摘要
[目的]观察不同浓度金属磨损颗粒对RAW264.7在体外分化成破骨细胞的影响,明确浓度与破骨细胞分化数量的关系。[方法]真空球磨法制备人工关节磨损颗粒:RANKL诱导RAW264.7体外分化成破骨细胞,通过TRAP染色,电镜扫描检测骨片吸收陷窝来鉴定破骨细胞;不同浓度人工关节磨损颗粒混悬液作用RAW264.7,并用RANKL诱导后第7d,TRAP染色后,光镜下计数破骨细胞数量。[结果]不同浓度磨损颗粒作用于RAW264.77d后,显微镜下计数破骨细胞数量,结果显示随着磨损颗粒混悬液浓度增加,RANKL诱导生成的破骨细胞增多,低、中、高浓度3组破骨细胞数均显著高于空白对照组(P<0.05),中、高浓度组破骨细胞数均显著高于低浓组(P<0.05),高浓度组破骨细胞数亦显著高于中浓组(P<0.05)。[结论](1)RAW264.7是一种较好的破骨前体细胞模型,RAW264.7诱导形成破骨细胞的方法简便易行,所获得破骨细胞均一性好;(2)人工关节金属磨损颗粒为RAW264.7细胞向具有骨质吸收功能的破骨细胞转化发挥正向作用,而且与混悬液的浓度有量效关系。
[Objective] To observe the effects of different conscentrations of metal wear particles on RAW 264.7 in vitro to differentiate into osteoclast cells,and clarify the relationship between the concentration of metal wear particles and the number of osteoclast differentiation.[Method]Preparation of wear particles of artificial joints were made by Vacuum Ball Milling.RANKL induced RAW 264.7 to differentiate into osteoclasts in vitro.Osteoclasts were treated with tartrate resistant acid phosphatase （TRAP） staining and bone resorption pits were detected by scanning electron microscop.Using different concentrations of artificial joint wear particles suspension co-culture with RAW 264.7 7 days after induction by RANKL,we counted the number of osteoclasts under light microscope after TRAP staining.[Result]The results showed that with the wear particles suspension concentration increasing.RANKL-induced osteoclast formation increased.The number of osteoclasts of the low,medium and high concentrations groups were significantly higher than in the control group （P0.05）.Osteoclast number of the medium and high concentration groups was significantly higher than in the low concentration group （P0.05）.Osteoclast number in high concentration group was significantly higher than in the medium concentration group （P0.05）.[Conclusion]（1）RAW 264.7 cells can be used as a preferable preosteoclast model.This method has good repeatability and feasibility.（2）Metal wear particles of artificial joints play a positive role in transformation of RAW 264.7 cells into osteoclasts which has bone resorption function and there is concentration effect of promotion osteoclast formation of the suspension of concentration.
出处
《中国矫形外科杂志》
CAS
CSCD
北大核心
2011年第4期316-319,共4页
Orthopedic Journal of China
基金
国家自然科学基金资助项目(编号:30760255)
关键词
人工关节
金属磨损颗粒
真空球磨
破骨细胞
joint prostheses
metal wear particles
vacuum ball milling
osteoclast
