钴纳米粒子和钴离子对成骨样细胞MG-63体外生长的影响  

Effects of cobalt nanoparticles and ions on osteoblast- like cells MG-63 in vitro

作　　者：杨乐[1] 徐长明[1] 茹江英[1] 章庆俊[1] 韩庆林[2] 刘璠[2] 

机构地区：[1]中国人民武装警察部队江苏省总队医院骨科,扬州225003 [2]南通大学附属医院骨科,南通226001

出　　处：《中国矫形外科杂志》2015年第6期541-547,共7页Orthopedic Journal of China

基　　金：国家自然科学基金资助项目(编号:21171743)

摘　　要：[目的]研究钴纳米粒子(Co-NPs)和钴离子(Co2+)对成骨细胞体外生长的影响,探讨金属磨损粒子与假体周围骨溶解的关系。[方法]分为三组:空白对照组、Co-NPs组、Co2+组。体外培养成骨样细胞MG-63,不同时间点观察细胞形态变化并计数,Real-time PCR检测Caspase3、ATM、BAX、Bcl-2、P21等凋亡基因的变化,ELISA方法检测培养上清中TNF-α和IL-6的表达情况,Real-time PCR检测细胞TNF-α和IL-6 mRNA水平的表达。ELISA检测细胞中碱性磷酸酶(ALP)的表达。[结果](1)与对照组相比,不同浓度Co2+组和高浓度Co-NPs组的细胞计数明显减少,高浓度Co2+组细胞形态发生变化;(2)Real-time PCR结果显示:相比于对照组,不同浓度Co2+组和高浓度Co-NPs组的MG-63细胞中凋亡相关基因Caspase3、ATM、BAX、P21表达上调,而BCl-2均未发生明显上下调变化;(3)ELISA结果显示:不同浓度Co2+组和高浓度Co-NPs组的MG-63细胞培养上清中TNF-α表达上调,而IL-6表达未发生明显变化;Real-time PCR的结果也显示TNF-α的mRNA水平上调,IL-6的mRNA水平未发生明显变化;(4)ALP检测结果显示:不同浓度Co2+和高浓度Co-NPs抑制ALP的表达。[结论]不同浓度Co2+和高浓度Co-NPs能抑制成骨细胞增殖和活性,促进其凋亡,同时促进其释放TNF-α。[Objective]To investigate the effect of cobalt nanoparticles and ions on human MG- 63 osteoblasts in vitro,to further explore the relationship of metal wear particles and osteolysis. [Method]Grouping： control group,Co- NPs group,and Co2 ＋group. The Osteoblast- like cell line MG- 63 was cultured in vitro,once Co- NPs,and Co2 ＋were added,changes in cell morphology were observed at different time point,real- time PCR was adopted to show the change of apoptotic genes Caspase3,ATM,BAX,Bcl- 2,P21,etc,and the expression of TNF- α and IL- 6 in the culture supernatant was detected by real- time PCR. The expression of alkaline phosphatase（ ALP） in the cells was detected by ELISA. [Result]（ 1） Compared with the control group,the cell count was obviously reduced both in different concentrations of Co2 ＋group and high concentration of Co-NPs group,the cell morphology was changed in high concentration of Co- NPs group.（ 2） Real- time PCR results showed that：compared with the control group,expression of apoptosis- related genes（ Caspase3、ATM、BAX、P21） in MG- 63 was increased in different concentrations of Co2 ＋group and high concentration of Co- NPs group,but no changes were found about Bcl- 2.（ 3） ELISA results showed that： the expression of TNF- α in the culture supernatant of MG- 63 was increased both in different concentrations of Co2 ＋group and high concentration of Co- NPs group,but no changes were found about IL- 6. The real- time PCR results also showed that： the level of mRNA was also increased in TNF- α,and the mRNA level did not change in IL- 6.（ 4） The results of ALP test showed that： different concentrations of Co2 ＋and high concentration of Co- NPs inhibited the expression of ALP. [Conclusion]Both different concentrations of Co2 ＋and high concentration of Co- NPs have the inhibition effects on proliferation and activity of osteoblast,and promotes the apoptosis and the release of TNF- α.

关 键 词：Co-NPs Co2+ 成骨细胞 凋亡 成骨 

分 类 号：R318.08[医药卫生—生物医学工程]