Both smooth muscle cells and endothelial cells play an important role in vascular wound healing. To elucidate the role of fructose-1,6-diphosphate, cell proliferation and cell migration studies were performed with human endothelial cells and rat smooth muscle cells. To mimic blood vessels, endothelial and smooth muscle cells were used in 1:10, 1:5, and 1:1 concentrations, respectively, mimicking large-, mid-, and capillary-sized blood vessels. Cell migration was studied with fetal bovine serum-starved cells. For cell proliferation assay, cells were plated at 30-50% confluency and then starved. The cells were incubated for 48 h with fructose-1,6-diphosphate at (per ml) 10 mg, 1 mg, 500 microg, 250 microg, 100 microg, and 10 microg, pulsed with tritiated-thymidine and incubated with 1 N NaOH for 30 min at room temperature, harvested, and counted. For migration assay, confluent cells were starved, wounded, and incubated for 24 h with same concentrations of fructose-1,6-diphosphate as in proliferation assay. The cells were fixed and counted. Smooth muscle cell proliferation was inhibited by fructose-1,6-diphosphate at 10 mg/ml. In the xenograft models of 1:10, 1:5, and 1:1 fructose-1,6-diphosphate inhibited proliferation at 10 mg/ml. In migration studies 10 mg fructose-1,6-diphosphate per ml was inhibitory to both cell types. In large-, mid-, and capillary-sized blood vessels, fructose-1,6-diphosphate inhibited proliferation of both cell types at 10 mg/ml. At the individual cell level, fructose-1,6-diphosphate is nonstimulatory to proliferation of endothelial cells while inhibiting migration, and it acts on smooth muscle cells by inhibiting both proliferation and migration.