金属离子Cu^(2+)、Fe^(3+)对水凝胶RADA16-NBD培养条件下成骨细胞生长与分化的影响  

Effect of Cu^(2+) and Fe^(3+) on osteoblast growth and differentiation in hydrogel RADA16-NBD

作　　者：石金明[1,2] 赵刚[2] 阮蔷 

机构地区：[1]佳木斯大学研究生学院,黑龙江省佳木斯市154002 [2]佳木斯大学第二附属医院正畸科,黑龙江省佳木斯市154002

出　　处：《中国组织工程研究》2016年第16期2347-2353,共7页Chinese Journal of Tissue Engineering Research

基　　金：佳木斯大学研究生科技创新项目(LZZ2014_027);黑龙江省自然科学基金项目(H201487)~~

摘　　要：背景:在自组装功能性RADA16-NBD水凝胶中进行成骨细胞MC3T3-E1体外培养已较成熟,且骨骼的正常代谢与多种金属离子(如铁、铜、锌、锰等)有关已被证实。目的:观察Cu^(2+)和Fe^(3+)对自组装功能性RADA16-NBD水凝胶培养下成骨细胞MC3T3-E1增殖、分化等相关方面的影响。方法:将成骨细胞与自组装功能性RADA16-NBD水凝胶进行三维培养,分3组干预,分别加入Cu^(2+)、Fe^(3+)、不含血清培养基(对照组),培养24,48,72 h,CCK-8法检测细胞增殖;培养1,3,5 d,检测细胞碱性磷酸酶活性;培养21 d时,观察细胞钙化结节情况;Transwill小室培养24 h,观察细胞迁移能力。结果与结论:①细胞增殖:Cu^(2+)组、Fe^(3+)组培养24,48 h的细胞增殖高于对照组(P<0.05,P<0.01);培养72 h时,3组细胞增殖无差异;②细胞碱性磷酸酶活性:Cu^(2+)组、Fe^(3+)组培养1,3,5 d的细胞碱性磷酸酶活性高于对照组(P<0.05),Cu^(2+)组培养3,15 d的细胞碱性磷酸酶活性高于Fe^(3+)组(P<0.05);③细胞迁移:Cu^(2+)组迁移细胞数量多于Fe^(3+)组(P<0.05);④结果表明:Cu^(2+)和Fe^(3+)均可促进MC3T3-E1成骨细胞的增殖、分化及迁移,但Cu^(2+)对成骨细胞的影响要优于Fe^3。BACKGROUND： It is a mature technology to culture MC3T3-E1 cells in the self-assembling peptide hydrogel, RADA16-NBD. Moreover, it is confirmed that a variety of metal ions, such as Fe, Cu, Zn, Mn, are involved in normal bone metabolism. OBJECTIVE： To observe the effect of Cu（2＋） and Fe（3＋） on the proliferation and differentiation of MC3T3-E1 cells cultured in the self-assembling peptide hydrogel, RADA16-NBD. METHODS： Osteoblasts cultured with RADA16-NBD were divided into three groups and respectively cultured in culture medium containing Cu（2＋）, Fe（3＋） or serum-free medium（control group）, respectively. After 24, 48 and 72 hours, cell proliferation was detected by cell counting kit-8. After 1, 3, 5 days, alkaline phosphatase activity was detected. At 21 days, formation of calcified nodules was observed. Cell migration ability of cells was observed at 24 hours of Transwill chamber culture. RESULTS AND CONCLUSION： Compared with the control group, the proliferative ability of cells cultured in the Cu（2＋）, Fe（3＋） groups was significantly higher（P〈0.05, P〈0.01）. At 72 hours of culture, there was no difference in the cell proliferation among the three groups. At 1, 3, 5 days of culture, the alkaline phosphatase activity in the Cu（2＋）, Fe（3＋） groups was significantly higher than that in the control group（P〈0.05）; while at 3 and 5 days of culture, the alkaline phosphatase activity in the Cu（2＋） group was significantly higher than that in the Fe（3＋） group（P〈0.05）. In addition, the number of migrated cells was higher in the Cu（2＋） group than the Fe（3＋） group（P〈0.05）. These findings indicate that both Cu（2＋） and Fe（3＋）, especially the former one, can promote MC3T3-E1 cell proliferation, differentiation and migration.

关 键 词：水凝胶 成骨细胞 细胞增殖 组织工程 生物材料 材料相容性 自组装 RADA16-NBD水凝胶 金属离子 三维培养 Transwill小室 黑龙江省自然科学基金 

分 类 号：R318[医药卫生—生物医学工程]