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. 2019;30(2):191-205.
doi: 10.3233/BME-191044.

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

Jin Cheng  1 , Cong Wang  1 , Yongquan Gu  1
Affiliations

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

Jin Cheng et al. Biomed Mater Eng. 2019.

Abstract

Background: 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.

Objective: To evaluate the effects of freeze-thaw cycles on porcine carotid arteries during decellularization and to develop a promising protocol for preparing ECM scaffolds.

Methods: 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.

Results: 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.

Conclusions: 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.

Keywords: Tissue engineering; decellularization; extracellular matrix; freeze-thaw; vascular grafts.

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