In this study, the capacity of bone morphogenetic protein 2 (BMP-2) gene-transfected bone marrow-derived mesenchymal stem cells (MSCs) in combination with nano-hydroxyapatite/collagen/poly(L-lactic acid) (nHAC/ PLA) to improve the repair of bone defects in rabbit was explored. MSCs from New Zealand White rabbits were cultured and injected with pIRES2-EGFPhBMP-2 or pIRES2-EGFP by electroporation. After the transfer efficiency was determined through the expression of EGFP, the MSCs were seeded on scaffolds to generate an in vitro 3D cell/scaffold construct. The adhesion and proliferation of the MSCs cultured in the scaffold was assessed by SEM. The cellular constructs obtained were allografted into the 15 mm critical-sized segmental bone defects in the radius of New Zealand White rabbits for 12 weeks. The bone regeneration was assessed by radiographical and histological analyses. In vitro, nHAC/PLA facilitated MSC adhesion and proliferation on the scaffold, and gene transfer efficiency reached a maximum of 35.5 ± 3.8%. In vivo, the implantation of BMP-2 transfected MSCs/nHAC/PLA construct significantly enhanced the formation of new bone in the segmental defect, compared to the control groups. This novel 3D BMP-2 transfected MSCs/nHAC/PLA construct has the potential for bone repair by genetic tissue engineering approach.
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