Fast decellularization process using supercritical carbon dioxide for trabecular bone

Decellularization is a process that consists on the removal of immunogenic cellular material from a tissue, so that it can be safely implanted as a functional and bioactive scaffold. Most decellularization protocols rely on the use of harsh chemicals and very long washing processes, leading to severe changes in the ultrastructure and loss of mechanical integrity. To tackle these challenges, supercritical carbon dioxide (scCO2) is herein proposed as an alternative methodology for assisting decellularization of porcine trabecular bone tissue and is combined, for the first time, with Tri(n-butyl) phosphate (TnBP). Histological and DNA analysis revealed that both TnBP and scCO2 were able to extract the DNA content from the scaffolds, being this effect more pronounced in treatments that used TnBP as a co-solvent. The combined protocol led to a decrease in DNA content by at least 90%, demonstrating the potential of this methodology and opening new possibilities for future optimizations.