Abstract
Purpose: Decellularized uterine scaffold, as a new achievement in tissue engineering, permits recellularization and regeneration of uterine tissues and supports pregnancy in a fashion comparable to the intact uterus. The main purpose of this study was using of different chemical methods to introduce an optimized protocol for decellularization of rat uterus.Method: We decellularized rat uteruses by four different protocols using sodium dodecyl sulfate (SDS) and triton-X100 with different doses and time incubations.We characterized the scaffolds through histopathological staining, DNA quantification, MTT assay, Blood compatibility assay, Field Emission Scanning Electron Microscopy (FESEM), and biomechanical studies.Results: Histology assessment showed that only in protocol 4, cell residues were completely removed. Masson’s trichrome staining demonstrates that in protocol P3 collagen bundles were decreased; however, no damage in the collagen bundles was observed by other protocols. Cell viabilities indirect MTT assays of all protocols were significantly higher than the native samples. The RBC hemolysis percent in the presence of prepared scaffolds from all 4 protocols was less than 2%. The mechanical properties of none of the protocols were significantly different from the native sample. Conclusion: Protocol 4 which used freeze-thawing before using detergent, was introduced as the optimized protocol due to complete removal of cell residue, preservation of the three-dimensional structure, complete removal of detergents, and preservation of the mechanical property of the scaffolds.