不同浓度破骨细胞分化因子和巨噬细胞集落刺激因子体外诱导大鼠破骨样细胞形成的研究  

Differentiation of osteoclast-like cells induced by using different concentrations of M-CSF and RANKL

作　　者：王晓庚[1] 刘文佳[1] 周洪[1] 李昂[2] 

机构地区：[1]西安交通大学医学院附属口腔医院正畸科,西安710004 [2]西安交通大学医学院附属口腔医院医学研究中心,西安710004

出　　处：《口腔医学》2008年第4期169-172,共4页Stomatology

基　　金：国家自然科学基金资助项目(30471911)

摘　　要：目的探讨破骨细胞分化因子和巨噬细胞集落刺激因子在大鼠破骨样细胞形成、分化中的作用,为进一步研究正畸牙齿移动的生物力学机制奠定基础。方法应用不同浓度的破骨细胞分化因子和巨噬细胞集落刺激因子对提取的SD大鼠骨髓单个核细胞进行诱导培养,分别于培养1、3、5、7 d利用TRAP染色、骨吸收陷窝检测等对单核及多核破骨样细胞的生成进行鉴定,对TRAP(+)的细胞进行计数和统计学分析。结果巨噬细胞集落刺激因子和破骨细胞分化因子对TRAP(+)的单核及多核破骨样细胞的诱导分化作用与时间有关:第3 d时逐渐增多,以第5 d和第7 d为最多;缺乏巨噬细胞集落刺激因子时,骨髓单个核细胞不能有效地分化为TRAP(+)的细胞;在破骨细胞分化因子浓度一定的条件下(50 ng/ml),TRAP(+)的细胞数与巨噬细胞集落刺激因子的浓度有关,当其浓度超过30 ng/ml时,TRAP(+)细胞数不再显著增加,曲线趋于平缓;当巨噬细胞集落刺激因子浓度一定的条件(50 ng/ml)下,TRAP(+)的细胞数与破骨细胞分化因子的浓度无关。结论以巨噬细胞集落刺激因子和破骨细胞分化因子作为诱导因子,对大鼠骨髓单个核细胞进行诱导培养,可获得TRAP(+)吸收陷窝(+)的破骨样细胞,且巨噬细胞集落刺激因子30 ng/ml、破骨细胞分化因子50 ng/ml时具有最强的生物学效应。Objective To investigate the effects of macrophage of colony-stimulating factor （M-CSF） and receptor activator of NF-κB ligand （RANKL） on the differentiation and formation of osteoclast-hke cells and to lay the foundation for the further study of biology mechanism for tooth movement.Methods The bone marrow mononuclear cells of rats were treated with different concentrations of M-CSF and RANKL. After having been cultured for 1,3,5 and 7 days,the number of tartrate-resistant acid phosphatase （TRAP） positive cells was measured by TRAP staining and pit staining.Results The effects of M-CSF and RANKL on the differentiation of osteoclastlike cells were related to time, the TRAP-positive mononuclear and multinucleated cells were formed on the third day, and the maximum number of TRAP-positive cells appeared on the fifth and seventh day. Without M-CSF, the bone marrow mononuclear cells couldn＇t differentiate to TRAP-positive cells. When the concentration of RANKL was 50 ng/ml, the number of TRAP-positive cells was related to the concentration of M-CSF; when the concentration of M-CSF exceeded 30 ng/ ml, the number of TRAP-positive cells didn＇ t increase significantly; when the concentration of M-CSF was 50 ng/ml, the number of TRAP-posi- tive cells had no relation to the concentration of RANKL. Conclusion With the cooperation of M-CSF, RANKL could induce TRAP-positive osteoclast formation in rat bone marrow culture, and the M-CSF 30 ng/ml, RANKL 50 ng/ml were the most effective.

关 键 词：破骨样细胞 体外培养 破骨细胞分化因子 巨噬细胞集落刺激因子 

分 类 号：R783.5[医药卫生—口腔医学]