Combination of freeze-thaw with detergents: A promising approach to the decellularization of porcine carotid arteries

Abstract

A tissue engineering technique based on use of the extracellular matrix (ECM) as a scaffold shows great potential for preparing small-caliber vascular grafts. Decellularization protocols are still not standardized for bioengineering. The effects of freeze-thaw cycles used for decellularization are unknown. To evaluate the effects of freeze-thaw cycles on porcine carotid arteries during decellularization and to develop a promising protocol for preparing ECM scaffolds. Porcine carotid arteries were decellularized with freeze-thaw cycles followed by three different chemical protocols. Histological analysis, scanning electron microscopy, mechanical tests and pore size measurement were performed to assess their effects on the ECM. The composition, structure, and mechanical properties were not significantly changed after freeze-thaw cycles, with the exception of endothelial cells loss. Freeze-thaw led to a porous structure within arteries. The use of Triton X-100 followed by sodium dodecyl sulfate (SDS) resulted in ECM scaffolds with well-preserved composition, structure, and mechanical properties, as well as with adequate porosity. As the initial step for decellularization, freeze-thaw had little impact on arteries. Decellularized porcine carotid arteries, prepared using freeze-thaw cycles followed by treatment with Triton X-100 and SDS, may serve as a promising biological scaffold as a tissue-engineered vascular graft.