Vesicular based delivery systems, such as liposomes or non-ionic surfactant vesicles (NIV), have been used to enhance the therapeutic outcome of drugs like amphotericin B (AmB). However, little is known about the use of NIV for pulmonary delivery. In this study, the antifungal AmB, the anticancer drug cisplatin and the bioluminescent substrate luciferin, were encapsulated into NIV (AmB-NIV, cis-NIV and luc-NIV) and nebulised using a passive vibrating-mesh nebuliser (I-neb) and two active vibrating-mesh nebulisers (Aeroneb Go and Aeroneb Lab). Vibrating mesh nebulisation decreased the entrapment efficiency of AmB-NIV by 60 % and vesicle size by 40 nm but no difference was observed between nebulisers. Both active vibrating-mesh nebulisers were superior at nebulising AmB and Aeroneb Lab was superior at nebulising cisplatin, generating aerosols with lower particle size compared to the passive vibrating-mesh nebuliser. However, the passive vibrating-mesh nebuliser generated a less disperse aerosol particle size cloud, with a lower aerodynamic size for luc-NIV. Differences in nebuliser performance were attributed to different physicochemical properties of NIV. NIV nebulisation presented a bimodal distribution when nebulised since the NIV carrier alone had a higher aerosol particle size compared to cis-NIV. Thus would provide rapid drug release due to unentrapped drug followed by a sustained release from the NIV carrier. All the NIV ingredients travelled as a single aggrupation when nebulised since they had similar MMAD values. Vibrating-mesh nebulisation is a successful strategy for pulmonary administration of AmB-NIV, cis-NIV and luc-NIV.