Construction of Artificial Ovaries with Decellularized Porcine Scaffold and Its Elicited Immune Response after Xenotransplantation in Mice

Abstract

Substitution by artificial ovary is a promising approach to restore ovarian function, and a decellularized extracellular matrix can be used as a supporting scaffold. However, biomimetic ovary fabrication and immunogenicity requires more investigation. In this study, we proposed an effective decellularization protocol to prepare ovarian scaffolds, which were characterized by few nuclear substances and which retained the extracellular matrix proteins. The ovarian tissue shape and 3-dimensional structure were well-preserved after decellularization. Electron micrographs demonstrated that the extracellular matrix fibers in the decellularized group had similar porosity and structure to those of native ovaries. Semi-quantification analysis confirmed that the amount of extracellular matrix proteins was reduced, but the collagen fiber length, width, and straightness did not change significantly. Granulosa cells were attached and penetrated into the decellularized scaffold and exhibited high proliferative activity with no visible apoptotic cells on day 15. Follicle growth was compromised on day 7. The implanted artificial ovaries did not restore endocrine function in ovariectomized mice. The grafts were infiltrated with immune cells within 3 days, which damaged the artificial ovary morphology. The findings suggest that immune rejection plays an important role when using artificial ovaries.

Extracellular matrix-derived scaffolds in constructing artificial ovaries for ovarian failure: a systematic methodological review

“…The most common source of dECM scaffolds was ovarian tissues (n = 8), followed by amniotic membranes (n = 2), greater omentum (n = 1), and peritoneal membranes (n = 1) ( Motamed et al , 2017 ; Sarabadani et al , 2021 ; Haghshenas et al , 2022 ). In seven studies, different regions of tissues (six ovaries and one amniotic membrane) were dissected and subsequently minced ( Supplementary Table S4 ) ( Laronda et al , 2015 ; Motamed et al , 2017 ; Hassanpour et al , 2018 ; Nikniaz et al , 2021 ; Chiti et al , 2022 ; Zheng et al , 2022 ; Wu et al , 2022b ). The greater omentum and amniotic membrane were directly cut into small fragments ( Fazelian-Dehkordi et al , 2022 ; Haghshenas et al , 2022 ).…”

“…Four studies used solo chemicals for decellularization, where two studies each used sodium dodecyl sulfate (SDS) and sodium lauryl ester sulfate (SLES), respectively. Seven studies used detergent combinations ( Nikniaz et al , 2021 ; Pennarossa et al , 2021 ; Sarabadani et al , 2021 ; Chiti et al , 2022 ; Fazelian-Dehkordi et al , 2022 ; Zheng et al , 2022 ; Wu et al , 2022b ). From the perspective of chemical types, ionic detergents (SDS, sodium deoxycholate, SLES) were the most commonly used reagents (n = 10) ( Laronda et al , 2015 ; Hassanpour et al , 2018 ; Alaee et al , 2021 ; Nikniaz et al , 2021 ; Pennarossa et al , 2021 ; Chiti et al , 2022 ; Haghshenas et al , 2022 ; Zheng et al , 2022 ; Wu et al , 2022b ), and five studies used non-ionic detergents, mainly Triton X-100 ( Nikniaz et al , 2021 ; Pennarossa et al , 2021 ; Chiti et al , 2022 ; Zheng et al , 2022 ; Wu et al , 2022b ).…”

“…Less frequently used detergents included ammonium hydroxide and 2-propanol. Trypsin/EDTA and DNase/RNase solution were equally frequently used enzymes following detergents (n = 6) ( Motamed et al , 2017 ; Hassanpour et al , 2018 ; Sarabadani et al , 2021 ; Chiti et al , 2022 ; Zheng et al , 2022 ; Wu et al , 2022b ). In three studies, the dECM scaffolds were further processed into hydrogels, which were easier to use and apply ( Supplementary Table S4 ) ( Chiti et al , 2022 ; Haghshenas et al , 2022 ; Zheng et al , 2022 ).…”

“…Decellularization refers to the removal of the cellular compartments while preserving the natural ECM with optimal porosity, stiffness and elasticity, thus yielding decellularized ECM (dECM) constructs ( Fig. 1 ) ( Saldin et al , 2017 ; Wu et al , 2022b ). Decellularization has been widely studied in bone, heart, dermal tissues, and small intestinal submucosa, in both basic research and clinical practice ( Bejleri and Davis, 2019 ; Xu et al , 2020 ; Datta et al , 2021 ).…”

“…( D ) H&E staining shows the absence of basophilic materials in the decellularized tissues. (A–D) Reprinted with permission from Wu et al (2022b ). ( E ) Representative images of thermal analysis characterization; the different slopes indicate the change of ECM ultrastructure or motifs after decellularization.…”