Bioactive FGF-2 in sterilized extracellular matrix

Retention of growth factor activity during disinfection, lyophilization, an d sterilization may be key to the development of safe, yet effective, human clinical products derived from biological sources. Traditional methods to reduce the antigenicity of biological materials used for wound healing are thought to destroy the bioactivity of growth factors contained within natur ally occurring extracellular matrices. A sterilized extracellular matrix wo und dressing (sECM) was examined for bioactive FGF-2 using a sandwich ELISA and an in-vitro bioassay. FGF-2 was present in sECM at a concentration as great as 97.9 +/- 11.7 ng/g. To detect bioactivity, the cell culture medium was incubated with sECM (sECM-CM) and was used as the growth medium for ra t pheochromocytoma (PC12) cells. After 48 hours, sECM-CM induced differenti ation in 22 percent of the cells versus eight percent of the positive contr ols. Addition of an FGF-2 neutralizing antibody reduced PC12 differentiatio n in the sECM-CM-treated cells to nine percent versus one percent in the co ntrol wells. These results indicate that both the presence and the bioactiv ity of FGF-2 are retained in sECM. Because FGF-2 is an important regulator of angiogenesis, wound healing, and regeneration, it is likely that bioacti ve FGF-2 retained in sECM contributes to the clinical successes observed wh en this material is used to treat hard-to-heal wounds.