Effects of different copper content of titanium-copper alloy on adhesion and migration of osteoblasts

LI Yue; ZHANG Yan-ting; HE Jing; MA Kai; WANG Min-ya; LI Sheng-yi; DENG Chun-fu; ZHANG Er-lin; ZHAO Baohong

doi:10.19538/j.kq.2017.06.008

Abstract：Objective To study the effect of different copper content of titanium-copper alloy on adhesion and migration of osteoblasts,and to find the optimal copper content.Methods The titanium-copper alloy with copper content of 2％,5％ and 10％ was prepared and named Ti-2wt％ Cu,Ti-5wt％ Cu and Ti-10wt％ Cu,respectively.The pure titanium material without copper was used as the control,Ti group.Mouse MC3T3-E1 osteoblasts were cultured on the surface of these materials.(1)Morphological observation of osteoblasts:the surface morphology of osteoblasts was observed;the cytoskeleton-actin staining was observed.(2)The proliferation of osteoblasts was detected by MTT assay.(3) Cell migration was analyzed by cell scratch test.SPSS 17.0 software was used for statistical analysis.Results (1)Under scanning electron microscope:cells showed more rapid and fuller adhesion and expansion on the Ti-5wt％Cu,Ti-10wt％Cu when the cells were seeded for 6 and 24 hours,but Ti-5wt％ Cu was more obvious.(2)By phalloidine staining:the cytoskeleton actin filaments were observed only in the Ti-10wt％Cu group when the cells were inoculated with phalloidin for 2h.The spreading area of Ti-5wt％Cu was the largest at 4h and 24h.(3)The MTT results showed:the number of osteoblasts on the surface of materials increased with the increase of copper content,that is,Ti-10wt％ Cu ＞ Ti-5wt％ Cu ＞ Ti-2wt％ Cu ＞ Ti,the difference being statistically significant (P ＜ 0.05).However,there was no significant difference between Ti-1Owt％ Cu and Ti-5wt％ Cu on the 7th day.(4)After 24 and 48 hours of scratches,the average width of scratches was Ti ＞ Ti-2wt％ Cu ＞ Ti-1Owt％ Cu＞Ti-5wt％ Cu and there was significant difference among the four groups (P ＜ 0.05).Conclusion In the range of copper content detected in this experiment,osteoblast adhesion,the formation of cytoskeleton and cell migration are the best in the range of 5％ of copper content.