Title : Development of 3D bioprinted lung airway model to study mucociliary clearance
Abstract:
Cystic fibrosis (CF) is a genetic disease caused by mutations in the gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR) protein. CFTR mainly serves as an ion channel enabling chloride and bicarbonate transport in the airways. Loss or dysfunction of CFTR channel activity at the apical membrane of airway epithelial cells causes dehydration of the periciliary and mucus layers leading to impaired mucociliary clearance and airway mucus plugging. It is crucial to develop robust in vitro models to understand effects of mutations on CFTR function. Current in vitro airway models rely heavily on transwell cultures that consist of an epithelial monolayer on inserts. The aim of this study was to design and develop a 3D lung airway model incorporating patient-derived primary cells and an extracellular matrix (ECM) to monitor patient specific CFTR function.
Patient-derived primary epithelial cells were isolated from CF patient and non-CF control lung explants at transplantation and CFTR function was monitored using short-circuit current assay. Mutant CFTR function was restored in the presence of FDA-approved drug Trikafta. To mimic the human lung airway, a multilayered tubular structure was fabricated through a 3D bioprinting process. Cell viability within the bioprinted lung airway was 92%, and the patient-derived epithelial cells were differentiated into mucous secreting cells and ciliated cells, forming an epithelial cell lining along the lumen side of the fabricated airway at day 30.
The model recapitulated a human epithelialized airway and holds promise for tissue regeneration given that interconnected microporous structure within the ECM provides a suitable substrate for cell survival, growth, migration, and differentiation. This 3D scaffold is anticipated to be a valuable novel approach to determine patient specific CFTR function and develop personalized treatment.
