Acceleration of distraction osteogenesis with recombinant human bone morphogenetic protein-2 transfected into normal mesenchymal cell.

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We made the plasmid vector coding fusion protein of green fluorescein protein and recombinant human bone morphogenetic protein-2(rhBMP-2).The plasmid vector were transfected into C2C12, and NIH3T3, fibroblast cell line of mouse with lipofection.Expression of mRNA of rhBMP-2 were increased.Transfected cells showed GFP expression with laser scanning microscopy, but no relation between expression of rhBMP-2 and GFP with immunohistological study.C2C12 cells transfected with GFP-rhBMP-2 shows increasing of ALP activity.Type I collagen and transfectd bone marrow stromal cells harvested from rat's femur, were transplantation into rat biceps femorins and 5mmfemoral bone defect made with external fixators.Rhoentogenographic study showed no bone formation.After plantation of transfected cells and collagen, lengthening of femur were done 0.5mm per day for 10days.The results were the same.Gene coding GFP-rhBMP-2 were transfected into normal mesenchymal cells, but did not show increaeing of production of fusion protein with immunohistochemically.Transfected cellsshowed slight increase of ALP activity under in vivocircumference, but dose not showed heterotopic bone formation or acceleration of bone formation.It needed to change the construction of GFP-rhBM-2 gene, or to swich the vector from plasmid to virus.