# Ameliorative effect of pomegranate peel extract nanoparticles and ovarian stem cells-derived exosomes on cyclophosphamide-induced premature ovarian insufficiency

## TL;DR

The study shows that pomegranate nanoparticles and ovarian stem cell exosomes can help reverse ovarian damage caused by chemotherapy in rats.

## Contribution

This study introduces a novel combination therapy using pomegranate nanoparticles and ovarian stem cell exosomes to treat chemotherapy-induced ovarian insufficiency.

## Key findings

- Treatment with pomegranate nanoparticles and exosomes improved ovarian follicle count and hormone levels in rats.
- The combination therapy reduced cyclophosphamide-induced ovarian degeneration and elevated estrogen while lowering FSH.
- Gene expression of FSHR, CYP19A1, and AMH was upregulated in treated groups compared to untreated POI rats.

## Abstract

Premature ovarian insufficiency (POI) is a fertility disorder impacting women under 40, characterized by an early deterioration of ovarian function, and is one of the major side effects caused by chemotherapy. Cyclophosphamide is a powerful chemotherapeutic agent used in various cancers; however, it inflicts substantial harm on other tissues, particularly the gonads, leading to temporary or permanent infertility.

Forty female albino rats (Rattus norvegicus) were divided into four groups. Group I (control group) received normal saline, then premature ovarian insufficiency was induced in the remaining groups by intraperitoneal injections of cyclophosphamide (CLP). After that, Group II received no treatment. Group III was administered a daily oral dose of pomegranate (Punica granatum) nanoparticles for one month. Group IV received a daily oral dose of pomegranates, as did group III, plus ovarian stem cell-derived exosomes via intraperitoneal injection twice weekly for one month. Rats were euthanized 30 days post-POI induction; blood was then collected to evaluate hormone levels, and sections of the ovaries were collected for histopathological analysis. Frozen sections were procured for gene expression and oxidative stress studies.

The hormonal assessment of groups indicated a notable reduction in estrogen (E2) level and an elevation of follicle-stimulating hormone (FSH) in group II compared to the control and treated groups. Additionally, the ovaries of group II exhibited pronounced degeneration of ovarian follicles, accompanied by the desquamation of granulosa cells. Gene expression study indicated a downregulation of FSHR, CYP19A1, and AMH in the same group. Rats in both groups III and IV exhibited an increased number of follicles, improved ovarian shape, a considerable elevation in blood E2, a marked decrease in serum FSH levels, and an up-regulation of the three examined genes.

The study aimed to assess the therapeutic efficacy of pomegranate (Punica granatum) peel extract nanoparticles alone and their synergistic effect with ovarian stem cell exosomes in reversing premature ovarian insufficiency (POI) caused by cyclophosphamide.

Treatment with (Punica granatum) nanoparticles and exosomes partially enhanced the structure and function of the ovaries, thereby alleviating the adverse effects of Cyclophosphamide.