Objective:
Decellularized uterine scaffold, as a new achievement in tissue engineering, enables recellularization andregeneration of uterine tissues and supports pregnancy in a fashion comparable to the intact uterus. The acellularmethods are methods preferred in many respects due to their similarity to normal tissue, so it is necessary to try tointroduce an acellularization protocol with minimum disadvantages and maximum advantages. Therefore, this studyaimed to compare different protocols to achieve the optimal uterus decellularization method for future in vitro and invivo bioengineering experiments.
Materials and Methods:
In this experimental study, rat uteri were decellularized by four different protocols (P) usingsodium dodecyl sulfate (SDS), with different doses and time incubations (P1 and P2), SDS/Triton-X100 sequentially(P3), and a combination of physical (freeze/thaw) and chemical reagents (SDS/Triton X-100). The scaffolds wereexamined by histopathological staining, DNA quantification, MTT assay, blood compatibility assay, FESEM, andmechanical studies.
Results:
Histology assessment showed that only in P4, cell residues were completely removed. Masson’s trichromestaining demonstrated that in P3, collagen fibers were decreased; however, no damage was observed in the collagenbundles using other protocols. In indirect MTT assays, cell viabilities achieved by all used protocols were significantlyhigher than the native samples. The percentage of red blood cell (RBC) hemolysis in the presence of prepared scaffoldsfrom all 4 protocols was less than 2%. The mechanical properties of none of the obtained scaffolds were significantlydifferent from the native sample except for P3.
Conclusion:
Uteri decellularized with a combination of physical and chemical treatments (P4) was the most favorabletreatment in our study with the complete removal of cell residue, preservation of the three-dimensional structure,complete removal of detergents, and preservation of the mechanical property of the scaffolds
