Abstract
The purpose of this study was to investigate the feasibility and advantages of constructing a novel tissue engineering bone, using β-tricalcium phosphate (β-TCP) and rat bone marrow mesenchymal stem cells (MSCs), modified with human bone morphogenetic protein 2 gene (hBMP2) and human vascular endothelial growth factor 165 gene (hVEGF165), through lentiviral transfection. Both genes were successfully co-expressed in the co-transfection group for up to eight weeks confirmed by enzyme-linked immunosorbent assay (ELISA). After seeding MSCs onto the scaffolds, scanning electron microscopy (SEM) observation showed that MSCs grew and proliferated well in co-transfection group at 7 and 14 days. There was no significant difference among all the groups in hoechst DNA assay for cell proliferation for 14 days after cell seeding (P > 0.05), but the highest alkaline phosphatase (ALP) activity was observed in the co-transfection group at 14 days after cell seeding (p < 0.01). These results demonstrated that it was advantageous to construct tissue engineering bone using β-TCP combined with MSCs lentivirally co-transfected with BMP2 and VEGF165, providing an innovative way for treating bone defects.
Highlights
As an integration of ex vivo gene therapy and tissue engineering approach, gene modified tissue engineered bone has been recently reported to be an attractive option to treat bone defects [1,2,3]
Since both bone morphogenetic protein 2 (BMP2) and vascular endothelial growth factor (VEGF) are involved in bone regeneration as osteogenic and angiogenic factors, it is possible that combined gene therapy of both genes in bone tissue engineering might have more significant effect on bone regeneration than single gene alone
There was no significant difference in the amount of BMP2 production between BMP group and BMP+VEGF group (P > 0.05), whereas no detectable
Summary
As an integration of ex vivo gene therapy and tissue engineering approach, gene modified tissue engineered bone has been recently reported to be an attractive option to treat bone defects [1,2,3]. Vascular endothelial growth factor (VEGF), the most potent angiogenic growth factor, has been specially chosen for several studies because vascularization appears to be a key factor in bone tissue engineering, especially for large and critical size bone defects [8,9]. There is concern that a single exposure to an exogenous growth factor may not induce an adequate osteogenic signal in many clinical situations with relatively limited bone healing potential because of compromised vascularity, limited bone stock, and abundant fibrous tissue [10]. To address these important questions, we constructed tissue engineering bone using β-tricalcium
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