Titanium particle‑mediated osteoclastogenesis may be attenuated via bidirectional ephrin‑B2/eph‑B4 signaling in vitro.
Titanium particle‑mediated osteoclastogenesis may be attenuated via bidirectional ephrin‑B2/eph‑B4 signaling in vitro.
复制标题
钛颗粒介导的破骨细胞生成可能通过体外双向肝配蛋白 B2/eph B4 信号减弱
DOI：
10.3892/ijmm.2018.3780
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发表时间：
2018-10
影响因子：
5.4
通讯作者：
Mao YQ
中科院分区：
文献类型：
--
作者：
Ge YW;Liu ZQ;Sun ZY;Yu DG;Feng K;Zhu ZA;Mao YQ
本研究探讨了EphB4信号转导通路在磨损颗粒介导的体外破骨细胞生成中的作用。用核因子受体激活剂κB配体(RANKL，50 ng/ml)诱导小鼠骨髓巨噬细胞分化为破骨细胞。用成骨细胞膜表面受体EphB4-Fc(4微克/毫升)刺激破骨细胞在钛(Ti)存在和不存在的情况下激活ePhin-B2配体。用抗酒石酸酸性磷酸酶(TRAP)染色检测破骨细胞数，鬼臼乙素染色检测破骨细胞骨架。采用骨坑吸收实验检测破骨细胞功能。用逆转录定量聚合酶链式反应和免疫印迹分析检测破骨细胞的形成。用ELISA检测炎症因子的产生。结果表明，在RANKL存在的条件下，钛显著促进骨髓基质细胞向成熟破骨细胞分化，并显著促进ePhin-B2、活化T细胞核因子1(NFATc1)、TRAP、Fos原癌基因、AP-1转录因子亚单位(C-fos)和基质金属蛋白酶9(MMP9)基因的表达。Phalloidin和TRAP染色显示，加入EphB4-Fc后，破骨细胞、成骨细胞、重塑、ePhin-B2、破骨细胞生成等关键词的数量、大小和细胞骨架元素均较未加入EphB4-FC的钛粒组明显减少。骨陷窝吸收实验显示，钛微粒+EphB4-FC组与钛粒组相比，吸收陷窝面积明显减少。EphB4-Fc组NFATc1、TRAP、C-fos和MMP9基因表达明显降低，而EphB4-Fc组5d后EphB4-Fc组EPhin-B2基因表达明显高于未加EphB4-Fc的钛颗粒组。钛颗粒通过双向信号抑制炎性细胞因子的产生。EphB4-Fc通过EphB4/EphB4双向信号通路抑制破骨细胞介导的钛颗粒的形成。这种双向信号通路的激活可能是治疗假体周围骨溶解的一种潜在的临床治疗方法。
The present study investigated the role of bidirectional ephrin-B2/erythropoietin-producing human hepatocellular receptor 4 (ephB4) signaling in the regulation of wear particle-mediated osteoclastogenesis in vitro. Mouse bone marrow macrophages (BMMs) were induced into osteoclasts by receptor activator of nuclear factor-κB ligand (RANKL, 50 ng/ml). EphB4-Fc, an osteoblast membrane surface receptor (4 µg/ml), was used to stimulate the ephrin-B2 ligand of osteoclasts in the presence and absence of titanium (Ti). Tartrate-resistant acid phosphatase (TRAP) staining was used to detect the number of osteoclasts, and phalloidin staining was used to examine the cytoskeletons of the osteoclasts. A bone pit absorption experiment was used to measure osteoclast function. Reverse transcription quantitative polymerase chain reaction and western blot analysis were used to examine osteoclastogenesis. ELISAs were used to detect the production of inflammatory factors. The data demonstrated that Ti significantly promoted the differentiation of BMMs into mature osteoclasts in the presence of RANKL and significantly promoted expression of the ephrin-B2, nuclear factor of activated T-cells 1 (NFATc1), TRAP, Fos proto-oncogene, AP-1 transcription factor subunit (C-FOS), and matrix metalloproteinase 9 (MMP9) genes. Phalloidin and TRAP staining revealed that following the addition of ephB4-Fc, the number, size and cytoskeletal elements of Key words: osteoclasts, osteoblasts, remodeling, ephrin-B2, osteoclastogenesis osteoclasts were significantly decreased compared with those in the titanium particle group without ephB4-Fc. Compared with the titanium particle group, the bone pit absorption experiment revealed significantly decreased absorption pit areas in the titanium particle+ephB4-Fc group. The expression of the NFATc1, TRAP, C-FOS and MMP9 genes was markedly decreased in the ephB4-Fc group; however, the expression of the ephrin-B2 gene was increased compared with the Ti particle group without ephB4-Fc after 5 days. Production of inflammatory cytokines was inhibited by Ti particles through bidirectional signals. Addition of ephB4-Fc inhibited the osteoclast-mediated formation of Ti particles via bidirectional ephrin-B2/ephB4 signaling. Activation of this bidirectional signaling pathway may be a potential clinical treatment for osteolysis surrounding prostheses.
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影响因子：
7.6
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通讯作者：
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