TY - JOUR T1 - Development and validation of an open-source, disposable, 3D-printed <em>in vitro</em> environmental exposure system for Transwell® culture inserts JF - ERJ Open Research JO - erjor DO - 10.1183/23120541.00705-2020 SP - 00705-2020 AU - Abiram Chandiramohan AU - Mohammedhossein Dabaghi AU - Jennifer A. Aguiar AU - Nicholas Tiessen AU - Mary Stewart AU - Quynh T. Cao AU - Jenny P. Nguyen AU - Nima Makhdami AU - Gerard Cox AU - Andrew C. Doxey AU - Jeremy A. Hirota Y1 - 2020/01/01 UR - http://openres.ersjournals.com/content/early/2020/11/26/23120541.00705-2020.abstract N2 - 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.FootnotesThis manuscript has recently been accepted for publication in the ERJ Open Research. It is published here in its accepted form prior to copyediting and typesetting by our production team. After these production processes are complete and the authors have approved the resulting proofs, the article will move to the latest issue of the ERJOR online. Please open or download the PDF to view this article.Conflict of interest: Dr. Chandiramohan has nothing to disclose.Conflict of interest: Dr. Dabaghi has nothing to disclose.Conflict of interest: Dr. Aguiar has nothing to disclose.Conflict of interest: Dr. Tiessen has nothing to disclose.Conflict of interest: Dr. Stewart has nothing to disclose.Conflict of interest: Dr. Cao has nothing to disclose.Conflict of interest: Dr. Nguyen has nothing to disclose.Conflict of interest: Dr. Makhdami has nothing to disclose.Conflict of interest: Dr. Cox has nothing to disclose.Conflict of interest: Dr. Doxey has nothing to disclose.Conflict of interest: Dr. Hirota has nothing to disclose. ER -