Table1.

Common physical decellularization methods and their influence on the immunogenicity of derived bioscaffolds

Method	Advantages	drawbacks	Ref.
Freeze and thaw cycles	↓ DAMP release via reducing detergent treatment time ↑ cell removal in tissues with dense mechanical barriers (i.e., osteochondral tissue)	Inefficient antigen removal	[56] [70] [71]
Non-thermal electroporation	↑ cell removal ↓ ECM damage and DAMP release	Cytotoxicity of some applied solvents	[67, 69]
High hydrostatic pressure	↑ cell membrane lysis at high pressures (above 150 MPa) ↓ pathogen-related immunogenicity via simultaneous sterilization at 900 MPa	Protein denaturation at pressures higher than 600 MPa Compromising the dECM mechanical properties	[72] [73] [74]
Mechanical sonication	Exploiting shear stress effect to lyse cell membrane ↑ efficacy of chemical and biologic agents	Disruption in ECM structural fibers ↑ exposing antigenic sites	[69, 72]
Mechanical agitation	↑ removal of immunogenic cell debris	Ineffective for removing immunogenic cell materials from large organs and dense tissues	[72] [75]
Perfusion	↑ delivery of chemical and biologic agents ↑ removal of antigens and immunogen cell debris	Only applicable in organs with innate vasculature Disrupting ECM at high flow rates	[76, 77]
Supercritical CO2	Non-cytotoxic nature Quick decellularization time Well preservation of ECM ↓ pathogen-related immunogenicity via simultaneous sterilization	ECM denaturation due to use of co-solvents	[56] [78]
Vacuum assistance	↑ DNA and α-gal epitope removal ↓ detergent treatment time ↓ ECM denaturation and DAMP release ↑ scaffold porosity and recellularization process	Insufficiency and need for chemical and enzymatic co-treatment	[79, 80]