Mitigation of cyclophosphamide-induced premature ovarian insufficiency in rats through bone marrow-derived mesenchymal stem cells and placental extract therapy: a comprehensive evaluation

Abstract

Objective

Premature ovarian insufficiency (POI) is a significant reproductive health concern characterized by the depletion of primary follicles, often resulting from exposure to genotoxic agents. This study aimed to explore the therapeutic potential of bone marrow-derived mesenchymal stem cells (BM-MSCs) and/or placental extracts (PE) in ameliorating POI induced by cyclophosphamide, a widely used chemotherapeutic agent.

Materials and methods

Thirty-five 8–10 weeks and weight 200 ± 20 g female albino Sprague–Dawley (SD) rats were evenly distributed into five groups: negative control (Group 1), positive control cyclophosphamide (CPA) (Group 2), stem cell therapy (CPA+MSCs) (Group 3), placental extract therapy (CPA+PE) (Group 4), and combination therapy (CPA+MSCs+PE) (Group 5). Rats in the CPA group received intraperitoneal (IP) cyclophosphamide injection (50 mg/kg), followed by daily (8 mg/kg) cyclophosphamide injection (14 days). Subsequently, the rats received 1 × 10⁶ MSCs via intravenous administration and/or 50 µL of PE, followed by sacrifice after 4 weeks. Flow cytometry was used to identify MSCs. Hormone levels (FSH, LH, E₂, and progesterone) were assessed using ELISA, and primordial follicles were quantified to evaluate primordial follicle reserves. Ovarian structure was histomorphologically evaluated, and PCNA immunohistochemistry was conducted. Morphometric measurements and statistical analyses were performed.

Results

Hormonal measurements revealed a decrease in E2 and progesterone levels, accompanied by an increase in FSH levels following cyclophosphamide treatment. However, after BM-MSC therapy, hormonal levels nearly returned to normal. In addition to BM-MSC therapy, PE treatment was also evaluated. PE administration resulted in partial restoration of hormonal balance, showing a mild increase in E2 and progesterone levels, with a slight reduction in FSH levels compared to the cyclophosphamide-treated group.

Histological examination revealed that cyclophosphamide caused significant loss of primordial follicles, stromal blood vessel damage, and substantial fibrosis. Interestingly, combination of MSCs and PE treatment showed some ameliorative effects on ovarian histology, with reduced fibrosis and slight preservation of ovarian follicles, although these changes were less pronounced than those observed with BM-MSC therapy alone.

Conclusion

MSCs therapy was more effective in restoring ovarian folliculogenesis, whereas combination with PE provided moderate protection against the histological and immunohistochemical alterations induced by cyclophosphamide.