脂多糖诱导钛种植体周围骨髓间质细胞炎症中的巨噬细胞游走抑制因子

Macrophage migration inhibitory factor in lipopolysaccharide-induced peri-implant inflammation of bone marrow mesenchymal stem cells

背景:目前大量研究证实,巨噬细胞游走抑制因子在多种生物活动中发挥着重要的作用,如肿瘤的发生发展等,近年来发现其在机体发生的炎症过程中也扮演者重要的角色,也取得了很多成绩,然而在口腔内牙种植体周围这一特殊的环境中,巨噬细胞游走抑制因子是否发挥着作用以及发挥着怎样的作用,尚不清楚。目的:观察巨噬细胞游走抑制因子在经过脂多糖诱导纯钛材料周围生长的骨髓间质细胞发生的炎症中发挥的作用。方法:首先将骨髓间质细胞接种于钛细胞培养爬片上,模拟口内种植体周围环境,实验分4组:对照组为不加任何刺激的骨髓间质细胞;脂多糖组利用脂多糖诱导骨髓间质细胞发生炎症;非特异性si RNA+脂多糖组利用脂多糖刺激转染过非特异性小干扰RNA的骨髓间质细胞;巨噬细胞游走抑制因子siR NA+脂多糖组利用脂多糖刺激转染过巨噬细胞游走抑制因子靶向小干扰RNA的骨髓间质细胞。结果与结论:实验通过流式细胞筛选技术,筛选出细胞表面蛋白CD29、CD90阳性率大于95%,而且CD45表达低于5%的骨髓间质细胞作为实验对象。CCK8实验结果提示,脂多糖与巨噬细胞游走抑制因子靶向小干扰RNA对骨髓间质细胞的增殖活性几乎无影响,然而与对照组相比,脂多糖可明显刺激骨髓间质细胞发生炎症反应,为对照组的15-20倍(P<0.01)。巨噬细胞游走抑制因子si RNA+脂多糖组白细胞介素1β、白细胞介素6、肿瘤坏死因子α表达低于脂多糖组(P<0.01)。结果证实,脂多糖可以促进种植体周围骨髓间质细胞的炎症反应,而且巨噬细胞游走抑制因子靶向小干扰RNA沉默可以在一定程度上抑制炎症反应的发生。

BACKGROUND：Currently, a large number of studies have confirmed that macrophage migration inhibitory factor plays an important role in a variety of biological activities, such as tumor development. In recent years, it also plays an important role in the inflammatory process, and has achieved a lot of results. However, it is unclear whether and how the macrophage migration inhibitory factor plays a role around the oral implant under oral environment. OBJECTIVE：To investigate the effect of macrophage migration inhibitory factor on the inflammation of bone marrow mesenchymal stem cels growing around the titanium implant. METHODS：First, bone marrow mesenchymal stem cels were seeded onto titanium cel culture disks to simulate the peri-implant environment in the mouth, and then, the cels were divided into four groups： control group, without any stimulation; lipopolysaccharide group, lipopolysaccharide-induced inflammation of bone marrow mesenchymal stem cels; non-specific smal interfering RNA （siRNA）＋lipopolysaccharide group, non-specific siRNA-transfected and lipopolysaccharide-induced cels; macrophage migration inhibitory factor siRNA＋lipopolysaccharide group, cels under the stimulation of lipopolysaccharide were transfected with＆nbsp;macrophage migration inhibitory factor. RESULTS AND CONCLUSION： Using flow cytometry, the cels expressing over 95% CD29 and CD90 as wel as less than 5% CD 45 were selected in the experiment. Cel counting kit-8 test showed that macrophage migration inhibitory factor siRNA＋lipopolysaccharide had no influence on the proliferation of bone marrow mesenchymal stem cels. Lipopolysaccharide significantly stimulated the inflammatory reactions of bone marrow mesenchymal stem cels, which was 15-20 times of the control group （P 〈 0.01）. However, compared with the lipopolysaccharide group, the levels of interleukin-1β, interluekin-6 and tumor necrosis factor-α were increased significantly after transfection with macrophage migration inhibitory factor siRNA＋lipopolysaccharide stimulation （P 〈 0.01）. These findings indicate that lipopolysaccharide can promote inflammation of bone marrow mesenchymal stem cels around the oral implant, but macrophage migration inhibitory factor siRNA can, to some extent, inhibit the occurrence of inflammation.