阿伦膦酸盐在不同分化阶段对破骨细胞生成及骨吸收功能的影响

Effect of Alendronate on Osteoclast Generation and Bone Absorbing Function at Different Stages of Differentiation

目的:研究阿伦膦酸盐作用在不同分化阶段对破骨细胞生成及骨吸收功能的影响。方法:体外分离骨髓单核细胞,用30μg/L M-CSF对其预诱导3d,然后将细胞分为A、B、C、D四组。A组(对照组),用50μg/L M-CSF+100μg/L RANKL进行破骨细胞诱导;B、C、D组除加入上述诱导因子外,在不同时间点加入0.1μmol/L阿伦膦酸盐(ALN),加入时间分别为:B组第0～2天加入,C组第3～4天加入,D组第5～6天加入。检测每组细胞正式诱导第6天TRAP染色情况及牙本质磨片吸收陷窝情况。结果:各组细胞均有TRAP阳性多核破骨细胞生成,并在牙本质磨片上形成吸收陷窝;但A组TRAP阳性多核细胞数目、吸收陷窝数目及陷窝面积最高,D组次之,B组最差。结论:阿伦膦酸盐在破骨细胞分化阶段作用越早,对破骨细胞生成的抑制效应越大,所形成破骨细胞骨吸收能力越差。

Objective：To study the effect of alendronate on osteoclast generation and bone absorbing function an different stages of osteoclast differentiation.Methods：Bone marrow mononuclear cells were isolated and underwent preinduction with 30ng/ml M-CSF for 3 days and then the cells were divided into 4 groups.Cells of group A（served as control） was underwent osteoclastogenic induction with 30ng/ml M-CSF and 50ng/ml RANKL.Despite of above osteoclastogenic induction agents,cells of group B,C,D were treated with 0.1uM alendronate at 0-2 day,3-4 day,5-6 day,respectively.Osteoclastogenesis was examined by TRAP staining 6 days after induction and dentin resorption lacunas were verified by scanning electron microscope（SEM）.Results：TRAP positive multinuclear cells were observed in all groups of cells and resorption lacunaes were formed in all groups.However,group A showed the most TRAP positive multinuclear cells and the largest resorption lacunaes among 4 groups,followed in turn by group D and group C,while group B had the least TRAP positive multinuclear cells and the smallest resorption lacunaes.Conclusion：The results of the study suggest that earlier alendronate is added during osteoclast differentiation,the more inhibition effects alendronate will exert on osteoclast generation and bone absorbing function.