Establishing a 3D Vascularized Tri‐Culture Model of the Human Airways via a Digital Light Processing Bioprinter
Bibliographic record
Abstract
The rise in chronic lung diseases globally and the corresponding lag in drug discovery in this field highlights the need for In Vitro models closely mimicking In Vivo lung tissue. Efforts to date have largely focused on In Vitro coculture models, often neglecting the pulmonary vasculature's role in lung physiology and lacking perfusability. To address this gap, we utilized digital light processing bioprinting to establish a complex three-dimensional (3D) vascularized tri-culture airway model. Models were generated using a photopolymerizable bioink consisting of 80% polyethylene glycol diacrylate (PEGDA) and 20% gelatin methacrylate (GelMa) and printed using the LUMENX+ bioprinter. Stiffness, diffusivity, and gel expansion were characterized. Models were printed with MRC-5 lung fibroblasts embedded in hydrogels, while EA.hy926 endothelial cells and 1HAEo- epithelial cells were seeded on the luminal surface and on the apical domain, respectively. Endothelialization was achieved by coating lumens with matrix proteins, followed by perfusion-based endothelial cell seeding and uniform distribution via rotating the model. Structural characterization, including immunofluorescence imaging, lactate dehydrogenase (LDH) viability, interleukin-6 and interleukin-8 quantification was performed following cigarette smoke extract (CSE) exposure. PEGDA/GelMa 80:20 hydrogels had a Young's modulus of 10.7 kPa, expanded by 101.5% in volume and 107% in weight after 24 h in phosphate-buffered saline, and turned completely blue following 12 h of exposure to 0.1% methylene blue. Immunofluorescence staining revealed an intact apical epithelial and luminal endothelial layer demonstrated by E-cadherin expression. Lung fibroblasts retained their spindle shape with dendritic extensions as shown by F-actin staining. Propidium iodide staining demonstrated 80-90% cell viability. Cigarette smoke exposure significantly increased IL-6 and IL-8 release, but not LDH release. A multiplex assay revealed distinct immune mediator profiles and clustering between co-cultures and tri-cultures at baseline, underscoring differences in intercellular communication. This study successfully engineered and characterized a 3D bioprinted vascularized tri-culture model that mimics human airways. The model is adaptable to future studies by incorporating additional cell types, primary cells, or modified designs and protocols.
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.001 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.001 | 0.001 |
| Insufficient payload (model declined to judge) | 0.002 | 0.001 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".