Development of an in vitro cytotoxicity model for aerosol exposure using 3D reconstructed human airway tissue; application for assessment of e-cigarette aerosol

Louise Neilson, Courtney Mankus, David Thorne, George Jackson, Jason DeBay, Clive Meredith British American Tobacco

Development of physiologically relevant test methods to analyse potential irritant effects to the respiratory tract caused by e-cigarette aerosols is required. This paper reports the method development and optimisation of an acute in vitro MTT cytotoxicity assay using human 3D reconstructed airway tissues and an aerosol exposure system. The EpiAirway™ tissue is a highly differentiated in vitro human airway culture derived from primary human tracheal/bronchial epithelial cells grown at the air–liquid interface, which can be exposed to aerosols generated by the VITROCELL smoking robot. Method development was supported by understanding the compatibility of these tissues within the VITROCELL system, in terms of airflow (L/min), vacuum rate (mL/min) and exposure time. Dosimetry tools (QCM) were used to measure deposited mass, to confirm the provision of e-cigarette aerosol to the tissues. EpiAirway™ tissues were exposed to cigarette smoke and aerosol generated from two commercial e-cigarettes for up to 6 h. Cigarette smoke reduced cell viability in a time-dependent manner to 12% at 6 h. E-cigarette aerosol showed no such decrease in cell viability and displayed similar results to that of the untreated air controls. Applicability of the EpiAirway™ model and exposure system was demonstrated, showing little cytotoxicity from e-cigarette aerosol and different aerosol formulations when compared directly with reference cigarette smoke, over the same exposure time.


e-cigarettes, smoking robot, EpiAirway (AIR-100), TEER, airway irritation

Materials Tested

e-cigarettes, aerosol, butyl methacrylate, heptyl butyrate, methyl stearate, olive oil, Triton X-100 (0.1%), cigarette smoke

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