing specific biomarkers using mJoint We will assess the contribution of and the interactions among the joint tissue components under normal and diseased conditions. Joint diseases with different etiologies (see Aim 2) will be simulated, tissue interactions analyzed, and potential biomarkers developed to predict joint health. Aim 4: Testing known drugs and screening candidate DMMs We will assess the efficacy of known and candidate DMMs using the mJoint disease models, including interleukin-4, NF-κB decoy oligonucleotides, statins, metalloproteinases inhibitors, and others, focusing also on the applicability of biomarkers identified in Aim 3. Aim 5: Testing potential of cell- based therapy The therapeutic efficacy of human MSCs and their products, such as exosomes and conditioned media, and other biologics, will be examined, in order to explore the scientific basis of the widely perceived utility of stem cell-based therapy for musculoskeletal disorders. In summary, the mJoint represents a high-utility in vitro platform to model synovial joint pathologies and to screen therapeutics for the treatment of joint diseases.