Evaluation the induction effect of fibrin scaffold with endothelial cells and melatonin on the angiogenesis, follicles and embryos quality on vitrified transplanted ovarian tissue

Abstract

One of the methods used to preserve fertility is freezing and transplantation of ovarian tissue. Since the delay in angiogenesis and damage caused by ischemia is the main problem in transplantation, researchers are looking for new approaches to accelerate angiogenesis and improve the quality of transplanted ovarian tissue. The present study was conducted with the aim of investigating the induction effect of fibrin scaffold along with endothelial cells and melatonin on the amount of angiogenesis, the quality of follicles and embryos on frozen transplanted ovarian tissue in rats. Methods: female Wistar rats were divided into 6 groups: control group, FT group: ovaries that were transplanted under the skin after freezing and thawing, FHT group: ovaries that were transplanted after freezing and thawing together with alginate-fibrin hydrogel were transplanted under the skin, FHT+Mel group, ovaries that were transplanted under the skin after freezing and thawing with alginate-fibrin hydrogel along with melatonin, FHT+ECs group, ovaries that after freezing and thawing encapsulated with alginate-fibrin hydrogel together with endothelial cells and transplanted under the skin, FHT + Mel+ ECs group of ovaries which after freezing and thawing encapsulated with alginate-fibrin hydrogel together with endothelial cells and Melatonin and transplanted under the skin. Ovaries were removed after 14 days and then RT-PCR method was used to check the expression of Ang I and Ang II genes and by IHC technique, VWF and α-SMA factors was evaluated, and Masson trichrome staining was usaged to evaluate the fibrotic areas and for evaluating the follicle quality H&E staining was done. Results: The statistical analysis of the data showed that the expression ratio of Ang I/Ang II genes in the FHT+Mel group increased significantly compared to the FT group. while other groups did not show any significant difference. Also, the expression of VWF and α-SMA factors in FHT+Mel and FHT+ECs groups showed a significant increase compared to FT. The histomorphological examination of the ovarian samples also showed that the integrity of the ovarian tissue and the number of follicles in the FT group were greatly lost. In the groups of hydrogel alone and hydrogel together with other compounds, although the integrity of the ovarian tissue was preserved, no significant change was observed in the number and quality of follicles. Mason trichrome staining also showed an increase in fibrotic areas in the FT group, while in the FHT, FHT+Mel and FHT+ECs groups, we saw a decrease in the fibrosis areas.