Rabbit bone marrow stromal cells transfected by adenovirus mediated bone morphogenetic protein-2 gene plus nano-hydroxyapatite to reconstruct tissue engineered bone in vitro

Abstract

Objective To investigate the effects of adenovirus-mediated human bone morphogenetic protein-2 expression vector(Ad-BMP-2) transfected with rabbit bone marrow stromal cells(rBMSCs) plus nano-hydroxyapatite(Nano-HA) in reconstruction of biomimetic artificial bone in vitro. Methods The adenovirus vector was constructed by GatewayTM technology. Nano-HA was prepared by Sol-Flocculation.rBMSCs transfected by Ad-BMP-2 were cultured in vitro. Immunohistochemistry, RT-PCR and Western Blot were used to detect the BMP-2 expression in the transfected cells. After culture for 48 h, the transfected cells were seeded onto the Nano-HA scaffold. Scanning electron microscopy was used to observe the growth and attachment of cells on the composite scaffold on the third and fifth days. The attached cells were also gathered on the third and fifth days for detection of the BMP-2 expressions by Western Blot. Results After transfection, hBMP-2 gene showed high expressions in mRNA and protein levels. Scanning electron mi-croscopy revealed even distribution and fine adherence of the cells in the composite scaffold. Western Blot showed high expression of BMP-2 in the transfected and compound cells. Conclusion The Ad-BMP-2 transfected with rBMSCs plus Nano-HA can successfully reconstruct the in vitro biomimetic artificial bone which can secrete growth factors stably. Key words: Bone morphogenetic proteins; Bone marrow cells; Tissue engineering; Aden-ovirus vector