title: Adenoviral Mediated Expression of BMP2 by Bone Marrow Stromal Cells Cultured in 3D Copolymer Scaffolds Enhances Bone Formation.
abstract: Selection of appropriate osteoinductive growth factors, suitable delivery method and proper supportive scaffold are critical for a successful outcome in bone tissue engineering using bone marrow stromal cells (BMSC). This study examined the molecular and functional effect of a combination of adenoviral mediated expression of bone morphogenetic protein-2 (BMP2) in BMSC and recently developed and characterized, biodegradable Poly(L-lactide-co-є-caprolactone){poly(LLA-co-CL)}scaffolds in osteogenic molecular changes and ectopic bone formation by using in vitro and in vivo approaches. Pathway-focused custom PCR array, validation using TaqMan based quantitative RT-PCR (qRT-PCR) and ALP staining showed significant up-regulation of several osteogenic and angiogenic molecules, including ALPL and RUNX2 in ad-BMP2 BMSC group grown in poly(LLA-co-CL) scaffolds both at 3 and 14 days. Micro CT and histological analyses of the subcutaneously implanted scaffolds in NOD/SCID mice revealed significantly increased radiopaque areas, percentage bone volume and formation of vital bone in ad-BMP2 scaffolds as compared to the control groups both at 2 and 8 weeks. The increased bone formation in the ad-BMP2 group in vivo was paralleled at the molecular level with concomitant over-expression of a number of osteogenic and angiogenic genes including ALPL, RUNX2, SPP1, ANGPT1. The increased bone formation in ad-BMP2 explants was not found to be associated with enhanced endochondral activity as evidenced by qRT-PCR (SOX9 and FGF2) and Safranin O staining. Taken together, combination of adenoviral mediated BMP-2 expression in BMSC grown in the newly developed poly(LLA-co-CL) scaffolds induced expression of osteogenic markers and enhanced bone formation in vivo.
authors.affiliation: Department of Clinical Dentistry, Faculty of Medicine and Dentistry, University of Bergen, Bergen, Norway.
authors.forename: Sunita
authors.initials: S
authors.lastname: Sharma
authors.affiliation: The Gade Laboratory for Pathology, Department of Clinical Medicine, University of Bergen, Bergen, Norway.
authors.forename: Dipak
authors.initials: D
authors.lastname: Sapkota
authors.affiliation: Department of Clinical Dentistry, Faculty of Medicine and Dentistry, University of Bergen, Bergen, Norway.
authors.forename: Ying
authors.initials: Y
authors.lastname: Xue
authors.affiliation: Department of Clinical Dentistry, Faculty of Medicine and Dentistry, University of Bergen, Bergen, Norway.|Department of Fibre and Polymer Technology, Royal Institute of Technology, Stockholm, Sweden.
authors.forename: Yang
authors.initials: Y
authors.lastname: Sun
authors.affiliation: Department of Fibre and Polymer Technology, Royal Institute of Technology, Stockholm, Sweden.
authors.forename: Anna
authors.initials: A
authors.lastname: Finne-Wistrand
authors.affiliation: Department of Medical Genetics and Molecular Medicine, Haukeland University Hospital, Bergen, Norway.
authors.forename: Ove
authors.initials: O
authors.lastname: Bruland
authors.affiliation: Department of Clinical Dentistry, Faculty of Medicine and Dentistry, University of Bergen, Bergen, Norway.
authors.forename: Kamal
authors.initials: K
authors.lastname: Mustafa
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D055396:Bone Morphogenetic Protein 2 / Q000235:genetics*; D002478:Cells, Cultured; D005822:Genetic Vectors; D057809:HEK293 Cells; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D011108:Polymers / Q000378:metabolism*
chemical_list: D055396:Bone Morphogenetic Protein 2; D011108:Polymers