A Tissue-Engineered Tracheobronchial In Vitro Co-Culture Model for Determining Epithelial Toxicological and Inflammatory Responses

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.

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Faculty Opinions recommendation of Tissue factor-bearing exosome secretion from human mechanically stimulated bronchial epithelial cells in vitro and in vivo.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.717968028.793467693 ◽

2012 ◽

Author(s):

Marina Pretolani

Keyword(s):

Epithelial Cells ◽

Tissue Factor ◽

Bronchial Epithelial Cells ◽

Bronchial Epithelial

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Tissue factor–bearing exosome secretion from human mechanically stimulated bronchial epithelial cells in vitro and in vivo

Journal of Allergy and Clinical Immunology ◽

10.1016/j.jaci.2012.05.031 ◽

2012 ◽

Vol 130 (6) ◽

pp. 1375-1383 ◽

Cited By ~ 61

Author(s):

Jin-Ah Park ◽

Asma S. Sharif ◽

Daniel J. Tschumperlin ◽

Laurie Lau ◽

Rachel Limbrey ◽

...

Keyword(s):

Epithelial Cells ◽

Tissue Factor ◽

Bronchial Epithelial Cells ◽

Bronchial Epithelial

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Expression of lipocortins in human bronchial epithelial cells: effects of IL-1β , TNF-α, LPS and dexamethasone

Mediators of Inflammation ◽

10.1155/s0962935196000300 ◽

1996 ◽

Vol 5 (3) ◽

pp. 210-217

Author(s):

M. M. Verheggen ◽

H. I. M. de Bont ◽

P. W. C. Adriaansen-Soeting ◽

B. J. A. Goense ◽

C. J. A. M. Tak ◽

...

Keyword(s):

Epithelial Cells ◽

Bronchial Epithelium ◽

Northern Blotting ◽

Bronchial Epithelial Cells ◽

Human Bronchial Epithelial Cells ◽

Bronchial Epithelial ◽

Annexin I ◽

Tnf Α

In this study, we investigated the expression of lipocortin I and II (annexin I and I in the human bronchial epithelium, bothin vivoandin vitro. A clear expression of lipocortin I and II protein was found in the epithelium in sections of bronchial tissue. In cultured human bronchial epithelial cells we demonstrated the expression of lipocortin I and II mRNA and protein using Northern blotting, FACScan analysis and ELISA. No induction of lipocortin I or II mRNA or protein was observed after incubation with dexamethasone. Stimulation of bronchial epithelial cells with IL-1β, TNF-α or LPS for 24 h did not affect the lipocortin I or II mRNA or protein expression, although PGE2and 6-keto-PGF1αproduction was significantly increased. This IL-1β- and LPS-mediated increase in eicosanoids could be reduced by dexamethasone, but was not accompanied by an increase in lipocortin I or II expression. In human bronchial epithelial cells this particular glucocorticoid action is not mediated through lipocortin I or II induction.

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Dose Response Of Continuous Positive Airway Pressure (CPAP) On Inflammatory Responses In Human Bronchial Epithelial Cells In Vitro

10.1164/ajrccm-conference.2012.185.1_meetingabstracts.a6457 ◽

2012 ◽

Author(s):

Mohammed D. AlAhmari ◽

Raymond J. Sapsford ◽

Jadwiga A. Wedzicha ◽

John R. Hurst

Keyword(s):

Continuous Positive Airway Pressure ◽

Epithelial Cells ◽

Dose Response ◽

Airway Pressure ◽

Positive Airway Pressure ◽

Inflammatory Responses ◽

Bronchial Epithelial Cells ◽

Human Bronchial Epithelial Cells ◽

Bronchial Epithelial

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LKB1/p53/TIGAR/autophagy-dependent VEGF expression contributes to PM2.5-induced pulmonary inflammatory responses

Scientific Reports ◽

10.1038/s41598-019-53247-6 ◽

2019 ◽

Vol 9 (1) ◽

Author(s):

Huan Xu ◽

Xiuduan Xu ◽

Hongli Wang ◽

Aodeng Qimuge ◽

Shasha Liu ◽

...

Keyword(s):

Epithelial Cells ◽

Inflammatory Responses ◽

Critical Role ◽

Bronchial Epithelial Cells ◽

Vegf Expression ◽

Bronchial Epithelial ◽

Autophagy Induction ◽

Pathway Activation ◽

Pm2.5 Exposure

Abstract One of the health hazards of PM2.5 exposure is to induce pulmonary inflammatory responses. In our previous study, we demonstrated that exposing both the immortalized and primary human bronchial epithelial cells to PM2.5 results in a significant upregulation of VEGF production, a typical signaling event to trigger chronic airway inflammation. Further investigations showed that PM2.5 exposure strongly induces ATR/CHK1/p53 cascade activation, leading to the induction of DRAM1-dependent autophagy to mediate VEGF expression by activating Src/STAT3 pathway. In the current study, we further revealed that TIGAR was another transcriptional target of p53 to trigger autophagy and VEGF upregulation in Beas-2B cells after PM2.5 exposure. Furthermore, LKB1, but not ATR and CHK1, played a critical role in mediating p53/TIGAR/autophagy/VEGF pathway activation also by linking to Src/STAT3 signaling cascade. Therefore, on combination of the previous report, we have identified both ATR/CHK1/p53/DRAM1- and LKB1/p53/TIGAR- dependent autophagy in mediating VEGF production in the bronchial epithelial cells under PM2.5 exposure. Moreover, the in vivo study further confirmed VEGF induction in the airway potentially contributed to the inflammatory responses in the pulmonary vascular endothelium of PM2.5-treated rats. Therefore, blocking VEGF expression or autophagy induction might be the valuable strategies to alleviating PM2.5-induced respiratory injuries.

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Deletion of Shp2 in bronchial epithelial cells impairs IL-25 production in vitro, but has minor influence on asthmatic inflammation in vivo

PLoS ONE ◽

10.1371/journal.pone.0177334 ◽

2017 ◽

Vol 12 (5) ◽

pp. e0177334

Author(s):

Zhangwei Qiu ◽

Jiesen Zhou ◽

Fang Liu ◽

Xuejun Qin ◽

Yuanrong Dai ◽

...

Keyword(s):

Epithelial Cells ◽

Bronchial Epithelial Cells ◽

Bronchial Epithelial ◽

Minor Influence

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IKBKE Upregulation is Positively Associated with Squamous Cell Carcinoma of the Lung In Vivo and Malignant Transformation of Human Bronchial Epithelial Cells In Vitro

Medical Science Monitor ◽

10.12659/msm.893815 ◽

2015 ◽

Vol 21 ◽

pp. 1577-1586 ◽

Cited By ~ 8

Author(s):

Hezuo Lv

Keyword(s):

Squamous Cell Carcinoma ◽

Epithelial Cells ◽

Cell Carcinoma ◽

Malignant Transformation ◽

Squamous Cell ◽

Bronchial Epithelial Cells ◽

Human Bronchial Epithelial Cells ◽

Bronchial Epithelial

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Bronchial epithelial cells exposed to isocyanates potentiate activation and proliferation of T-cells

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.1990.259.4.l320 ◽

1990 ◽

Vol 259 (4) ◽

pp. L320-L327 ◽

Cited By ~ 8

Author(s):

S. Mattoli ◽

S. Miante ◽

F. Calabro ◽

M. Mezzetti ◽

A. Fasoli ◽

...

Keyword(s):

T Cell ◽

Epithelial Cells ◽

Inflammatory Responses ◽

Cell Damage ◽

Bronchial Epithelial Cells ◽

Bronchial Epithelial ◽

Hla Dr ◽

Immunological Mechanisms ◽

Chemotactic Factors

Bronchial epithelial cells release chemotactic factors for lymphocytes and express HLA-DR antigens. Thus they may contribute to the T-cell-mediated inflammatory responses involved in a number of pulmonary diseases such as asthma. In this study, the in vitro exposure of human bronchial epithelial cells to toluene 2,4-diisocyanate (TDI), an inflammatory and asthmogenic stimulus presumed to act at least in part through immunological mechanisms, provoked cell damage followed by proliferation of the cells that survived the injury. At the time of the proliferative response, epithelial cells released factors that upregulated the activation and proliferation of T lymphocytes presensitized by antigen receptor triggering. The T-cell activating factors were interleukin (IL) 1- and 6-like substances, as demonstrated by the ability of specific antisera to inhibit most of the biological effect, and by the ability of recombinant IL-1 and IL-6 to reproduce it. Appreciable amounts of immunoreactive IL-1 and IL-6 were indeed recovered in the supernatants of TDI-exposed epithelial cells. The release of these cytokines may represent an important mechanism by which epithelial cells respond to some environmental stimuli and contribute to the persistence of inflammatory responses in the airways.

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