Surfactant is a perfect carrier for direct pulmonary delivery of steroids. In order to preserve surfactant properties, steroid interference with surface tension should be carefully evaluated. We evaluated the in vitro effect of dexamethasone (Dexa), budesonide (Bude) and betamethasone (Beta) on the surfactant surface tension lowering properties of a synthetic surfactant. The surface tension properties of synthetic surfactant Exosurf Neonatal (Exo) were evaluated in vitro by a Fisher surface tensiometer at 37℃. Commercially available Dexa, Beta and Bude solutions had lower surface tension (p<0.001) compared with water or normal saline, yet they had higher surface tension compared with Exo (p<0.001). Direct addition of the liquid form of Dexa, Beta and Bude to Exo changed the surface tension in a dose-dependent manner. Within the concentrations of 0.375 and 0.625 mg Dexa/5 ml Exo (0.075 ml to 0.125 ml Dexa/5 ml Exo) there was a dramatic rise in surface tension. Within the concentrations of 1 and 3.4 mg Beta/5 ml Exo (0.25 ml to 0.85 ml/5 ml Exo) a sigmoid rise in surface tension was found. Between the doses of 0.5 to 1.5 mg Bude/5 ml Exo (1 ml to 3 ml Bude/5 ml Exo) a gradual rise in surface tension was found. We concluded that the addition of high concentration steroid solutions might affect the surface tension of Exo. Below the concentration of 0.375 mg Dexa/5 ml Exo, 1 mg Beta/5 ml Exo and 0.5 mg Bude/5 ml Exo the surface tension lowering properties of Exo were not significantly altered.