Breathing on chip: Dynamic flow and stretch accelerate mucociliary maturation of airway epithelium in vitro

Janna C Nawroth; Doris Roth; Annemarie van Schadewijk; Abilash Ravi; Tengku Ibrahim Maulana; Christiana N Senger; Sander van Riet; Dennis K Ninaber; Amy M de Waal; Dorothea Kraft; Pieter S Hiemstra; Amy L Ryan; Anne M van der Does

doi:10.1016/j.mtbio.2023.100713

Breathing on chip: Dynamic flow and stretch accelerate mucociliary maturation of airway epithelium in vitro

Mater Today Bio. 2023 Jun 27:21:100713. doi: 10.1016/j.mtbio.2023.100713. eCollection 2023 Aug.

Authors

Janna C Nawroth^{1

2

3

4}, Doris Roth², Annemarie van Schadewijk⁵, Abilash Ravi⁵, Tengku Ibrahim Maulana², Christiana N Senger¹, Sander van Riet⁵, Dennis K Ninaber⁵, Amy M de Waal⁶, Dorothea Kraft³, Pieter S Hiemstra⁵, Amy L Ryan^{1

7}, Anne M van der Does⁵

Affiliations

¹ Hastings Center for Pulmonary Research, Division of Pulmonary, Critical Care and Sleep Medicine, Department of Medicine, University of Southern California, Los Angeles, CA, USA.
² Emulate Inc., Boston, MA, USA.
³ Helmholtz Pioneer Campus and Institute for Biological and Medical Imaging, Helmholtz Zentrum München (GmbH), Neuherberg, Germany.
⁴ Chair of Biological Imaging at the Central Institute for Translational Cancer Research (TranslaTUM), School of Medicine, Technical University of Munich, Munich, Germany.
⁵ PulmoScience Lab, Department of Pulmonology, Leiden University Medical Center, Leiden, the Netherlands.
⁶ Department of Infectious Diseases, Leiden University Medical Center, Leiden, the Netherlands.
⁷ Department of Stem Cells and Regenerative Medicine, University of Southern California, Los Angeles, CA, USA.

Abstract

Human lung function is intricately linked to blood flow and breathing cycles, but it remains unknown how these dynamic cues shape human airway epithelial biology. Here we report a state-of-the-art protocol for studying the effects of dynamic medium and airflow as well as stretch on human primary airway epithelial cell differentiation and maturation, including mucociliary clearance, using an organ-on-chip device. Perfused epithelial cell cultures displayed accelerated maturation and polarization of mucociliary clearance, and changes in specific cell-types when compared to traditional (static) culture methods. Additional application of airflow and stretch to the airway chip resulted in an increase in polarization of mucociliary clearance towards the applied flow, reduced baseline secretion of interleukin-8 and other inflammatory proteins, and reduced gene expression of matrix metalloproteinase (MMP) 9, fibronectin, and other extracellular matrix factors. These results indicate that breathing-like mechanical stimuli are important modulators of airway epithelial cell differentiation and maturation and that their fine-tuned application could generate models of specific epithelial pathologies, including mucociliary (dys)function.

Keywords: Airflow; Airway epithelium; Mechanobiology; Mucociliary clearance; Organs-on-chips.