Drynda Andreas, Singh Gurpal, Buchhorn Gottfried H, Awiszus Friedemann, Ruetschi Marcel, Feuerstein Bernd, Kliche Stefanie, Lohmann Christoph H
Department of Orthopaedic Surgery, Otto-von-Guericke University, Magdeburg, Germany.
J Biomed Mater Res A. 2015 Jun;103(6):1940-8. doi: 10.1002/jbm.a.35330. Epub 2014 Sep 17.
CXCR4, the chemokine receptor for CXCL12, also known as SDF-1 (stromal cell derived factor-1), has been shown to play a pivotal role in bone metastasis, inflammatory, and autoimmune conditions but has not been investigated in periprosthetic osteolysis. We co-cultured osteoblast-like cells with increasing concentrations of metallic (Co-35Ni-20Cr-10Mo and Co-28Cr-6Mo) and Co-ions simulating wear debris. Real-time polymerase chain reaction (RT-PCR) and Western blotting were used to quantify gene and protein expression of CXCR4. The expression of tumor necrosis factor-alpha (TNF-α) and the effects of AMD3100 (bicyclam) on both CXCR4 and TNF-α expression among these cells was investigated. RT-PCR showed an increase in CXCR4 mRNA (7.5-fold for MG63 and 4.0-fold for SaOs-2 cells) among cells co-cultured with metal alloy particles. Western blotting showed a time-dependent increase in protein expression of CXCR4. The attempted blockade of CXCR4 by its known competitive receptor agonist AMD3100 led to a significant inhibition TNF-α mRNA expression. Immunohistochemistry showed CXCR4 positivity among patients with failed metal-on-metal hip replacements and radiographic evidence of osteolysis. Our data collectively suggest that the CXCR4 chemokine is upregulated in a dose- and time-dependent manner in the presence of metallic wear debris.
CXCR4是趋化因子CXCL12的受体,CXCL12也被称为基质细胞衍生因子-1(SDF-1),已被证明在骨转移、炎症和自身免疫性疾病中起关键作用,但尚未在假体周围骨溶解中进行研究。我们将成骨样细胞与浓度不断增加的模拟磨损颗粒的金属(钴-35镍-20铬-10钼和钴-28铬-6钼)及钴离子进行共培养。采用实时聚合酶链反应(RT-PCR)和蛋白质印迹法对CXCR4的基因和蛋白表达进行定量分析。研究了肿瘤坏死因子-α(TNF-α)的表达以及AMD3100(双环胺)对这些细胞中CXCR4和TNF-α表达的影响。RT-PCR结果显示,与金属合金颗粒共培养的细胞中CXCR4 mRNA增加(MG63细胞增加7.5倍,SaOs-2细胞增加4.0倍)。蛋白质印迹法显示CXCR4蛋白表达呈时间依赖性增加。用其已知的竞争性受体激动剂AMD3100阻断CXCR4可显著抑制TNF-α mRNA表达。免疫组织化学显示,金属对金属髋关节置换失败且有骨溶解影像学证据的患者中CXCR4呈阳性。我们的数据共同表明,在存在金属磨损颗粒的情况下,CXCR4趋化因子以剂量和时间依赖性方式上调。