Application of Detergents or High Hydrostatic Pressure as Decellularization Processes in Uterine Tissues and Their Subsequent Effects on In Vivo Uterine Regeneration in Murine Models

Abstract

Infertility caused by ovarian or tubal problems can be treated using In Vitro Fertilization and Embryo Transfer (IVF-ET); however, this is not possible for women with uterine loss and malformations that require uterine reconstruction for the treatment of their infertility. In this study, we are the first to report the usefulness of decellularized matrices as a scaffold for uterine reconstruction. Uterine tissues were extracted from Sprague Dawley (SD) rats and decellularized using either sodium dodecyl sulfate (SDS) or high hydrostatic pressure (HHP) at optimized conditions. Histological staining and quantitative analysis showed that both SDS and HHP methods effectively removed cells from the tissues with, specifically, a significant reduction of DNA contents for HHP constructs. HHP constructs highly retained the collagen content, the main component of extracellular matrices in uterine tissue, compared to SDS constructs and had similar content levels of collagen to the native tissue. The mechanical strength of the HHP constructs was similar to that of the native tissue, while that of the SDS constructs was significantly elevated. Transmission electron microscopy (TEM) revealed no apparent denaturation of collagen fibers in the HHP constructs compared to the SDS constructs. Transplantation of the decellularized tissues into rat uteri revealed the successful regeneration of the uterine tissues with a 3-layer structure 30 days after the transplantation. Moreover, a lot of epithelial gland tissue and Ki67 positive cells were detected. Immunohistochemical analyses showed that the regenerated tissues have a normal response to ovarian hormone for pregnancy. The subsequent pregnancy test after 30 days transplantation revealed successful pregnancy for both the SDS and HHP groups. These findings indicate that the decellularized matrix from the uterine tissue can be a potential scaffold for uterine regeneration.

Effect of bovine pericardial extracellular matrix scaffold niche on seeded human mesenchymal stem cell function

“…Conversely, SDS scaffolds continue to leach components which are toxic to 96.5% of cells, even after 7 d of washing. Previous reports of the effects of SDS-decellularization on resultant scaffold recellularization potential have shown mixed results, with some groups showing positive findings and others describing negative results32333435363738394041424344. The possible reasons for the differences in recellularization potential reported for SDS-decellularized scaffolds include: (1) Different tissues and organs may alter the ability to remove SDS from resultant scaffolds.…”

STAT3 accelerates uterine epithelial regeneration in a mouse model of decellularized uterine matrix transplantation

“…For decellularization of mouse uteri, the collected uterine fragments were treated by SDS (Wako), as we described previously (3). Briefly, the tissue fragments from donor mice were immersed in 1% of SDS with PBS solution at room temperature.…”

“…We recently reported a novel technique of uterine decellularization in a rat model (3). Notably, the rat uterus was partially reconstructed after the transplantation of uterine scaffold decellularized by the treatment of SDS or high hydrostatic pressure.…”

Methods of tissue decellularization used for preparation of biologic scaffolds and in vivo relevance

“…Ionic detergents are used in each of 131 the published protocols. Bhrany et al [39] used an SDS-based solu- 132 tion while Marzaro et al [24], Ozeki et al [40], Totonelli et al [41], 133 and Keane et al [23] used a 4% sodium deoxycholate solution as 134 the main decellularization agent. Each of the protocols also 135 required an endonuclease (DNase and/or RNase) to remove resid- 136 ual nucleic acids.…”