Effect of preoperative simvastatin treatment on transplantation of cryopreserved-warmed mouse ovarian tissue quality

Abstract

After the ovarian tissue (OT) transplantation, the ischemia-reperfusion injury causes depletion and apoptosis of follicle. Recent reports stated that simvastatin reduces ischemic damage. Therefore, we used the mouse whole ovarian vitrification and autotransplantation models to investigate the effects of simvastatin. Five-week-old B6D2F1 mice were randomly divided into four groups. Three groups were given simvastatin orally (5 mg/kg) before ovariectomy, either 2 hours before (2H Tx) or once a day for 3 or 7 days. The control group was given saline 2 hours before ovariectomy. All ovaries were cryopreserved by vitrification, held in liquid nitrogen for 1 week before being warmed, and autotransplanted. The grafts were collected for analysis on 2, 7, or 21 days after transplantation. Ovarian follicle morphology and apoptosis were assessed by hematoxylin and eosin staining and terminal deoxynucleotidyl transferase dUTP nick end labeling assay. Vessel integrity in ovary was evaluated by immunohistochemistry using anti-CD31 antibody. Serum FSH level was measured to estimate the transplanted ovarian reserve. The proportion of morphologically normal (G1) follicles at 7 and 21 days and the percentage of CD31 (+) tissue at 21 days was significantly higher in the 2H Tx group than that in the control group. In addition, the 2H Tx group showed a significantly increased intact primordial follicle ratio at 2 and 21 days after OT transplantation. Administration of simvastatin 2 hours before ovariectomy could improve the quality after transplantation of cryopreserved mouse OT.

Keywords: Autotransplantation; Fertilty preservation; Ovary; Simvastatin; Vitrification.