Empagliflozin reduces the adverse effects of diabetes mellitus on testicular tissue in type 2 diabetic Rats: A stereological and biochemical study.

Empagliflozin as an antioxidant decreases blood glucose and insulin resistance in type 2 diabetes mellitus. Base on the empagliflozin antioxidant properties we decided to investigate the its effects on the testis histological changes through stereological techniques and biochemical evaluations in T2 diabetes mellitus rats. Rats were divided into: control, diabetes mellitus (DM, streptozotocin + nicotinamide) and diabetes mellitus + empagliflozin (DM + EMPA, 10 mg/kg/day) groups. 56 days after inducing diabetes mellitus testis histological changes and serum biochemical factors along with the level of Bax, Bcl2 and Nrf2 genes expression in the testicular tissue were assessed. A significant decrease in the mean total volume of testis and its components, the level of Bcl2 and Nrf2 gene expression (p < 0.001) along with a significant increase in the level of IL-6, TNF-α, MDA, Bax gene expression were observed in the DM group compared to the control group (p < 0.001). In the DM + EMPA group, the mean total volume of testis and its components, the level of Bcl2 gene expression (p< 0.01) and Nrf2 (p < 0.001) significantly increased whereas the mean level of IL-6 (p < 0.01), TNF-α (p < 0.001), MDA (p < 0.001), Bax (p < 0.001) gene expression significantly decreased compared to the DM group. Our results showed that empagliflozin, by improving the antioxidant defense system, can reduce testicular inflammation and apoptosis and partly prevent the adverse effects of diabetes mellitus on testicular tissue.

Kisspeptin decreases the adverse effects of human ovarian vitrification by regulating ROS-related apoptotic occurrences.

Kisspeptin is characterized as a neuropeptide with a pivotal function in female and male infertility, and its antioxidant properties have been demonstrated. In this study, the effects of kisspeptin on the improvement of the vitrification and thawing results of human ovarian tissues were investigated. In this work, 12 ovaries from patients who underwent hysterectomy were collected laparoscopically, and then 32 samples from each of their tissues were taken. Haematoxylin and eosin (H&E) staining was performed to check the normality of the ovarian tissue and, subsequently, the samples were allocated randomly into four groups, including: (1) fresh (control), (2) vitrification, (3) vitrified + 1 µM kisspeptin, and (4) vitrified + 10 µM kisspeptin groups. After vitrification, thawing, and tissue culture processes, H&E staining for tissue quality assessment, terminal deoxynucleotidyl transferase dUTP nick end labelling assay for apoptosis evaluation, and malondialdehyde (MDA), superoxide dismutase (SOD), and ferric reducing ability of plasma tests for oxidative stress appraisal were carried out. Our histological results showed incoherency of ovarian tissue morphology in the vitrification group compared with other groups. Other findings implicated increased apoptosis rate and MDA concentration and reduced SOD activity and total antioxidant capacity (TAC) in the vitrification group compared with the control group (P < 0.05). Moreover, decreased apoptosis rate and MDA concentration, and increased TAC and SOD function were observed in the vitrification with kisspeptin groups (1 µM and 10 µM) compared with the vitrified group (P < 0.05). Our reports express that kisspeptin is an effective agent to overcome the negative effects of vitrification by regulating reactive oxygen species-related apoptotic processes.

The effect of platelet lysate on mouse ovarian structure, function and epigenetic modifications after autotransplantation.

RESEARCH QUESTION: What are the effects of platelet lysate on structure, function and epigenetic modifications of heterotopically transplanted mouse ovarian tissues? DESIGN: Mice were divided into three groups (n = 17 per group): control (mice with no ovariectomy, grafting or treatment), autograft and autograft plus platelet lysate (3 ml/kg at the graft sites). Inflammatory markers, serum malondialdehyde (MDA) concentration and total antioxidant capacity were assessed on day 7 after transplantation. Twenty-eight days after transplantation, stereological and hormonal analyses were conducted. Chromatin immunoprecipitation and quantitative real-time polymerase chain reaction were also used to quantify the epigenetic modifications of maturation genes, parallel to their expression. RESULTS: The total volume of the ovary, cortex and medulla, and the number of different types of follicles, the concentration of interleukin (IL)-10, progesterone and oestradiol and total antioxidant capacity significantly decreased in the autograft group compared with the control group (P < 0.001); these parameters significantly increased in the autograft plus platelet lysate group compared with the autograft group (P < 0.001). The concentrations of tumour necrosis factor alpha, IL-6 and MDA increased significantly in the autograft group compared with the control group (P < 0.001); in the autograft plus platelet lysate group, these parameters significantly decreased compared with the autograft group (P < 0.001). In the autograft plus platelet lysate group, the expression levels of Gdf-9 (P < 0.0021), Igf-1 (P < 0.0048) and Igf-2 (P < 0.0063) genes also increased along with a lower incorporation of MeCP2 in the promoter regions (P < 0.001) compared with the autograft group. CONCLUSIONS: Platelet lysate can contribute to follicular survival by improving folliculogenesis and increasing the expression of oocyte maturation genes.

Effect of Myo-inositol on sperm quality and biochemical factors in cryopreserved semen of patients with Asthenospermia.

In this study, the influence of myoinositol (MYO) as an antioxidant on the inhibition of the negative impacts of cryopreservation on sperm quality in men with Asthenospermia was investigated. In this prospective study, each semen sample from 25 cases was separated into three groups: Fresh, Control (with freezing medium), Myoinositol (2 mg/ml). According to the World Health Organization criteria (WHO) (2010), total motility, progressive sperm motility, viability, normal morphology, and DNA integrity were assessed. In addition, the hypo-osmotic swelling (HOS) test and mitochondrial membrane potential (MMP) were used. Total antioxidant capacity (TAC), malondialdehyde (MDA), and antioxidant enzyme activity were determined by the ELISA method. In contrast to the fresh samples, lipid peroxidation, DNA integrity damage, DNA fragmentation, HOST, and MMP had significant enhancement in the control samples. Sperm quality was significantly decreased (p < 0.05). Mean percentage viability, normal morphology, total motility, progressive motility, and DNA integrity were significantly enhanced in the MYO group in comparison to the control group (p < 0.05). The MDA and TAC levels and DNA damage in the MYO group were significantly lower compared to the control group (p < 0.05). The findings confirm that sperm quality in patients with Asthenospermia is improved by the administration of 2 mg/ml of myoinositol together with the freezing medium after sperm cryopreservation.

L-Carnitine improves follicular survival and function in ovarian grafts in the mouse.

CONTEXT: Ovarian tissue transplantation is performed to preserve fertility in patients undergoing chemotherapy and radiotherapy. However, the ischemia-reperfusion injury which occurs after the ovarian tissue transplantation causes follicular depletion and apoptosis. l -Carnitine has antioxidant and anti-inflammation properties. AIMS: Therefore, we aimed to investigate the beneficial effect of l -carnitine on mouse ovaries following heterotopic autotransplantation. METHODS: Mice were randomly divided into three groups (six mice per group): control, autografted and autografted+l -carnitine (200mg/kg daily intraperitoneal injections). Seven days after ovary autografting, the serum levels of malondialdehyde (MDA), total antioxidant capacity, tumor necrosis factor alpha (TNF-α), interleukin (IL)-6 and IL-10 were measured. Ovary histology, serum concentrations of progesterone and estradiol were also measured 28days after autotransplantation. Data were analysed using one-way analysis of variance (ANOVA) and Tukey test, and the means were considered significantly different at P Key results: In the autografted+l -carnitine group, the total volume of the ovary, the volume of the cortex, the number of follicles, the serum concentrations of IL-10, estradiol and progesterone significantly increased compared to the autografted group. In the autografted+l -carnitine group, serum concentrations of IL-6, TNF-α and MDA were significantly decreased compared to the autografted group. CONCLUSIONS: Our results indicated that l -carnitine can ameliorate the consequences of ischemia-reperfusion on the mice ovarian tissue following autotransplantation. IMPLICATIONS: l -carnitine improves the structure and function of transplanted ovaries.

Evaluating the therapeutic effect and toxicity of theophylline in infertile men with asthenoteratozoospermia: a double-blind, randomized clinical trial study.

Theophylline as a cyclic adenosine monophosphate (cAMP) phosphodiesterase inhibitor (cAMP-PDEI) elevates cAMP levels. We aimed to evaluate the therapeutic effect and toxicity of theophylline on the sperm parameters, oxidative stress (OS), and inflammation in asthenoteratozoospermic men. Sixty asthenoteratozoospermic patients were divided into groups of placebo and theophylline (200 mg/day). After 3 months of oral treatment, sperm parameters, viability, and DNA fragmentation were analyzed by the CASA system, eosin nigrosin staining, sperm DNA fragmentation kit, respectively. The seminal plasma level of reactive oxygen species (ROS) of neat semen samples, malondialdehyde (MDA), total antioxidant capacity (TAC), tumor necrosis factor alpha (TNF-α), and interleukin-10 (IL-10) was assessed. Data were analyzed statistically using the independent samples t-test and the paired t-test and the means were considered significantly different at p < 0.05. Sperm motility, viability, and the number of sperms with normal morphology and the seminal plasma level of TAC and IL-10 and also sperm DNA fragmentation increased significantly in the theophylline group compared to the placebo. The MDA, TNF-α, and ROS levels decreased significantly in the theophylline group compared to the placebo. Theophylline improved sperm parameters, reduced OS and inflammation, but also created genotoxicity and increased sperm DNA fragmentation. Therefore, to benefit from the desired effects of theophylline and inhibit the toxicity of it in the treatment of men with asthenoteratozoospermia, it is suggested to be used simultaneously with another antioxidant to protect sperm DNA from fragmentation.

The effect of melatonin on testis histological changes and spermatogenesis indexes in mice following treatment with dexamethasone.

Dexamethasone is a common medicine that is capable of causing malformation in the male reproductive system. The aim of this study was to investigate the effect of melatonin on testis histological changes and Spermatogenesis indexes in adult mice following treatment with dexamethasone. Adult male NMRI mice were divided randomly into four groups: control, dexamethasone (i.p injections, 7 mg/kg/day), dexamethasone + melatonin and melatonin (i.p injections, 20 mg/kg/day). After 7 days of treatment, the right testes were studied stereologically and the left testes were used to measure the daily sperm production (DSP). The serum levels of malondialdehyde (MDA), testosterone and total antioxidant capacity (TAC) were also measured. The left caudal epididymis was used to analyze sperm parameters. Data were analyzed using one way ANOVA and means were considered significantly different at p < 0.05. A significant decrease in the testis volume, seminiferous tubules volume, the number of spermatocytes, round and long spermatids, Spermatogenesis indexes, sperm parameters such as motility, count, viability, tail length and DSP, serum testosterone level, TAC and the body weight was found in the dexamethasone group compared to the control. Meanwhile a significant swelling of the interstitial tissue and a significant increase in the MDA level was found in the dexamethasone group compared to the control. The above parameters reached the control level in the dexamethasone + melatonin group. Melatonin can compensate for the adverse effects of dexamethasone on sperm parameters and the histology of the seminiferous tubules in mice.

The effect of Quercetin on the quality of sperm parameters in frozen-thawed semen of patients with Asthenospermia.

The study has aimed to investigate the effect of Quercetin, as a potent antioxidant, on preventing the negative effects of freezing process on sperm quality of patients with Asthenospermia. Semen sample from 25 patients was randomly divided into three groups; fresh, control and Quercetin (50 µM). Seven days after freezing, samples were thawed at ambient temperature. Total motility, progressive sperm motility, normal morphology, viability and DNA integrity were evaluated according to WHO criteria, Papanicolaou, eosin- nigrosine and acridine orange staining respectively. In addition, the health of sperm membrane and mitochondrial membrane potential was assessed by HOS test and rhodamine staining. MDA and antioxidant enzyme activity were also evaluated using ELISA method. In contrast to the fresh group, the mean level of MDA and DNA damage had significant increase in the control group and decreased significantly sperm quality (p ≤ 0.001). The mean percentage of total motility and progressive motility, normal morphology, viability and antioxidant enzyme activity had significant increase in the Quercetin group than the control group. In the Quercetin group, the MDA level and DNA damage also had significant reduction in comparison with the control group (p ≤ 0.001). Therefore, the Quercetin supplementation improves the quality of cryopreserved human semen.

Hysterosalpingography in the assessment of proximal tubal pathology: a review of congenital and acquired abnormalities.

Tubal and peritoneal disease are the main causes of infertility. Tubal pathology can be either congenital malformation or acquired, proximal or distal, unilateral or bilateral and transient or permanent. Several imaging methods such as laparoscopy, fluoroscopy, saline infusion sonography, and hysterosalpingography (HSG) have been used in the assessment of tubal and peritoneal pathology. Although laparoscopy is the modality of choice for investigating tubal patency and pelvic structure in many infertility centers, HSG is usually the initial diagnostic method for infertility workup because of its ease of performance, accuracy, and minimal risk of complications. This method provides useful information about size, contour, and anatomy of the inner surface of the fallopian tubes and is the gold standard for evaluation of tubal lumen. Tubal and peritubal pathology show various imaging manifestations on HSG. This review illustrates the radiographic features of congenital and acquired structural abnormalities of the proximal tubal pathology and along with etiology of proximal obstruction or occlusion will be described.

Comparing the effect of sitagliptin and metformin on the oocyte and embryo quality in classic PCOS patients undergoing ICSI.

BACKGROUND: Insulin resistance plays a major role in the pathogenesis of polycystic ovary syndrome (PCOS). Therefore, there is a growing interest in the use of insulin sensitizer drugs in the treatment of PCOS. Research in recent years has shown that sitagliptin has been reported to improve ovarian cycles and ovulation in PCOS patients. AIMS: We aimed to compare the effects of metformin and sitagliptin on PCOS individuals undergoing ICSI. METHODS: Sixty PCOS patients were divided into 3 groups: metformin, sitagliptin, and placebo group. Treatment was carried out 2 months before the start of the ovulation cycle and continued until the day of oocyte aspiration. The serum levels of total testosterone, estradiol, and fasting insulin along with the total number of retrieved, normal and abnormal MII, and fertilized oocytes, the number of transferred embryos (grades I, II and III), and biochemical and clinical pregnancy rates as well as the ovarian hyperstimulation syndrome (OHSS) were evaluated. RESULTS: There was a significant reduction in the serum levels of Insulin and total testosterone in the treated groups compared with the placebo. The number of mature and normal MII oocytes increased significantly in the treated groups compared with the placebo. Moreover, the number of immature oocytes decreased significantly and the number of grade I embryos increases significantly in the sitagliptin group compared with the placebo group. CONCLUSION: We conclude that sitagliptin can improve the maturation of oocytes and embryos' quality more effectively than metformin, in PCOS patients undergoing ICSI. TRIAL REGISTRATION: Trial registration is NCT04268563 ( https://clinicaltrials.gov ).

Sitagliptin/Metformin: A New Medical Treatment in Polycystic Ovary Syndrome.

Metformin has long been used in the treatment of polycystic ovarian syndrome (PCOS). Recently, sitagliptin has been reported to improve ovarian cycles and ovulation in PCOS. We suggest that a combination of sitagliptin and metformin can be more effective than either treatment alone in improving different aspects of PCOS.

Melatonin improves the structure and function of autografted mice ovaries through reducing inflammation: A stereological and biochemical analysis.

Melatonin has anti-oxidant, anti-inflammatory and anti-apoptotic properties. We aimed to investigate the effect of melatonin on the structure and function of mice ovaries following autograft transplantation. NMRI mice were divided into: control, autografted + saline, autografted + melatonin (20 mg/kg/day i.p. injection for 1 day before until 7 days after transplantation). 28 days post transplantation, ovary compartments were studied stereologically. Follicle apoptosis and the level of progesterone and estradiol were also measured. The inflammation, serum MDA concentration and total antioxidant capacity were also assessed on day 7 post transplantation. The total volume of the ovary, cortex and medulla (P < 0.05) and the number of different types of follicles (P < 0.001), the concentration of IL-10, progesterone and estradiol (P < 0.001) and TAC (P < 0.01) significantly decreased in the autografted + saline group compared to the control. The levels of IL-6 (P < 0.01), TNF-α, MDA and the apoptotic rate (P < 0.001) increased significantly in the autografted + saline group compared to the control, while the total volume of the ovary, cortex and medulla (P < 0.05) and the number of different types of follicles (P < 0.001), the concentration of IL-10, progesterone and estradiol (P < 0.001) and TAC (P < 0.01) significantly increased in the autografted + melatonin group compared to the autografted + saline group. The levels of IL-6 (P < 0.01), TNF-α, MDA and the apoptotic rate (P < 0.001) decreased significantly in the autografted + melatonine group compared to the autografted + saline group. In the autografted + melatonin group, the localization of CD31-positive cells in the theca layer was similar to the control group. Melatonin can improve the structure and function of the grafted ovary.

Improvement of the folliculogenesis by transplantation of bone marrow mesenchymal stromal cells in mice with induced polycystic ovary syndrome.

BACKGROUND: Many studies have reported that inflammation and oxidative stress are involved in the pathogenesis of polycystic ovary syndrome (PCOS). Bone marrow mesenchymal stromal cells (BM-MSCs) have anti-oxidant and anti-inflammation properties. In this study, we investigate the beneficial effect of stem cell therapy on folliculogenesis in mice with induced PCOS METHODS: Mouse model of PCOS was performed through daily injection of testosterone enanthate (1 mg/100 g/body weight subcutaneous (s.c).) for a period of 5 weeks. Naval Medical Research Institute (NMRI) mice (21 days old) were divided into three groups: control, PCOS and PCOS + BM-MSCs. BM-MSCs were labeled with Hoechst 33342 (0.5 µg/mL) and then injected into the mice (106/animal, via the tail vein) at 1 and 14 days after PCOS confirmation. Mice were humanely killed at 2 weeks after last transplantation. Ovarian stereological studies were done. Follicle-stimulating hormone (FSH), Luteinizing hormone (LH), testosterone, interleukin (IL)-6 and tumor necrosis factor (TNF)-α serum levels were measured. The levels of malondialdehyde (MDA) and total antioxidant capacity (TAC) in serum were analyzed. Apoptotic index for ovarian follicles was assessed using Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL). CD31 expression in ovarian vessels was assessed with the immunohistochemistry. RESULTS: There was a significant increase in the total volume of ovary, cortex, number of antral follicles, volume of oocyte and zona pellucida thickness, and there was a significant decrease in the primary and preantral follicles number in the PCOS + BM-MSCs group compared with the PCOS group. There was a significant increase in the serum level of FSH and TAC and a significant decrease in the serum level of testosterone, LH, MDA and percentage of TUNEL-positive apoptotic cells in the PCOS + BM-MSCs group in comparison with the PCOS group. DISCUSSION: BM-MSC transplantation improves folliculogenesis in mice with induced PCOS. BM-MSC therapy can be an operative treatment for PCOS via anti-inflammatory, anti-oxidant and anti-apoptotic properties.

Adipose-derived mesenchymal stromal cell transplantation at the graft site improves the structure and function of autografted mice ovaries: a stereological and biochemical analysis.

BACKGROUND: Ovarian tissue autografting is a fertility restoration technique that is frequently used in young women with cancer who undergo radio/chemotherapy. A limiting factor in this technique is ischemia-reperfusion (I/R) damage. Because adipose-derived mesenchymal stromal cells (ADMSCs) protect different ischemic tissues against I/R damage, we examined the effect of ADMSC transplantation at the graft site in mice ovary autografting. METHOD: Mice were divided into three groups: control, autograft and autograft + ADMSCs. Seven days after ovary autografting and ADMSC transplantation, serum superoxide dismutase (SOD) activity, total antioxidant capacity, serum concentrations of malondialdehyde (MDA), tumor necrosis factor alpha (TNFα), interleukin (IL)-6 and IL-10 were measured. After 28 days, ovary histology, serum concentrations of progesterone and estradiol and apoptosis rate were also estimated. At 1-3 and 28 days post-ovary autografting and ADMSC transplantation, angiogenesis was detected. The results were analyzed using one-way analysis of variance (ANOVA) and Tukey test, and the means were significantly different at P ≤ 0.05. RESULT: In the autograft + ADMSCs group, the total volume of the ovary, cortex and medulla (P ≤ 0.001), the number of follicles, SOD activity, IL-10 (P ≤ 0.001) and progesterone and estradiol (P ≤ 0.01) concentrations significantly increased compared with the autograft group. Apoptosis rate, IL-6, TNFα and MDA concentrations in the autograft + ADMSCs group were lower than the autograft group (P ≤ 0.001). The angiogenesis was accelerated and the localization of CD31-positive cells in the cortex was similar to the control group following ADMSC transplantation. DISCUSSION: ADMSC transplantation enhances the structure and function of grafted ovary.

The effect of Verapamil on ischaemia/reperfusion injury in mouse ovarian tissue transplantation.

One of the challenges that must be overcome during ovarian tissue transplantation is Ischaemia/Reperfusion Injury (IRI). The most important hypothesis explaining the cellular events in I/R processes are calcium overload and oxygen free radicals constitute. Here, we study the effect of verapamil on IRI, and consequently on follicle survival during ovarian transplants in an autograft model. Female mice were randomly assigned into three groups in order to ovarian autotransplantation as follow: Group 1 (Control group), Group 2 (Transplanted group) and Groups 3 (Transplanted + Verapamil group). The grafted ovaries were collected at 3, 7 and 14 days after transplantation for evaluation of follicle content and morphology, apoptosis and Malondialdehyde (MDA) concentration. The results showed that verapamil treatment significantly preserved primordial follicular reserve and reduced the number of degenerated follicles compared to the transplanted group (P < 0.05). MDA levels were significantly higher on the 14th day after transplantation, in group 2 than in group 3. In conclusion, verapamil treatment is effective for the preservation of the follicular pool and reducing tissue damage induced by transplantation of ovarian tissue.

Protective antioxidant effects of N-acetylcysteine against impairment of spermatogenesis caused by paranonylphenol.

Paranonylphenol (p-NP) is an environmental pollutant that causes oxidative stress. The purpose of this study was to evaluate the protective effect of N-acetylcysteine (NAC) as an antioxidant on sperm parameters and testis in mice after treatment with p-NP. Adult mice were randomly divided into four groups (n = 6, each group) including 1-control, 2- p-NP (250 mg kg-1  day-1 ), 3- NAC (150 mg kg-1  day-1 ) and 4- p-NP + NAC. After 35 days of oral treatment, the mean of spermatogenic index (p < 0.02), sperm count (p < 0.01), daily sperm production (p < 0.01), sperm tail length (p < 0.02), progressive movement (p < 0.04), normal morphology (p < 0.04) and viability (p < 0.01) of spermatozoa and also serum testosterone level (p < 0.04) were significantly reduced in p-NP group when compared to other groups. While the count of the positive TUNEL cells in the seminiferous tubules (p < 0.01) and level of the malondialdehyde (MDA) in testis (p < 0.02) and serum (p < 0.01) significantly increased. In the histopathologic assay in the p-NP group, apoptosis, atrophy, oedema, reduction in sperm density in lumens and vacuoles were observed. The findings of this study indicate that NAC as a potent antioxidant be able to compensate the adverse effects of p-NP in spermatogenesis, testis and levels of testosterone and MDA in the p-NP + NAC group significantly compared to the p-NP group.

Ovary stereological features and serum biochemical factors following induction of polycystic ovary syndrome with testosterone enanthate in mice: An experimental study.

BACKGROUND: Polycystic ovary syndrome (PCOS) is an endocrine disorder featured by insulin resistance and hyperandrogenism. Testosterone enanthate can induce PCOS in mice models. OBJECTIVE: We investigated the ovary stereological features along with the oxidative stress and inflammatory factors in mice following PCOS induction using testosterone enanthate. MATERIALS AND METHODS: Twelve female NMRI mice (3 wk old) were divided into 2 groups (n=6/each): Control and PCOS. PCOS was induced through daily injections of testosterone enanthate (1 mg/100g subcutaneous s.c for 5 wk). Finally, ovaries were studied stereologically. The serum levels of the follicle-stimulating hormone, luteinizing hormone, testosterone, interleukin-6, and tumor necrosis factor-α were measured using ELISA kit. Serum levels of Malondialdehyde and the antioxidant capacity were measured relatively using thiobarbituric acid and ferric reducing antioxidant power assay. RESULTS: The mean total volume of ovary and the mean volume of cortex (p<0.001), volume of oocyte in the preantral (p=0.011) and antral follicle (p=0.015), thickness of zona pellucida (p=0.016), the number of antral follicles (p=0.012), the serum levels of follicle-stimulating hormone (p<0.001) and the antioxidant capacity (p=0.020) reduced significantly in the PCOS group compared to the control. The number of primary (p=0.017) and preantral (p=0.006) follicles and the serum levels of testosterone (p<0.001), Luteinizing hormone (p=0.002), Malondialdehyde, Interleukin 6 and Tumor necrosis factor-α (p<0.001) showed a significant increase in the PCOS group compared to the control. CONCLUSION: Testosterone enanthate induced PCOS causes stereological features in the ovary, increases the oxidative stress and inflammatory markers in mice.

N-Acetylcysteine Compared to Metformin, Improves The Expression Profile of Growth Differentiation Factor-9 and Receptor Tyrosine Kinase c-Kit in The Oocytes of Patients with Polycystic Ovarian Syndrome.

BACKGROUND: Paracrine disruption of growth factors in women with polycystic ovarian syndrome (PCOS) results in production of low quality oocyte, especially following ovulation induction. The aim of this study was to investigate the effects of metformin (MET), N-acetylcysteine (NAC) and their combination on the hormonal levels and expression profile of GDF-9, BMP-15 and c-kit, as hallmarks of oocyte quality, in PCOS patients. MATERIALS AND METHODS: This prospective randomized, double-blind, placebo controlled trial aims to study the effects of MET, NAC and their combination (MET+NAC) on expression of GDF-9, BMP-15 and c-kit mRNA in oocytes [10 at the germinal vesicle (GV) stage, 10 at the MI stage, and 10 at the MII stage from per group] derived following ovulation induction in PCOS. Treatment was carried out for six weeks, starting on the third day of previous cycle until oocyte aspiration. The expression of GDF9, BMP15 and c-kit were determined by quantitative real time polymerase chain reaction (RT-qPCR) and western blot analysis. Data were analyzed with one-way ANOVA. RESULTS: The follicular fluid (FF) level of c-kit protein significantly decreased in the NAC group compared to the other groups. Significant correlations were observed between the FF soluble c-kit protein with FF volume, androstenedione and estradiol. The GDF-9 expression in unfertilized mature oocytes were significantly higher in the NAC group compared to the other groups (P<0.001). Similar difference was not observed between the MET, NAC+MET and control groups. The c-kit expression in unfertilized mature oocytes were significantly lower in the NAC group compared to the other groups (P<0.001). Similar difference was not observed between the MET, NAC+MET and control groups (Registration number: IRCT201204159476N1). CONCLUSION: We concluded that NAC can improve the quality of oocytes in PCOS.

Impact of pre-incubation time of silk fibroin scaffolds in culture medium on cell proliferation and attachment.

Cell behaviours such as proliferation and attachment can be affected by the length of pre-incubation period of the scaffolds in the culture medium for long term. The aim of this study was to investigate the long term pre-incubation of 3D silk fibroin scaffolds in complete culture medium on cell attachment and proliferation. After the preparation of silk fibroin scaffolds by the technique of freeze drying, the scaffolds were pre-incubated in complete culture medium for 2 d, 6 d or 10 d before apical papilla stem cells (SCAP) seeding. Modifications of the scaffold surface and wettability were examined by FE-SEM and water contact angle, respectively. Results showed a decrease both in roughness and water contact angle as pre-incubation time increases. DNA measurement after 18h and 10 d cell seeding showed a significant increase of DNA concentration which represents better attachment and proliferation with pre-incubation time increase. Qualitative examination, live&dead assay or H&E staining method after 30h and 10 d cell seeding respectively, indicated that pre-incubation of scaffolds has time dependent effect on cell proliferation and attachment. This suggests that improvement of cell attachment and proliferation may be mediated by differences in the amount of wettability (decreased water contact angle) after exposure of scaffold to culture medium for long term which, in turn, causes more protein adsorption in the surface of silk fibroin scaffold (decreased roughness).

Stereological study on the effect of vitamin C in preventing the adverse effects of bisphenol A on rat ovary.

BACKGROUND: Bisphenol A (BPA), an environmental pollutant, can generate free radicals which damages the reproductive system. Vitamin C is an antioxidant which may prevent the adverse effects of free radicals. OBJECTIVE: The aim was to investigate the effect of vitamin C on the ovary tissue in rats treated with BPA. MATERIALS AND METHODS: In this experimental study, 24 female Wistar rats (200±20 gr) were randomly divided into 4 groups (n=6): control, BPA (60 µg/Kg/day), vitamin C (150 mg/Kg/day) and BPA + vitamin C and orally treated for 20 days. The left ovaries were taken out, fixed for tissue processing and studied using stereological methods. Data were analyzed with SPSS using one-way ANOVA, and the means were considered significantly different at (p<0.05). RESULTS: The total volume of ovary and cortex (p<0.01), medulla (p<0.05), the volume of corpus luteum (p<0.001) and the mean number of antral follicles (p<0.001) significantly reduced in BPA group compared with control, while the number of atretic follicles increased (p<0.05). The volume of oocyte (p<0.01) and its nucleus (p<0.001) in the antral follicles and the thickness of zona pellucida (ZP) in the secondary (p<0.05) and antral (p<0.001) follicles significantly decreased in BPA group compared with controls. The above parameters in the BPA + vitamin C group were compensated to control level. CONCLUSION: Vitamin C can be used as a potential antioxidant in the case of BPA toxication.