TABLE 1.

A systematic summary of decellularization methods, their mechanisms of actions, and related references.

Decellularization technique		Mechanisms of action	References
Physical treatments	Free-thaw cycle	Thermal shock generated from repeated freezing and thaw cycles ruptures membranes	Rabbani et al. (2021)
	High hydrostatic pressure	Cold isostatic pressure disrupts cellular membranes within tissues	Funamoto et al. (2010)
	Electroporation	Microsecond- durational electrical pulses to enhance the cellular membrane permeability drastically	Sano et al. (2010)
	Supercritical fluid	Gases/liquids exist above critical pressure/temperature eliminate cell compartments within tissues	Di Maio et al. (2021)
	Subcritical fluid	Liquefied (subcritical) DME extracts lipids of tissues	Kanda et al. (2021)
	Immersion and agitation	Immerse tissues in chambers with decellularizing agents, and agitate tissues with a magnetic plate, ultrasound source, shaker, or an agitator attached to the end of the chamber.	Syed et al. (2014)
	Perfusion	The fluid (agent solution) passage through the circulation system to tissues/organs	He and Callanan, (2013), Corridon et al. (2017), Xu et al. (2017)
Chemical treatments	Acids and bases	Bases hydrolyze proteins to promote cellular debasement and acids lead to denaturation and protein function loss	Mendoza-Novelo et al. (2011), Angelova et al. (2018)
	Surfactants (ionic, non-ionic and zwitterionic)	Lyse cell membranes through protein crystallization, destabilizing, denaturing, targeting lipid-lipid interactions	Koley and Bard (2010)
	Chaotropes	Chaotropic agents disrupt the hydrogen bonding networks, van der Waals forces, and hydrophobic effects	Salvi et al. (2005)
	Osmotic stress	Hypertonic/hypotonic solution-induced osmotic stress/shock responses disrupt cell stability and interfere with the interaction between DNA and proteins	Fu et al. (2014)
Biologic treatments	Enzymatic approach	Enzyme-based processes that disrupt the bonds and interactions between nucleic acids and interacting cells through the disruptions of neighboring proteins and other cellular components.	Rieder et al. (2004), Fu et al. (2014)
	Combination of physical/chemical/enzymatic treatments	Physical, chemical, and enzymatic treatments are combined to optimize vascular decellularization	Row et al. (2017)