Effect of fibroblast growth factor receptors 1 dominant negative on mineralization of bone marrow stromal cells in bone induction culture

Abstract

Objective To investigate the mineralization impact of fibroblast growth factor receptors 1 dominant negative (FGFR1DN) on osteogenic induction culture of bone marrow stromal cells (BMSCs). Methods The 3rd generation BMSCs were divided into 4 equal groups (n=24). FGFR1-DN group was transfected by pcDNA3.1(+)-FGFR1DN, FGFR1 group by pcDNA3.1(+)-FRFR1, blank load group by pcDNA3.1(+)-blank vehicle and non-transfection group by nothing. After successful transfection was con-firmed when the cells were in the logarithmic phase, osteogenic induction culture was conducted continuously for 21 days. Mineralized nodule formation was observed by alizarin red staining. The amount of mineralized material was calculated according to the standard curve of alizarin red concentration. Results Continuous osteogenic induction for 21 days showed on the bottom of the hole visible round opaque calcified nodules after alizarin red staining. The BMSCs in the FGFR1-DN group induced in the logarithmic growth phase by os-teoblasts exhibited significantly increased osteogenic capacity while those in the FGFR1 group displayed di-minished osteogenic capacity. The concentration of alizarin red was the highest in the FGFR1-DN group (1.33±0.19), the lowest in the FGFR1 group (1.00±1.17), and moderate in the blank load group (1.20±0.16) and non-transfection group (1.17±0.17), showing significant between-group differences (P< 0.05). Conclusions FGFR1-DN can promote cell proliferation in the early differentiation of BMSCs and also mineralization of osteoblasts in bone induction culture after the logarithmic growth phase. This may provide a hint for local gene treatment of bone defects. Key words: Bone marrow cells; Fibroblast growth factor receptor 1; Dominant negative; Mineralization