Lin Zixuan, Li Zhong, Li Eileen N, Li Xinyu, Del Duke Colin J, Shen He, Hao Tingjun, O'Donnell Benjamen, Bunnell Bruce A, Goodman Stuart B, Alexander Peter G, Tuan Rocky S, Lin Hang
Department of Orthopaedic Surgery, Center for Cellular and Molecular Engineering, University of Pittsburgh School of Medicine, Pittsburgh, PA, United States.
Xiangya School of Medicine, Central South University, Changsha, China.
Front Bioeng Biotechnol. 2019 Dec 17;7:411. doi: 10.3389/fbioe.2019.00411. eCollection 2019.
Osteoarthritis (OA) is a chronic disease mainly characterized by degenerative changes in cartilage, but other joint elements such as bone are also affected. To date, there are no disease-modifying OA drugs (DMOADs), owing in part to a deficiency of current models in simulating OA pathologies and etiologies in humans. In this study, we aimed to develop microphysiological osteochondral (OC) tissue chips derived from human induced pluripotent stem cells (iPSCs) to model the pathologies of OA. We first induced iPSCs into mesenchymal progenitor cells (iMPCs) and optimized the chondro- and osteo-inductive conditions for iMPCs. Then iMPCs were encapsulated into photocrosslinked gelatin scaffolds and cultured within a dual-flow bioreactor, in which the top stream was chondrogenic medium and the bottom stream was osteogenic medium. After 28 days of differentiation, OC tissue chips were successfully generated and phenotypes were confirmed by real time RT-PCR and histology. To create an OA model, interleukin-1β (IL-1β) was used to challenge the cartilage component for 7 days. While under control conditions, the bone tissue promoted chondrogenesis and suppressed chondrocyte terminal differentiation of the overlying chondral tissue. Under conditions modeling OA, the bone tissue accelerated the degradation of chondral tissue which is likely via the production of catabolic and inflammatory cytokines. These findings suggest active functional crosstalk between the bone and cartilage tissue components in the OC tissue chip under both normal and pathologic conditions. Finally, a selective COX-2 inhibitor commonly prescribed drug for OA, Celecoxib, was shown to downregulate the expression of catabolic and proinflammatory cytokines in the OA model, demonstrating the utility of the OC tissue chip model for drug screening. In summary, the iPSC-derived OC tissue chip developed in this study represents a high-throughput platform applicable for modeling OA and for the screening and testing of candidate DMOADs.
骨关节炎(OA)是一种主要以软骨退行性变化为特征的慢性疾病,但其他关节成分如骨骼也会受到影响。迄今为止,尚无改善病情的骨关节炎药物(DMOADs),部分原因是当前模型在模拟人类骨关节炎病理和病因方面存在不足。在本研究中,我们旨在开发源自人诱导多能干细胞(iPSC)的微生理骨软骨(OC)组织芯片,以模拟骨关节炎的病理过程。我们首先将iPSC诱导为间充质祖细胞(iMPC),并优化了iMPC的软骨诱导和成骨诱导条件。然后将iMPC封装到光交联明胶支架中,并在双流生物反应器中培养,其中上游流为软骨生成培养基,下游流为成骨培养基。分化28天后,成功生成了OC组织芯片,并通过实时RT-PCR和组织学确认了其表型。为了创建骨关节炎模型,使用白细胞介素-1β(IL-1β)对软骨成分进行了7天的刺激。在对照条件下,骨组织促进软骨生成并抑制上层软骨组织的软骨细胞终末分化。在模拟骨关节炎的条件下,骨组织加速了软骨组织的降解,这可能是通过产生分解代谢和炎性细胞因子实现的。这些发现表明,在正常和病理条件下,OC组织芯片中的骨组织和软骨组织成分之间存在活跃的功能相互作用。最后,一种常用于骨关节炎的选择性COX-2抑制剂塞来昔布,在骨关节炎模型中显示出可下调分解代谢和促炎细胞因子的表达,证明了OC组织芯片模型在药物筛选中的实用性。总之,本研究中开发的iPSC衍生的OC组织芯片代表了一个高通量平台,适用于模拟骨关节炎以及筛选和测试候选DMOADs。