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Generation and Grafting of Tissue-engineered Vessels in a Mouse Model
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Layer-specific cell differentiation in bi-layered vascular grafts under flow perfusion
Iris Pennings1,2, Eline E van Haaften3,4, Tomasz Jungst5
1Department of Oral and Maxillofacial Surgery & Special Dental Care, UMC Utrecht, Utrecht University, Utrecht, The Netherlands.
Biofabrication
|September 26, 2019
Summary
This study developed a novel bioreactor to guide cell differentiation in bioengineered vascular grafts. The system successfully created native-like neovessels by maintaining distinct cell layers for improved vascular tissue engineering.
Area of Science:
- Biomaterials Science
- Tissue Engineering
- Regenerative Medicine
Background:
- Bioengineered vascular grafts offer an alternative to traditional substitutes but face challenges in achieving biomimetic cellular organization.
- Inducing and maintaining multiple cell phenotypes within a layered structure is crucial for functional neovessel regeneration.
Purpose of the Study:
- To develop and validate a perfusable two-compartment bioreactor for maturing bioengineered vascular grafts.
- To investigate the induction and maintenance of specific cell phenotypes (endothelial cells and vascular smooth muscle cells) in a layered, biomimetic vascular graft structure.
Main Methods:
- Vascular grafts were fabricated using electrospinning and melt electrowriting, followed by co-culture of endothelial colony forming cells and multipotent mesenchymal stromal cells (MSCs).
- A two-compartment bioreactor provided cell-specific media to the luminal and outer layers of the grafts under flow perfusion.
- Cell differentiation was assessed by analyzing specific gene markers (e.g., COX-2, KLF2, eNOS, αSMA, calponin) and cellular morphology.
Main Results:
- The bioreactor successfully induced and maintained endothelial cells (ECs) in the luminal layer and differentiated multipotent mesenchymal stromal cells (MSCs) into vascular smooth muscle-like cells (vSMCs) in the outer layer.
- ECs exhibited flow responsiveness, and MSCs differentiated into vSMC-like cells with appropriate markers (αSMA, calponin) and extracellular matrix production (collagen IV, elastin) without affecting the EC layer.
- The distinct cellular layers created a barrier, ensuring effective separation of cell-specific media in the bioreactor compartments.
Conclusions:
- The developed two-compartment bioreactor system effectively promotes layer-specific cell differentiation and organization in bioengineered vascular grafts.
- This system facilitates the creation of biomimetic neovessels with native-like cellular structures, advancing vascular tissue engineering.
- The approach holds significant potential for investigating and improving the development of functional bioengineered vascular grafts.
