Colloidal drug carriers could improve the therapy of numerous airway diseases. However, it remains unclear to what extent nanoscale particulate matter affects the biophysical function of the essential surface-active lining layer of the lungs, especially under predisposed conditions of airway diseases. Accordingly, the current study investigated the impact of defined polymer nanoparticles on impaired lung surfactants. Admixtures of plasma proteins (albumin and fibrinogen) to Curosurf led to a controllable decrease in surface activity (i.e., adsorption and minimal surface tension of >25 and >5 mN/m, respectively), which served as models for dysfunctional lung surfactants. Next, Curosurf preincubated with plasma proteins was challenged with negatively- and positively charged poly(lactide) nanoparticles. Negatively charged nanoparticles significantly perturbed the biophysical function of impaired Curosurf in a,dose-dependent manner, most likely due to a binding of essential surfactant components. By contrast, addition of positively charged nanoparticles led to no further loss of surface activity, but a remarkable depletion of plasma protein content. Once adsorbed to the surface of polymer nanoparticles, plasma proteins were hindered to displace relevant surfactant components from the air/liquid interface. Overall, the current study indicated that, depending on their physicochemical properties, colloidal drug carriers could compromise the biophysical function of impaired lung surfactants. Notably, a positive surface charge represents a parameter for the rationale design of polymer nanomedicines causing negligible adverse events on an impaired surface-active lining layer in the lungs.