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Enhanced Re-Endothelialization of Decellularized Rat Lungs
Collin T Stabler1, Luiz C Caires1,2, Mark J Mondrinos1
11 Department of Bioengineering, College of Engineering, Temple University , Philadelphia, Pennsylvania.
Tissue Engineering. Part C, Methods
|March 4, 2016
Summary
Optimizing lung scaffold re-endothelialization with endothelial cells (ECs) using a supine position enhances cell distribution and vascular function for lung engineering and transplantation.
Area of Science:
- Regenerative Medicine
- Tissue Engineering
- Vascular Biology
Background:
- Decellularized lung scaffolds offer promise for lung transplantation and disease modeling.
- Efficient re-endothelialization of the pulmonary vasculature is critical for recellularized lung scaffold success.
- Current methods face challenges in achieving homogeneous endothelial cell (EC) seeding.
Purpose of the Study:
- To optimize the re-endothelialization of decellularized rodent lung scaffolds using rat lung microvascular endothelial cells (ECs).
- To investigate the impact of lung posture during cell seeding on EC distribution and retention.
- To assess the functional recovery of the endothelial barrier post-seeding and perfusion culture.
Main Methods:
- Decellularized rat lungs were seeded with ECs via the pulmonary artery in either supine or upright positions.
- Cell distribution and retention were quantified across lung lobes and vessel diameters.
- EC integrin binding to extracellular matrix components (collagen, fibronectin) was examined.
- A custom leakage assay evaluated endothelial barrier function after in vitro maturation.
Main Results:
- The supine position significantly improved homogeneous EC seeding and retention, particularly in apex regions and right upper/left lobes.
- Greater EC retention was observed in larger proximal vessels (100-5000 μm) with the supine posture.
- EC adhesion depended on integrin binding to residual extracellular matrix proteins like collagen and fibronectin.
- Perfusion culture led to EC spreading and partial restoration of endothelial barrier function.
Conclusions:
- Lung posture during EC seeding is a critical factor for achieving uniform vascularization in engineered lung scaffolds.
- The supine position enhances EC distribution and retention in the pulmonary vasculature.
- Restoring endothelial barrier function is achievable through optimized seeding and perfusion culture, paving the way for improved lung engineering strategies.
