Development of an enhanced liver scaffold recellularization using fibronectin

Area of Science:

• Biomaterials Science
• Regenerative Medicine
• Tissue Engineering

Background:

• Decellularized liver scaffolds show promise for regenerative medicine but face challenges like poor cell adhesion and vascularization.
• Fibronectin (FN) is known to improve scaffold functionality, but its specific application in liver scaffolds is underexplored.
• Current limitations hinder the clinical translation of bioengineered liver grafts.

Purpose of the Study:

• To develop and evaluate a perfusion-assisted fibronectin (FN) coating technique for decellularized liver scaffolds.
• To enhance endothelial and hepatocyte cell adhesion and integration within the scaffolds.
• To improve the overall biocompatibility and reduce thrombogenicity of liver scaffolds.

Main Methods:

• Decellularized rat liver scaffolds were coated with fibronectin (FN) using a perfusion-assisted technique.
• Endothelial cells (EA.hy926) and hepatocytes (HepG2) were seeded onto the scaffolds.
• Scaffolds were incubated in a bioreactor, and cell adhesion, proliferation, viability, and scaffold functionality were assessed.
• Ex vivo blood perfusion tests were conducted to evaluate thrombogenicity.

Main Results:

• FN coating significantly improved endothelial cell adhesion in vascular structures and increased hepatocyte coverage.
• Enhanced hepatocyte proliferation and cell viability were confirmed via immunohistochemistry and TUNEL/RT-qPCR analyses.
• Ex vivo perfusion tests showed reduced thrombogenicity, indicating improved biocompatibility and endothelialization.

Conclusions:

• Perfusion-assisted FN functionalization is an effective strategy to enhance liver scaffold biocompatibility, cell integration, and vascularization.
• This approach addresses key barriers in developing functional bioengineered liver grafts.
• The study provides a promising method for advancing liver tissue engineering and potential transplantation models.