Effect Of CPPF/CPC On OPG/RANKL Of Human Osteoblasts In Vitro

Objective: To preliminary study the effect of CPPF/CPC on OPG/RANKL ofcultured human osteoblasts in vitro, and to provide theoretical basis of clinic application ofCPPF/CPC.Methods:1. Mixed the CPPF and CPC according to mass ratio of1∶9, and addedcuring liquid by solid-to-liquid ratio of3∶1, then poured into a cylindrical mold withdiameter of3cm,and thickness of3mm.2.Cultured the human osteoblasts MG63and modified the cell concentration to1X105,then inoculated the cultured cells with two groups. the first group inoculated withcell culture plate composed of self-made CPPF/CPC called artificial bone group andsecond group inoculated with common6-well culture plate called Control Group. Thenumber of cell was measured by Count plate method after cells were cultured for0h,6h,12h,24h,36h,48h of the two groups.3. RT-PCR were used to detecte the mRNA expression of the OPG and RANKL after cellscultured for36h.4. Western blot were used to detecte the protein expression of the OPG and RANKL aftercells cultured for36h.Results:1.After cell cultured for of6h,12h,24h, the cell number of artificial bonegroup were significantly lower than control group (p<0.05), but there had no differencewith two groups after cultured for36h,or48h(p>0.05). 2.After cell cultured for36h, the expression of OPG and RANKL at the mRNA level ofartificial bone group were significantly elevated than the Control Group (p<0.05).3.After cell cultured for36h, the expression of OPG and RANKL at the protein level ofartificial bone group were significantly elevated than the Control Group (p<0.05), but therewere no significant differences in the ratio of OPG/RANKL。Conclusion:1. CPPF/CPC(1:9) did not change CPC that not conducive to theperformance of adhesion, proliferation of bone cells.2. conparted with CPC，after CPPF/CPC(1:9) artificial bone implanted into the body of thebone formation and absorption process was improved obviously, and the absorptionprocess faster than osteogenesis process.3. CPPF/CPC(1:9) composite artificial bone have good osteoconductivity bone inductionand bone conduction, is expected to become the bone repair and substitute material forhuman body.