金属离子Cu2+、Fe3+对水凝胶RADA16-NBD培养条件下成骨细胞生长与分化的影响

摘要

BACKGROUND:It is a mature technology to culture MC3T3-E1 cels 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 Cu2+and Fe3+ on the proliferation and differentiation of MC3T3-E1 cels 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 Cu2+, Fe3+ or serum-free medium (control group), respectively. After 24, 48 and 72 hours, cel proliferation was detected by cel counting kit-8. After 1, 3, 5 days, alkaline phosphatase activity was detected. At 21 days, formation of calcified nodules was observed. Cel migration ability of cels was observed at 24 hours of Transwil chamber culture. RESULTS AND CONCLUSION:Compared with the control group, the proliferative ability of cels cultured in the Cu2+, Fe3+ groups was significantly higher (P < 0.05,P < 0.01). At 72 hours of culture, there was no difference in the cel proliferation among the three groups. At 1, 3, 5 days of culture, the alkaline phosphatase activity in the Cu2+, Fe3+ 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 Cu2+ group was significantly higher than that in the Fe3+ group (P < 0.05). In addition, the number of migrated cels was higher in the Cu2+ group than the Fe3+ group (P < 0.05). These findings indicate that both Cu2+ and Fe3+, especialy the former one, can promote MC3T3-E1 cel proliferation, differentiation and migration.%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Cu2+和Fe3+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Cu2+、Fe3+、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；Transwil小室培养24 h，观察细胞迁移能力。　　结果与结论：①细胞增殖：Cu2+组、Fe3+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：Cu2+组、Fe3+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)，Cu2+组培养3，15 d的细胞碱性磷酸酶活性高于Fe3+组(P<0.05)；③细胞迁移：Cu2+组迁移细胞数量多于Fe3+组(P<0.05)；④结果表明：Cu2+和Fe3+均可促进MC3T3-E1成骨细胞的增殖、分化及迁移，但Cu2+对成骨细胞的影响要优于Fe3。