TNF-α suppression and osteoprotegerin overexpression inhibits wear debris-induced inflammation and osteoclastogenesis in vitro
TNF-α suppression and osteoprotegerin overexpression inhibits wear debris-induced inflammation and osteoclastogenesis in vitro
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DOI：
10.5301/ijao.5000445
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发表时间：
2015-10-01
影响因子：
1.7
通讯作者：
Song, Keguan
中科院分区：
文献类型：
--
作者：
Peng, Li;Zhang, Haowei;Song, Keguan
目的：假体周围骨质溶解，涉及RANK/RANKL/骨保护素（OPG）和TNF-α/NF κ B信号传导，有助于骨吸收和炎症。我们构建了慢病毒载体来抑制TNF-α和增强OPG表达，并评估了它们对破骨细胞/成骨细胞共培养系统中磨损碎屑诱导的炎症和破骨细胞生成的影响。我们用Lenti-negative对照转导小鼠成骨细胞MC 3 T3-E1（Lenti-NC）、Lenti-OPG或Lenti-siTNF-α-OPG和具有Lenti-NC、Lenti-TNF-α siRNA或Lenti-siTNF-α-OPG的鼠巨噬细胞/单核细胞RAW 264.7细胞。然后，通过酶联免疫吸附试验评估TNF-α和OPG蛋白水平。我们在transwell小室中在0.1 mg/mL Ti颗粒存在下共培养转导的MC 3 T3-E1和RAW264.7细胞，以研究TNF-α抑制减少磨损碎屑诱导的炎症的能力。我们还通过RT-PCR评估了TNF-α、IL-1 β、IL-6和OPG的mRNA水平，以及通过抗酒石酸酸性磷酸酶评估了破骨细胞生成。Lenti-siTNF α-OPG改善了MC 3 T3-E1/RAW264.7共培养物中Ti颗粒诱导的TNF-α、IL-1 β、IL-6的表达，同时提高了OPG的mRNA和蛋白水平，减少抗酒石酸酸性磷酸酶（TRAP）（+）细胞的比例。Lenti-siTNF alpha-OPG在体外可抑制磨损碎屑诱导的炎症反应和破骨细胞生成，并且可以代表用于治疗或预防磨损颗粒诱导的骨质溶解的有希望的治疗候选物。
Purpose: Periprosthetic osteolysis, involving RANK/RANKL/osteoprotegerin (OPG) and TNF-alpha/NF kappa B signaling, contributes to bone resorption and inflammation. We constructed lentivirus vectors to inhibit TNF-alpha and enhance OPG expression and assessed their impacts on wear debris-induced inflammation and osteoclastogenesis in an osteoclast/osteoblast coculture system.Methods: We transduced mouse osteoblastic MC3T3-E1 cells with Lenti-negative control (Lenti-NC), Lenti-OPG or Lenti-siTNF alpha-OPG, and murine macrophage/monocyte RAW264.7 cells with Lenti-NC, Lenti-TNF-alpha siRNA or Lenti-siTNF alpha-OPG. Then, TNF-alpha and OPG protein levels were evaluated by enzyme-linked immunosorbent assay. We cocultured transduced MC3T3-E1 and RAW264.7 cells in transwell chambers in the presence of 0.1 mg/mL Ti particles to investigate the capacity of TNF-alpha inhibition to reduce wear debris-induced inflammation. We also assessed mRNA levels TNF-alpha, IL-1 beta, IL-6 and OPG by RT-PCR as well as osteoclastogenesis by tartrate-resistant acid phosphatase.Results: Lenti-siTNF alpha-OPG ameliorated Ti-particle-induced expression of TNF-alpha, IL-1 beta, IL-6 in MC3T3-E1/RAW264.7 cocultures, while enhancing mRNA and protein levels of OPG, and reducing the fraction of tartrate-resistant acid phosphatase (TRAP)(+) cells.Conclusions: Lenti-siTNF alpha-OPG can inhibit the wear debris-induced inflammatory responses and osteoclastogenesis in vitro, and may represent a promising therapeutic candidate for the treatment or prevention of wear particle-induced osteolysis.