Abstract:

BACKGROUND: Vascular endothelial growth factor has an important role in bone repair by promoting angiogenesis, and by stimulating and affecting hemotaxis, proliferation, survival, and activity of major skeletal cells, including osteoblasts, osteoclasts, and chondrocytes.
OBJECTIVE: To optimize the conditions of transfection of recombinant pEGFP/hVEGF165 plasmid into rabbit osteoblasts by liposome and to provide experimental basis for constructing new seed cells for tissue engineered bone.
METHODS: Isolation, culture and identification of rabbit osteoblasts was conducted. Using different amounts and ratio of plasmid and liposome, in the 24-well plate, recombinant plasmid pEGFP/hVEGF165 was transfected into rabbit osteoblasts by liposome. Forty-eight hours after transfection, the transfention ratio was detected by inverted fluorescence microscope.
RESULTS AND CONCLUSION: The interaction between liposomes and plasmid could obviously affect transfention ratio. And under the conditions of plasmid liposome 2.5 μL and plasmid 1.2 μg in the 24-well culture plates, there was the highest transfection efficiency (56%). The optimal transfection conditions of recombinant plasmid pEGFP/hVEGF165 into rabbit osteoblasts in vitro were liposome 2.5 μL and plasmid 1.2 μg.