Towards an in-vitro multi-cellular human airways model for evaluating the effects of Electronic cigarettes

Pranav Vasanthi Bathri Narayanan, Laura J. Leslie, James E. Brown, Lindsay J. Marshall

Research output: Contribution to conferencePoster

Abstract

Despite being poor representations of human airways architecture, innumerable animals, especially rodents, have been used in cigarette smoking studies. The advent of Electronic-cigarettes (ECs) could herald a further escalation, with data from in-vivo experiments already increasingly published. The current study aims to demonstrate the application of a human in-vitro model for evaluation of EC, providing alternatives to the outdated in-vivo models. The human airways model consists of relevant cell types that would be directly encountered during vaping. Human bronchial epithelial cells and pulmonary fibroblasts were co-cultured at air-liquid interface (ALI) under conditions that promote mucociliary differentiation, tight junction formation and mucin production. An in-house built smoking machine was used to deliver vapour from a commercially available EC (ECV) or whole cigarette smoke (WCS) to the co-culture model according to ISO standard. This methodology closely mimics human smoking behaviour, as opposed to enforced nasal inhalation in rodent smoking/vaping model. 24h post exposure, XTT cell viability analysis showed that WCS caused a significant decrease (p<0.0001) in cell viability (<70%) compared to control cells exposed to air only. ECV on the other hand did not have a significant impact on cell viability, thus suggesting low cytotoxicity. This difference in effect correlates with a number of existing in-vitro and in-vivo ECV/WCS studies, illustrating that the current model is a relevant, more realistic platform for EC studies compared to animal models. Further, such an airways model resembling in-vivo physiology can be used to study COPD progression and development, a condition difficult to replicate in rodents.
LanguageEnglish
PagesP20
Publication statusPublished - 2016
EventAnimal Replacement Science Conference 2016: Advances, Awareness, Applications - London, United Kingdom
Duration: 9 Dec 2016 → …

Conference

ConferenceAnimal Replacement Science Conference 2016
CountryUnited Kingdom
CityLondon
Period9/12/16 → …

Fingerprint

Smoking
Smoke
Tobacco Products
Rodentia
Cell Survival
Air
Tight Junctions
Mucins
Coculture Techniques
Nose
Chronic Obstructive Pulmonary Disease
Inhalation
Animal Models
Fibroblasts
Epithelial Cells
In Vitro Techniques
Electronic Cigarettes
Lung
Vaping

Cite this

Vasanthi Bathri Narayanan, P., Leslie, L. J., Brown, J. E., & Marshall, L. J. (2016). Towards an in-vitro multi-cellular human airways model for evaluating the effects of Electronic cigarettes. P20. Poster session presented at Animal Replacement Science Conference 2016, London, United Kingdom.
Vasanthi Bathri Narayanan, Pranav ; Leslie, Laura J. ; Brown, James E. ; Marshall, Lindsay J. / Towards an in-vitro multi-cellular human airways model for evaluating the effects of Electronic cigarettes. Poster session presented at Animal Replacement Science Conference 2016, London, United Kingdom.
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abstract = "Despite being poor representations of human airways architecture, innumerable animals, especially rodents, have been used in cigarette smoking studies. The advent of Electronic-cigarettes (ECs) could herald a further escalation, with data from in-vivo experiments already increasingly published. The current study aims to demonstrate the application of a human in-vitro model for evaluation of EC, providing alternatives to the outdated in-vivo models. The human airways model consists of relevant cell types that would be directly encountered during vaping. Human bronchial epithelial cells and pulmonary fibroblasts were co-cultured at air-liquid interface (ALI) under conditions that promote mucociliary differentiation, tight junction formation and mucin production. An in-house built smoking machine was used to deliver vapour from a commercially available EC (ECV) or whole cigarette smoke (WCS) to the co-culture model according to ISO standard. This methodology closely mimics human smoking behaviour, as opposed to enforced nasal inhalation in rodent smoking/vaping model. 24h post exposure, XTT cell viability analysis showed that WCS caused a significant decrease (p<0.0001) in cell viability (<70{\%}) compared to control cells exposed to air only. ECV on the other hand did not have a significant impact on cell viability, thus suggesting low cytotoxicity. This difference in effect correlates with a number of existing in-vitro and in-vivo ECV/WCS studies, illustrating that the current model is a relevant, more realistic platform for EC studies compared to animal models. Further, such an airways model resembling in-vivo physiology can be used to study COPD progression and development, a condition difficult to replicate in rodents.",
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Vasanthi Bathri Narayanan, P, Leslie, LJ, Brown, JE & Marshall, LJ 2016, 'Towards an in-vitro multi-cellular human airways model for evaluating the effects of Electronic cigarettes' Animal Replacement Science Conference 2016, London, United Kingdom, 9/12/16, pp. P20.

Towards an in-vitro multi-cellular human airways model for evaluating the effects of Electronic cigarettes. / Vasanthi Bathri Narayanan, Pranav; Leslie, Laura J.; Brown, James E.; Marshall, Lindsay J.

2016. P20 Poster session presented at Animal Replacement Science Conference 2016, London, United Kingdom.

Research output: Contribution to conferencePoster

TY - CONF

T1 - Towards an in-vitro multi-cellular human airways model for evaluating the effects of Electronic cigarettes

AU - Vasanthi Bathri Narayanan, Pranav

AU - Leslie, Laura J.

AU - Brown, James E.

AU - Marshall, Lindsay J.

PY - 2016

Y1 - 2016

N2 - Despite being poor representations of human airways architecture, innumerable animals, especially rodents, have been used in cigarette smoking studies. The advent of Electronic-cigarettes (ECs) could herald a further escalation, with data from in-vivo experiments already increasingly published. The current study aims to demonstrate the application of a human in-vitro model for evaluation of EC, providing alternatives to the outdated in-vivo models. The human airways model consists of relevant cell types that would be directly encountered during vaping. Human bronchial epithelial cells and pulmonary fibroblasts were co-cultured at air-liquid interface (ALI) under conditions that promote mucociliary differentiation, tight junction formation and mucin production. An in-house built smoking machine was used to deliver vapour from a commercially available EC (ECV) or whole cigarette smoke (WCS) to the co-culture model according to ISO standard. This methodology closely mimics human smoking behaviour, as opposed to enforced nasal inhalation in rodent smoking/vaping model. 24h post exposure, XTT cell viability analysis showed that WCS caused a significant decrease (p<0.0001) in cell viability (<70%) compared to control cells exposed to air only. ECV on the other hand did not have a significant impact on cell viability, thus suggesting low cytotoxicity. This difference in effect correlates with a number of existing in-vitro and in-vivo ECV/WCS studies, illustrating that the current model is a relevant, more realistic platform for EC studies compared to animal models. Further, such an airways model resembling in-vivo physiology can be used to study COPD progression and development, a condition difficult to replicate in rodents.

AB - Despite being poor representations of human airways architecture, innumerable animals, especially rodents, have been used in cigarette smoking studies. The advent of Electronic-cigarettes (ECs) could herald a further escalation, with data from in-vivo experiments already increasingly published. The current study aims to demonstrate the application of a human in-vitro model for evaluation of EC, providing alternatives to the outdated in-vivo models. The human airways model consists of relevant cell types that would be directly encountered during vaping. Human bronchial epithelial cells and pulmonary fibroblasts were co-cultured at air-liquid interface (ALI) under conditions that promote mucociliary differentiation, tight junction formation and mucin production. An in-house built smoking machine was used to deliver vapour from a commercially available EC (ECV) or whole cigarette smoke (WCS) to the co-culture model according to ISO standard. This methodology closely mimics human smoking behaviour, as opposed to enforced nasal inhalation in rodent smoking/vaping model. 24h post exposure, XTT cell viability analysis showed that WCS caused a significant decrease (p<0.0001) in cell viability (<70%) compared to control cells exposed to air only. ECV on the other hand did not have a significant impact on cell viability, thus suggesting low cytotoxicity. This difference in effect correlates with a number of existing in-vitro and in-vivo ECV/WCS studies, illustrating that the current model is a relevant, more realistic platform for EC studies compared to animal models. Further, such an airways model resembling in-vivo physiology can be used to study COPD progression and development, a condition difficult to replicate in rodents.

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M3 - Poster

SP - P20

ER -

Vasanthi Bathri Narayanan P, Leslie LJ, Brown JE, Marshall LJ. Towards an in-vitro multi-cellular human airways model for evaluating the effects of Electronic cigarettes. 2016. Poster session presented at Animal Replacement Science Conference 2016, London, United Kingdom.