Fujikawa Y, Sabokbar A, Neale S D, Itonaga I, Torisu T, Athanasou N A
Nuffield Department of Orthopaedic Surgery, University of Oxford, Oxford, UK.
Bone. 2001 Mar;28(3):261-7. doi: 10.1016/s8756-3282(00)00453-1.
Macrophage-colony stimulating factor (M-CSF) is an essential requirement for human osteoclast formation, but its effect on the proliferation and differentiation of circulating osteoclast precursor cells is unknown. Other growth factors and cytokines are also known to support/stimulate osteoclast formation from mouse marrow precursors, but it is not certain whether these factors similarly influence human osteoclast formation. In this study, human monocytes were cocultured with osteoblast-like UMR-106 cells on coverslips and dentine slices for up to 21 days in the presence of 1,25 dihydroxyvitamin D(3) (10(-7) mol/L), dexamethasone (10(-8) mol/L), and various concentrations of either M-CSF or other humoral factors (interleukin [IL]-1beta, IL-3, IL-6, and IL-11; tumor necrosis factor-alpha [TNF-alpha]; and granulocyte macrophage [GM]-CSF). The effect on osteoclast formation was assessed by tartrate-resistant acid phosphatase (TRAP) and vitronectin receptor staining and lacunar bone resorption. The results of time-course and proliferation studies showed that M-CSF stimulated both the proliferative and differentiation stages of human osteoclast formation from circulating osteoclast precursors in a dose-dependent manner. A high concentration of M-CSF (100 ng/mL) did not inhibit osteoclast formation. IL-3 and GM-CSF were also capable of stimulating human osteoclast formation, although these growth factors were much less potent than M-CSF. IL-3- and GM-CSF-stimulated osteoclast formation was inhibited by an antibody specific for human M-CSF. Osteoclast formation and lacunar resorption was not seen when either TNF-alpha, IL-1beta, IL-6 (+ soluble IL-6 receptor), or IL-11 was substituted for M-CSF during coculture. These results confirm that M-CSF is essential for human osteoclast formation from circulating mononuclear precursors, and also shows that IL-3 and GM-CSF may support osteoclast differentiation via the stimulation of M-CSF production by human monocytes.
巨噬细胞集落刺激因子（M-CSF）是人类破骨细胞形成的必要条件，但其对循环破骨细胞前体细胞增殖和分化的影响尚不清楚。其他生长因子和细胞因子也已知可支持/刺激小鼠骨髓前体细胞形成破骨细胞，但不确定这些因子是否同样影响人类破骨细胞的形成。在本研究中，将人单核细胞与成骨样UMR-106细胞在盖玻片和牙本质切片上共培养长达21天，培养环境中存在1,25-二羟基维生素D(3)（10(-7) mol/L）、地塞米松（10(-8) mol/L）以及不同浓度的M-CSF或其他体液因子（白细胞介素[IL]-1β、IL-3、IL-6和IL-11；肿瘤坏死因子-α[TNF-α]；以及粒细胞巨噬细胞[GM]-CSF）。通过抗酒石酸酸性磷酸酶（TRAP）和玻连蛋白受体染色以及陷窝骨吸收来评估对破骨细胞形成的影响。时间进程和增殖研究结果表明，M-CSF以剂量依赖的方式刺激循环破骨细胞前体细胞形成人类破骨细胞的增殖和分化阶段。高浓度的M-CSF（100 ng/mL）并未抑制破骨细胞的形成。IL-3和GM-CSF也能够刺激人类破骨细胞的形成，尽管这些生长因子的效力远低于M-CSF。IL-3和GM-CSF刺激的破骨细胞形成被针对人类M-CSF的特异性抗体所抑制。在共培养期间，当用TNF-α、IL-1β、IL-6（+可溶性IL-6受体）或IL-11替代M-CSF时，未观察到破骨细胞形成和陷窝吸收。这些结果证实，M-CSF对于从循环单核前体细胞形成人类破骨细胞至关重要，并且还表明IL-3和GM-CSF可能通过刺激人单核细胞产生M-CSF来支持破骨细胞分化。
