Ultrasound-mediated Microbubble Destruction Enhances Bone Morphogenetic Protein-2 Gene Expression In The Rat Bone Marrow Mesenchymal Stem Cells

Objective:To explore whether ultrasound-mediated microbubble destruction can enhance the expression efficiency of plasmid-GV147-rhBMP2-GFP for bone morphogenetic protein-2(BMP-2)in the rat bone marrow mesenchymal stem cells.Methods:BMSCs of 8- week old male Sprague-Dawley rats were separated and cultured. In the 3rd passage, the BMSCs were divided into three groups:group A was without any treatment, group B was given GV147-rhBMP2-GFP plasmid, group C was given GV147-rhBMP2-GFP plasmid and microbubbles plus ultrasonic irradiation. About forty-eight hours after transfection, the transient expression situation and the growth condition of cells was observed under fluorescence microscopy. RT—PCR was taken to evaluate the mRNA expression of rhBMP-2, and Western blot analysis was taken to evaluate the protein expression of BMP-2 respectively.Using SPSS13.0 software for statistical analysis of the data.Results:(1) Green fluorescence was observed in group B and C by fluorescence microscopy,while which was negative in group C.The green fluorescence of group C is more than that of group B.(2) RT—PCR assay showed that the mRNA expression of rhBMP-2 ingroup C was more than that in group B. By the way,Western-bolt assay showed that the expression of BMP-2 in group C was more than that in group B.Conclusions:Ultrasound-mediated microbubble destruction could enhance the transfection and expression efficiency of rhBMP-2 gene in the rat bone marrow mesenchymal stem cells. It will be a new gene therapy method for bone fracture.