Tamaki Yasunobu, Sasaki Kan, Sasaki Akiko, Takakubo Yuya, Hasegawa Hiroshi, Ogino Toshihiko, Konttinen Yrjö T, Salo Jari, Takagi Michiaki
Department of Orthopaedic Surgery, Yamagata University School of Medicine, Yamagata 990-9585, Japan.
J Biomed Mater Res B Appl Biomater. 2008 Jan;84(1):191-204. doi: 10.1002/jbm.b.30861.
Total hip replacement can be complicated by periprosthetic osteolysis. Monocytes/macrophages play a major role in the formation of the foreign body granulomas induced by wear debris. We hypothesized that periprosthetic monocytes/macrophages do not only accelerate inflammatory and osteoclast-mediated osteolytic processes, but also resorb periprosthetic bone directly by themselves. This study was designed to evaluate the osteolytic potential in vitro of monocytes/macrophages derived from bone marrow.
Monocytes/macrophages were produced by filtration of rat bone marrow cells, followed by culture in the presence of macrophage-colony stimulating factor (M-CSF). Monocyte/macrophage properties were ascertained using immunocytochemistry and phagocytic activity. Osteolytic cytokines and extracellular matrix degrading proteinases were quantified at the mRNA level.
Adherent cell fraction was immunoreactive for the monocyte/macrophage specific marker CD68 and active in the phagocytosis of carbon particles up to 72 h. They also showed immunoreactivity to cathepsin K, IL-1beta, IL-6, and M-CSF, but mostly did not react to TRAP. mRNA levels of osteolytic cytokines and extracellular matrix degrading proteinases were enhanced, but that of RANKL were not. Monocytes/macrophages resorbed dentine discs and carbonated calcium phosphate was very actively resorbed after stimulation with titanium particles.
Harvested bone marrow cells expressed monocyte/macrophage phenotype, but not osteoclastic markers. The capacity of these cathepsin-K-positive phagocytic cells to resorb dentine discs and carbonated calcium phosphate in vitro suggests a direct role of monocytes/macrophages in bone resorption and periprosthetic osteolysis. The finding supports our hypothesis and previous histomorphometric observations on the presence of such osteolytic macrophages in vivo around loosening prosthesis.
全髋关节置换术可能会因假体周围骨溶解而变得复杂。单核细胞/巨噬细胞在磨损颗粒诱导的异物肉芽肿形成中起主要作用。我们推测，假体周围的单核细胞/巨噬细胞不仅会加速炎症和破骨细胞介导的骨溶解过程，还会直接自身吸收假体周围的骨组织。本研究旨在评估源自骨髓的单核细胞/巨噬细胞的体外溶骨潜能。
通过过滤大鼠骨髓细胞产生单核细胞/巨噬细胞，然后在巨噬细胞集落刺激因子（M-CSF）存在的情况下进行培养。使用免疫细胞化学和吞噬活性确定单核细胞/巨噬细胞的特性。在mRNA水平上对溶骨细胞因子和细胞外基质降解蛋白酶进行定量。
贴壁细胞部分对单核细胞/巨噬细胞特异性标志物CD68具有免疫反应性，并且在长达72小时内对碳颗粒的吞噬作用活跃。它们对组织蛋白酶K、IL-1β、IL-6和M-CSF也表现出免疫反应性，但大多对抗酒石酸酸性磷酸酶（TRAP）无反应。溶骨细胞因子和细胞外基质降解蛋白酶的mRNA水平升高，但核因子κB受体活化因子配体（RANKL）的mRNA水平未升高。单核细胞/巨噬细胞吸收牙本质盘，在用钛颗粒刺激后，碳酸磷酸钙被非常活跃地吸收。
收获的骨髓细胞表达单核细胞/巨噬细胞表型，但不表达破骨细胞标志物。这些组织蛋白酶K阳性吞噬细胞在体外吸收牙本质盘和碳酸磷酸钙的能力表明单核细胞/巨噬细胞在骨吸收和假体周围骨溶解中起直接作用。这一发现支持了我们的假设以及先前关于松动假体周围体内存在这种溶骨巨噬细胞的组织形态计量学观察结果。
