Freezing and bioreactor in the low-concentration detergents: A novel approach in the decellularization of small-diameter arteries

Abstract

Using decellularized small-diameter vascular bypass substitutes (<6 mm) is an efficient method for bypass grafting. A solution containing 0.5% SDS (weight/volume) is commonly used for extended periods to generate acellular tissues. However, this solution causes damage to the microfibril structure and alters the mechanical forces. Hence, the objective of this study is to reduce the concentration of SDS to preserve the structure and achieve efficient decellularization. The study employs a diluted solution of 0.3% SDS (weight/volume) to treat fresh and frozen swine small-diameter arteries, utilizing physical methods such as freezing and thawing. The effectiveness of cell removal was evaluated using histological analysis and the remaining DNA content of the sample. Furthermore, the acellular circuit also assesses the cytotoxicity and proliferation of HUVECs to gauge their safety. Through the use of 0.3% SDS, a bioreactor system, and freezing-thawing, the pig arteries are successfully decellularized, resulting in residual DNA levels of less than 50 ng/mg dry weight. This process does not cause any major changes to the biomechanical or structural properties of the arteries. The acellular samples exhibit no toxicity on the L929 cell line and promote the growth of HUVECs at their highest rate on the fourth day. This allows for the placement of acellular vascular grafts to evaluate physiological processes within the animal body. This is an important requirement in clinical blood vessel transplantation.