Development of Reliable and High-Throughput Human Biomimetic Cartilage and Bone Models to Explore Senescence and Personalized Osteoarthritis Treatment Options

IF 2.3 3区 医学 Q2 ORTHOPEDICS
Ilja Boone, Evelyn Houtman, Margo Tuerlings, Jim J. van den Berg, Johannes Lehmann, Peter L. J. de Keizer, Rob G. H. H. Nelissen, Ingrid Meulenbelt
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Abstract

To facilitate effective preclinical testing of senescence treatments for osteoarthritis (OA), we have created reliable biomimetic and high-throughput models using aged human joint tissues. Moreover, concerns regarding scalability led to the concurrent development of a high-throughput human in vitro senescence cartilage organoid model. Osteochondral explants and cells for the cartilage organoid model were isolated from patients undergoing joint replacement surgery due to OA. To induce senescence, explants and organoids were subjected to radiation and/or mechanical loading. Samples were harvested; gene expression of relevant senescent and cartilage genes was measured using RT-qPCR, and protein expression was evaluated using histology. A general senescence phenotype was induced by the perturbations, as shown by senescence-associated β-galactosidase staining. In-depth gene expression analysis revealed that hyperphysiological mechanical loading upregulated gene expression of IL8 and SERPINE1, representing aspects of a senescence-associated secretory phenotype (SASP) profile. Irradiation upregulated CDKN1A, encoding p21, and downregulated LMNB1, representing a cell cycle arrest profile with the absence of a SASP response. Combining the two perturbations showed upregulation of CDKN1A, IL8, and SERPINE and downregulation of LMNB1, representing a complementary senescence model. The high-throughput human in vitro cartilage organoid senescence model showed similar effects to the irradiation explant model. In this study, we present a variety of senescence models of human aged chondrocytes that allows for rapid initial screening of anti-senescence compounds in high-throughput, as well as in-depth, characterization of post-mitotic aged chondrocytes prone to OA pathophysiology. This research advances the development of essential therapeutics for OA.

Abstract Image

开发可靠和高通量的人体仿生软骨和骨模型,探索衰老和个性化骨关节炎治疗方案。
为了促进骨关节炎(OA)衰老治疗的有效临床前测试,我们使用衰老的人类关节组织创建了可靠的仿生和高通量模型。此外,对可扩展性的关注导致了高通量人类体外衰老软骨类器官模型的同步发展。骨软骨外植体和软骨类器官模型的细胞是从骨性关节炎患者的关节置换手术中分离出来的。为了诱导衰老,外植体和类器官受到辐射和/或机械负荷。采集样本;RT-qPCR检测衰老相关基因和软骨相关基因的表达,组织学检测蛋白表达。如衰老相关的β-半乳糖苷酶染色所示,扰动诱导了一般的衰老表型。深入的基因表达分析显示,高生理机械负荷上调了IL8和SERPINE1的基因表达,代表了衰老相关分泌表型(SASP)的各个方面。辐照上调编码p21的CDKN1A,下调LMNB1,代表了缺乏SASP应答的细胞周期阻滞谱。结合这两种扰动显示CDKN1A、IL8和SERPINE上调,LMNB1下调,代表互补衰老模型。高通量体外人软骨类器官衰老模型与辐照外植体模型效果相似。在这项研究中,我们提出了多种人类衰老软骨细胞的衰老模型,这些模型允许高通量快速初始筛选抗衰老化合物,以及深入表征易发生OA病理生理的有丝分裂后衰老软骨细胞。这项研究促进了骨性关节炎基本治疗方法的发展。
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来源期刊
Journal of Orthopaedic Research®
Journal of Orthopaedic Research® 医学-整形外科
CiteScore
6.10
自引率
3.60%
发文量
261
审稿时长
3-6 weeks
期刊介绍: The Journal of Orthopaedic Research is the forum for the rapid publication of high quality reports of new information on the full spectrum of orthopaedic research, including life sciences, engineering, translational, and clinical studies.
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