The effects of Tween 20, Tween 80, and Cremophor EL, surface active agents which are used for dispersion of water-insoluble substances, on vascular responsiveness were investigated using rat aortic rings. In high concentrations all these agents produced persistent contractions in aortic rings independent of the presence of endothelium. These contractions were not influenced by inhibitors of known endogenous contractile mediators. Incubation with these agents caused a concentration-dependent inhibition of the endothelium-dependent relaxant responses to acetylcholine in intact precontracted aortic rings. Endothelium-independent relaxations produced by sodium nitroprusside were not inhibited, but rather potentiated in the presence of Tween 80 (10(-1) ml/l). On the other hand, Tween 80 inhibited the contractile effects of 5-hydroxytryptamine, phenylephrine, and bradykinin significantly. The data suggests that these substances affect both endothelial cells and vascular smooth muscle.