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Treatment | Main role | Example | Advantages | Disadvantages |
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Mechanical | Force | Removal of tissue layers | Scrapping with the use of the scalpel | Some tissues like bladder and intestinal submucosa have natural planes of dissection and therefore mechanical force can be used to delaminate the tissue layers | Not all tissues can be mechanically treated, because it can disturb the ECM structure |
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Physical | Temperature |
Cell membrane lysis | Freezing cycles | Effective when combined with full decellularization protocol | Applied alone, would not remove cells completely |
Pressure | Hydrostatic pressure | Effective for tissues which do not have densely organized ECM, such as liver and lungs | Used alone, would not completely remove the cells |
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Chemical | Acids and alkalis | Protein denaturation, solubilisation of cells elements, initial nucleic acid infraction | Acetic acid, peracetic acid, hydrochloric acid, sulphuric acid, paracetic acid ammonium hydroxide, sodium azide, sodium deoxycholate | Effective | Not selective, possible alternation of ECM constituents: collagen, GAG, growth factors |
Hypertonic/hypotonic solutions | Cell disruption by the osmotic shock, disruption of DNA-protein interaction | Tris/HCl | Efficient | Do not effectively remove the cellular residues |
Ionic/nonionic detergents | Destroy DNA-protein interactions, lipids, and lipoproteins | Triton X-100 SDS | Effective, destroy lipid-lipid and lipid-protein interaction but keep protein-protein interactions | Possible protein denaturation, loss of GAGs, laminin, and fibronectin |
solvents | Dehydratation cells lysis lipids removal | Alcohol, glycerol, acetone | Effective | Possible ECM constituent destruction |
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Enzymatic | Enzymes | Cells rupture destruction of peptide bonds destruction of nucleic acids | Trypsin, DNase/RNase | Targets the residues of nucleic acids | May remain in the tissue with unknown amount and intensify the immune response of the host tissues |
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