Comparing various protocols of human and bovine ovarian tissue decellularization to prepare extracellular matrix-alginate scaffold for better follicle development in vitro

Abstract

Background Nowadays, the number of cancer survivors is significantly increasing as a result of efficient chemo/radio therapeutic treatments. Female cancer survivors may suffer from decreased fertility. In this regard, different fertility preservation techniques were developed. Artificial ovary is one of these methods suggested by several scientific groups. Decellularized ovarian cortex has been introduced as a scaffold in the field of human fertility preservation. This study was carried out to compare decellularization of the ovarian scaffold by various protocols and evaluate the follicle survival in extracellular matrix (ECM)-alginate scaffold. Results The micrographs of H&E and DAPI staining confirmed successful decellularization of the ovarian cortex in all experimental groups, but residual DNA content in SDS-Triton group was significantly higher than other groups (P < 0.05). SEM images demonstrated that complex fiber network and porosity structure were maintained in all groups. Furthermore, elastin and collagen fibers were observed in all groups after decellularization process. MTT test revealed higher cytobiocompatibility of the SDS-Triton-Ammonium and SDS-Triton decellularized scaffolds compared with SDS groups. Compared to the transferred follicles into the sodium alginate (81%), 85.9% of the transferred follicles into the decellularized scaffold were viable after 7 days of cultivation (P = 0.04). Conclusion Although all the decellularization procedures was effective in removal of cells from ovarian cortex, SDS-Triton-Ammonium group showed less residual DNA content with higher cytobiocompatibility for follicles when compared with other groups. In addition, the scaffold made from ovarian tissues decellularized using SDS-Triton-Ammonium and sodium alginate is suggested as a potential 3D substrate for in vitro culture of follicles for fertility preservation.

Three-Dimensional Modelling of Ovarian Cancer: From Cell Lines to Organoids for Discovery and Personalized Medicine

“…A handful of studies have shown high biocompatibility with ovarian cell types grown in reconstituted dECM hydrogels and scaffolds. A mixture of sodium alginate with decellularized murine ovarian tissue supported in vitro follicle survival ( Nikniaz et al, 2021 ). Preliminary studies of hydrogels derived from decellularized porcine ovarian ECM highlighted the effect of ECM stiffness on ovarian follicle development, with stiffer matrices reducing oocyte viability and triggering premature follicle release ( Buckenmeyer et al, 2016 ), though this effect has not been investigated with ovarian cancer cells.…”

Recent scaffold‐based tissue engineering approaches in premature ovarian failure treatment

“…Alginate hydrogel Nikniaz et al (2021) cultured ovarian follicles in decellularized ovarian scaffold using alginate hydrogel, examined follicle survival and examined the different decellularization protocols to compare optimized tissue degradation biocompatibility and DNA residues. In this study, the ECM scaffold matrix does not show a significant difference in the decellularized structure of the scaffold; however, the method for remaining DNA elimination is different, and cell viability can be affected by different decellularization methods (Berkholtz et al, 2006).…”

“…The results of this study are interpreted according to the following issues: (a) This study was performed in laboratory conditions for 7 days and needed further studies, (b) Decellularization of the ovarian cortex by SDS‐Triton‐Ammonium has less residual DNA than other methods and is also suitable for follicle culture, (c) Preantral follicles in combination with ECM and alginate survive and grow. (d) Preantral follicles survive and grow with ECM and alginate (Nikniaz et al., 2021).…”

Future Challenges and Opportunities of Extracellular Matrix Hydrogels in Female Reproductive Medicine

“…ECM products that are obtained from decellularization of female reproductive organs have salient roles in the field of reproductive medicine, with research focusing mainly on those that are derived from the uterus [ 51 , 64 , 65 , 66 , 67 , 68 , 69 , 70 , 71 , 72 , 73 , 74 , 75 , 76 , 77 , 78 , 79 , 80 ], ovary [ 50 , 81 , 82 , 83 , 84 , 85 , 86 , 87 , 88 , 89 , 90 , 91 , 92 ], cervix [ 93 ], vagina [ 94 , 95 ], and placenta [ 96 , 97 , 98 , 99 , 100 , 101 , 102 , 103 , 104 , 105 ]. The variable methods that are used for decellularization are dependent on the tissue type and its characteristics.…”