Effect of r-Mt-Cpn10 on human osteoblast cells

To observe the effect of recombinant mycobacterium tuberculosis heat shock protein 10 (r-Mt-Cpn10) on human osteoblast proliferation, cell cycle, alkaline phosphatase, calcium nodules and the expression of Receptor Activator of Nuclear Factor KB Ligand (RANKL) and Osteoprotegerin (OPG).

Osteoblasts were cultured in the medium with different concentration of r-Mt-Cpn10. No drug was added to the medium in the control group. The effect of r-Mt-Cpn10 on osteoblast proliferation was detected by MTT. The 3rd generation of osteoblasts was taken and detected the effect on the activity of osteoblasts secreted alkaline phosphatase on 1, 3, 5, 7 and 9 d of cell culture. The effects of different concentrations of r-Mt-Cpn10 on the expression of RANKL and OPG were detected.

The r-Mt-Cpn10 blocked osteoblasts in the G2/M phase and G1 to S phase. Compared with the control group, the r-Mt-Cpn10 with different concentrations inhibited the proliferation and alkaline phosphatase activity of osteoblast (P<0.05), the number of calcium nodules formation was significantly reduced. The r-Mt-Cpn10 increased the expression of RANKL in a dose-dependent manner and reduced the expression of OPG (P<0.01).

The inhibition of r-Mt-Cpn10 on the osteoblast proliferation and alkaline phosphatase activity was achieved by osteoblasts arrest in G2/M phase and G1 to S phase, it can also regulate the expression of RANKL and OPG which affecting local bone metabolic balance.