Number Of Primordial Follicles Research Articles

P-624 Description of antral follicle count in a population of healthy French women based on age, ultrasound technique, and clinical factors

Abstract Study question How is the age-specific reference range for antral follicle count in healthy French women correlated with clinical markers ? Summary answer AFC declines linearly with age is correlated with weight, hormonal contraception, cycles duration, but not BMI, smoking or infertility history. What is known already AFC is related to the number of primordial follicles within the ovaries and is a widely used marker for baseline assessment of ovarian reserve, but there are no standardized norms for AFC, and major differences between the nomograms published in recent years notably due to the improvement of ultrasound techniques. Moreover, most published data are based on infertile women. Several factors are postulated to have influence on AFC of which ethnic origin, yet the largest cohorts published recently are Asian and therefore do not apply to a Caucasian population. Study design, size, duration In this retrospective study we used data from 640 women who had undergone a Fertility Check Up in our center, from 2018 to 2023. This concept, unique in France, offers all women of reproductive age who wish to undergo a complete ultrasound examination to check for infertility risk factors, whatever their parental plans. Participants/materials, setting, methods 640 healthy French women undergoing a Fertility Check Up were recruited. All participants completed a questionnaire about demographics, medical and surgical history, obstetrical records, familial medical history and any usage of hormonal contraception. AFC was conducted via transvaginal ultrasound using a three-dimensional acquisition, by one of three trained operators, utilizing a Voluson E10 ultrasound machine (GE Healthcare). Operators employed either the sonoAVC option (semi-automatic technique) or a manual two-dimensional counting of the acquired 3D-dimensional volume. Main results and the role of chance The distribution of AFC by age were modelized by a numerical and a graphic nomograms. AFC demonstrated a linear decline with a decrease of 1.11 follicle (IC95% 0.98 - 1.24) per year of age. Age accounted for 12.5% of the variation (R2) in AFC. Linear regression analysis showed a significant negative correlation between AFC and women’s weight (p = 0.02), but not with BMI (p = 0.23), and a significant impact of hormonal contraception (p = 0.001), with an overall reduction of 6.6 follicles in AFC compared with the women without any hormonal contraception (IC95% 4.6 - 8.6). Short menstrual cycles were associated with lower AFC (p < 0.001), when long cycles and amenorrhea were significantly associated with higher AFC (p < 0.01). There was no impact of age of menarche, smoking, family history of POI, or of a history of infertility of more than one year. Multivariate analysis showed that AFC was significantly higher with 3D-manual versus automated-3D follicular counting, the difference averaged 3.59 follicles, reducing as AFCs became smaller. Limitations, reasons for caution We present data from Caucasian women using our ultrasound machines, but in practice, each sonographer must familiarize himself with the capabilities of his own machine. Furthermore, our study does not answer the question of AFC evolution over time, for a given patient. Wider implications of the findings Some questions remain unanswered, including the longitudinal trajectory of AFC within an individual and the impact of ethnic origin. Future longitudinal studies reporting the evolution of AFC with age and involving women of diverse ethnic backgrounds are needed. Ultimately, this study highlights the impact of ultrasound equipment and technique. Trial registration number Not applicable

Primary oocytes with cellular senescence features are involved in ovarian aging in mice

In mammalian females, quiescent primordial follicles serve as the ovarian reserve and sustain normal ovarian function and egg production via folliculogenesis. The loss of primordial follicles causes ovarian aging. Cellular senescence, characterized by cell cycle arrest and production of the senescence-associated secretory phenotype (SASP), is associated with tissue aging. In the present study, we report that some quiescent primary oocytes in primordial follicles become senescent in adult mouse ovaries. The senescent primary oocytes share senescence markers characterized in senescent somatic cells. The senescent primary oocytes were observed in young adult mouse ovaries, remained at approximately 15% of the total primary oocytes during ovarian aging from 6 to 12 months, and accumulated in aged ovaries. Administration of a senolytic drug ABT263 to 3-month-old mice reduced the percentage of senescent primary oocytes and the transcription of the SASP factors in the ovary, in addition, led to increased numbers of primordial and total follicles and a higher rate of oocyte maturation. Our study provides experimental evidence that primary oocytes, a germline cell type that is arrested in meiosis, become senescent in adult mouse ovaries and that senescent cell clearance reduced primordial follicle loss and mitigated ovarian aging phenotypes.

Use of Mesenchymal Stem Cells in Experimental Ovarian Damage.

Bisphenol-A (BPA) has a well-proven deleterious effect on the hypothalamicpituitary- gonadal axis. The current study investigated the therapeutic potentials of mesenchymal stem cells (MSCs) in a murine model of BPA-induced ovarian damage. Fifty adult female rats were divided into: Group 1; control group, Group IIa, IIb: rats were given oral gavage of BPA (25 and 50 mg/Kg body weight respectively) on a daily basis for 15 days, and Group IIIa, IIIb; rats were intravenously treated with of MSCs (106 cells) after receiving the last dose of BPA as in group II. Plasma and ovarian tissue levels of Malondialdehyde (MDA) and gonadal axis hormones were assessed. Apoptosis was evaluated by TUNNEL assay and by apoptosis markers (FAS, FASL, Caspase 3, SLTM). A histological examination of ovarian tissue was also conducted. BPA resulted in a significant elevation in plasma levels of LH, FSH, and ovarian tissue levels of MDA and a significant decrease in estradiol and progesterone. All genetic and protein markers of apoptosis were elevated in BPA treated group with decreased oestrogen receptor expression in the ovarian tissue. Increased apoptotic cells were confirmed by TUNEL assay. A high dose of BPA was able to increase the number of atretic follicles in the ovarian tissue whereas the numbers of primordial, primary, secondary and Graafian follicles were decreased. All the laboratory and histological abnormalities were ameliorated by treatment with MSCs. The antioxidant and anti-apoptotic effects of MSCs could possibly explain the ability of this therapeutic modality to ameliorate BPA-induced-ovarian damage.

Morphological assessment of the ovarians after single and fractional local electron iradiation

BACKGROUND: When malignant neoplasms of the pelvic organs are irradiated, healthy ovarian tissues can get into the irradiation area. So, among all physico-chemical factors, ionizing radiation is the most common cause of ovarian failure, which has a negative impact on fertility. Conducting research in this area is especially important in connection with the active introduction of electron therapy in the protocols for the treatment of malignant neoplasms of the small pelvis with the need to find ways to prevent and treat post-radiation ovarian lesions. In addition, one of the main tasks of modern radiobiology is the creation of experimental animal models in order to reveal the mechanisms of radiation exposure with subsequent extrapolation of the results obtained to humans in order to level the side effects of radiation therapy and select optimal doses. AIM: The aim of the study was a morphofunctional assessment of the ovaries after local electron irradiation in single and fractional modes. MATERIALS AND METHODS: Wistar rats (n = 30) were divided into three groups: I — control (n = 10); II (n = 10) — subjected to a single local irradiation with electrons at a dose of 2 Gy; III (n = 10) — subjected to fractional local irradiation with electrons in a total dose of 20 Gy. RESULTS: After a single local irradiation with electrons at a dose of 2 Gy, multiple hemorrhages and a decrease in the number of growing follicles with a discontinuous theca layer, which were unevenly distributed over its volume, were noted in the ovary. A statistically significant difference in the number of follicles was revealed: a decrease in the number of primordial, primary, secondary and tertiary follicles and an increase in atretic follicles. The most pronounced difference in the number of follicles between the studied groups was found in the group of fractional electron irradiation at a dose of 20 Gy: the smallest number of primordial and the largest number of atretic follicles with signs of post-radiation fibrosis. CONCLUSIONS: The most profound damage to the ovary develops after exposure to fractional electron irradiation at a total dose of 20 Gy compared with a single exposure to ionizing radiation at a dose of 2 Gy: a reduced number of follicles, a decrease in the area and thickness of the cortical substance, as well as the thickness of the ovarian tunica, in combination with the growth of the connective tissue.

Indole-3-carbinol ameliorates ovarian damage in female old mice through Nrf2/HO-1 pathway activation

Ovarian aging leads to infertility and birth defects. We aimed to clarify the role of Indole-3-carbinol (I3C) in resistance to oxidative stress, apoptosis, and fibrosis in ovarian aging. I3C was administered via intraperitoneal injection for 3 weeks in young or old mice. Immunohistochemistry; Masson, Sirius red, and TUNEL staining; follicle counting; estrous cycle analysis; and Western blotting were used for validating the protective effect of I3C against ovarian senescence. Human granulosa-like tumor cell line and primary granulosa cells were used for in vitro assay. The results indicated that I3C inhibited ovarian fibrosis and apoptosis while increasing the number of primordial follicles. Mechanistic studies have shown that I3C promoted the nuclear translocation of nuclear factor-erythroid 2-related factor (Nrf2) and upregulated the expression of heme oxygenase 1 (HO-1). Additionally, I3C increased cell viability and decreased lactate dehydrogenase, malondialdehyde, reactive oxygen species and JC-1 levels. Furthermore, the antioxidant effect of I3C was found to be dependent on the activation of Nrf2 and HO-1, as demonstrated by the disappearance of the effect upon inhibition of Nrf2 expression. In conclusion, I3C can alleviate the ovarian damage caused by aging and may be a protective agent to delay ovarian aging.

Study on mechanism of regulation of sex hormones in treatment of hyperprolactinemia-induced ovulatory disorder infertility with Bushen Culuan Formula

Transcriptomics was used to investigate the mechanism of action of Bushen Culuan Formula in the treatment of infertility caused by hyperprolactinemia(HPRL), and animal experiments were carried out to verify the results. After establishing an animal model of HPRL-induced infertility, the mice were divided into normal group, model group, Bushen Culuan Formula groups with high-, medium-, and low-doses, and bromocriptine group, and they were observed in terms of the estrous cycle, gonadal index, serum sex hormones, morphology of ovary and mammary gland, follicle count, and fertility. The results showed that the Bushen Culuan Formula could effectively restore the estrous cycle, down-regulate the levels of prolactin(PRL), follicle-stimulating hormone(FSH), and luteinizing hormone(LH), up-regulate the level of estradiol(E_2), increase the number of primordial follicles and sinus follicles, and improve the ovulation rate and fertility of mice. Through RNA sequencing combined with biosignature analysis, Bushen Culuan Formula may regulate the metabolism of lipids, antioxidant enzymes, and other substances in the cells of the ovary and pituitary gland through the signaling pathways of cAMP-PKA, Kiss-1/GPR54, and Hippo and exert therapeutic effects. The results of animal experiments showed that Bushen Culuan Formula could up-regulate serum dopamine(DA) level and pituitary DRD2 expression, down-regulate hypothalamus and ovary cAMP levels, as well as protein expressions of the pituitary gland and ovary PKA, CREB, and p-CREB, and treat HPRL-induced infertility by regulating the cAMP-PKA signaling pathway.

Female offspring of mice perinatally exposed to benzophenone-3 showed early subfertility linked to a poor oocyte stockpile.

Previously, we found that the ultraviolet filter benzophenone-3 (BP3) causes fetal growth restriction in mice when is applied when implantation occurs (first week of gestation). However, whether BP3 can affect gestation and fertility after implantation period is unknown. We aimed to study the effects on reproductive physiology of the offspring caused by perinatal exposure to BP3. C57BL/6 pregnant mice were dermally exposed to 50mg BP3/kg bw.day or olive oil (vehicle) from gestation day 9 (gd9) to postnatal day 21 (pnd1). We observed no differences in mother's weights, duration of gestation, number of pups per mother, onset of puberty or sex ratio. The weights of the pups exposed to benzophenone-3 were transiently lower than those of the control. Estrous cycle was not affected by perinatal exposure to BP3. Besides, we performed a fertility assessment by continuous breeding protocol: at 10weeks of age, one F1 female and one F1 male mouse from each group was randomly chosen from each litter and housed together for a period of 6months. We noticed a reduction in the number of deliveries per mother among dams exposed to BP3 during the perinatal period. To see if this decreased fertility could be associated to an early onset of oocytes depletion, we estimated the ovarian reserve of germ cells. We found reduced number of oocytes and primordial follicles in BP3. In conclusion, perinatal exposure to BP3 leads to a decline in the reproductive capacity of female mice in a continuous breeding protocol linked to oocyte depletion.

Laparoscopic medulla-sparing ovarian tissue biopsy for cryopreservation: step-by-step surgical technique

To describe a laparoscopic technique for ovarian tissue biopsy (OTB) for fertility preservation. In the last years, the demand for fertility preservation has grown because of the increasing survival rates among patients with cancer and the rising awareness of the importance of quality of life after gonadotoxic therapy. Among fertility-sparing approaches, ovarian tissue cryopreservation is a valid strategy to preserve ovarian endocrine and reproductive function in prepubertal and postpubertal women who will undergo gonadotoxic cancer treatments. Currently, there is no universal consensus regarding ovarian tissue retrieval technique for fertility preservation. Step-by-step description of the surgical technique with narrated video footage. Academic tertiary hospital. Patients with a high risk of premature ovarian insufficiency, usually due to gonadotoxic treatments, who undergo OTB for fertility preservation were included in the study. In this video, we present the clinical case of a 28-year-old patient affected by Hodgkin lymphoma who underwent laparoscopy for OTB before chemotherapy. After exposing the chosen ovary, an incision at the tubal pole of the ovary is made with scissors. Through section and dissection, a large cortical biopsy of the ovary is performed without removing and avoiding any damage to the medulla. At the end of the procedure, hemostasis was achieved with selective coagulation using bipolar coagulation. Step by step educational video. The post-operative course was uneventful and the patient was discharge 24 hours after surgery. Standardization of a step-by-step laparoscopic technique can provide an effective method to optimize ovarian tissue removal while minimizing tissue injury. Medulla-sparing ovarian biopsy allows retrieval of only the cortical part of the ovary, maximizing the number of primordial follicles obtained without damaging the vascular supply of the ovary contained within the medulla. Primordial follicles are resistant to cryoinjury owing to their relatively inactive metabolism, and they are usually found at approximately 0.8 mm below the surface of the cortex. This technique could also reduce the back-table processing time of the ovarian tissue before cryopreservation. One disadvantage could be the difficulty of the technique compared to an oophorectomy because it requires a skilled surgeon that can easily find the cleavage plane between the medulla and the cortex, even in patients submitted to previous chemoradiotherapy or during gonadotropin-releasing hormone analogue therapy.

Ulipristal (UPA) effects on rat ovaries: Unraveling follicle dynamics, ovulation inhibition, and safety implications for prolonged use

Ulipristal (UPA), a selective progesterone receptor modulator, has both agonistic and antagonistic effects on progesterone receptors. UPA suppresses ovulation by inhibiting the luteinizing hormone (LH) surge from the pituitary gland; however, the direct effect of UPA on ovarian tissue remains poorly studied. In the present study, we examined the effects of UPA on the ovaries of rats. Rats were treated for 28 days with UPA, and the effects of UPA on ovarian tissue were examined histologically and the expression of antioxidant genes and cell death markers were also investigated. UPA treatment increased the number of primordial follicles at each treatment group, primordial follicles increased at all dose levels, but the size/magnitude of the effect decreased with the increasing dose. The number of primary and antral follicles tended to increase with increasing UPA levels. Furthermore, the decrease in primary follicle number could be attributed to the exhaustion of follicles, but the examination of proliferation markers, oxidative stress markers, and cell death markers revealed no remarkable toxic effects on ovarian tissues. These results suggest that UPA treatment promotes follicle development at each stage but inhibits ovulation by suppressing the LH surge, resulting in an increase in atretic follicles or unruptured luteinized cysts. These results suggest that UPA may not have both toxic effects on the ovary and a direct local effect on ovarian follicles, but we should be careful about the effects of prolonged UPA treatment in patients with uterine fibroids on their future fecundity.

Multiple treatments with human embryonic stem cell-derived mesenchymal progenitor cells preserved the fertility and ovarian function of perimenopausal mice undergoing natural aging

ObjectivesCurrently, no approved stem cell-based therapies for preserving ovarian function during aging. To solve this problem, we developed a long-term treatment for human embryonic stem cell-derived mesenchymal progenitor cells (hESC-MPCs). We investigated whether the cells retained their ability to resist ovarian aging, which leads to delayed reproductive senescence.Materials and methodsIn a middle-aged female model undergoing natural aging, we analyzed whether hESC-MPCs benefit the long-term maintenance of reproductive fecundity and ovarian reservoirs and how their transplantation regulates ovarian function.ResultsThe number of primordial follicles and mice with regular estrous cycles were increased in perimenopausal mice who underwent multiple introductions of hESC-MPCs compared to age-matched controls. The estradiol levels in the hESC-MPCs group were restored to those in the young and adult groups. Embryonic development and live birth rates were higher in the hESC-MPC group than in the control group, suggesting that hESC-MPCs delayed ovarian senescence. In addition to their direct effects on the ovary, multiple-treatments with hESC-MPCs reduced ovarian fibrosis by downregulating inflammation and fibrosis-related genes via the suppression of myeloid-derived suppressor cells (MDSCs) produced in the bone marrow.ConclusionsMultiple introductions of hESC-MPCs could be a useful approach to prevent female reproductive senescence and that these cells are promising sources for cell therapy to postpone the ovarian aging and retain fecundity in perimenopausal women.

51 A preliminary study of the effect of dietary cottonseed on the follicular reserve of does artificially infected with Haemonchus contortus.

Abstract Numerous studies have reported the negative effect of cottonseed gossypol on the ovarian reserve of the follicles, impairing female fertility. Injuries resulting in the depletion of primordial follicles can lead to permanent infertility. At birth, female mammals possess a fixed number of ovarian primordial follicles, which develop into primary, secondary, and antral follicles in a continuous and irreversible process. In this study, we evaluated the potential effects of dietary cottonseed gossypol in combination with parasitic infection on ovarian follicular reserve in meat goats. A total of 40 Boer x Spanish cross does were randomly subjected to cottonseed (CS) and parasite (P: Haemonchus contortus) in a 2x2 factorial arrangement of does (n = 8) per treatment groups (CS-/P-; CS+/P-; CS-/P+, and CS+/P+). Does were artificially infected with Haemonchus contortus for 2 d, and after 25 d, they began shedding parasite eggs. Thereafter, does were fed a hay diet supplemented with cottonseed (0.5% BW; CS+) or concentrate (CS-), to meet the nutritional requirements. Ten weeks later, animals were slaughtered, and ovaries of each treatment group (n = 4) were fixed with formalin and subjected to a standard procedure of histology staining (Periodic acid–Schiff and Hematoxylin counterstaining). Cuts of one left and one right ovary of two does were placed on microscope slides, and 48 slides were subjected to a microscope visualization (EVOS FL Auto; 20x magnification). Primordial follicles were classified as healthy (stained nucleus), unhealthy (pycnotic), and undetermined. Data were analyzed with ANOVA 1 and expressed as mean ± sem. P < 0.05 indicates a significant difference. A total of 20,562 primordial follicles corresponding to 10,410 (50.6%) healthy; 3,729 (18.1%) unhealthy; and 6,909 (33.6%) undetermined were examined; of which 26%, 20%, 31%, and 23% of total primordial follicles belonged to the CS-/P-, CS-/P+, CS+/P-, and CS+/P+ treatment groups, respectively. A significantly (P < 0.05) greater number of follicles were counted on left vs. right ovary sections (11,584 vs. 8,978) corresponding to 56.3% vs. 43.7% of total follicle counts. Averages of 121 ± 6 vs. 101±6 follicles were calculated per left and right ovary sections. Diets containing cottonseed (CS+/P-) increased the average number of follicles per ovary section, while parasite infections significantly (P < 0.05; CS-/P+ and CS+/P+) decreased these numbers. Regardless of the treatment group, the proportions of healthy follicles remained comparable between treatment groups. We concluded that dietary cottonseed and parasitic infections inversely affect ovarian follicle reserve, while the proportions of healthy follicles were unaffected across treatment groups. Nevertheless, higher microscopic magnification is needed to better characterize the large proportion of undetermined follicles.

The Effects of Vitamin C on Histological Changes Induced by Acrylamide on Rat Ovary

Background and Aims: Acrylamide (ACR) adverse effects have been expressed in various tissues. Vitamin C protects the tissue from oxidative damage. The goal of this study was to examine the protective effects of vitamin C on histopathological changes induced by ACR on rat ovaries.
 Materials and Methods: In this experimental study, 28 adult female Wistar rats were used. The animals (n=7) were divided into 4 groups (1) Control group, (2) ACR (10 mg/kg) group, (3) vitamin C (200 mg/kg), (4) ACR [10 mg/kg) + vitamin C (200 mg/kg) daily for 35 days. After the last oral dose, the ovaries were removed placed in the tissue processor, and molded. To determine the volume of the ovary and the number of follicles, we used the Cavalieri principle and the physical dissector, respectively.
 Results: The results showed ACR increased the volume of the ovary (p ≤ 0.05), the volume of the medulla (p ≤ 0.01), and the decrease in the volume of the cortex (p ≤ 0.01) in comparison with the control group. These parameters were improved in the ACR + vitamin C group. The total number of primordial, primary, secondary, and graafian follicles showed a significant reduction in the ACR group compared to the other groups. The total number of follicles was increased in the ACR + vitamin C group.
 Conclusion: Based on this study, we concluded that vitamin C can improve histopathological changes of the ovary in ACR-exposed rats

Relationship between plasma anti-Müllerian hormone concentrations in female Holstein calves immediately after birth and milk yield and composition in dams during early to mid-gestation

The energy status of dams during the early stages of gestation may affect ovarian reserve. It was hypothesized that greater milk yield and composition during the early stages of gestation may suppress the number of primordial follicles in the fetuses of dairy cattle. This study aimed to evaluate the anti-Müllerian hormone (AMH), used as an indicator of ovarian reserve, in female Holstein calves immediately after birth. It also examined the relationship between the AMH concentrations and the milk yield and composition of dams during early pregnancy. A total of 85 female Holstein calves (born to first-calving cows [nulliparous heifers], n = 31; second-calving [primiparous] cows, n = 22; and third- or subsequent-calving [multiparous] cows, n = 32) were examined. Blood samples were collected from the female calves after birth and before the first colostrum feeding for the analysis of plasma AMH concentrations. Data on daily milk yield and monthly milk composition during the 1st to 6th month of pregnancy were additionally collected from milking dams. The plasma AMH concentrations ranged from 0.106 to 1542.3 pg/mL, with an average of 291.6 ± 30.5 pg/mL. There was no significant difference observed among the plasma AMH concentrations in female calves born from nulliparous heifers, primiparous cows, multiparous cows. Primiparous cows exhibited numerous negative correlations between the plasma AMH concentrations of their female calves immediately after birth and the daily milk yield and milk composition from the 1st to the 4th month of pregnancy; there was, however, no correlation observed in multiparous cows. Overall, there were large individual variations observed in the plasma AMH concentrations immediately after birth. Both milk yield and the amount of milk components strongly influenced the plasma AMH concentrations in calves immediately after birth in primiparous cows in the growth phase. These factors, conversely, did not affect the plasma AMH concentrations of calves in multiparous cows. The results of this study suggest that high-producing pregnant primiparous cows devote more energy to their growth and lactation, potentially compromising the reproductive potential of their offspring. Thus, improving the feeding management of primiparous cows with the goal of increasing the fertility of their progeny will help high-yielding dairy cows reproduce more successfully.

Plasma-derived from human umbilical cord blood restores ovarian function and improves serum reproductive hormones levels in mice with premature ovarian insufficiency (POI) through cytokines and growth factors.

Premature ovarian insufficiency (POI) patients experience a decline in ovarian function and a reduction in serum reproductive hormones, leading to a significant impact on the outcomes of assisted reproductive technology. Despite the absence of an effective clinical treatment to restore fertility in POI patients, recent research has indicated that cord blood plasma (CBP) derived from human umbilical cord blood (hUCB) may offer therapeutic benefits for various degenerative diseases. The primary aim of this study is to explore approaches for enhancing ovarian function and serum reproductive hormones through the administration of CBP in a murine model. Initially, hUCB was utilized to obtain CBP (CBP), which was subsequently analyzed for cytokine and growth factor profiles in comparison to adult blood plasma (ABP) by use of flow cytometry. Subsequently, POI mouse models were established through the induction of 4-vinylcyclohexene diepoxide, followed by the injection of CBP into the tail. At 7, 14, and 21 days posttreatment, mouse ovaries and blood were collected, and their estrus cycle, body weight, and ovarian weights were evaluated using precise electronic balance. Finally, ovarian morphology and follicle number were assessed through HE staining, while serum levels of anti-Müllerian hormone (AMH), estradiol (E2) and follicle-stimulating hormone (FSH) were determined by ELISA. Our study revealed that individuals with CBP exhibited significantly lower concentrations of proinflammatory cytokines, including IL-β (p < 0.01) and IL-2 (p < 0.05), while displaying elevated levels of anti-inflammatory cytokines and chemokines, such as IL-2, IL-4, IL-6, IL-8, IL-12P70, IL-17A, IP-10, interferon-γ, and tumor necrosis factor-α (p < 0.01). Furthermore, CBP demonstrated remarkably higher levels of growth factors, including transforming growth factor-β1, vascular endothelial growth factor, and insulin-like growth factor-1 (p < 0.01) than ABP. Notably, our investigation also revealed that CBP restored the content of serum reproductive hormones, such as AMH, E2, and FSH (p < 0.05), and increased the number of primordial and primary follicles (p < 0.01) and decreased the number of luteal and atretic follicles (p < 0.01) in vivo. Our findings suggested that CBP-secreted cytokines and growth factors could be restored POI ovarian function, enhanced serum reproductive hormones and rescued follicular development in vivo. These findings further support the potential of CBP as a promising strategy in clinical applications for POI related infertility.

Protective effect of Luffa cylindrica fermentation liquid on cyclophosphamide-induced premature ovarian failure in female mice by attenuating oxidative stress, inflammation and apoptosis

Premature ovarian failure (POF) is a leading cause of women’s infertility without effective treatment. The purpose of this study was to investigate the protective effects of Luffa cylindrica fermentation liquid (LF) on cyclophosphamide (CTX) -induced POF in mice and to preliminarily investigate the underlying mechanisms. Thirty-two Balb/c mice were divided into four groups randomly. One group served as the control, while the other three received CTX injections to establish POF models. A 14-day gavage of either 5 or 10 μL/g LF was administered to two LF pretreatment groups. To analyze the effects of LF, the ovarian index, follicle number, the levels of serum sex hormones, superoxide dismutase (SOD), glutathione (GSH), malondialdehyde (MDA), inflammatory factors, and apoptosis of the ovarian cells were measured. The effects of LF pretreatment on the expression of TLR4/NF-κB and apoptosis pathways were also evaluated. We found that LF pretreatment increased the ovarian index and the number of primordial and antral follicles while decreasing those of atretic follicles. LF pretreatment also increased the serum levels of estradiol (E2) and anti-Müllerian hormone (AMH), while decreasing those of luteinizing hormone (LH) and follicle-stimulating hormone (FSH). Furthermore, LF pretreatment increased the levels of SOD and GSH in the ovaries, while decreasing those of MDA, tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β). LF administration reduced the amount of TUNEL+ ovarian cells and the levels of TLR4 and NF-κB P65 protein expression. In conclusion, LF has antioxidant, anti-inflammatory as well as anti-apoptotic effects against CTX-induced POF, and the inhibition of TLR4/NF-κB and apoptosis pathways may be involved in its mechanisms.Graphical

Comparative effects of the antioxidant glutathione with metformin and Diane-35 on hormonal, metabolic, and inflammatory indicators in a DHEA-induced PCOS rat model

Objective Comparison of hormonal, metabolic and inflammatory markers of glutathione with metformin and Diane-35 in a rat model of PCOS induced by dehydroepiandrosterone. Methods Twenty-five female rats were randomized into four groups. Group 1 was administered a subcutaneous dose of 0.2 ml saline/day. Group 2 was given 0.2 ml of 1% carboxymethyl cellulose (CMC)/day orally for 28 days. A PCOS model was established with DHEA in rats. Group 3 was given 4.5 mg/kg/day of Diane-35 orally dissolved in 1% CMC for 28 days. Group 4 was given 300 mg/kg/day of metformin orally dissolved in 1 ml of saline for 28 days, and Group 5 was administered 100 mg/kg of glutathione intraperitoneally on days 35, 42, and 49. On day 56, the rats were sacrificed. Serum markers and follicle count were examined. Results Serum IL-6, hs-CRP, insulin, testosterone, SHBG, and MDA values were significantly lower in the glutathione group than in the PCOS group (p = 0.0006, p = 0.023, p = 0.0082, p = 0.0007, p = 0.0048, and p < 0.0001, respectively). The number of all follicles was similar between the control and glutathione groups (p < 0.05). When we compared the other groups with the PCOS group, the number of primary, secondary, atretic, and cystic follicles was significantly lower in the metformin and glutathione groups. The number of primordial and antral follicles was significantly higher than in the PCOS group. Conclusions Glutathione plays anti-inflammatory and antioxidant roles, similar to metformin, by lowering serum IL-6, insulin, testosterone, CRP, and MDA levels; decreasing atretic/cystic follicle count; and improving antral follicle count and folliculogenesis in PCOS patients.

Sleep deprivation causes gut dysbiosis impacting on systemic metabolomics leading to premature ovarian insufficiency in adolescent mice

Sleep deprivation causes gut dysbiosis impacting on systemic metabolomics leading to premature ovarian insufficiency in adolescent mice

Method of Isolation and In Vitro Culture of Primordial Follicles in Bovine Animal Model.

The mammalian ovary is a substantial source of oocytes arranged into follicles at various stages of folliculogenesis, from the primordial to the ovulatory ones. Primordial follicles constitute the most abundant source of gametes inside the mammalian ovary at any given time.The isolation of a high number of primordial follicles, together with the development of protocols for in vitro follicle growth, would provide a powerful tool to fully exploit the female reproductive potential and boost the rescue and restoration of fertility in assisted reproduction technologies in human medicine, animal breeding, and preservation of threatened species. However, the most significant limitation is the lack of efficient methods for isolating a healthy and homogeneous population of viable primordial follicles suitable for in vitro culture. Here, we provide a fast and high-yield strategy for the mechanical isolation of primordial follicles from limited portions of the ovarian cortex in the bovine animal model.

Effect of age on follicular dynamics of goat ovaries

Reproductive aging is influenced by a gradual decline in both the quantity and quality of oocytes found within ovarian follicles. In mammals, fertility relies heavily on the health of oocyte reserves, the dynamics of follicular development, and the functioning of the ovaries. This study aimed to investigate the impact of age on the follicular dynamics of goat ovaries. A total of 40 ovaries of Black Bengal goats were obtained from a local slaughterhouse and sorted into four age groups: 6-12 months, 13-15 months, 18-21 months, and 22-24 months. Subsequently, the ovaries were treated, and follicles were assessed through visually and histologically. The findings revealed that nearly all parameters, including weight, length, and width, of both ovaries in the 18-21 months age group were insignificantly different (p&gt;0.05) from the other age groups. Histologically, the number of normal primordial and primary follicles was also insignificantly higher (p&gt;0.05) in goats aged 18-21 months compared to the other age groups. Conversely, the quantity of secondary follicles significantly increased (p&lt;0.05) with age, but in terms of antral follicles, a quadratic trend was observed, where they increased up to a certain age and then declined. Additionally, the results showed that the number of degenerated primary and antral follicles was significantly higher (p&lt;0.05) in the ovaries of 22–24-month-old goats, while the number of degenerated secondary follicles was insignificantly higher (p&gt;0.05) compared to the remaining age groups. In conclusion, age had an effect on the follicular dynamics of goat ovaries, suggesting that ovaries from goats aged 18-21 months, sourced from slaughterhouses, hold potential for quality follicles and oocytes that can be valuable for further research. Bangladesh Journal of Animal Science 52 (4): 88-97

Effect of age on follicular dynamics of goat ovaries

Reproductive aging is influenced by a gradual decline in both the quantity and quality of oocytes found within ovarian follicles. In mammals, fertility relies heavily on the health of oocyte reserves, the dynamics of follicular development, and the functioning of the ovaries. This study aimed to investigate the impact of age on the follicular dynamics of goat ovaries. A total of 40 ovaries of Black Bengal goats were obtained from a local slaughterhouse and sorted into four age groups: 6-12 months, 13-15 months, 18-21 months, and 22-24 months. Subsequently, the ovaries were treated, and follicles were assessed through visually and histologically. The findings revealed that nearly all parameters, including weight, length, and width, of both ovaries in the 18-21 months age group were insignificantly different (p&gt;0.05) from the other age groups. Histologically, the number of normal primordial and primary follicles was also insignificantly higher (p&gt;0.05) in goats aged 18-21 months compared to the other age groups. Conversely, the quantity of secondary follicles significantly increased (p&lt;0.05) with age, but in terms of antral follicles, a quadratic trend was observed, where they increased up to a certain age and then declined. Additionally, the results showed that the number of degenerated primary and antral follicles was significantly higher (p&lt;0.05) in the ovaries of 22–24-month-old goats, while the number of degenerated secondary follicles was insignificantly higher (p&gt;0.05) compared to the remaining age groups. In conclusion, age had an effect on the follicular dynamics of goat ovaries, suggesting that ovaries from goats aged 18-21 months, sourced from slaughterhouses, hold potential for quality follicles and oocytes that can be valuable for further research. Bangladesh Journal of Animal Science 52 (4): 88-97