Žigon-Branc Sara, Barlič Ariana, Knežević Miomir, Jeras Matjaž, Vunjak-Novakovic Gordana
Dept. of Biomedical Engineering, Columbia University, New York, USA.
Educell Cell Therapy Service Ltd, Trzin, Slovenia.
Biotechnol Prog. 2018 Jul;34(4):1045-1058. doi: 10.1002/btpr.2629. Epub 2018 Mar 31.
Inflammation plays a major role in progression of rheumatoid arthritis, a disease treated with antagonists of tumor necrosis factor-alpha (TNF-α) and interleukin 1β (IL-1β). New in vitro testing systems are needed to evaluate efficacies of new anti-inflammatory biological drugs, ideally in a patient-specific manner. To address this need, we studied microspheroids containing 10,000 human osteoarthritic primary chondrocytes (OACs) or chondrogenically differentiated mesenchymal stem cells (MSCs), obtained from three donors. Hypothesizing that this system can recapitulate clinically observed effects of anti-inflammatory drugs, spheroids were exposed to TNF-α, IL-1β, or to supernatant containing secretome from activated macrophages (MCM). The anti-inflammatory efficacies of anti-TNF-α biologicals adalimumab, infliximab, and etanercept, and the anti-IL-1β agent anakinra were assessed in short-term microspheroid and long-term macrospheroid cultures (100,000 OACs). While gene and protein expressions were evaluated in microspheroids, diameters, amounts of DNA, glycosaminoglycans, and hydroxiproline were measured in macrospheroids. The tested drugs significantly decreased the inflammation induced by TNF-α or IL-1β. The differences in potency of anti-TNF-α biologicals at 24 h and 3 weeks after their addition to inflamed spheroids were comparable, showing high predictability of short-term cultures. Moreover, the data obtained with microspheroids grown from OACs and chondrogenically differentiated MSCs were comparable, suggesting that MSCs could be used for this type of in vitro testing. We propose that in vitro gene expression measured after the first 24 h in cultures of chondrogenically differentiated MSCs can be used to determine the functionality of anti-TNF-α drugs in personalized and preclinical studies. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:1045-1058, 2018.
炎症在类风湿性关节炎的发展过程中起主要作用，类风湿性关节炎是一种用肿瘤坏死因子-α（TNF-α）和白细胞介素1β（IL-1β）拮抗剂治疗的疾病。需要新的体外测试系统来评估新型抗炎生物药物的疗效，理想情况下是以患者特异性的方式。为满足这一需求，我们研究了含有10000个人类骨关节炎原代软骨细胞（OAC）或软骨分化间充质干细胞（MSC）的微球体，这些细胞来自三名供体。假设该系统可以重现抗炎药物的临床观察效果，将微球体暴露于TNF-α、IL-1β或含有活化巨噬细胞分泌组的上清液（MCM）中。在短期微球体和长期大球体培养（100000个OAC）中评估了抗TNF-α生物制剂阿达木单抗、英夫利昔单抗和依那西普以及抗IL-1β药物阿那白滞素的抗炎效果。在微球体中评估基因和蛋白质表达，在大球体中测量直径、DNA、糖胺聚糖和羟脯氨酸的含量。测试药物显著降低了TNF-α或IL-1β诱导的炎症。抗TNF-α生物制剂在添加到炎症微球体后24小时和3周时效力的差异具有可比性，表明短期培养具有较高的可预测性。此外，从OAC和软骨分化的MSC生长的微球体获得的数据具有可比性，这表明MSC可用于此类体外测试。我们建议，在软骨分化的MSC培养的最初24小时后测量的体外基因表达可用于在个性化和临床前研究中确定抗TNF-α药物的功能。©2018美国化学工程师学会生物技术进展，34:1045 - 1058，2018。
