Systematic analysis of the influence of enzymatic and chemical detergents on structure, biomechanics and biocompatibility of decellularized vascular grafts

The aim of this study was to systematically compare standard detergents for the generation of Decellularization Vascular Grafts (DVG) in terms of their influence on vascular key characteristics. The most common enzymatic and chemical detergents for decellularization were identified from literature, standardized and included: i) Trypsin, ii) Sodium Dodecyl Sulfate (SDS)- and iii) Triton X-100. All protocols were applied to porcine vessels and the manufactured DVG were analyzed for histological, ultrastructural morphology and biomechanical characteristics. Further, DVG were seeded with Human Umbilical Vein Endothelial Cells (HUVEC) and cultured in a bioreactor to investigate biocompatibility after decellularization. Anti-Coagulation properties were assessed by the Chandler Loop model and a platelet-activation-assay. The Trypsin and SDS treatment were the most effective protocols in terms of tissue clearance but both impaired the ultrastructural integrity of the vessel wall in contrast to the Triton X-100 treatment. Moreover, biomechanical characteristics in the test stand did not differ significantly across the applied protocols but treatment of DVG with Trypsin was associated with a reduced Young's modulus and injuries in the vessel wall in a pulsatil flow model after 30 d. Moreover, coagulation was decreased in the Trypsin-treated group and was slightly increased in the SDS group but no significant difference towards the control group was noted. DVG after Triton X-100 treatment were the only ones capable for successful cell seeding. The here presented experimental data emphasized the main advantages and disadvantages of the most common enzymatic and chemical detergents for the manufacturing of DVG.