Table 1.

Different physical methods used for animal tissue decellularization

Agent/method	Application	Effect on the ECM	Reference
Thermal shock	The formation of water crystals inside the cell destroys the cell membrane	Water crystals can destroy the ECM	[8,9,10,11,12,13,14,15,16,17]
Mechanical pressure	Pressure can disrupt tissue and cells	Pressure can damage the ECM components	[4,8,10,19,20,21,22,23]
Electroporation	The pulsed electric fields destroy the cell membrane	The pulsed electric field can destroy the ECM. The electrodes are relatively small and cover a limited area	[4,24,25,26]
Perfusion	It facilitates the distribution of the chemical agent and the removal of cellular substances	Pressure induced by perfusion can destroy the ECM	[32,43,44,45,46,47,48,49,50,51,52,53,54,55,56,57]
Pressure gradient	It facilitates the distribution of the chemical agent and the removal of cellular substances. Pressure can disrupt the cell	The pressure gradient can destroy the ECM	[4,8,32,33]
Supercritical fluids	It facilitates the distribution of the chemical agent and the removal of cellular substances	The pressure required to apply the supercritical fluid phase can destroy the ECM	[10,38,39,40]
Ultrasonic waves	High-power waves are capable of disrupting intermolecular bonds, disrupting the cell membrane, and removing its internal components	Uncontrolled cavitation can damage the structure and mechanical properties of the tissue. Structural fibers may have transverse connections	[27,28,29,30,31]
Immersion and agitation	It causes cell death and often facilitates the distribution of chemical agents and the removal of cellular substances	Severe stirring or the use of ultrasound to cause turbulence can damage the ECM	[42]
Vacuum-assisted	This method facilitates decellularization by allowing more agents to reach cells	A high negative pressure could have adverse effects on the ECM	[34,35,36,37]

ECM – Extracellular matrix