Table 2.

Decellularization Tissue Sterilization and Preservation methods for developing dECM biomaterials/grafts for clinical use.

Category	Treatment/Technique	Effect on Tissue	Ref.
Sterilization Methods
	Gamma and Electron irradiation	-Radiation based destruction of existing microbes -Destruction of DNA and prevention of microorganism replication, however, can cause damage to ECM collagen network	[277,278]
	Ethylene oxide (EtO)	-Irreversible alkylating agent -Prevents replication of microorganisms by damaging DNA -Suppresses cellular metabolism and division and inactivates many bacteria and viruses	[58,60]
	Supercritical carbon dioxide (scCO2)	-Penetrates ECM and inactivates microorganisms present with minimal structural disruption -Relatively non-toxic, attractive method for the creation of constructs with high biocompatibility	[279,280]
Preservation Methods
	Lyophilization	-Vacuum sublimation process for water removal through sublimation of ice -Protein stabilization at room temperature for long term storage -Required subsequent reconstitution step	[65,262]
	Cryopreservation	-Slow-rate freezing or snap freezing -Requires use of cryoprotective agents to mitigate damaging effects of ice crystal formation -Stabilizes material for long term storage by preventing degradation of biological molecules	[281]
	Antibiotics and Antimycotics in −20/-80 °C	-Inactivates bacteria through specific intracellular targeting and destruction -Slows chemical processes and degradation for short-long term storage	[282]