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.

Erna G Santoso,Keita Yoshida,Yutaka Osuga,Osamu Yoshino,Katsuko S Furukawa,Shigeru Saito,Takashi Ushida,Yasushi Hirota,Akio Kishida,Masanori Aizawa,Masaya Yamamoto

doi:10.1371/journal.pone.0103201

Erna G Santoso, Keita Yoshida + Show 9 more

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https://doi.org/10.1371/journal.pone.0103201

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Journal: PloS one	Publication Date: Jul 24, 2014
Citations: 118	License type: CC BY 4.0

Affiliation: Tokyo Medical and Dental University

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.

Highlights

Infertility is often associated with the inability of women to either conceive or maintain pregnancy
The sample processed with 0.1% sodium dodecyl sulfate (SDS) for 1 hour (Fig. 2B) showed the most residual cells in the epithelial and stromal layers compared to the other samples
We focused on decellularized tissues for segmental uterine reconstruction; these methods are applicable for larger samples such as the whole uterus

Summary

Introduction

Infertility is often associated with the inability of women to either conceive or maintain pregnancy. A common infertility treatment is assisted reproductive technology (ART), which can work for ovulatory and tubal disorders but not for uterine abnormalities. Gynecological malignancies such as cervical and endometrial cancers often lead to the removal of the uterus by hysterectomy, which prevents the women from carrying out future pregnancies. A current solution for this type of infertility includes gestational surrogacy [11]. Gestational surrogacy is often preferred as it carries the lowest risk and highest success rate. It is impossible to prevent the possibility of accidents such as complications and death of the surrogate mother during childbirth. The conception and childbirth impose physical and mental stresses on the surrogate mother. By using surrogacy, the intended mother would not be able to experience the childbearing process

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