Ovary-derived Decellularized Extracellular Matrix-based Bioink for Fabricating 3D Primary Ovarian Cells-laden Structures for Mouse Ovarian Failure Correction

Abstract

Fertility preservation is becoming a clinical duty in practice. Three-dimensional (3D) bioprinting technology is potentially realize ovarian morphological repair and reproductive endocrine function rebuild. There is no published work on 3D bioprinting ovary using a decellularized extracellular matrix (dECM)-based bioink, though dECM is the preferred matrix choice for an artificial ovary. The study aimed to explore swine ovarian dECM-based bioink to fabricate 3D primary ovarian cells (POCs)-laden structures for mouse ovarian failure correction. In this study, the ovarian dECM was converted to dECM-based bioink by dECM solution mixed with a seaweed gelatin blend solution of bioink that was characterized using scanning electron microscopy, circular dichroism, rheology, hematoxylin and eosin staining, and immunohistochemistry. The 3D scaffolds were, then, printed with or without POCs by the extrusion 3D bioprinter. The laden POCs viability was detected with the live/dead assay kit. A female castrated mouse model was established, and the mice were treated with five different methods. The results revealed that the 3D scaffold encapsulating POCs group had more positive signals in neoangiogenesis, cell proliferation and survival than the 3D scaffold group, and ensured sex hormone secretion. Meanwhile, the expression of germ cells in the 3D scaffold encapsulating POCs group was more intensely than the non-printed hydrogel encapsulating POCs group. The work shows that the 3D bioprinting ovary employing ovarian dECM-based bioink is a promising approach for ovarian failure correction.

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 ).…”

“…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 ). Regarding the operation duration, most studies completed decellularization within several days, except for those that used intact ovaries, where the operation spanned >3 weeks ( Laronda et al , 2015 ; Hassanpour et al , 2018 ).…”

“…Raman spectrum was calculated in one study to analyze the DNA and ECM ( Alaee et al , 2021 ). Additionally, three studies tested the rheological properties of the dECM hydrogels ( Chiti et al , 2022 ; Haghshenas et al , 2022 ; Zheng et al , 2022 ). Eight studies routinely examined cytotoxicity and biocompatibility to rule out unwanted adverse effects ( Motamed et al , 2017 ; Hassanpour et al , 2018 ; Nikniaz et al , 2021 ; Pennarossa et al , 2021 ; Fazelian-Dehkordi et al , 2022 ; Haghshenas et al , 2022 ; Zheng et al , 2022 ; Wu et al , 2022b ).…”