Perspectives on the extracellular matrix in inflammatory bowel disease and bowel decellularization protocols

Table 1 A summary of some of the agents used in the decellularisation process, including the type of agent, concentrations applied, treatment times, characteristics and effects on the tissue, and the preservation of the extracellular matrix

Agent	Type	Concentration	Time	Characteristics and effects	Preservation of the ECM
Sodium dodecyl sulfate	Ionic detergent	0.05%-2%	12-24 hours	Highly effective at removing cells, solubilises cell membranes; can degrade ECM (especially glycosaminoglycans)	May degrade ECM, including collagen and glycosaminoglycans
Triton X-100	Non-ionic detergent	0.1%-1% (v/v)	Variable (e.g., 24 hours)	Less damaging to the ECM, effective in removing cell debris in thick tissues	Good preservation of the ECM structure
Ammonium hydroxide	Alkaline	0.1% (v/v)	Overnight	Used in combination with Triton X-100, aids decellularisation	Preserves villous structures and ECM
Deionized water	Osmotic agent	N/A	Variable	Causes hyposmotic shock in cells, contributing to cell removal	Helps remove cellular content
Sodium deoxycholate	Ionic detergent	Variable (e.g., 0.05%)	Variable	Removes cells effectively, but can compromise the MEC	May degrade glycosaminoglycans, impacting the ECM
Methanol/chloroform	Degreaser	1:1 (v/v)	Variable	Used to remove lipids before decellularisation	Helps remove cellular content
Trypsin/EDTA	Digestive enzymes	0.05%/0.05%	Variable	Used for cellular digestion, in combination with detergents	Helps remove cellular content
KI (potassium iodine)	Additional agent	1.0 M	Variable	Used in combination with Triton X-100 for decellularisation	Helps remove cellular content
Peracetic acid	Sterilising	Variable	Variable	Used to sterilise tissue after decellularisation	Preserves the structure of the mesentery and vascular channels
DNAse	Enzyme	Variable	Variable	Removes residual DNA after decellularisation	No direct effect on ECM, focuses on DNA removal

ECM: Extracellular matrix; N/A: Not applicable.