Effect of protection of enoxaparin against experimental ischemia/reperfusion injury in the rat ovary on in vitro fertilization outcomes

SUMMARY OBJECTIVE: The study aimed to investigate the protection of enoxaparin (E) against experimental ischemic (I) and ischemic-reperfusion (I/R) injury in rat ovaries on in vitro fertilization outcomes. METHODS: In total, 56 adult female Sprague-Dawley albino rats were randomly assigned to 6 groups of 8 animals each: Sham, Ischemia, I/R, Sham+E, I+E, and I/R+E. Ischemia groups were subjected to bilateral adnexal torsion for 3 h. In contrast, I/R and I/R+E groups received subsequent detorsion for 3 h. Enoxaparin (0.5 mg/kg s.c.) was administered 30 min prior to ischemia (I+platelet-rich plasma) or reperfusion (I/R+I+platelet-rich plasma). Ovaries were stimulated through intraperitoneal injection of 150-300 internal units IU/kg pregnant mare serum gonadotropin. Anti-Müllerian hormone levels were measured before and after surgery in all groups. RESULTS: When the number of metaphase II oocytes was evaluated, statistically significant differences were observed between the I and I+E (p=0.001) and I/R and I/R+E (p=0.000) groups. When both I and I+E groups and I/R and I/R+E groups were compared, it was found that E application increased the number of fertilized oocytes. The number of embryos on the second day was higher in the I/R+E group than that in the I/R group. Statistically significant differences were found in the number of grade 1 embryos between the I/R and I/R+E groups (p=0.003). In comparing anti-Müllerian hormone values within the group, the highest decrease was observed in the I and I/R groups. CONCLUSION: Enoxaparin effectively minimizes ovarian damage and preserves ovarian reserve following ovarian torsion.

Effect of protection of enoxaparin against experimental ischemia/reperfusion injury in the rat ovary on in vitro fertilization outcomes.

OBJECTIVE: The study aimed to investigate the protection of enoxaparin (E) against experimental ischemic (I) and ischemic-reperfusion (I/R) injury in rat ovaries on in vitro fertilization outcomes. METHODS: In total, 56 adult female Sprague-Dawley albino rats were randomly assigned to 6 groups of 8 animals each: Sham, Ischemia, I/R, Sham+E, I+E, and I/R+E. Ischemia groups were subjected to bilateral adnexal torsion for 3 h. In contrast, I/R and I/R+E groups received subsequent detorsion for 3 h. Enoxaparin (0.5 mg/kg s.c.) was administered 30 min prior to ischemia (I+platelet-rich plasma) or reperfusion (I/R+I+platelet-rich plasma). Ovaries were stimulated through intraperitoneal injection of 150-300 internal units IU/kg pregnant mare serum gonadotropin. Anti-Müllerian hormone levels were measured before and after surgery in all groups. RESULTS: When the number of metaphase II oocytes was evaluated, statistically significant differences were observed between the I and I+E (p=0.001) and I/R and I/R+E (p=0.000) groups. When both I and I+E groups and I/R and I/R+E groups were compared, it was found that E application increased the number of fertilized oocytes. The number of embryos on the second day was higher in the I/R+E group than that in the I/R group. Statistically significant differences were found in the number of grade 1 embryos between the I/R and I/R+E groups (p=0.003). In comparing anti-Müllerian hormone values within the group, the highest decrease was observed in the I and I/R groups. CONCLUSION: Enoxaparin effectively minimizes ovarian damage and preserves ovarian reserve following ovarian torsion.

The Effectiveness of Granulocyte Colony-Stimulating Factor (G-CSF) Against Experimental Ischemia-Reperfusion Injury in Rat Ovaries and Its Effect on In Vitro Fertilization Outcomes.

Local ischemic damage resulting from ovarian torsion is a common cause of decreased follicular activity and infertility. It was aimed to evaluate the effect of granulocyte colony-stimulating factor (G-CSF) protection on in vitro fertilization (IVF) results against experimental ischemic (I) and ischemic-reperfusion (I/R) injury in rat ovaries. The study consisted of 35 adult female Sprague-Dawley albino rats (sham, ischemia, I/R, I + G-CSF, and I/R + G-CSF) randomly assigned to 5 groups, each containing 7 rats. While bilateral adnexal torsion was applied to the ischemia groups for 3 h, detorsion was applied to the reperfusion groups. Intraperitoneal 100 IU/kg G-CSF was administered 30 min prior to ischemia (I + G-CSF) or reperfusion (I/R + G-CSF). After ovulation induction (intraperitoneal 150-300 IU/kg PMSG), the oocytes were collected and IVF was applied. Statistically significant differences were observed between the collected oocyte numbers in I and I + G-CSF, I/R, and I/R + G-CSF groups (P values were P = 0.001 for the I-I + G-SCF groups and P = 0.003 for I/R-IR + G-CSF, respectively). An increase in the number of MII oocytes obtained was observed in the I + G-CSF and I/R + G-CSF groups compared to the I and I/R groups. Grade 1 and grade 2 embryo numbers were statically different between the I/R and I/R + G-CSF groups (P values were P = 0.023 for grade 1 embryos and P = 0.045 for grade 2 embryos, respectively). G-CSF treatment was found to be effective in reducing I- and I/R-induced ovarian damage.

The protective effect of cilostazol on experimental ischemia/reperfusion injury in rats ovaries on in vitro fertilization outcomes.

Objective: Ovarian torsion decreases ovarian reserve because of ischemic and reperfusion damage it causes. In this study, we investigated the protective effect of cilostazol (CIL) on experimental ischemia (I) and ischemic-reperfusion (I/R) damage in rat ovaries with in vitro fertilization (IVF) results. Materials and Methods: Forty-eight adult female Sprague-Dawley albino rats were randomly assigned to 6 groups with 8 animals in each group: Sham (S), I, I/R, S + CIL, I + CIL and I/R + CIL. The I groups were subjected to bilateral adnexal torsion for 3 h, while the I/R and I/R + CIL groups received subsequent detorsion for 3 h. Twenty-two mg/kg of CIL was given via oral gavage 30 min before surgery on the I (I+ CIL) or reperfusion (I/R + CIL) groups. Oocytes were collected before the IVF procedure and after ovulation induction with 150-300 IU/kg pregnant mare serum gonadotropin. Results: The metaphase oocytes reached their highest value of 4.73±0.96 in the S+ CIL group and reached their lowest value of 0.51±0.55 in the I/R group. There were statistically significant differences in the number of second-day embryos among the I, I+ CIL, and I/R and I/R+ CIL groups (p=0.000). When the groups were compared in terms of Anti-Müllerian hormone change, the highest decrease was observed in the I and I/R groups. Conclusion: CIL pretreatment before surgery has a protective effect against I and I/R in rats with ovarian torsion.

Protective role of melatonin against testicular damage caused by polymicrobial sepsis in adult rats.

BACKGROUND: This study aimed to investigate the possible protective effects of melatonin (MEL) against the damage to testicular tissue in rats caused by polymicrobial sepsis as a result of cecal ligation and perforation (CLP). METHODS: In this study, 21 male Wistar albino rats were used. The rats were randomly divided into three groups (n=7): Sham Control (Group 1), CLP (Group 2), and CLP + MEL (Group 3). Sepsis was created using the CLP method. MEL was administered intraperitoneally in two equal doses of 10 mg/kg at 30 min before and 6 h after perforation. Tissue sections taken from paraffin blocks were stained with hematoxylin and eosin (H and E) and examined histopathologically under a light microscope. Intracellular H2O2 and apoptosis evaluations were carried out using the flow cytometric method. RESULTS: Sepsis caused a significant reduction in all sperm parameters. There was a significant decrease in sperm density, motility and cell numbers with normal morphology (p<0.05). Intracellular H2O2 level and apoptotic cell percentages increased in sperm cells in the CLP group. MEL treatment was found to significantly reduce sperm abnormalities, testicular damage, intracellular H2O2 levels, and apoptosis. CONCLUSION: This study showed that melatonin administration could be a potential treatment option to reduce acute testicular tissue damage due to sepsis.

Effect of Propolis on Precocious Puberty in Female Rats

Objective: Nutrition and exposure to various chemicals, including environmental pollution, insecticides, and plant phytoestrogens (having oestrogen-like effects), are environmental factors that affect puberty onset. The aim of this study was to identify the effects of propolis, which has been reported to have oestrogenic effects, on precocious puberty and the reproductive system in prepubertal female rats (ovary, endometrium, breast). Methods: Thirty-four 25-day-old, prepubertal, female Sprague-Dawley rats were included. Rats were randomly divided into the propolis (n=17) and control groups (n=17). The primary endpoint was the number of rats that developed vaginal opening, a sign of puberty, at 12-day follow-up. In addition, the effect of propolis on ovary, uterus and breast tissue was evaluated histologically. Results: Vaginal patency occurred earlier (about 7.5 days sooner) in the propolis group and all animals in the propolis group had vaginal patency by day 12. The number of ovarian follicles (in all follicles), endometrial thickness, and mammary gland secretory gland area were significantly higher in the propolis group than in the control group (all p<0.001). In addition, Ki-67 activity in the endometrium, breast tissue and ovary was more intense in the propolis group compared to the control group (all p<0.001). Conclusion: Propolis triggers precocious puberty in female rats, possibly by interacting with the oestrogen receptor. The mechanism of action of propolis should be considered before prescribing it. In addition, further studies are needed to explore the mechanism of action of propolis and to determine the component of propolis that triggers puberty.

The effect of protection of platelet-rich plasma against experimental ischemia/reperfusion injury in the rat ovary on in vitro fertilization outcomes.

OBJECTIVE: Ovarian torsion is a common cause of local ischemic damage, reduced follicular activity and infertility. This study aimed to investigate how well platelet-rich plasma (PRP) protects against experimental ischemic (I) and ischemia-reperfusion (I/R) injury in rat ovaries and its effect on in vitro fertilization (IVF) outcomes. METHOD: Fifty-six adult female Sprague-Dawley albino rats were randomly assigned to six groups of eight animals each: Sham, Ischemia, I/R, Sham + PRP, I + PRP, and I/R + PRP. The remaining eight animals were used to prepare the PRP. The ischemia groups were subjected to bilateral adnexal torsion for 3 h, while the I/R and I/R + PRP groups received subsequent detorsion for 3 h. Intraperitoneal (i.p.) PRP was administered 30 min prior to ischemia (I + PRP) or reperfusion (I/R + PRP). The ovaries were stimulated through an intraperitoneal injection of 150-300 internal units of IU/kg PMSG. After ovulation induction, oocytes were taken from the ovaries, and IVF was performed. RESULTS: The number of MII oocytes reached the highest number with 4.63 ± 0.74 in the S group and had the lowest number with 0.50 ± 0.53 in the I/R group. There were statistically significant differences for the number of embryos obtained on the second day between the I and I + PRP groups and the I/R and I/R + PRP groups (p = 0.000). In comparing anti-Müllerian hormone 1 (AMH1) and AMH2 values within the group, the highest decrease was observed in the I and I/R groups. CONCLUSION: PRP is effective in minimizing ovarian damage and preserving ovarian reserves following ovarian torsion.

Protective Effects of Platelet-rich plasma for in vitro Fertilization of Rats with Ovarian Failure Induced by Cyclophosphamide.

OBJECTIVE: Premature ovarian insufficiency (POI) contributes significantly to female infertility. Cyclophosphamide (CYC has adverse effects on folliculogenesis. Platelet-rich plasma (PRP) is an autologous product rich in many growth factors. We evaluated the protective effect of PRP on in vitro fertilization in female rats with CYC-induced ovarian damage. METHODS: Twenty-eight adult female Sprague-Dawley rats were randomly divided into four groups. Group 1 (control-sodium chloride 0.9%; 1 mL/kg, single-dose intraperitoneal [IP] injection); group 2 (CYC), 75 mg/kg, single-dose IP injection and sodium chloride 0.9% (1 mL/kg, single-dose IP injection); group 3 CYC plus PRP, CYC (75 mg/kg, single-dose and PRP (200 µl, single-dose) IP injection); and group 4 (PRP, 200 µl, single-dose IP injection). RESULTS: In the comparisons in terms of M1 and M2 oocytes, it was observed that the CYC group presented a significantly lower amount than the control, CYC/PRP, and PRP groups. (for M1, p = 0.000, p = 0.029, p = 0.025; for M2, p = 0.009, p = 0.004, p = 0.000, respectively). The number of fertilized oocytes and two-celled good quality embryos was found to be statistically significant between the CYC and control groups, CYC + PRP and PRP groups (p = 0.009, p = 0.001, p = 0.000 for oocytes, respectively. For embryos; p = 0.016, p = 0.002, p = 0.000). CONCLUSION: Platelet-rich plasma can protect the ovarian function against damage caused by CYC, and, in addition, it improves oocyte count and the development of embryos as a result of oocyte stimulation during the IVF procedure.

OBJETIVO: A insuficiência ovariana prematura (POI) contribui significativamente para a infertilidade feminina. A ciclofosfamida (CYC) tem efeitos adversos na foliculogênese. O plasma rico em plaquetas (PRP) é um produto autólogo rico em muitos fatores de crescimento. Avaliamos o efeito protetor do PRP na fertilização in vitro em ratas com lesão ovariana induzida por CYC. MéTODOS: Vinte e oito ratas Sprague-Dawley adultas foram divididas aleatoriamente em quatro grupos. Grupo 1 (controle - cloreto de sódio 0,9%; 1 mL/kg, injeção intraperitoneal [IP] em dose única); grupo 2 (CYC), 75 mg/kg, injeção IP de dose única e cloreto de sódio 0,9% (1 mL/kg, injeção ip de dose única); grupo 3 CYC + PRP, CYC (75 mg/kg, dose única e PRP (200 µl, dose única) injeção IP); e grupo 4 (PRP, 200 µl, injeção IP de dose única). RESULTADOS: Nas comparações em termos de ovócitos M1 e M2, observou-se que o grupo CYC apresentou uma quantidade significativamente menor que os grupos controle, CYC/PRP, e PRP. (Para M1, p = 0,000, p = 0,029, p = 0,025; para M2, p = 0,009, p = 0,004, p = 0,000, respectivamente). O número de oócitos fertilizados e embriões bicelulares de boa qualidade foi considerado estatisticamente significativo entre os grupos CYC e controle, CYC + PRP e grupos PRP (p = 0,009, p = 0,001, p = 0,000 para oócitos, respectivamente. Para embriões, p = 0,016, p = 0,002, p = 0,000). CONCLUSãO: O PRP pode proteger a função ovariana contra os danos causados pelo CYC e, além disso, proporciona melhora na contagem de oócitos e no desenvolvimento de embriões como resultado da estimulação ovariana durante o procedimento de fertilização in vitro.

The effect of Propofol versus Dexmedetomidine as anesthetic agents for oocyte pick-up on in vitro fertilization (IVF) outcomes.

This study aimed to evaluate the effects of propofol and dexmedetomidine over different timescales on the IVF outcomes for transvaginal oocyte retrieval (TVOR). Twenty-four rats included in the study were divided into two main groups and three subgroups were subjected to the ovulation induction process. Group 1 was administered propofol (100 mg/kg i.v.) and group 2 were administered dexmedetomidine (25 µg/kg i.p.) The oviduct collection procedure was completed within 15 min for subgroup Pro15min, Dex15min (n = 4), within 16 to 30 min for subgroup Pro30min, Dex30min (n = 4) and within 31 to 60 min for subgroup Pro60min, Dex60min (n = 4) after euthanasia. The total number of oocytes was counted. After in vitro fertilization, the number and quality of embryos were evaluated. The number of pups born were evaluated after embryo transfer. The embryo number, quality and pup count decreased as the administration time for propofol increased (p < 0.05). No statistically significant difference was found between the dexmedetomidine subgroups for embryo number, quality and pup count(p > 0.05). As the exposure time to propofol increased, the number and quality of embryos obtained, and the pup count, decreased. The use of dexmedetomidine had no negative impacts on the number of embryos, their quality or the number of pups.

Decreased ovarian reserve and ovarian morphological alterations in female rat offspring exposed to a ketogenic maternal diet

SUMMARY OBJECTIVE: This study evaluates the effects of a ketogenic diet on morphology and follicle reserve. METHOD: Sixteen Sprague-Dawley rats were randomized into two groups: standard diet group (n=8) and ketogenic diet group (n=8). Rats were time mated. Dams were permitted to deliver spontaneously. The animals were monitored for the onset of puberty. All the rats were weighed and anesthetized, serum anti-Müllerian hormone level was measured, and the oviducts were removed. The morphological characteristics of follicles were determined and total ovarian volumes were calculated. RESULTS: The mean ovarian volume was statistically significantly lower in the ketogenic diet group compared to the standard diet group (14.41±0.99 mm3 versus 18.89±1.28 mm3) (p=0.000). The mean number of antral follicles was 13.63±1.80 in the standard diet group and 4.462±0.760 in the ketogenic diet group. The mean ovarian weight of the ketogenic diet group was significantly lower than that of the standard diet group (0.42±0.06 g versus 0.815±107 g). The mean anti-Müllerian hormone levels were significantly higher in the standard diet group compared to the ketogenic diet group (1.023±4.75 ng/mL versus 0.69±0.07 ng/mL) (p=0.000). The mean percentage of staining of Ki-67 was 35.28±4.75 in the standard diet group and 16.98±3.33 in the ketogenic diet group (p=0.000). CONCLUSION: Maternal ketogenic diet reduces ovarian follicular reserve in female offspring and has important implications for maintaining reproductive potential at a population level.

The Short-Term Effect of Eugenol on the Prevention of Experimentally Induced Myringosclerosis in a Rat Model.

OBJECTIVE: The aim of our study was to assess the possible short-term effects of topical and oral eugenol on the suppression of experimentally developed myringosclerosis (MS). METHODS: Four groups of seven male Wistar albino rats were used in the study. The tympanic membranes (TMs) of all subjects were myringotomized, and group 1 was given no treatment (as control group), group 2 received saline, group 3 had received topical eugenol and group 4 received oral eugenol. RESULTS: In macroscopic evaluation the control and saline groups showed much more MS compared to the topical and oral eugenol groups which had statistically significantly less changes (p<0.05). Fibrosis and inflammation regarding the lamina propria (LP) of the eardrums of the topical and oral eugenol groups were significantly less than those of the control and saline groups (p<0.001). In microscopic evaluation, TMs were found to be thicker in the control and saline groups (p<0.001). CONCLUSION: Our study showed that the application of topical and oral forms of eugenol reduced fibrosis and prevented the advancement of MS in the LP of the TMs in the short-term. More studies with different extracts are needed to investigate the efficacy of phytotherapeutic agents for preventing MS development following myringotomy.

Decreased ovarian reserve and ovarian morphological alterations in female rat offspring exposed to a ketogenic maternal diet.

OBJECTIVE: This study evaluates the effects of a ketogenic diet on morphology and follicle reserve. METHOD: Sixteen Sprague-Dawley rats were randomized into two groups: standard diet group (n=8) and ketogenic diet group (n=8). Rats were time mated. Dams were permitted to deliver spontaneously. The animals were monitored for the onset of puberty. All the rats were weighed and anesthetized, serum anti-Müllerian hormone level was measured, and the oviducts were removed. The morphological characteristics of follicles were determined and total ovarian volumes were calculated. RESULTS: The mean ovarian volume was statistically significantly lower in the ketogenic diet group compared to the standard diet group (14.41±0.99 mm3 versus 18.89±1.28 mm3) (p=0.000). The mean number of antral follicles was 13.63±1.80 in the standard diet group and 4.462±0.760 in the ketogenic diet group. The mean ovarian weight of the ketogenic diet group was significantly lower than that of the standard diet group (0.42±0.06 g versus 0.815±107 g). The mean anti-Müllerian hormone levels were significantly higher in the standard diet group compared to the ketogenic diet group (1.023±4.75 ng/mL versus 0.69±0.07 ng/mL) (p=0.000). The mean percentage of staining of Ki-67 was 35.28±4.75 in the standard diet group and 16.98±3.33 in the ketogenic diet group (p=0.000). CONCLUSION: Maternal ketogenic diet reduces ovarian follicular reserve in female offspring and has important implications for maintaining reproductive potential at a population level.

Effect of follicular fluid NO, MDA and GSH levels on in vitro fertilization outcomes.

OBJECTIVE: To investigate the biochemical markers such as nitric oxide (NO), malondialdehyde (MDA) and reduced glutathione (GSH), indicators of the oxidative status of the follicle, to predict the outcome of in vitro fertilization. MATERIAL AND METHODS: Follicular aspirates of dominant follicles were collected during oocyte retrieval. Biochemical analyses of NO, MDA and GSH were performed on all aspirates. RESULTS: When the successful and unsuccessful pregnancy groups were compared in terms of NO, MDA and GSH, follicular fluid MDA was significantly higher (p=0.001) and follicular fluid NO level was significantly lower (p=0.039) in the pregnant group. Correlation analysis between oxidative stress and IVF parameters showed that MDA had a positive weak correlation with the number of grade 1 embryos (r=0.271, p=0.033) and fertilization rate (r=0.263, p=0.039). ROC curve analysis found that malondialdehyde has an area under the curve of 0.74 and can predict pregnancy with high sensitivity. CONCLUSION: As malondialdehyde was significantly different in pregnant and non-pregnant women and had a good sensitivity profile in predicting pregnancy, it may be considered a marker for predicting IVF success.

In vitro maturation may prevent the cancellation of in vitro fertilization cycles in poor responder patients: A case report.

In vitro maturation (IVM) is a promising technique that is used for the maturation of immature oocytes in laboratory conditions and preferred for use in patients with a diagnosis of polycystic ovary syndrome (PCOS) as an alternative to conventional in vitro fertilization (IVF) treatment. In this report, we present a case who surprisingly showed insufficient response to gonadotrophin stimulation during IVF treatment and whose cycle was retrieved from cancellation by using the in vitro maturation technique. As a result, we conclude that IVM may be a good option not only for PCOS patients, but also for poor responders.