Study on transfection of bone morphogenetic protein‐2 gene into rabbit bone marrow stromal cells and composition to nanohydroxyapatite for constructing tissue engineered bones

Abstract

A study is presented on effects of the adenovirus-mediated human bone morphogenetic protein-2 (Ad-BMP-2) expression vector's transfection into rabbit bone marrow stromal cells (rBMSCs) and the subsequent composition to nanohydroxyapatite (Nano-HA) for the construction of biomimetic artificial bone in vitro. The adenovirus vector was constructed by Gateway TM technology, and the rBMSCs transfected by Ad-BMP-2 were cultured in vitro. Immunohistochemistry, reverse transcriptase-polymerase chain reaction and Western blot were used to detect the bone morphogenetic protein-2 (BMP-2) expression. After culture for 48 h, the transfected cells were seeded onto the Nano-HA scaffold. Scanning electron microscopy (SEM) was used to observe the growth and attachment of cells on the composite scaffold on the third and fifth days. After transfection, the hBMP-2 gene showed high expressions in mRNA and protein levels. SEM revealed even distribution and fine adherence of the cells in the composite scaffold. The Western blot showed high expression of BMP-2 in the transfected and compound cells. The target cells that were transfected with the adenovirus vector that contained the genes, were bonded to the Nano-HA scaffold, and therefore the cell carrier complexes became stable and effectively secreted growth factors, and the biomimetic artificial bones were successfully constructed in vitro.