Acidic fibroblast growth factor attenuates the cytotoxic effects of peroxynitrite in primary human osteoblast precursors.

Abstract

Skeletal injury and associated ischemia and inflammation induce the generation of pro-oxidants such as peroxynitrite (ONOO-), which has been demonstrated to induce apoptosis in several cell lines. Fibroblast growth factor (FGF-1) is important for coordinating osteogenesis and angiogenesis of osseous repair. In vitro studies were performed examining the effect of FGF-1 on human osteoblast progenitor stromal stem (HSS) cell proliferation, differentiation, and response to ONOO-. HSS cells were isolated and growth kinetics determined in the presence and absence of FGF-1. The effect of FGF-1 on HSS cell expression of osteoblast-specific osteopontin and osteocalcin mRNA and protein was examined by reverse transcriptase polymerase chain reaction and Western blot techniques. To determine the sensitivity of HSS cells to ONOO- in the absence and presence of FGF-1 pretreatment, cells were exposed to varying concentrations of the oxidant and examined for cell death using quantitative fluorescence staining with fluorescein diacetate and propidium diacetate. Treatment of HSS cells with FGF-1 significantly enhanced cellular growth rates by 5 days (4.6 x 105 cells/mL vs. 3.1 x 105 cells/mL) and induced expression of both osteopontin and osteocalcin mRNA and protein. Exposure of HSS cells to ONOO- resulted in a dose- and time-dependent delayed cell death that was more characteristic of apoptosis than necrosis. Pretreatment of HSS cells with FGF-1 prevented ONOO- mediated apoptosis. In vitro, treatment of HSS cells with FGF-1 stimulates cell growth and induces expression of differentiation markers specific to osteoblasts. FGF-1 treatment renders osteoblast precursors resistant to the cytotoxic effects of ONOO-. These results suggest that FGF-1 promotes the progression of bone repair mechanisms by increasing the population of osteoblasts and imparting protection to the cell line from the hostile inflammatory environment.