Glycosaminoglycans constitute an important part of the ECM structure for providing biochemical cues to guide cell behaviour24. Numerous studies have been conducted where their presence has been beneficial for directing differentiation of cells. Murphy et al.24 have emphasized the importance of GAG in increasing differentiation of mesenchymal stem cells towards a chondrogenic lineage. The role of intrinsic ECM properties including GAG content influencing cell behaviour has been demonstrated by Reilly et al. also25. Whilst it’s important to retain GAG for influencing stem cell activity, a reduction in content may also have a benefit as it reduces the density of the ECM and increases pore size26. This would allow for greater cellular infiltration if recellularization is attempted. Therefore the ideal scaffold for cartilage should have a balance of reduced GAG whilst not completely depleting its level so that cell differentiation can be guided. This has certainly been reflected by the scaffold produced in the trypsin protocol within this study where GAG has been reduced but not completely lost. Utomo et al.15 have reported an almost similar outcome for decellularization of ear cartilage with a 75.3% reduction compared to an 88% for the trypsin protocol at day 14 in this study.