Title : A novel technique for decellularization of human esophagus for 3D bioprinting
Abstract:
The clinical treatment for esophageal defects is a huge challenge as it leads to poor quality of patient life, where the conduit tissue taken from the patients’ stomach, jejunum or colon is implanted in the damaged region. The treatment comes along with many post operative complications including fistula, leakage and bleeding. The aim of this study was to optimize a technique for decellularization of human esophagus to remove cellular components and utilize the decellularized human extra cellular matrix (dhEM) proteins for fabrication of 3D bioprinted scaffolds that promises to be a potential alternative to existing implants. Full length of human esophagus was obtained from cadaveric samples and cleaned with phosphate buffer saline to remove the blood components. About 100 mg of the esophagus is homogenized for 5 minutes using EGTA-EDTA buffer, proteases inhibitors and sodium dodecyl sulphate. Further the esophagus is centrifuged and the supernatant containing dhEM was characterized for DNA content (using fluorometer), sulphated glycosaminoglycans (sGAGs) and elastin (ELISA), and the retention of proteins was analyzed by Bicinchoninic acid assay and SDS-PAGE. Further, the dhEM was mixed with gelatin methacrylate along with mouse fibroblasts and 3D bioprinted into tubular scaffolds. The retention of major ECM proteins like sGAGs and elastin are essential to influence cell adhesion and proliferation that enhances the regenerative potential of 3D bioprinted scaffold. The optimized decellularization process was found to be successful by removing cellular contents and retaining the major ECM proteins in human esophagus. This technique confirms the feasibility of dhEM for 3D bioprinting.
Keywords: 3D bioprinting, esophagus, bioinks, regeneration
