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Adenoviral BMP-2 gene transfer in mesenchymal stem cells: in vitro and in vivo bone formation on biodegradable polymer scaffolds
Authors:Partridge Kris  Yang Xuebin  Clarke Nicholas M P  Okubo Yasunori  Bessho Kazuhisa  Sebald Walter  Howdle Steven M  Shakesheff Kevin M  Oreffo Richard O C
Institution:University Orthopaedics, University of Southampton, General Hospital, Southampton SO16 6YD, United Kingdom.
Abstract:The aim of this study was to determine the feasibility of adenoviral gene transfer into primary human bone marrow osteoprogenitor cells in combination with biodegradeable scaffolds to tissue-engineer bone. Osteoprogenitors were infected with AxCAOBMP-2, a vector carrying the human BMP-2 gene. Alkaline phosphatase activity was induced in C2C12 cells following culture with conditioned media from BMP-2 expressing cells, confirming successful secretion of active BMP-2. Expression of alkaline phosphatase activity, type I collagen and mineralisation confirmed bone cell differentiation and maintenance of the osteoblast phenotype in extended culture for up to 6 weeks on PLGA porous scaffolds. In vivo implantation of adenoviral osteoprogenitor constructs on PLGA biodegradeable scaffolds, using diffusion chambers, also demonstrated bone cell differentiation and production of bone tissue. The maintenance of the osteoblast phenotype in extended culture and generation of mineralised 3-D scaffolds containing such constructs indicate the potential of such bone tissue engineering approaches in bone repair.
Keywords:osteoprogenitor  adenoviral gene-transfer  biodegradable polymer  dl-lactic- co-÷glycolic acid)" target="_blank">poly(dl-lactic- co-÷glycolic acid)  PLGA  tissue engineering
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