Investigation of multiple whole smoke dosimetry techniques using a VITROCELL®VC10® smoke exposure system

AbstractAccessible in vitro models recapitulating the human airway that are amenable to study whole cannabis smoke exposure are needed for immunological and toxicological studies that inform public health policy and recreational cannabis use. In the present study, we developed and validated a novel 3D printed In Vitro Exposure System (IVES) that can be directly applied to study the effect of cannabis smoke exposure on primary human bronchial epithelial cells.Using commercially available design software and a 3D printer, we designed a four-chamber Transwell® insert holder for exposures to whole smoke. Software was used to model gas distribution, concentration gradients, velocity profile and shear stress within IVES. Following simulations, primary human bronchial epithelial cells cultured at air-liquid interface on Transwell® inserts were exposed to whole cannabis smoke. Following 24 hours, outcome measurements included cell morphology, epithelial barrier function, lactate dehydrogenase (LDH) levels, cytokine and gene expression.Whole smoke delivered through IVES possesses velocity profiles consistent with uniform gas distribution across the four chambers and complete mixing. Airflow velocity ranged between 1.0-1.5 μm s−1 and generated low shear stresses (<< 1 Pa). Human airway epithelial cells exposed to cannabis smoke using IVES showed changes in cell morphology and disruption of barrier function without significant cytotoxicity. Cannabis smoke elevated IL-1 family cytokines and elevated CYP1A1 and CYP1B1 expression relative to control.IVES represents an accessible, open-source, exposure system that can be used to model varying types of cannabis smoke exposures with human airway epithelial cells grown under air-liquid interface culture conditions.

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Simultaneous mainstream-sidestream smoke exposure systems II. The rat exposure system

Toxicology ◽

10.1016/0300-483x(85)90128-3 ◽

1985 ◽

Vol 35 (1) ◽

pp. 13-24 ◽

Cited By ~ 57

Author(s):

R.B. Griffith ◽

S. Standafer

Keyword(s):

Smoke Exposure ◽

Exposure System ◽

Sidestream Smoke

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Development and validation of a mouse model of contemporary cannabis smoke exposure

ERJ Open Research ◽

10.1183/23120541.00107-2021 ◽

2021 ◽

pp. 00107-2021

Author(s):

Matthew F. Fantauzzi ◽

Steven P. Cass ◽

Joshua J.C. McGrath ◽

Danya Thayaparan ◽

Peiyao Wang ◽

...

Keyword(s):

Mouse Model ◽

Lung Tissue ◽

Alveolar Macrophages ◽

Immune Cell ◽

Smoke Exposure ◽

Exposure Model ◽

Exposure System ◽

Immune Mediator ◽

Pharmacologically Active ◽

The Impact

Cannabis is widely used for both recreational and medicinal purposes. Inhalation of combusted cannabis smoke is the most common mode of drug consumption, exposing the lungs to the pharmacologically active ingredients, including tetrahydrocannabinol (THC) and cannabidiol (CBD). While the relationship between cannabis smoke exposure and compromised respiratory health has yet to be sufficiently defined, previous investigations suggest that cannabis smoke may dysregulate pulmonary immunity. Presently, there exists few pre-clinical animal models that have been extensively validated for contemporary cannabis smoke exposure. To address this need, we developed a mouse model with readouts of total particular matter, serum cannabinoid and carboxyhemoglobin levels, lung cellular responses, and immune mediator production.Using a commercially available smoke exposure system and a cannabis source material of documented THC/CBD composition, we exposed mice to a total particulate matter of 698.89 (sd=66.09) µg·L−1 and demonstrate increases in serum cannabinoids and carboxyhemoglobin. We demonstrate that cannabis smoke modulates immune cell populations and mediators in both male and female BALB/c mice. This modulation is highlighted by increases in airway and lung tissue macrophage populations, including tissue-resident alveolar macrophages, monocyte-derived alveolar macrophages, and interstitial macrophage subpopulations. No changes in airway or lung tissue infiltration of neutrophils were observed. Immune mediator analysis indicated significant upregulation of MDC, TARC, and VEGF within the lung tissue of cannabis smoke exposed mice. This accessible and reproducible smoke exposure model provides a foundation to explore the impact of chronic cannabis exposures and/or co-exposures with pathogens of clinical relevance, such as influenza.

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Development, qualification, validation and application of the Ames test using a VITROCELL® VC10® smoke exposure system

Toxicology Reports ◽

10.1016/j.toxrep.2018.04.003 ◽

2018 ◽

Vol 5 ◽

pp. 542-551 ◽

Cited By ~ 4

Author(s):

Kathy Fowler ◽

Wanda Fields ◽

Victoria Hargreaves ◽

Lesley Reeve ◽

Betsy Bombick

Keyword(s):

Ames Test ◽

Smoke Exposure ◽

Exposure System

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Development and validation of an open-source, disposable, 3D-printed in vitro environmental exposure system for Transwell® culture inserts

ERJ Open Research ◽

10.1183/23120541.00705-2020 ◽

2020 ◽

pp. 00705-2020

Author(s):

Abiram Chandiramohan ◽

Mohammedhossein Dabaghi ◽

Jennifer A. Aguiar ◽

Nicholas Tiessen ◽

Mary Stewart ◽

...

Keyword(s):

Epithelial Cells ◽

Airway Epithelial Cells ◽

Smoke Exposure ◽

Airway Epithelial ◽

Exposure System ◽

Human Airway ◽

3D Printed ◽

Air Liquid Interface ◽

Human Airway Epithelial Cells

Accessible in vitro models recapitulating the human airway that are amenable to study whole cannabis smoke exposure are needed for immunological and toxicological studies that inform public health policy and recreational cannabis use. In the present study, we developed and validated a novel 3D printed In Vitro Exposure System (IVES) that can be directly applied to study the effect of cannabis smoke exposure on primary human bronchial epithelial cells.Using commercially available design software and a 3D printer, we designed a four-chamber Transwell® insert holder for exposures to whole smoke. COMSOL® Multiphysics software was used to model gas distribution, concentration gradients, velocity profile and shear stress within IVES. Following simulations, primary human bronchial epithelial cells cultured at air-liquid interface on Transwell® inserts were exposed to whole cannabis smoke using a modified version of the Foltin Puff procedure. Following 24 h, outcome measurements included cell morphology, epithelial barrier function, lactate dehydrogenase (LDH) levels, cytokine and gene expression.Whole smoke delivered through IVES possesses velocity profiles consistent with uniform gas distribution across the four chambers and complete mixing. Airflow velocity ranged between 1.0–1.5 µm s−1 and generated low shear stresses (≪ 1 Pa). Human airway epithelial cells exposed to cannabis smoke using IVES showed changes in cell morphology and disruption of barrier function without significant cytotoxicity. Cannabis smoke elevated IL-1 family cytokines and elevated CYP1A1 and CYP1B1 expression relative to control, validating IVES smoke exposure impacts in human airway epithelial cells at a molecular level.The growing legalisation of cannabis on a global scale must be paired with research related to potential health impacts of lung exposures. IVES represents an accessible, open-source, exposure system that can be used to model varying types of cannabis smoke exposures with human airway epithelial cells grown under air-liquid interface culture conditions.

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