Technique for decellularization and characterization of rat esophageal extracellular matrix for potential application as a 3D scaffold.

Abstract

Conventional treatment for esophageal defects involves surgical removal of the defect area and implant conduit tissues. There exist morbidities and mortalities associated with the treatment including fistula and leakage leading to compromise in quality of life. The aim of this study was to optimize a method for complete decellularization of rat esophagus and to solubilize the decellularized extracellular matrix (dECM) proteins to evaluate in vitro properties for scaffold fabrication. For decellularization, rat esophagi were decellularized using 3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate (CHAPS) and sodium dodecyl sulfate (SDS) buffers for 6 h and overnight, respectively. Post decellularization, the tissue was characterized for DNA, glycosaminoglycans, and elastin quantification; H&E and Masson's trichrome staining; scanning electron microscopy; and SDS-PAGE to evaluate the quantity and quality of the obtained dECM. DNA quantification and histological analysis revealed complete decellularization, while the retention of sGAGs and elastin showed the presence of extracellular proteins in the tissue. The SEM analysis revealed proper orientation of the extracellular matrix and significant proteins were retained in the dECM, which will enhance the regenerative potential. The decellularized tissues were biocompatible, exhibited no toxicity and were also soluble, which can be adapted for scaffold fabrication.