Abstract:

BACKGROUND: It has become one of the most promising fields for the application of growth factor in gene treatment. Bone morphogenetic protein (BMP) can induce mesenchymal cells into osteoblasts and new bone formation, which plays an important role in bone healing.
OBJECTIVE: To realize the continuous expression of BMP partly and to promote cell proliferation and differentiation through transfecting mesenchymal 3T3-E1 cells (MC3T3-E1) in vitro with BMP-2 via the chitosan-based carrier.
METHODS: Chitosan-BMP-2 complex was prepared by complex coacervation method. The ability of chitosan combined with BMP-2 was tested by agarose gel electrophoresis. The expression plasmid carrying BMP-2 gene was induced into osteoblasts by using chitosan nanoparticle vector transfection technique. The transfection efficiency was evaluated. Cell proliferation and intracellular alkaline phosphatase activity were tested by MTT assay.
RESULTS AND CONCLUSION: Electrophoresis diagram showed that chitosan could well combined with BMP-2 plasmid. BMP-2 target gene could be induced into osteoblasts successfully by chitosan, and the tranfection efficiency was about 35%. MTT assay results exhibited that alkaline phosphatase activity was increased. These findings suggest that chitosan nanoparticles carrier system can successfully coupled BMP-2 target gene and effectively transfected MC3T3-E1, as well as promote MC3T3-E1 proliferation and differentiation.