Abstract
Bone morphogenetic protein 2 (BMP2) and basic fibroblast growth factor (bFGF) have been shown to exhibit a synergistic effect to promote bone repair and healing. In this study, we constructed a novel adenovirus with high coexpression of BMP2 and bFGF and evaluated its effect on osteogenic differentiation of goat bone marrow progenitor cells (BMPCs). Recombinant adenovirus Ad-BMP2-bFGF was constructed by using the T2A sequence. BMPCs were isolated from goats by density gradient centrifugation and adherent cell culture, and were then infected with Ad-BMP2-bFGF or Ad-BMP2. Expression of BMP2 and bFGF was detected by ELISA, and alkaline phosphatase (ALP) activity was detected by an ALP assay kit. In addition, von Kossa staining and immunocytochemical staining of collagen II were performed on BMPCs 21 days after infection. There was a high coexpression of BMP2 and bFGF in BMPCs infected with Ad-BMP2-bFGF. Twenty-one days after infection, ALP activity was significantly higher in BMPCs infected with Ad-BMP2-bFGF than in those infected with Ad-BMP2. Larger and more mineralized calcium nodules, as well as stronger collagen II staining, were observed in BMPCs infected with Ad-BMP2-bFGF than in those infected with Ad-BMP2. In summary, we developed a novel adenovirus vector Ad-BMP2-bFGF for simultaneous high coexpression of BMP2 and bFGF, which could induce BMPCs to differentiate efficiently into osteoblasts.
Highlights
Several studies have shown that bone morphogenetic protein (BMP) and basic fibroblast growth factor induce bone formation in ectopic and orthotopic sites in vivo [1,2]
Twenty-one days after infection, alkaline phosphatase (ALP) activity was significantly higher in bone marrow progenitor cells (BMPCs) infected with Ad-Bone morphogenetic protein 2 (BMP2)-basic fibroblast growth factor (bFGF) than in those infected with Ad-BMP2
The results showed that infection by AdBMP2-bFGF led to a small increase in cell viability, but there were no significant differences in cell viability between BMPCs infected with Ad-BMP2-bFGF or AdBMP2 and control (Figure 4)
Summary
Several studies have shown that bone morphogenetic protein (BMP) and basic fibroblast growth factor (bFGF) induce bone formation in ectopic and orthotopic sites in vivo [1,2]. Systemic coadministration of BMP and bFGF in preclinical models has been shown to stimulate bone deposition in skeletal tissues [3,4]. BMP2 is an important stimulator of bone formation by controlling the proliferation and differentiation of osteoblasts [5,6]. A recent study reported that BMP2 could modulate osteogenic differentiation of adipose-derived stem cells [7]. The osteogenic activity of bone marrow stromal cells (BMSCs) induced by both BMP2 and bFGF was greater than that induced by either bFGF or BMP2 alone [9]
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