Follicle survival and oxidative stress in human ovarian tissue xenografts after oral antioxidant treatment: N-acetylcysteine and FERTILIX

Objective To improve ovarian tissue transplantation effectiveness by limiting oxidative stress after transplantation, thus increasing follicle survival. Design Experimental laboratory-controlled study. Immunodeficient mice were transplanted with frozen–thawed human ovarian cortex pieces, and received 7 days of oral treatment. The grafts were recovered 7 days and 4 weeks after transplantation. Subjects A total of 2–4 ovarian tissue fragments per patient were donated by the patients (n = 26) undergoing ovarian tissue cryopreservation for fertility preservation and transplanted to immunodeficient mice. Exposure Xenotransplanted mice received 7 days of oral treatment with either 150 mg/kg N-acetylcysteine (NAC) (NAC 150), 300 mg/kg NAC (NAC 300), FERTILIX, or Control (vehicle). Main Outcome Measured Follicle density 1 and 4 weeks after transplantation, malondialdehyde (MDA) immunohistochemistry assay and expression of antioxidant, mitochondrial, and apoptotic key genes. Results After 7 days of transplantation, ovarian xenografts showed no significant differences in antioxidant genes (HMOX and CATALASE), mitochondrial stress genes (TFAM and HSP10), oxidative stress gene (ALDH3A2), BAX/BCL2 ratio, or malondialdehyde-positive areas (Control: 7.54% ± 2.23%; NAC 150: 4.99% ± 1.23%; NAC 300: 5.91% ± 1.19%; and FERTILIX: 10.30% ± 2.63%). However, follicle densities (follicles/mm ³, mean ± SEM) showed a modest, nonsignificant increase in the NAC 150 (13.2 ± 2.85) and FERTILIX (20 ± 7.79) groups compared with controls (8.5 ± 2.85) after 4 weeks of grafting. Conclusion These findings suggest that orally administered FERTILIX and NAC have little to no efficacy in reducing oxidative stress or improving follicle survival in xenotransplanted human ovarian tissue.