Enhanced decellularization efficiency in combination of low-concentration chemical and physical methods on the human umbilical arteries

Abstract

Background

Cardiovascular diseases are the leading cause of mortality worldwide, with coronary artery bypass grafting being the most effective treatment for severe cases. While autografts are preferred, donor veins are often limited. Human umbilical arteries (hUAs) show promise as an alternative. However, to make vascular graft by decellularization, a traditional chemical method can damage tissue structure and function.

Objective

This study aims to evaluate the shortening of treatment time and the hUA decellularization efficiency of the perfusion bioreactor systems.

Methods

hUAs were perfused with 1% Triton X-100 for 6 h, followed by two different concentrations of (0.5% and 1%) SDS for 18 h, and subsequently subjected to a washing procedure. The decellularization process was evaluated using histological staining and DNA quantification, along with tests for cytotoxicity, cell adhesion and proliferation.

Results

The 0.5% SDS protocol proved most effective, reducing residual DNA to below 50 ng/mg of dry weight while preserving collagen structure. It showed no cytotoxicity to L929 cells, SEM analysis confirmed human umbilical vein endothelial cell (HUVEC) attachment and CCK-8 testing showed promoted HUVECs proliferation.

Conclusion

The decellularization protocol of perfusing through 1% TX for 6 h and 0.5% SDS for 18 h through the perfusion bioreactor system is efficient in the intact hUAs tissue. This sets the stage for future in vivo studies and potential clinical applications.