Table 4.

List of enzymatic treatments used for decellularization.

Method	Mechanism	Disadvantages	Reference
Nuclease	Catalyze the hydrolysis of the interior or terminal bonds of RNA and DNA, and aid in the complete removal of residual nucleic acids	Induce severe distortion of ECM structure.	[67,100,101,117,126,128,129,134]
		Hard to be removed incompletely, and can impede recellularization and transplantation
Trypsin	Cleave peptide bonds on the carboxyl-side of arginine and lysine	Hard to achieve sufficient decellularization.	[130,133,[135], [136], [137], [138], [139], [140]]
		Need lengthy incubation time.
		Disrupt elastin and collagen structure
Dispase	Cleave specific peptides (collagen Ⅳ and fibronectin) in the basement membranes	Damage basement membranes and ECM	[67]
Lipase	Catalyze the hydrolysis of lipids and aids in delipidation	Hard to remove all lipids	[141,142]
Phospholipase	Hydrolyze phospholipid components of cells and solubilizes cells	Significantly reduce GAG content	[143,144]