Effect of combined application of bFGF and inorganic polyphosphate on bioactivities of osteoblasts and initial bone regeneration

Basic fibroblast growth factor (bFGF) and inorganic polyphosphate (poly(P)) have been recognized as therapeutic agents that enhance bone regeneration. It has also been shown that poly(P) may enhance the mitogenic activity of bFGF. The purpose of this study is to evaluate the combined effect of bFGF and poly(P) on bioactivities of osteoblasts and initial bone regeneration in vitro and in vivo. MC3T3-E1 cells were treated with bFGF, poly(P) or bFGF+poly(P), then subjected to cell proliferation assay, alkaline phosphatase (ALP) activity measurement, quantitative real-time reverse transcription-polymerase chain reaction and Alizarin S Red staining. In an in vivo study, bFGF-, poly(P)- and bFGF+poly(P)-modified interconnected porous hydroxyapatite (IPHA) complexes were fabricated, and placed into the femurs of rabbits to evaluate new bone formation histologically and histomorphometrically. The highest enhancement of cell proliferation were observed in those treated with bFGF+poly(P) on days 5 and 7. Cells treated with bFGF+poly(P) also exhibited increased ALP activity on days 5 and 10, up-regulated mRNA levels of osteocalcin and osteopontin, and enhanced calcification when compared to the non-treated cells. In vivo, the highest bone formation ratio was observed in bFGF+poly(P)-modified IPHA complexes. This study indicated that co-application of bFGF and poly(P) may provide enhanced bone formation by modulating cell proliferation and the mineralization process. It is anticipated that a combined application of bFGF and poly(P) can provide a novel method for bone regeneration in clinical use.