Table 2.
List of physical treatments used for decellularization.
| Method | Mechanism | Disadvantages | Reference |
|---|---|---|---|
| Freeze-thaw cycles | Disrupt cell membranes and cause cell lysis by forming intracellular ice crystals | Disrupt ECM microstructure by ice crystals. | [42,43,[72], [73], [74], [75]] |
| Require subsequent treatments for the removal of cellular contents | |||
| Immersion and agitation | Rupture tissues and cells, and isolate cells from basement membranes | Alter ECM architecture. | [42,50,65,66,70,72] |
| Need optimal standards for mechanical force and exposure time | |||
| Perfusion | Remove cells from ECM and help the removal of cellular components and debris within the organ's natural vasculature | Cannot process tissues without innate vasculature. | [14,[49], [50], [51], [52],57,[76], [77], [78], [79], [80], [81]] |
| Sophisticated to operate the perfusion devices | |||
| Scraping | Mechanically remove cells from ECM surface | Damage underlying basement membranes. | [82] |
| Need standards for controlling mechanical force | |||
| Sonication | Generate acoustic cavitation bubbles, induce shear stress effect, and thus rupture the cell membrane; Ease and assist the penetration of agents by emitting the vibration, and help cellular debris removal | High power or longer duration of sonication could disrupt the main structural fibers and produce adverse effects on the vascular tissues | [[83], [84], [85]] |
| Supercritical fluids | Facilitate chemical exposure, and lead to cell removal | Not yet widely used | [[86], [87], [88]] |
| Pressure gradient/Convective flow | Burst cells, and aid in the delivery of solutions to force cellular components and soluble proteins out of tissues | Make solutions difficult to penetrate ECM due to the constantly high pressure | [[89], [90], [91], [92], [93]] |
| Pressurization | Result in cell lysis | Denature the proteins, and disrupt the ECM structure by formed ice crystals. | [51,94,95] |
| Expensive to operate | |||
| Electroporation | Cause micropore formation in cell membranes and lead to cell lysis | Hard to process large-sized tissues. | [[96], [97], [98], [99]] |
| Used solvents can be toxic |