Osteogenesis induced in goat bone marrow progenitor cells by recombinant adenovirus coexpressing bone morphogenetic protein 2 and basic fibroblast growth factor

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.

Mesenchymal stromal cells for bone sarcoma treatment: Roadmap to clinical practice

“…Contemporary engineering and regenerative medicine techniques [165] using molecular [166] , [167] , [168] and mechanical “guidance” [169] , [170] , [171] can differentiate ADMSCs into various tissue types. Previous animal studies (157,155) demonstrated the regenerative ability of SCs that were applied either intravenously or topically, loaded on a different type of scaffolds [172] , [173] , [100] .…”

Expression of osteoprotegerin and receptor activator for the nuclear factor‐κB ligand in XACB/LV‐bFGF/MSCs transplantation for repair of rabbit femoral head defect necrosis

“…13,14 At the molecular level, OPG, RANK, and RANKL play an important role in this process. 15 The OPG/RANK/RANKL system has been identified as an important pathway for bone metabolism in recent years, 9 and has been widely used as a major marker for bone metabolism research. [16][17][18] OPG, also known as the broken bone cell inhibitory factor, is a growth factor receptor that belongs to the tumor necrosis factor (TNF) receptor family.…”

“…8 In the induction of bone marrow stromal cells into osteoblasts or bone progenitor cells in the process, the commonly used cytokines are basic fibroblast growth factor (bFGF). 9 Osteoprotegerin (OPG) and the nuclear factor-κB receptor activator ligand (RANKL) as a key factor in osteoclast differentiation and bone resorption 10 can directly affect the bone cell function, and is a common pathway for bone resorption. In this study, Osteoprotegerin and nuclear factor-kappa B were treated by xenogeneic antigens of cancellous bone (XACB)/lentiviral (LV)-bFGF)/MSCs transfected with LV-transfected bFGF-transfected MSCs to explore the therapeutic effect of tissue engineered bone on necrosis of femoral head and provide experimental evidence for the clinical application of bone tissue engineering in the future.…”

“…Bone marrow stromal cells are pluripotent stem cells that can differentiate into osteoblasts and osteoblasts have excellent potential for use . In the induction of bone marrow stromal cells into osteoblasts or bone progenitor cells in the process, the commonly used cytokines are basic fibroblast growth factor (bFGF) . Osteoprotegerin (OPG) and the nuclear factor‐κB receptor activator ligand (RANKL) as a key factor in osteoclast differentiation and bone resorption can directly affect the bone cell function, and is a common pathway for bone resorption.…”