The Effect Of Ticlopidine On Cultured Human Endothelial Cells: Possible Influence On Extracellular Matrix Formation

Endothelial cells derived from human umbilical cord vein were seeded and grown to confluence in culture medium TC 199 with 20% foetal calf serum without other growth factors supplement. Cells were then detached utilizing a trypsin-EDTA solution, harvested in fresh culture media containing different concentrations of Ticlopidine, poured on glass coverslips placed on 35 mm Petri dishes and cultured for six days without further medium replacement. Control dishes were prepared using fresh medium without Ticlopidine.

Treated cells revealed a decreased tendency to adhere to the substratum as well as morphological changes in comparison with controls when examined by phase contrast microscopy. This effect was more evident with higher concentration (0.33 mg/ml) of Ticlopidine, while lower concentrations (0.05 mg/ml) caused morphological changes only. Indirect immunofluorescence for fibronectin utilizing monospecific antibodies, revealed a different fluorescent pattern if compared with controls. In particular a less pronounced intracellular granular fluorescence and a delay in the appearance of the fibrillar extracellular matrix.