Table1 Common physical decellularization methods and their influence on the immunogenicity of derived bioscaffolds

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]