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  • Evaluation of two decellularization methods in the development of a whole-organ decellularized rat liver scaffold.

Evaluation of two decellularization methods in the development of a whole-organ decellularized rat liver scaffold.

Liver international : official journal of the International Association for the Study of the Liver (2013-01-11)
Haozhen Ren, Xiaolei Shi, Liang Tao, Jiangqiang Xiao, Bing Han, Yue Zhang, Xianwen Yuan, Yitao Ding
ABSTRACT

Hepatic tissue engineering is considered as a possible alternative to liver transplantation for end-stage liver disease. Several methods of decellularization of xenogeneic liver are available to produce three-dimensional organ scaffolds for engineering liver tissues. However, rare studies have examined and compared the effectiveness of different methods on the structure and composition of intact decellularized liver extracellular matrix. Two decellularization methods were adopted herein. Their effects on collagen, elastin, glycosaminoglycans (GAGs), hepatocyte growth factor (HGF) content and influence to the function of hepatocytes cultured in scaffolds were examined and compared. The complete tissue decellularization was successfully achieved after treatment with sodium dodecyl sulphate (SDS) and Triton X-100. The total absence of nuclear structures and removal of viable cells were confirmed by haematoxylin-eosin staining and scanning electron microscopy. Collagen was preserved after both treatments. However, the elastin content decreased to about 20% and 60%, the GAGs content decreased to about 10% and 50% and the HGF content decreased to about 20% and 60% of the native liver level after SDS and Triton X-100 treatment respectively. The Triton X-100-treated scaffolds were much superior than SDS-treated scaffolds in supporting liver-specific function, including albumin secretion (P = 0.001), urea synthesis (P = 0.002), ammonia elimination (P = 0.007) and mRNA expression levels of drug metabolism enzymes. This study suggested that liver extracellular matrix scaffolds constructed using perfusion of Triton X-100 as described herein might provide a more effective and ideal material for the usage in tissue engineering and regenerative medicine approaches.

MATERIALS
Product Number
Brand
Product Description

Octoxinol 10, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
Triton X-305 solution, 70% in H2O
Sigma-Aldrich
Triton X-102
Sigma-Aldrich
Triton X-100 solution, BioUltra, for molecular biology, ~10% in H2O
Sigma-Aldrich
Triton X-100, laboratory grade
Sigma-Aldrich
Triton X-100, BioXtra
Sigma-Aldrich
Triton X-100, for molecular biology
Sigma-Aldrich
Triton X-100, peroxide- and carbonyl-free
Sigma-Aldrich
Triton X-45