The Effects of CGRP and rhBMP-2 on the Proliferation and ALP Activity of Rabbit Osteoblast-like Cells

Objective To study the synergistic effects of calcitonin gene-related peptide(CGRP)and recombinant human bone morphogenetic protein-2(rhBMP-2)on the generation and activity of rabbit osteoblast-like cells obtained from rabbit bone marrow stromal cells by induced mineralization.Method The osteoblast-like cells were obtained from rabbit bone marrow stromal cells by induced mineralization and subcultured to the 3rd passage used for the experiment.The cells were seeded separately into 96 well plates with approximately 2×10~6cells/well and incubated in different mediums.The samples were derided into 6 groups:①Group A,blank group,②Group B,0.05 ng/ml CGRP,③Group C,5 ng/ml CGRP,④Group D,100 ng/ml rhBMP-2,⑤Group E,5 ng/ml CGRP+100ng/ml rhBMP-2,⑥GroupF,0.05ng/ml CGRP+100 ng/ml rhBMP-2.The proliferation of osteoblast-like cells were measured by MTT and the activity of ALP studied by staining method.Result The optical density(OD)values of cells proliferation were higher in 100ng/ml rhBMP,0.05ng/ml CGRP+100ng/ml rhBMP-2 and 5ng/ml CGRP+100ng/ml rhBMP-2 groups than in normal control(p0.05),with the highest in 0.5ng/mlCGRP+100ng/ml rhBMP-2,and also higher in first three groups than in normal control for ALP activity(p0.05), but no obvious differences between first three groups(p0.05).Conclusion The combination of rhBMP-2 and CGRP has synergistic effect on the proliferation of rabbit osteoblast-like cells,but not for ALP.