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A tissue-engineered tracheobronchial in vitro co-culture model for determining epithelial toxicological and inflammatory responses

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journal contribution
posted on 2021-08-05, 14:25 authored by Luis Soriano, Tehreem Khalid, Fergal O'BrienFergal O'Brien, Cian O'LearyCian O'Leary, Sally-Ann CryanSally-Ann Cryan
Translation of novel inhalable therapies for respiratory diseases is hampered due to the lack of in vitro cell models that reflect the complexity of native tissue, resulting in many novel drugs and formulations failing to progress beyond preclinical assessments. The development of physiologically-representative tracheobronchial tissue analogues has the potential to improve the translation of new treatments by more accurately reflecting in vivo respiratory pharmacological and toxicological responses. Herein, advanced tissue-engineered collagen hyaluronic acid bilayered scaffolds (CHyA-B) previously developed within our group were used to evaluate bacterial and drug-induced toxicity and inflammation for the first time. Calu-3 bronchial epithelial cells and Wi38 lung fibroblasts were grown on either CHyA-B scaffolds (3D) or Transwell® inserts (2D) under air liquid interface (ALI) conditions. Toxicological and inflammatory responses from epithelial monocultures and co-cultures grown in 2D or 3D were compared, using lipopolysaccharide (LPS) and bleomycin challenges to induce bacterial and drug responses in vitro. The 3D in vitro model exhibited significant epithelial barrier formation that was maintained upon introduction of co-culture conditions. Barrier integrity showed differential recovery in CHyA-B and Transwell® epithelial cultures. Basolateral secretion of pro-inflammatory cytokines to bacterial challenge was found to be higher from cells grown in 3D compared to 2D. In addition, higher cytotoxicity and increased basolateral levels of cytokines were detected when epithelial cultures grown in 3D were challenged with bleomycin. CHyA-B scaffolds support the growth and differentiation of bronchial epithelial cells in a 3D co-culture model with different transepithelial resistance in comparison to the same co-cultures grown on Transwell® inserts. Epithelial cultures in an extracellular matrix like environment show distinct responses in cytokine release and metabolic activity compared to 2D polarised models, which better mimic in vivo response to toxic and inflammatory stimuli offering an innovative in vitro platform for respiratory drug development.


Science Foundation Ireland (SFI) Centre for Research in Medical Devices (CURAM)

European Regional Development Fund, grant number 13/RC/2073



The original article is available at https://www.mdpi.com

Published Citation

Soriano L, Khalid T, O'Brien FJ, O'Leary C, Cryan SA. A tissue-engineered tracheobronchial in vitro co-culture model for determining epithelial toxicological and inflammatory responses. Biomedicines. 2021;9(6):631.

Publication Date

2 June 2021

PubMed ID



  • Amber (Advanced Material & Bioengineering Research) Centre
  • Anatomy and Regenerative Medicine
  • School of Pharmacy and Biomolecular Sciences
  • Tissue Engineering Research Group (TERG)
  • CURAM Centre for Research in Medical Devices

Research Area

  • Respiratory Medicine
  • Cancer
  • Chemistry and Pharmaceutical Sciences
  • Immunity, Infection and Inflammation
  • Biomaterials and Regenerative Medicine




  • Published Version (Version of Record)