Genemaras Amaris A, Ennis Hayley, Bradshaw Brad, Kaplan Lee, Huang C-Y Charles
1 Department of Biomedical Engineering, College of Engineering, University of Miami, Coral Gables, FL, USA.
2 Department of Orthopedics, Division of Sports Medicine, University of Miami Miller School of Medicine, Miami, FL, USA.
Cartilage. 2018 Jul;9(3):293-303. doi: 10.1177/1947603516684589. Epub 2016 Dec 27.
Objective Early intervention therapies targeting inflammation and cell death during the acute phase of cartilage injury have the potential to prevent posttraumatic osteoarthritis. The objective of this study was to investigate the effects of interleukin receptor antagonist protein (IRAP), hyaluronan (HA), dexamethasone (DEX), and mesenchymal stem cell (MSC) treatment on the expression of established genetic markers for matrix degradation, apoptosis, and inflammation in articular cartilage during the acute phase of injury. Design A custom impact device was used to create replicable injury ex vivo to intact porcine knee joint. One hour after impact, IRAP, HA, DEX, or MSCs was intra-articularly injected. At 8 hours postinjury, cartilage and meniscus samples were harvested for genetic expression analysis. Expression of miR-27b, miR-140, miR-125b, miR-16, miR-34a, miR-146a, miR-22, ADAMTS-4, ADAMTS-5, MMP-3, IL-1β, and TNF-α was analyzed by real-time polymerase chain reaction. Results At 8 hours postinjury, expression of ADAMTS-4, ADAMTS-5, MMP-3, IL-1β, and TNF-α in cartilage was significantly decreased in IRAP- and DEX-treated joints as compared to nontreated injured joints, whereas only IRAP upregulated expression of miR-140, miR-125b, miR-27b, miR-146a, and miR-22 in cartilage. HA and MSC treatments had no significant effects on catabolic and inflammatory gene expression in cartilage. However, HA treatment significantly upregulated expression of all miRNAs except miR-16. In addition, the treatments tested also exhibited significant influences on meniscus. Conclusions This study provides a valuable starting point for further research into potential targets for and efficacy of various early intervention strategies that may delay or prevent the progression of posttraumatic osteoarthritis after acute cartilage injury.
目的 针对软骨损伤急性期炎症和细胞死亡的早期干预疗法有可能预防创伤后骨关节炎。本研究的目的是调查白细胞介素受体拮抗剂蛋白(IRAP)、透明质酸(HA)、地塞米松(DEX)和间充质干细胞(MSC)治疗对损伤急性期关节软骨中已确定的基质降解、细胞凋亡和炎症相关基因标志物表达的影响。
设计 使用定制的冲击装置在体外对完整的猪膝关节造成可复制的损伤。冲击后1小时,关节腔内注射IRAP、HA、DEX或MSC。损伤后8小时,采集软骨和半月板样本进行基因表达分析。通过实时聚合酶链反应分析miR-27b、miR-140、miR-125b、miR-16、miR-34a、miR-146a、miR-22、ADAMTS-4、ADAMTS-5、MMP-3、IL-1β和TNF-α的表达。
结果 损伤后8小时,与未治疗的损伤关节相比,IRAP和DEX治疗的关节软骨中ADAMTS-4、ADAMTS-5、MMP-3、IL-1β和TNF-α的表达显著降低,而只有IRAP上调了软骨中miR-140、miR-125b、miR-27b、miR-146a和miR-22的表达。HA和MSC治疗对软骨中的分解代谢和炎症基因表达没有显著影响。然而,HA治疗显著上调了除miR-16外所有miRNA的表达。此外,所测试的治疗方法对半月板也有显著影响。
结论 本研究为进一步研究各种早期干预策略可能延迟或预防急性软骨损伤后创伤后骨关节炎进展的潜在靶点和疗效提供了有价值的起点。