Effect of etidronate on COX-2 expression and PGE2 production in macrophage-like RAW 264.7 cells stimulated by titanium particles
Effect of etidronate on COX-2 expression and PGE2 production in macrophage-like RAW 264.7 cells stimulated by titanium particles
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DOI：
10.1007/s00776-007-1180-8
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发表时间：
2007-11-01
影响因子：
1.7
通讯作者：
Kurosaka, Masahiro
中科院分区：
文献类型：
--
作者：
Suzuki, Yoshihiro;Nishiyama, Takayuki;Kurosaka, Masahiro
背景资料。全关节置换术最常见的失败是伴骨溶解的无菌性松动。以往的报道表明，前列腺素E-2(PGE(2))由吞噬磨损碎屑的巨噬细胞分泌，可诱导假体周围骨溶解。许多临床研究报道，双磷酸盐治疗可以减少全关节置换术后假体周围的骨丢失和松动。双膦酸盐是一种具有减少骨吸收能力的合成化合物。此外，据报道，一些双膦酸盐通过减少促炎细胞因子的分泌而具有抗炎作用。然而，双膦酸盐减少假体周围骨吸收的机制仍不清楚。本研究的目的是探讨依替膦酸盐(EHDP)抑制假体周围骨吸收的机制之一。将0.001、0.01、0.1、1、10、100mU M的乙二磷酸与1 mg/ml的钛颗粒共同作用于巨噬细胞样RAW 264.7细胞，培养24 h后，提取细胞总基因，用逆转录聚合酶链式反应检测环氧合酶-2的表达。收集培养上清液，用双抗体夹心法测定前列腺素E(2)、白介素1β(IL-1β)、白介素6(IL-6)、肿瘤坏死因子-α(TNF-α)的含量。分析表明，EHDP呈剂量依赖性抑制COX-2的表达和PGE(2)的产生。100 mU M的EHDP可抑制细胞产生PGE(2)，接近未刺激细胞的水平。EDP还能抑制培养上清液中IL-1β、IL-6和TNF-α的产生。促炎症细胞因子的阻断作用可能是一种减少种植体周围骨吸收的机制。
Background. The most common failure of total joint replacement is aseptic loosening in association with osteolysis. Previous reports have shown that prostaglandin E-2 (PGE(2)) secreted from macrophages that phagocytosed wear debris induced periprosthetic osteolysis. Many clinical studies have reported that bisphosphonate therapy reduced periprosthetic bone loss and loosening of the implants after total joint replacements. Bisphosphonates are synthetic compounds with the ability to decrease bone resorption. In addition, some bisphosphonates have been reported to have anti-inflammatory effects by reducing the secretion of pro-inflammatory cytokines. However, the mechanism of bisphosphonates that reduces periprosthetic bone resorption remains unclear. The purpose of this study was to investigate one of the mechanisms by which etidronate (EHDP) inhibits periprosthetic bone resorption.Methods. Macrophage-like RAW 264.7 cells were treated with EHDP at concentrations of 0.001, 0.01, 0.1, 1, 10, and 100 mu M together with the titanium particles at a concentration of 1 mg/ml. After a 24-h culture period, total mRNA was isolated and reverse transcription-polymerase chain reaction (RT-PCR) was done to examine the expression of cyclooxygenase-2 (COX-2). The supernatants were also collected and production of PGE(2), interleukin-1 beta (IL-1 beta), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-alpha) were quantified using an enzyme-linked immunosorbent assay (ELISA).Results. Analyses showed that COX-2 expression and PGE(2) production were suppressed by EHDP in a dose-dependent manner. By 100 mu M of EHDP, PGE(2) production of the cells was suppressed approximately to the level of the nonstimulated cells. Production of IL-1 beta, IL-6, and TNF-alpha in the supernatant was also suppressed by EHDP.Conclusions. The blockage effect of pro-inflammatory cytokines is a possible etidronate mechanism that reduces bone resorption around implants.