M-CSF、RANKL浓度及M-CSF预诱导对破骨细胞生成影响的研究  

Effects of M-CSF Concentration,RANKL Concentration and M-CSF Preinduction on Osteoclastogenesis

作　　者：董伟[1] 于静[1] 戚孟春[1] 白宇宏[1] 梁锐英[1] 陈槐卿[2] 

机构地区：[1]华北煤炭医学院口腔系,唐山063000 [2]四川大学华西医学中心生物医学工程研究室,成都610041

出　　处：《生物医学工程学杂志》2010年第6期1336-1340,共5页Journal of Biomedical Engineering

摘　　要：研究破骨细胞(OC)诱导因子不同浓度及诱导方式对破骨细胞生成及骨吸收功能的影响。体外分离骨髓单个核细胞,将细胞分为四组:A组,加入30 ng/ml巨噬细胞集落刺激因子(M-CSF)+50 ng/ml核因子κB受体激活因子配体(RANKL);B组,加入50 ng/ml M-CSF+100 ng/ml RANKL;C组,用30 ng/ml M-CSF进行预诱导,3 d后加入30 ng/ml M-CSF+50 ng/ml RANKL进行正式诱导;D组,用30 ng/ml M-CSF进行预诱导,3 d后加入50ng/ml M-CSF+100 ng/ml RANKL进行正式诱导。检测每组细胞正式诱导第6 d抗酒石酸酸性磷酸酶(TRAP)染色情况及正式诱导第9 d牙本质磨片吸收陷窝情况。各组细胞均有TRAP阳性多核破骨细胞出现,并在牙本质磨片上形成吸收陷窝,但B组和D组中TRAP阳性多核细胞数目、吸收陷窝数目及陷窝面积分别显著高于A组和C组;同时,C组和D组分别显著高于A组和B组。结果表明,较高诱导因子浓度(50 ng/ml M-CSF+100ng/mlRANKL)及用M-CSF预诱导,更有利于破骨细胞的分化及骨吸收能力的提高。This study was directed to the effects of macrophage-colony stimulating factors（M-CSF） concentration,recerptor activator of nuclear factor κB ligand（RANKL） concentration and M-CSF preinduction on osteoclastogenesis and the related resorption function.Bone marrow mononuclear cells were isolated and were divided into 4 groups.Group A underwent osteoclastogenic induction with the use of 30ng/ml M-CSF and 50ng/ml RANKL,while Group B received 50ng/ml M-CSF and 100ng/ml RANKL treatment.Both C and D Group underwent preinduction with the use of 30ng/ml M-CSF for 3days,and then they were treated with 30ng/ml M-CSF and 50 ng/ml RANKL,50ng/ml M-CSF and 100ng/ml RANKL,respectively.Osteoclastogenesis was examined by TRAP staining 6days after induction,and dentin resorption lacunae were detected by Scanning Electron Microscope 9 days after induction.TRAP positive multinuclear cells were observed in all groups of cells,and resorption lacunae were formed in all of them.However,more TRAP positive multinuclear cells were observed and more large resorption lacunae were deteced in groups B and D than in groups A and C,respectively.The number of TRAP positive cells,number of resorption lacunae and lacuna areas in groups C and D were also greater than those in groups A and B,respectively.Higher concentration of M-CSF and RANKL and preinduction with M-CSF may benefit osteoclastogenesis and increase resorption function of osteoclast.

关 键 词：破骨细胞 巨噬细胞集落刺激因子 核因子κB受体激活因子配体 抗酒石酸酸性磷酸酶 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]