依那西普对钛颗粒诱导小鼠骨溶解及破骨细胞增殖的影响

Effects of etanercept on Ti particle-induced osteolysis and osteoclastogenesis in mice

目的:已证实肿瘤坏死因子α在人工关节无菌性松动和类风湿关节炎的发病机制中均发挥重要的作用。依那西普作为肿瘤坏死因子α拮抗剂在治疗类风湿关节炎中疗效显著。拟通过实验观察依那西普对钛颗粒诱导小鼠骨溶解及破骨细胞增殖的影响,为依那西普防治人工关节无菌性松动提供实验依据。方法:实验于2007-02/2007-08在宁夏医学院生殖及遗传学重点实验室和宁夏医学院附属医院病理学实验室完成。①60只BALB/C小鼠随机分为6组,每组10只。分别为空白对照组,单纯钛处理组,20,50,100,200μg/kg依那西普+钛处理组。实验过程中对动物处置符合动物伦理学要求。依那西普由美国Wyeth公司提供(生产批号:P321686);钛金属颗粒由美国Zimmer公司提供。②建立钛颗粒骨吸收模型,其中空白对照组行原位缝合,不置入钛颗粒。依那西普各组分别在第0,2,4,6,8天腹腔注射不同剂量的依那西普(20,50,100,200μg/kg)。10d后取出颅骨,行常规苏木精-伊红染色,了解骨吸收情况。抗酒石酸酸性磷酸酶染色计算破骨细胞数量。结果:苏木精-伊红染色结果为单纯钛处理组骨吸收明显增加,骨小梁结构紊乱及骨质疏松,破骨细胞的形态变大,数目变多;依那西普干预组上述表现减轻,尤以200μg/kg剂量为明显。与空白对照组相比,单纯钛处理组可明显增加骨溶解(P<0.001)及破骨细胞生成(P<0.001);与单纯钛处理组相比,依那西普各组可减少骨溶解(P<0.05)及破骨细胞生成(P<0.05),尤以200μg/kg剂量最为明显(P<0.05),抗酒石酸酸性磷酸酶染色与苏木精-伊红染色结果相符。结论:依那西普可抑制钛颗粒诱导的骨溶解及破骨细胞的增殖,有望成为预防人工关节无菌性松动的药物。

AIM： It has been demonstrated that tumor necrosis factor- α （TNF- α ） plays an important role in the pathogenesy of artificial joint aseptic loosening and rheumatoid arthritis. Etanercept, antagon of TNF- α, shows notable effect in treating rheumatoid arthritis. In this study, we investigated the influence of etanercept on Ti particles-induced ostoolysis and osteoclastogenesis in mice and evaluate the feasibility of etanercept for treatment of artificial joint aseptic loosening. METHODS： The experiment was performed at the Key Laboratory of Genesiology and Genetics, and Pathological Laboratory of Hospital, Ningxia Medical College from February to August 2007. （1）Sixty BALB/C mice were randomly divided into 6 groups （n =10）： blank control group, Ti （Zimmer, USA） treatment group, and 20, 50, 100, 200 μ g/kg etanercept （Wyeth, USA, No. P321686） plus Ti treatment group. （2）Bone resorption models of Ti panicles were established in all groups exception blank control group （in situ suture with no Ti particles）. Different doses of etanercept （20, 50, 100, 200 μg/kg） were intraperitoneally injected into etanercept groups on days 0, 2, 4, 6, and 8, respectively. Ten days later, mice cranial bones were harvested for HE staining to observe bone resorption. Number of osteoclast was counted using acid phosphatase （TRAP）. RESULTS： HE staining showed that bone resorption in Ti treatment group was increased obviously and the structure of bone trabecula was disorderly and osteoporotic; the volume of osteoclast was enlarged and number was increased. The findings were reduced in etanercept groups, especially in 200 μ g/kg group. Compared with blank control group, Ti particles significantly increased osteolysis （P 〈 0.001） and osteoclastogenesis （P 〈 0.001）; compared with Ti particles group, etanercept effectively inhibited Ti particles induced osteolysis （P 〈 0.05） and osteoclastogenesis （P 〈 0.05）, especially in 200 μ g/kg group （P 〈 0.05）. The result of TRAP was accorded with HE staining. CONCLUSION： Etanercept can inhibit Ti particles-induced osteolysis and osteoclastogenesis in vivo and is promising to be a therapeutic candidate for the prevention of artificial joint aseptic loosening.