Papathanasiou Ioanna, Michalitsis Sotirios, Hantes Michael E, Vlychou Marianna, Anastasopoulou Lydia, Malizos Konstantinos N, Tsezou Aspasia
Laboratory of Cytogenetics and Molecular Genetics, University of Thessaly, Faculty of Medicine, Biopolis, 41500, Larissa, Greece.
Department of Orthopaedic Surgery, University of Thessaly, Faculty of Medicine, Biopolis, 41500, Larissa, Greece.
BMC Musculoskelet Disord. 2016 Jan 13;17:21. doi: 10.1186/s12891-016-0871-8.
Anterior cruciate ligament (ACL) tear is considered a risk factor for osteoarthritis development. The purpose of our study was to investigate the expression levels of the apoptotic enzyme caspase 3, pro-inflammatory cytokines interleukin-1β (IL-1β) and interleukin-6 (IL-6) and degrading enzyme matrix metalloproteinase 13 (MMP-13), all indicative of cartilage degeneration and osteoarthritis development in patients' chondrocytes after ACL rupture.
We investigated the correlation between grade of cartilage degradation and time from injury or patients' age. IL-1β, IL-6 and MMP-13 mRNA expression levels were investigated in normal (n = 4) and chondrocytes from patients with ACL rupture (n = 33) using real-time polymerase chain reaction (PCR). Moreover, MMP-13 and caspase-3 protein expression levels were evaluated by western blot analysis. Trend analysis and correlation coefficient were performed to derive the relations between gene expression (MMP13, IL-6, IL-1β) and grading of cartilage defects and between gene expression (MMP13, IL-6, IL-1β) and patients' age, respectively.
Correlations were established between grade of cartilage degradation and time from injury. MMP-13, IL-6, IL-1β and caspase 3 expression levels were significantly upregulated in chondrocytes from ACL-deficient knee compared to normal. Among the patients with ACL-deficient knees, a significant upregulation of MMP-13 was observed in patients with ACL-rupture > 18 months from the time of injury to arthroscopy compared to patients with ACL-injury up to 18 months, whereas IL-6 and IL-1β expression was higher in chondrocytes from patients with more than 10 months ACL injury compared to those that underwent surgery within the first 10 months after injury. Νο association was observed between IL-1β, IL-6 and MMP-13 expression levels and cartilage defects or patients' age.
Our results showed that increased levels of apoptotic, inflammatory and catabolic factors in chondrocytes are associated with time from injury and could contribute to cartilage degradation and osteoarthritis development after ACL rupture.
前交叉韧带(ACL)撕裂被认为是骨关节炎发展的一个危险因素。我们研究的目的是调查凋亡酶半胱天冬酶3、促炎细胞因子白细胞介素-1β(IL-1β)和白细胞介素-6(IL-6)以及降解酶基质金属蛋白酶13(MMP-13)的表达水平,这些指标均表明ACL断裂后患者软骨细胞中软骨退变和骨关节炎的发展情况。
我们研究了软骨降解程度与受伤时间或患者年龄之间的相关性。使用实时聚合酶链反应(PCR)研究了正常软骨细胞(n = 4)和ACL断裂患者软骨细胞(n = 33)中IL-1β、IL-6和MMP-13 mRNA的表达水平。此外,通过蛋白质印迹分析评估MMP-13和半胱天冬酶-3蛋白的表达水平。分别进行趋势分析和相关系数分析,以得出基因表达(MMP13、IL-6、IL-1β)与软骨缺损分级之间以及基因表达(MMP13、IL-6、IL-1β)与患者年龄之间的关系。
建立了软骨降解程度与受伤时间之间的相关性。与正常情况相比,ACL损伤膝关节软骨细胞中MMP-13、IL-6、IL-1β和半胱天冬酶3的表达水平显著上调。在ACL损伤膝关节患者中,与受伤至关节镜检查时间≤18个月的ACL损伤患者相比,受伤时间>18个月的ACL断裂患者中观察到MMP-13显著上调,而受伤超过10个月患者的软骨细胞中IL-6和IL-1β表达高于受伤后前10个月内接受手术患者。未观察到IL-1β、IL-6和MMP-13表达水平与软骨缺损或患者年龄之间存在关联。
我们的结果表明,软骨细胞中凋亡、炎症和分解代谢因子水平升高与受伤时间相关,可能导致ACL断裂后软骨退变和骨关节炎发展。