Cell-mediated and direct gene therapy for bone regeneration

Wang

Experimental and Therapeutic Medicine

et al. 2013

Bone marrow-derived mesenchymal stem cells (MSCs) are dominant seed cell sources for bone regeneration. Bone morphogenetic proteins (BMPs) initiate cartilage and bone formation in a sequential cascade. Vascular endothelial growth factor (VEGF) is an essential coordinator of extracellular matrix remodeling, angiogenesis and bone formation. In the present study, the effects of the vascular endothelial growth factor 165 (VEGF165) and bone morphogenetic protein 2 (BMP2) genes on bone regeneration were investigated by the lentivirus-mediated cotransfection of the two genes into rat bone marrow-derived MSCs. The successful co-expression of the two genes in the MSCs was confirmed using quantitative polymerase chain reaction (qPCR) and western blot analysis. The results of alizarin red and alkaline phosphatase (ALP) staining at 14 days subsequent to transfection showed that the area of staining in cells transfected with BMP2 alone was higher than that in cells transfected with BMP2 and VEGF165 or untransfected control cells, while the BMP2 + VEGF165 group showed significantly more staining than the untransfected control. This indicated that BMP2 alone exhibited a stronger effect in bone regeneration than BMP2 in combination with VEGF165. Similarly, in inducing culture medium, the ALP activity of the BMP2 + VEGF165 group was notably suppressed compared with that of the BMP2 group. The overexpression of VEGF165 inhibited BMP2-induced MSC differentiation and osteogenesis in vitro. Whether or not local VEGF gene therapy is likely to affect bone regeneration in vivo requires further investigation.

Section: Discussionmentioning

confidence: 99%

Effects of BMP2 and VEGF165 on the osteogenic differentiation of rat bone marrow-derived mesenchymal stem cells

Wang

Experimental and Therapeutic Medicine

et al. 2013

J Biomedical Materials Res

“…[1][2][3][4][5][6] Successful bone regeneration may require at least three important components: osteogenic progenitor cells that can undergo effective osteogenic differentiation and produce extracellular matrix, biological factors that are osteoinductive and able to induce or enhance new bone ingrowth, and scaffolding materials that are biocompatible and osteoconductive. [4][5][6] Osteogenic differentiation is a sequential cascade that recapitulates most of the molecular events occurring during skeletal development. 7 Mesenchymal stem cells (MSCs) are commonly used osteoblast progenitor cells for bone formation.…”

Section: Introductionmentioning

confidence: 99%

Characterization of scaffold carriers for BMP9‐transduced osteoblastic progenitor cells in bone regeneration

Shui

Zhang

Liang

et al. 2013

Successful bone tissue engineering at least requires sufficient osteoblast progenitors, efficient osteoinductive factors, and biocompatible scaffolding materials. We have demonstrated that BMP9 is one of the most potent factors in inducing osteogenic differentiation of mesenchymal progenitors. To facilitate the potential use of cell-based BMP9 gene therapy for bone regeneration, we characterize the in vivo osteoconductive activities and bone regeneration potential of three clinically-used scaffold materials, type I collagen sponge, hydroxyapatite-tricalcium phosphate (HA-TCP) and demineralized bone matrix (DBM), using BMP9-expressing C2C12 osteoblastic progenitor cells. We find that recombinant adenovirus-mediated BMP9 expression effectively induces osteogenic differentiation in C2C12 cells. Although direct subcutaneous injection of BMP9-transduced C2C12 cells forms ectopic bony masses, subcutaneous implantation of BMP9-expressing C2C12 cells with collagen sponge or HA-TCP scaffold yields the most robust and mature cancellous bone formation, whereas the DBM carrier group forms no or minimal bone CONFLICT OF INTERESTThe authors state that they have no conflicts of interest. HHS Public Access

“…Successful bone regeneration may require at least three important components: osteogenic progenitor cells that can undergo effective osteogenic differentiation and produce extracellular matrix, biological factors that are osteoinductive and able to induce or enhance new bone ingrowth, and scaffolding materials that are biocompatible and osteoconductive 4–6 . Osteogenic differentiation is a sequential cascade that recapitulates most of the molecular events occurring during skeletal development 7 .…”

Section: Introductionmentioning

confidence: 99%

Characterization of scaffold carriers for BMP9-transduced osteoblastic progenitor cells in bone regeneration

Shui

Zhang

Liang

et al. 2013

J. Biomed. Mater. Res.

Successful bone tissue engineering at least requires sufficient osteoblast progenitors, efficient osteoinductive factors, and biocompatible scaffolding materials. We have demonstrated that BMP9 is one of the most potent factors in inducing osteogenic differentiation of mesenchymal progenitors. To facilitate the potential use of cell-based BMP9 gene therapy for bone regeneration, we characterize the in vivo osteoconductive activities and bone regeneration potential of three clinically-used scaffold materials, type I collagen sponge, hydroxyapatite-tricalcium phosphate (HA-TCP) and demineralized bone matrix (DBM), using BMP9-expressing C2C12 osteoblastic progenitor cells. We find that recombinant adenovirus-mediated BMP9 expression effectively induces osteogenic differentiation in C2C12 cells. Although direct subcutaneous injection of BMP9-transduced C2C12 cells forms ectopic bony masses, subcutaneous implantation of BMP9-expressing C2C12 cells with collagen sponge or HA-TCP scaffold yields the most robust and mature cancellous bone formation, whereas the DBM carrier group forms no or minimal bone masses. Our results suggest that collagen sponge and HA-TCP scaffold carriers may provide more cell-friendly environment to support the survival, propagation, and ultimately differentiation of BMP9-expressing progenitor cells. This line of investigation should provide important experimental evidence for further pre-clinical studies in BMP9-mediated cell based approach to bone tissue engineering.