Optimization and evaluation of the decellularization of a biohybrid material by supercritical CO2 treatment

Abstract

Recently, decellularized extracellular matrix (ECM) has been reported to provide important components to be used as a scaffold for cell delivery on damaged tissues after ischemic heart diseases. Among different decellularization techniques, the supercritical fluids are being increasingly used in living and synthetic tissues, offering a fast and less toxic decellularization process to further biological applications. The supercritical carbon dioxide is the most commonly candidate to replace or compensate the large use of toxic detergents like the Sodium Dodecyl Sulfate (SDS) due to its extraction properties. However, in this work, the synergetic effect between both SDS and supercritical CO2 was used to decellularize the biohybrid material developed by the Laboratory of Vascular Translational Science coated with gelatin to mimic the ECM and with 14 and 7 days (G14 and G7) of cell culture, for later use as a cardiac graft. After confocal analysis of the fluorescent labeled biopolymers after treatments, it was seen the potentialized decellularization effect of SDS 0.5% + scCO2 only in the G14 since almost no DNA was found. DNA quantifications were performed within the immersion liquid (SDS) and the biopolymers after treatment to check the amount of DNA that was managed to remove. Finally, this condition appeared to have a decellularization effect in a much shorter time span than the standard decellularization process and with a much smaller amount of solvent.