Bone Morphogenetic Protein-2 (BMP-2) and Vascular Endothelial Growth Factor (VEGF) Transfection to Human Periosteal Cells Enhances Osteoblast Differentiation and Bone Formation

Abstract

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.

A simple rocker‐induced mechanical stimulus upregulates mineralization by human osteoprogenitor cells in fibrous scaffolds

“…Briefly, removed tissue of approximately 1 × 1 cm 2 was rinsed with phosphate buffered saline (PBS; OXOID Limited, Hampshire, England) containing 100 mg/ml P/S, cut into smaller pieces and added to 0.25% collagenase type II in EM without fetal calf serum and incubated at 37 °C for 3 h (Samee et al, 2008). After this time, the cells were centrifuged, the supernatant removed and the cells placed into a 25‐cm 2 tissue‐culture flask in 2 ml of fresh EM.…”

“…Harvesting bone marrow MSC (hBMSC) from the iliac crest incurs less pain at the operation site, fewer arterial and nerve injuries, and a lower risk of infection compared with harvesting of traditional iliac crest bone grafts (Samee et al, 2008). Tissue engineering strategies using MSCs from patients have shown promise for reconstruction of a cleft defect at the alveolar ridge region; resulting in a successful repair and support of tooth eruption over the following 2 years (Hibi, Yamada, Ueda, & Endo, 2006).…”

Emerging Ideas: Treatment of Precollapse Osteonecrosis Using Stem Cells and Growth Factors

“…Combined use of VEGF and BMP-2 [19,24,34,35,37,40] and BMP-4 [20,25,36] showed synergistic effects in osteogenesis. It has been shown that combined use of VEGF with one of the most potent BMPs, BMP-6 [23,26], enhances osteogenesis and angiogenesis of mouse bone marrow stem cells in vitro and in vivo [8], however, the effectiveness of this therapy for ON has not been determined.…”

“…In addition to its effects on angiogenesis, VEGF plays an important role in bone growth and repair by interacting with other growth factors [35,36]. However, overexpression of VEGF actually can adversely affect osteogenesis and decrease bone formation [25,[34][35][36][37]. In our in vitro study, the optimal ratio of VEGF and BMP-6 will be determined so that the undesired effects of VEGF can be avoided in the in vivo study.…”

“…Stem cell-mediated bone regeneration augmented with combined use of VEGF and BMPs has been reported but only for BMP-2 [19,24,34,35,37,40] and BMP-4 [20,25,36]. It has been shown that VEGF enhances BMP-6-induced osteogenesis of mouse stem cells in vitro and in vivo, however, the effectiveness of this therapy for ON has not been determined [8].…”