Adipose-derived stem cells transfected by adenoviral vector mediated human transforming growth factor-β1 gene with β-tricalcium phosphate scaffold in vitro

Abstract

Objective To observe the biological behaviors of cultured adipose-derived stem cells (ADSCs) transfected by adenoviral vector-mediated human transforming growth factor-β1 (Ad-hTGF-β1) combined with β-tricalcium phosphate (β-TCP) scaffold and investigate the feasibility of them for cartilage tissue engineering. Methods The adenoviral plasmid vector containing hTGF-β1 gene was transfected into ADSCs. The ADSCs were seeded onto the β-TCP scaffold and the effects of adhesion and morphological changes were observed under the phase-contrast microscopy and scanning electron microscopy (SEM). Reverse transcription-polymerase chain reaction (RT-PCR), Western blotting and immunohistochemistry were applied to detect the expression of collagen Ⅱ in the β-TCP scaffold. Results The SEM results showed the porous β-TCP scaffold had macro- and micro-porous strutures with the porosity being 65%. The ADSCs transfected by Ad-hTGF-β1 were successfully cultured in vitro. The induced cells adhered to the surface of the β-TCP scaffold and proliferated well. RT-PCR, Western blotting and immunohistochemistry revealed that Collagen Ⅱ was detected after mixed culture. Conclusion The porous β-TCP scaffold with excellent property may be a good matrix for ADSCs transfected by Ad-hTGF-β1, and can be used for cartilage engineering. Key words: Adipose-derived stem cell;  β-tricalcium phosphate;  Scaffold;  Tissue engineering