Effect of Cu2+ and Fe3+ on osteoblast growth and differentiation in hydrogel RADA16-NBD / 中国组织工程研究

ABSTRACT

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.