A Combination of Physical and Chemical Treatments Is More Effective in The Preparation of Acellular Uterine Scaffolds

Document Type : Original Article


1 Department of Immunology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran

2 Department of Tissue Engineering, School of Medicine, Shahroud, University of Medical Sciences, Shahroud, Iran

3 Sexual Health and Fertility Research Center, University of Medical Sciences, Shahroud, Iran


Decellularized uterine scaffold, as a new achievement in tissue engineering, enables recellularization and
regeneration of uterine tissues and supports pregnancy in a fashion comparable to the intact uterus. The acellular
methods are methods preferred in many respects due to their similarity to normal tissue, so it is necessary to try to
introduce an acellularization protocol with minimum disadvantages and maximum advantages. Therefore, this study
aimed to compare different protocols to achieve the optimal uterus decellularization method for future in vitro and in
vivo bioengineering experiments.

Materials and Methods:
In this experimental study, rat uteri were decellularized by four different protocols (P) using
sodium 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 were
examined by histopathological staining, DNA quantification, MTT assay, blood compatibility assay, FESEM, and
mechanical studies.

Histology assessment showed that only in P4, cell residues were completely removed. Masson’s trichrome
staining demonstrated that in P3, collagen fibers were decreased; however, no damage was observed in the collagen
bundles using other protocols. In indirect MTT assays, cell viabilities achieved by all used protocols were significantly
higher than the native samples. The percentage of red blood cell (RBC) hemolysis in the presence of prepared scaffolds
from all 4 protocols was less than 2%. The mechanical properties of none of the obtained scaffolds were significantly
different from the native sample except for P3.

Uteri decellularized with a combination of physical and chemical treatments (P4) was the most favorable
treatment 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.


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