Toxicology in Vitro
Available online 12 July 2015
In Press, Corrected Proof — Note to users
Development of an in vitro cytotoxicity model for aerosol exposure using 3D reconstructed human airway tissue; application for assessment of e-cigarette aerosol
doi:10.1016/j.tiv.2015.05.018
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Open Access
Highlights
•
There are currently no standards for the assessment of aerosol emissions in vitro.
•
There is a need to develop in vitro biological techniques for the analysis of aerosols.
•
EpiAirway™ tissue model can distinguish between known irritants and non-irritants.
•
E-cigarette aerosols showed little to no cytotoxicity compared with cigarette smoke.
•
Using an aggressive puffing regime, e-cigarette aerosol showed little cytotoxicity.
Abstract
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.
Keywords
Available online 12 July 2015
In Press, Corrected Proof — Note to users
Development of an in vitro cytotoxicity model for aerosol exposure using 3D reconstructed human airway tissue; application for assessment of e-cigarette aerosol
- Louise Neilsona, , ,
- Courtney Mankusb,
- David Thornea,
- George Jacksonb,
- Jason DeBayb,
- Clive Mereditha
doi:10.1016/j.tiv.2015.05.018
Get rights and content
Open Access
Highlights
•
There are currently no standards for the assessment of aerosol emissions in vitro.
•
There is a need to develop in vitro biological techniques for the analysis of aerosols.
•
EpiAirway™ tissue model can distinguish between known irritants and non-irritants.
•
E-cigarette aerosols showed little to no cytotoxicity compared with cigarette smoke.
•
Using an aggressive puffing regime, e-cigarette aerosol showed little cytotoxicity.
Abstract
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.
Keywords
- Cytotoxicity;
- E-cigarettes;
- Aerosol;
- EpiAirway™;
- In vitro;
- Human airway
