title: Bone morphogenetic protein-2 (BMP-2) and vascular endothelial growth factor (VEGF) transfection to human periosteal cells enhances osteoblast differentiation and bone formation.
abstract: Periosteum has been demonstrated to contain mesenchymal progenitor cells differentiating to osteoblasts, and both bone morphogenetic protein-2 (BMP-2) and vascular endothelial growth factor (VEGF) may play important roles in cell-based approaches to bone regeneration. The purpose of this study was to evaluate the feasibility and efficacy of BMP-2 and/or VEGF on periosteal cell differentiation to osteoblasts in vitro and ectopic bone formation in vivo. Human periosteum-derived cells were transfected with BMP-2, VEGF, BMP-2 + VEGF, or vehicle as a control by non-viral gene transfer and then cultured and implanted to nude mice intramuscularly. Real-time polymerase chain reaction analysis of the culture revealed that transgenes for BMP-2 and BMP-2 + VEGF induced more mRNA expression of alkaline phosphatase, collagen type I, and osteocalcin than VEGF and vehicle treatments; additionally, alizarin red S staining, alkaline phosphatase staining, and alkaline phosphatase activity were significantly higher in the BMP-2 + VEGF transgene than in the other versions. After implantation, ectopic bone was observed at 4 weeks and greatly increased at 8 weeks in all groups. In particular, the combination of BMP-2 and VEGF formed significantly more bone at 4 weeks, and VEGF transfection resulted in more blood vessels relative to the conditions without VEGF. Thus, VEGF might enhance BMP2-induced bone formation through modulation of angiogenesis.
authors.affiliation: Section of Oral Implantology and Regenerative Dental Medicine, Department of Masticatory Function Rehabilitation, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo, Japan. mayu.irm@tmd.ac.jp
authors.forename: Mayurach
authors.initials: M
authors.lastname: Samee
authors.forename: Shohei
authors.initials: S
authors.lastname: Kasugai
authors.forename: Hisatomo
authors.initials: H
authors.lastname: Kondo
authors.forename: Keiichi
authors.initials: K
authors.lastname: Ohya
authors.forename: Hitoyata
authors.initials: H
authors.lastname: Shimokawa
authors.forename: Shinji
authors.initials: S
authors.lastname: Kuroda
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism / Q000502:physiology; D000818:Animals; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000235:genetics*; D002454:Cell Differentiation / Q000235:genetics; D002469:Cell Separation; D004247:DNA / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D051379:Mice; D008819:Mice, Nude; D010006:Osteoblasts / Q000502:physiology*; D015675:Osteocalcin / Q000096:biosynthesis / Q000235:genetics; D010011:Osteocytes / Q000502:physiology*; D010012:Osteogenesis / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D016212:Transforming Growth Factor beta / Q000235:genetics*; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics*
chemical_list: C525718:BMP2 protein, human; C525719:Bmp2 protein, mouse; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D016212:Transforming Growth Factor beta; D042461:Vascular Endothelial Growth Factor A; D015675:Osteocalcin; D004247:DNA; D000469:Alkaline Phosphatase