Preparation and characterization of a rat uterine decellularized scaffold

Infertility is a special reproductive health defect. For women, congenital uterine malformations, extensive adhesions in the uterine cavity, and hysterectomy are associated with infertility. Uterine transplantation is technically feasible, but its clinical application and development are limited by donor shortages and immune rejection. Thus, uterine tissue engineering research has promising prospects. This study sought to explore the ideal perfusion strategy and evaluation process for successfully preparing natural uterine decellularized scaffolds using decellularized perfusion technology to provide a good platform for uterine tissue engineering research.

Female Sprague-Dawley rats were selected. Eluents, including TritonX-100 supplemented with sodium dodecyl sulfate, were perfused into the uterus through the uterine artery after physical freezing, thawing, and enzymatic hydrolysis. After decellularization, each scaffold was evaluated by general observation, methylene blue staining, hematoxylin and eosin staining, immunohistochemical staining, quantitative analysis of genomic DNA, collagen detection and identification, cytokine content determination, transmission electron microscopy (TEM), and scanning electron microscopy (SEM).

After perfusion, a transparent uterine scaffold was established, and the histological examination and TEM showed that it contained no cell residue. The DNA content was shown to be less than 5% that of the normal uterus. Methylene blue staining and SEM showed that the vascular network and spatial structure were intact. Immunohistochemical staining and collagen quantification showed that the extracellular matrix components of the scaffold were completely preserved. In addition, the enzyme-linked immunosorbent assay results showed that the cytokines, including epidermal growth factor, basic fibroblast growth factor, and transforming growth factor beta, had been retained in the decellularized scaffold, and still showed some biological activity.

A decellularized uterine scaffold was successfully established, and its physical and chemical properties were preserved; consequently, it may be used as an alternative platform for uterine tissue engineering research.