Raman spectroscopy results demonstrated, the amount of collagen types I and IV, GAG, and fibronectin was not significantly different in decellularized scaffold compared with native group but the elastin protein level was significantly decreased ( P < 0.001). The treated group with 1% SDS for 72 h showed the morphology similar to native control in having the minimum level of DNA and well preserved ECM. The MTT assay was applied for assessments of cytotoxicity of scaffold. The ultrastructure and porosity of decellularized scaffold were studied by scanning electron microscopy (SEM). The amount of tissues collagen types I and IV, fibronectin, glycosaminoglycans (GAGs), and elastin was analyzed by Raman spectroscopy. The nucleus by Acridine orange and extracellular matrix (ECM) using Masson's trichrome, Alcian blue, and periodic acid–Schiff staining were studied. After analysis the morphology and DNA content of tissues the group with better morphology and lower DNA content was selected for further assessments. The freeze–thaw cycles in combination with treatment by Triton X-100 and four concentrations of sodium dodecyl sulfate (SDS 0.1, 0.5, 1, and 1.5%) with two exposure times (24 and 72 h) were applied for tissues decellularization. The present study was aimed to compare different decellularization protocols for human endometrial fragments.
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