Influence on the osteogenic activity of the human bone marrow mesenchymal stem cells transfected by liposome-mediated recombinant plasmid pIRES-hBMP2-hVEGF165 in vitro.
Bone marrow mesenchymal stem cells (BMSCs) is an attractive option for use as seed cells in tissue engineering strategies. Bone morphogenetic proteins (BMPs) to be involved in the formation of various tissue types including bone, cartilage, tendon, and ligament. Vascular endothelial growth factor (VEGF) is a promising reagent for inducing angiogenesis. Constructed a coexpressing vector of human bone morphogenetic protein 2 (hBMP-2) and vascular endothelial growth factor 165 (hVEGF165). The vectors were transfected into proliferated BMSCs isolated from healthy adult bone marrow. The expression of hBMP-2 and hVEGF165 genes of BMSCs were assayed by Western-blot, and the level of alkaline phosphatase activities of BMSCs was determined by RT-PCR analysis of osteocalcin mRNA expression. The levels of collagen I by immunohistochemical staining were also determined.BMSCs transfected with reconstructed plasmid pIRES-hBMP-2-VEGF165 could secret a high level of BMP-2 and hVEGF165. The production of type I collagen, the activity of alkaline phosphatase, and the expression of osteocalcin mRNA were also significantly improved in the transfected BMSCs, compared with the control group.The exogenous hBMP-2 and hVEGF165 genes can be expressed constitutively in the transfected BMSCs, and the lineage-committed differentiation abilities of BMSCs containing combination of genes BMP-2 and VEGF165 are enhanced.
Guo-ping
Wu
W
Department of Plastic and Burns Surgery, The Affiliated Hospital of Luzhou Medical College, Luzhou, Sichuan, People's Republic of China. drgpwu@yahoo.com.cn <drgpwu@yahoo.com.cn>
Xiao-chuan
He
H
Zhi-hui
Yang
Y
Li
Guo
G
D000328:Adult; D015153:Blotting, Western; D016026:Bone Marrow Transplantation / Q000379:methods*; D055396:Bone Morphogenetic Protein 2 / Q000235:genetics* / Q000378:metabolism; D024042:Collagen Type I / Q000235:genetics; D015870:Gene Expression / Q000235:genetics; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D007124:Immunoenzyme Techniques; D008081:Liposomes; D045164:Mesenchymal Stem Cell Transplantation / Q000379:methods*; D015675:Osteocalcin / Q000235:genetics; D010012:Osteogenesis / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D011995:Recombination, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection / Q000379:methods*; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics* / Q000378:metabolism
C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; D024042:Collagen Type I; D008081:Liposomes; D012333:RNA, Messenger; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D015675:Osteocalcin