Table 6 Description, advantages, and disadvantages of decellularized tissue preparation techniques [116]

Chemical methods

Sodium dodecyl sulfate (SDS), Triton X-100, sodium deoxycholate.

Involve the use of various chemical agents which disrupt cell membranes and solubilize cellular proteins, facilitating the release of cellular components from tissues while preserving the ECM.

Relatively straightforward

Efficiently remove cellular material.

The choice of chemical agents and their concentration must be carefully optimized to avoid damaging the ECM and compromising its structural and functional integrity.

Physical methods

Freeze–thaw cycles, agitation, sonication, hydrodynamic forces.

Involve mechanical forces or physical treatments to remove cellular components. For example, freeze–thaw cycles cause cell membranes to rupture and cellular contents to be released, while agitation methods utilize mechanical stirring or shaking to dislodge cells from the tissue surface.

Do not require chemical agents that may affect the ECM composition.

Can be challenging to completely remove all cellular remnants, including intracellular proteins and nucleic acids.

Biological methods

Nucleases (DNase, RNase) to degrade nucleic acids, proteases (such as trypsin or collagenase) to break down proteins, and glycosidases to remove glycosaminoglycans.

Utilize enzymes to selectively degrade cellular components, facilitating cell removal.

Offer selectivity in cellular component removal and can be tailored to specific tissues or ECM compositions.

Careful optimization of enzymatic concentrations, incubation times, and temperature is necessary to ensure efficient cell removal while preserving the integrity of the ECM.