Systematic in vitro comparison of decellularization protocols for blood vessels

Area of Science:

• Biomaterials Science
• Tissue Engineering
• Regenerative Medicine

Background:

• Decellularization of native blood vessels creates 3D scaffolds for vascular grafting.
• Varied decellularization protocols hinder direct comparison and optimization.
• Porcine vena cava is a suitable model for studying vascular scaffolds.

Purpose of the Study:

• To systematically compare decellularization and recellularization efficacy of five protocols on porcine vena cava.
• To evaluate the impact of detergents (SDS, SDC, CHAPS, TritonX-100) and DNase on extracellular matrix (ECM) properties.
• To identify an optimized protocol for enhanced recellularization and reduced process time.

Main Methods:

• Perfusion bioreactor setup for porcine vena cava decellularization.
• Utilized detergents: sodium dodecyl sulfate (SDS), sodium deoxycholate (SDC), CHAPS, and TritonX-100.
• Assessed decellularization, ECM integrity, mechanical strength, and recellularization with HUVECs.

Main Results:

• All tested protocols efficiently decellularized blood vessels.
• TritonX-100 protocols, especially the novel one, showed the highest recellularization efficacy with HUVECs.
• The novel TritonX protocol reduced process time and ECM stiffness.
• SDS, SDC, and CHAPS protocols exhibited lower recellularization potential.
• DNase application was crucial for complete decellularization in TritonX protocols.

Conclusions:

• Decellularization of blood vessels is achievable with various detergents.
• TritonX-100 treated ECM supports superior endothelial cell recellularization compared to SDS, SDC, and CHAPS.
• A novel TritonX-based protocol offers an improved and efficient method for generating vascular scaffolds.