Evaluation of Decellularization in Umbilical Cord Artery

Abstract

Major achievements in creating decellularized whole tissue scaffolds have drawn considerable attention to decellularization as a promising approach for tissue engineering. Developing a tissue-engineered small-diameter (≤2 mm) vascular graft, using decellularized human umbilical arteries (hUAs), for reconstructive surgery is a challenging task. Polymers used in the past, proved unsuitable due to serious adverse effects and autologous vessels are available only in 40% of patients. In this study, histological and proteomic analysis was performed to evaluate the efficiency of two decellularization protocols. In decellularization protocol A, hUAs were incubated in 3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate (CHAPS) and sodium dodecyl sulfate (SDS) followed by incubation in alpha minimal essential medium (α-MEM) with foetal bovine serum (FBS) while in decellularization protocol B the hUAs were incubated in Hypotonic Tris and SDS followed by incubation in nuclease solution. Histological analysis of decelullarised hUA with both protocols revealed good preservation of extracellular cell matrix (ECM) proteins and immunofluorescent staining detected collagen I and fibronectin. The DNA content within the hUAs after decellularization with protocol A was 6.2% and with protocol B 17.3%. Proteomic analysis identified cytoplasmic enzymes such as, dehydrogenase X, α-enolase and peptidyl-prolyl cis-trans isomerase A only in native samples, while, cytoskeletal proteins such as a-actin, filamin and ECM proteins like collagens were found both in native and decellularised hUA. In conclusion, both decellularization protocols effectively removed the cellular material while the ECM remained intact. Future studies are warranted to elucidate the specific effects of altered structure–function relationships on the overall fate of decellularized hUAs.