Table 3.

List of chemical treatments used for decellularization.

Method	Typical Agents	Mechanism	Disadvantages	Reference
Ionic detergents	SDS,	Solubilize cytoplasmic membranes, nucleic membranes and nucleic acids, and dissociate deoxyribonucleic acid (DNA) from protein	Damage ECM structure and collagen integrity.	[36,50,51,58,114,115]
	SDC,		Reduce the contents of glycosaminoglycan (GAG) and growth factors
	Triton X-200
Non-ionic detergents	Triton X-100	Solubilize cell membranes and dissociate DNA from proteins (especially suitable for cell-derived dECM) without disrupting protein-protein interactions	Alter the ECM architecture.	[43,50,51,57,58,107,108,115,116]
			Reduce laminins/fibronectin content.
			Remove cells incompletely
Zwitterionic detergents	CHAPS,	Solubilize cell membranes and dissociate DNA from proteins by disrupting DNA-protein, lipid-lipid, lipid-protein interactions	Disrupt and dissociate proteins in the ECM	[100,[117], [118], [119], [120]]
	SB-10,
	SB-16
Acids	Peracetic acid,	Disrupt cell membranes, solubilize cytoplasmic organelles, induce cell lysis, denature proteins and catalyze hydrolytic degradation of biomolecules	Damage ECM architecture.	[76,105,114,121]
	Hydrochloric acid		Affect intracellular molecules such as GAG
Bases	Ammonium hydroxide	Solubilize cytoplasmic components, disrupt nucleic acids and catalyze hydrolytic degradation of biomolecules	Influence GAG content, collagen and growth factors.	[57,61,105]
			Weaken mechanical properties
Hypertonic solutions	Sodium chloride	Rupture cell membranes and lyse cells by osmotic shock	Difficult to remove DNA remnants	[42,118]
Hypotonic solutions	Tris-HCl	Induce cell lysis, and detach DNA from proteins	Difficult to remove DNA remnants	[116,118,122]
Organic solvents	Ethanol, Methanol, Acetones	Extract the lipid of the ECM and cause tissue dehydration, leading to cell membranes solubilization and cell lysis	Cross-link and precipitate proteins, including collagen.	[35,56,73,[123], [124], [125]]
Chelators	EDTA, EGTA	Dissociate cells and ECM by binding divalent metal ions necessary for cell adhesion	Disrupt protein-protein interactions, and denature proteins in ECM	[75,118,119,125]