Comparison of mouse ovarian follicular development and gene expression in the presence of ovarian tissue extract and sodium selenite: An experimental study.

Background: Ovarian tissue extract (OTE) and sodium selenite (SS) enhance the growth and maturation of preantral follicles in a dose-dependent manner. Objective: The present study was designed to bring more information regarding the mechanism of OTE and SS on the mRNA expression of follicle-stimulating hormone receptors (FSHR) and the proliferation cell nuclear antigens (PCNA) of in vitro matured isolated follicles. Materials and Methods: The tissue extract was prepared from adult ovaries. The preantral follicles (n = 266) were isolated from 12-16-day-old mice and cultured in the control, experimental I (10 ng/ml SS), and experimental II (OTE) groups for 12 days. The follicular diameter, survival, and maturation rates, also, the production of 17-ß-estradiol and progesterone, and the follicular expression of PCNA and FSH receptor genes were analyzed. Results: The survival rate of follicles in the SS-treated group (84.58%) was significantly higher than that OTE (75.63%; p = 0.023) and control (69.38%; p = 0.032) groups. The mean diameter of culture follicles in experimental group I (403.8 µm) and experimental group II (383.97 µm) increased significantly in comparison with the control group (342.05 µm; p = 0.032). The developmental rate of follicles, percentages of antrum formation, released metaphase II oocytes (p = 0.027; p = 0.019 respectively), production of hormones and the expression of 2 studied genes were significantly increased in both experimental groups in compare with control group (p = 0.021; p = 0.023 respectively). Conclusion: The OTE and SS have a positive effect on development of mouse preantral follicles via over-expression of FSHR and PCNA genes.

Dual Oxidase 1, 2 Gene Expression in Women With Polycystic Ovary Syndrome (PCOS).

Objective: Dual oxidases (DUOX1, DUOX2) belong to the NADPH oxidase (NOX) family, which produce H2O2 necessary for thyroid hormone biosynthesis. This study aims to evaluate gene expression for DUOX1, DUOX2 in PCOS patients and its relation with thyroid hormone and magnesium levels. Materials and methods: Totally 88 cases were studied including 24 people with PCOS and hypothyroidism, 44 people with PCOS and normal thyroid function, and 20 hypothyroid patients without PCOS. In comparison 40 healthy controls in the age group of 16-35 years matched for age group and BMI were evaluated. Using Vegaro syringe 5 cc of blood was sampled from all 128 people and after RNA extraction and cDNA synthesis using Real-Time PCR technique, the expression level of DUOX1 and DUOX2 genes was investigated. Results: The results of hormonal tests showed that there is a significant difference between the level of T4, T3, and TSH hormones in hypothyroid patients with or without PCOS in comparison to the control group. Regarding the level of Mg, the results showed that there is a significant difference between the levels of Mg in PCOS group with or without hypothyroidism in comparison to the control group. Gene expression results showed that the relative changes of DUOX1 gene expression in different groups compared to the control group were significantly reduced P<0.05. In the polycystic group with hypothyroidism, the gene expression level showed a decrease compared to the normo-thyroid polycystic group and the hypothyroid non-PCO group, which was statistically significant P<0.05. Conclusion: According to the results of the present study and the previous studies that have been published in the field of Duox1, it can be assumed that the reduction of Duox1 expression can interfere with the oxidative stress system. Further studies with other molecular techniques may help to understand the exact action mechanism of these genes.

Effects of Cold Plasma on Staphylococcus Aureus.

Objective: To investigate the effect of cold plasma on Staphylococcus aureus destruction at different treatment times. Materials and methods: Staphylococcus aureus was cultured on 4 plates of LB Agar medium each at 1.5 × 103 CFU / mL (colony-forming unit per milliliter) and one group was selected as the control group and the other 3 groups were treated with plasma for 5, 7 and 10 minutes. They were incubated for 24 hours at 37 °C. Finally, the number of colonies formed was counted. Results: It was shown that treatment with cold atmospheric plasma significantly reduced bacterial colonies and in comparison to the control plate with a colony count of 1.5 × 103 CFU/mL treatment with air plasma for 10 minutes decreased the Pseudomonas colony count to zero. Conclusion: It was observed that the cold atmospheric plasma jet device manufactured in atomic Energy Organization of Iran can significantly kill bacteria in a short time. Increasing the duration of treatment significantly reduces bacterial colonies.

The Effect of COVID-19 Pandemic on Outcome of Assisted Reproductive Technology: A Report from a Single Infertility Center.

Objective: COVID-19 pandemic has affected all aspects of human life including social, economic, healthy behaviors and even individual relationships. This study aimed to investigate the effect of corona virus outbreak on assisted reproductive technology (ART) outcome. Materials and methods: In this retrospective and prospective cohort, 260 ART cycles of ovum pick up (OPU), fresh embryo transfer (ET) and frozen embryo transfer (FET) were evaluated in 223 women (from December 2019 to February 2020) and during COVID-19 outbreak (February and July 2020) in an infertility center. Primary and secondary outcomes of ART cycles including chemical and clinical pregnancy rate were evaluated. Results: The mean±SD (standard deviation) age of women was 34.17±6.56 years. Chemical and clinical pregnancy rates were 23.91% (33/138) per embryo transfer and 75.8% (25/33) per positive pregnancy test, respectively while ongoing pregnancy was seen only in 69.7% (23/33) of those with positive pregnancy test. Spontaneous abortion rate was 15.15% (5/33) per laboratory pregnancy. COVID-19 symptoms were reported in 2.83% and 15.38% of women during and after ART cycles, respectively. Conclusion: It seems that COVID-19 pandemic has not negative effect on outcome of ART cycles except for cancelation rate due to COVID-19 that increased at the beginning of COVID-19 outbreak as it was unknown at that time and awareness was limited.

Examining the Effect of Chamomile on Clinical Symptoms and Hormonal Parameters Among Patients With Polycystic Ovarian Syndrome.

Objective: Polycystic ovarian syndrome is the most common cause of infertility and endocrine disorder among women due to anovulation. The aim of the present study is to investigate the effect of chamomile on oligomenorrhea and hirsutism symptoms as well as hormonal parameters among patients suffering from polycystic ovarian syndrome. Materials and methods: The present study is a randomized clinical trial performed on 70 patients with diagnosis of polycystic ovarian syndrome according to Rotterdam criteria referring to the infertility clinic of a university hospital. The patients were randomly assigned into intervention and control groups, with the former receiving two chamomile capsules 500 mg for three months, and the latter receiving two placebo capsules for 3 months. Both groups were evaluated in terms of laboratory parameters (FBS, LDL, and testosterone) on the third day of first and third cycles. The collected data were analyzed by SPSS 20. Results: The mean age, body mass index, marital status, history of infertility, and pregnancy rate showed no significant difference between the two groups. In the chamomile group, clinical symptoms of hirsutism (p<0.001) and oligomenorrhea (p=0.048) decreased following the treatment, but significant difference was found between the two groups only in hirsutism symptoms (p=0.028). Regarding the parameters of FBS (p=0.06), HDL (p=0.224), cholesterol (p=0.99), triglyceride (p=0.106), testosterone (p=0.894), and LDL (p=0.61), no significant difference was observed between the two groups. Nevertheless, following the treatment, testosterone decreased in both placebo (p=0.005) and chamomile (p=0.001) groups. Conclusion: overall, the chamomile therapeutic regimen has relatively been able to mitigate the clinical symptoms and testosterone levels in patients suffering from polycystic ovarian syndrome. Use of chamomile plant as a simple, inexpensive, and effective measure can be suggested for improving and treating patients with PCOS after confirmation by further studies.

The first report of clonidine in vivo/in vitro effects on infertile women with polycystic ovary syndrome (in vivo/in vitro study).

The sympathetic nervous system (SNS) is hyperactive in women with polycystic ovary syndrome (PCOS). This study was designed in two sections: in vivo/in vitro with clonidine as the alpha-2 adrenoceptor (ADR-α2) agonist for modulating this hyperactivity. Eighty women with PCO participated in this randomised clinical trial (in vivo). A clonidine (0.1 mg) tablet was given twice a day for two months. Polycystic ovary morphology (PCOM) and pregnancy rate were the main outcome measurements. In the candidates for in vitro fertilisation (IVF), clonidine was added to the culture medium during IVF for two study groups (PCO-clonidine/PCO-without) and two control groups (egg donors-clonidine/egg donors-without). Our results showed that the pregnancy rate significantly was higher in the study group (p = .002). The mRNA expression of ADR-α1 and ADR-ß2 in PCO was higher than control group (p value <.001). But ADR-α1 expression in PCO-clonidine group decreased (p value = .042), the same as ADR-α2 expression. The intensity of this effect showed a pattern for ADR-α1

In Vitro Implantation Model Using Human Endometrial SUSD2+ Mesenchymal Stem Cells and Myometrial Smooth Muscle Cells.

OBJECTIVE: This study evaluated a novel in vitro implantation model using human endometrial mesenchymal stem cells (EMSCs), SUSD2+, and myometrial smooth muscle cells (SMCs) that were co-cultured with mouse blastocysts as the surrogate embryo. MATERIALS AND METHODS: In this experimental study, SUSD2+ MSCs were isolated from human endometrial cell suspensions (ECS) at the fourth passage by magnetic-activated cell sorting. The ECS and SUSD2+ cells were separately co-cultured with human myometrial muscle cells for five days. After collection of mouse blastocysts, the embryos were placed on top of the co-cultured cells for 48 hours. The interaction between the embryo and the cultured cells was assessed morphologically at the histological and ultrastructural levels, and by expression profiles of genes related to implantation. RESULTS: Photomicrographs showed that trophoblastic cells grew around the embryonic cells and attached to theECS and SUSD2+ cells. Ultrastructural observations revealed pinopode and microvilli-like structures on the surfaces of both the ECS and SUSD2+ cells. Morphologically, the embryos developed to the egg-cylinder stage in both groups. Gene expression analysis showed no significant differences between the two groups in the presence of an embryo, but an increased expression of αV was detected in SUSD2+ cells compared to ECS cells in the absence of an embryo. CONCLUSION: This study showed that SUSD2+ cells co-cultured with SMCs could interact with mouse embryos. The co-cultured cells could potentially be used as an implantation model.

An efficient protocol for decellularization of the human endometrial fragments for clinical usage.

The present study was aimed to compare different decellularization protocols for human endometrial fragments. The freeze-thaw cycles in combination with treatment by Triton X-100 and four concentrations of sodium dodecyl sulfate (SDS; 0.1, 0.5, 1, and 1.5%) with two exposure times (24 and 72 h) were applied for tissues decellularization. After analysis the morphology and DNA content of tissues the group with better morphology and lower DNA content was selected for further assessments. The nucleus by Acridine orange and extracellular matrix (ECM) using Masson's trichrome, Alcian blue, and periodic acid-Schiff staining were studied. The amount of tissues collagen types I and IV, fibronectin, glycosaminoglycans (GAGs), and elastin was analyzed by Raman spectroscopy. The ultrastructure and porosity of decellularized scaffold were studied by scanning electron microscopy (SEM). The MTT assay was applied for assessments of cytotoxicity of scaffold. The treated group with 1% SDS for 72 h showed the morphology similar to native control in having the minimum level of DNA and well preserved ECM. Raman spectroscopy results demonstrated, the amount of collagen types I and IV, GAG, and fibronectin was not significantly different in decellularized scaffold compared with native group but the elastin protein level was significantly decreased (P < 0.001). SEM micrographs also showed a porous and fiber rich ECM in decellularized sample similar to the native control. This combined protocol for decellularization of human endometrial tissue is effective and it could be suitable for recellularization and clinical applications in the future.

Impact of endometriosis on reproductive health: an integrative review.

Endometriosis is a debilitating chronic disease that affects 10% of the general female population, with a frequency of 38% in infertile women. This review aims to provide an overview of the impact of endometriosis on different aspects of reproductive health. A literature search was performed in four international databases, including PubMed, Science Direct, Scopus, and Google scholar, from January 1995 to December 2019 to identify the potentially relevant articles. We included all relevant articles published in English and identified 46 studies on health issues in women with endometriosis. The research suggests that women with endometriosis are at risk for physical, psychosocial, social, and sexual disturbances and obstetrical complications. More research is needed to clarify better understanding of endometriosis complications specifically obstetrical consequences.

Psychological well-being of infertile women and its relationship with demographic factors and fertility history: a cross-sectional study.

BACKGROUND: Infertility leads to a wide range of psychological injuries that may reduce psychological well-being. This study aimed to determine the psychological well-being of infertile women and its relation with demographic factors and fertility history. METHODS: This cross-sectional study was conducted on 300 infertile women referred to three infertility centres, Tehran, Iran. The sampling was continuous. We collected data from a self-generated demographic and fertility questionnaire and Ryff's Psychological Well-being Scale (PWB). Data analysis was done by independent t-test, one way ANOVA. The significance level was set at P < 0.05. RESULTS: The results showed that there was no significant relationship between demographic variables including age, occupation of each couple, spousal's education, economic status and place of residence with PWB, but the mean score of PWB was significantly different in women's educational levels (P = 0.03). There was also a significant difference between the mean score of PWB among different groups in the duration of marriage (P = 0.01). Fertility characteristics variables include the duration of infertility, duration of treatment of infertility, and current treatment were not the relation with PWB. However, the mean score of PWB in the number of IVF (P = 0.003) and the failed IVF pregnancies (P = 0.01) had a significant statistical difference. CONCLUSION: The results showed that PWB related to several variables. Paying attention to these variables can help in the preparation and development of counseling or educational programs.

Exploring the impact of endometriosis on women's lives: A qualitative study in Iran.

AIM: Considering the wide impact of endometriosis on various aspects of health, this study aimed to explore the impact of endometriosis on Iranian women's lives. DESIGN: The present study used a qualitative approach and conventional content analysis. METHODS: Twenty patients suffering from endometriosis referring to Imam Khomeini Hospital in Tehran took part in this study. The sampling was done purposefully until data saturation. After deep semi-structured interviews, the content analysis of the interviews was done according to the steps proposed by Zhang and Wildemuth. RESULTS: The mean age of the participants was 34.53 (SD: 5.81) years (range: 23-43) with duration of illness of 6.30 ± 5.86 years. Their educational level varied from high school to university, and most of them were employed. Analysis of the data from participants' experiences led to the formation of 5 categories under the titles "Physical suffering, instability of marital life, mental disorder, disruption in social life and self-care" and 11 subcategories. CONCLUSION: The findings of the present study showed that endometriosis can adversely affect women's lives including physical, sexual, psychological and social problems. Although in some cases patients adopt different strategies for self-care, the need for more support is felt.

Endometriosis reproductive health questionnaire (ERHQ): A self-administered questionnaire to measure the reproductive health in women with endometriosis.

INTRODUCTION: Endometriosis is a disabling disease impacting on different aspects of reproductive health. The present study aims to develop and evaluate the psychometric properties of a questionnaire to measure the reproductive health in women with endometriosis. MATERIAL AND METHODS: This study used an exploratory mixed-methods design and included three phases. In the first phase, a sample of 20 women with endometriosis participated in semi-structured interviews. In the second phase, item pools were generated. In the third phase, face validity, content validity, and construct validity were assessed. To estimate the reliability of the tool internal consistency and test-retest methods were used. RESULTS: sixty items were included in the psychometric evaluation stage of the scale. After the assessment of the content validity ratio (CVR), content validity index (CVI), and exploratory factor analysis, 15 items were omitted, leaving 35 items in the final scale. The exploratory factor analysis revealed four factors: physical problems, psychological problems, counteracting strategies, and instability of marital life. The reliability, according to Cronbach's alpha was 0.809, and the external reliability, as evaluated by the test-retest method and the intraclass correlation, was 0.825. CONCLUSION: ERHQ is a new, valid, and reliable patient-generated instrument to measure the reproductive health in women with endometriosis. It can be used by researchers and health providers to provide a better understanding of the impact of endometriosis on different aspects of reproductive health over time and to meet the needs of patients living with this condition.

Comparison of Dydrogesterone and GnRH Antagonists for Prevention of Premature LH Surge in IVF/ICSI Cycles: A Randomized Controlled Trial.

Objective: To compare the effect of dydrogesterone and Gonadotropin releasing hormone (GnRH) antagonists on prevention of premature luteinizing hormone (LH) surge and pregnancy outcomes in infertile women undergoing Invitro fertilization/ Intra cytoplasmic sperm injection (IVF/ICSI). Materials and methods: In a Randomized controlled trial (RCT), two-hundred eligible women undergoing in vitro fertilization (IVF) /intracytoplasmic sperm injection (ICSI) treatment were randomly assigned into two groups. Human menopausal gonadotropin (HMG) was administered for controlled ovarian stimulation (COS) in both groups. Intervention group (group 1) received 20 mg dydrogesterone from day 2 of menstrual cycle till trigger day and control group (group2) received GnRH antagonist from the day that leading follicle reached 13 mm in diameter till trigger day. Serum levels of LH, estradiol and progesterone were measured on the trigger day. The primary outcome measure was the incidence of a premature LH surge, and the secondary outcomes investigated were the chemical and clinical pregnancy rates in the first FET cycles. Results: There were no significant differences in patients' age, BMI, AMH levels, previous IVF cycle, and cause of infertility between the two groups. None of the patients in two groups experienced a premature luteinizing hormone surge. The numbers of retrieved oocytes, the MII oocytes and good quality embryos, were significantly higher in the intervention group than antagonist group (p < 0.05). The overall chemical pregnancy rate in intervention group (43/91: 46.2%) and control group (45/91: 49.5%) (p = 0.820) was similar. Meanwhile, the clinical pregnancy rate was similar between groups too. Conclusion: Regarding the cost, efficacy and easy usage of dydrogestrone, it may be reasonable to use it as an alternative to GnRH antagonist for the prevention of premature LH surge.

Concerns of Infertile Women Candidates for Egg Donation: A Qualitative Study.

Objective: Women with premature ovarian insufficiency, menopause, gonadal dysgenesis, and genetic disorders as well as those with a history of recurrent in vitro fertilization failure may benefit from the assisted fertility techniques. These women experience concerns that directly affect their married life especially in egg donation treatment. This study was conducted to investigate the reproductive health concerns of infertile women who were candidates for egg donation. Materials and methods: This qualitative content analysis was conducted in 17 infertile women who were referred to Imam Khomeini Hospital Infertility Clinic from July 2018 to March 2019. The participants were selected through purposeful sampling. Data were collected through individual in-depth semi-structured interviews. A conventional qualitative content analysis approach was adopted for data analysis using the MAXQDA12 software. Results: The results of data analysis showed five themes, including threatened married life, lack of supportive situation, religious beliefs, psychosocial damage, and damaged feminine identity. Conclusion: Infertility is usually accompanied by a great psychological sadness referred to as "stress of infertility". Assisted reproductive techniques (ART) are also a source of stress for patients; therefore, it has a significant impact on the marital relationship and mental health of infertile women. Hence, there is a need for psychological support from the treatment team to reduce woman reproductive health concerns.

Supplement Usage Pattern in a Group of COVID- 19 Patients in Tehran.

Objective: The coronavirus disease 19 (COVID-19) is a highly transmittable and pathogenic viral infection, causes severe acute respiratory syndrome and was spread throughout the world in early 2020. The effects of vitamin and micronutrient supplements on the prevention and treatment of COVID- 19 seems challenging in scientific considerations. On the other side generally, experts warn against over-consumption of these supplements. Materials and methods: This study aimed to investigate the vitamin and micronutrient supplementation usage pattern in past history of patients with COVID-19 via a cross-sectional inquiry. Totally 510 patients referring to the infectious disease clinic of Imam Khomeini Hospital in Tehran from March 2020 to May 2020 were recruited. The inclusion criterion was suspected patients for COVID-19 based on clinical findings and CT scans of the lung. The infected patients included both inpatients (171) and outpatients (339). Demographic information, clinical signs, and the supplement pattern use were collected through a questionnaire and the data were statistically analyzed. Results: Vitamin D3 intake was reported in 30% (103 patients) of outpatients and 16.5% (28 patients) of hospitalized patients, which is statistically significant (P=0.001). It shows that, the frequency of vitamin D3 consumption in the outpatient group was higher than inpatient group. This significant difference has also been shown in zinc consumption, in 29 patients (9%) outpatients versus 4 patients (2%) inpatients were reported (P=0.007). Multi nominal regression showed that vitamin D3 intake has a supportive effect and reduces the risk of exacerbation and worsening of the disease. (OR=0.291; 95% CI 0.102-.0834, P=0.022). Conclusion: According to the results of the present study and the findings of other studies, considering the supportive effect of vitamin D3 in reducing the severity of infectious diseases; Clinical trials with an appropriate sample size are recommended to investigate the functional role of this vitamin in improving viral diseases of the respiratory tract.

Quality of Life Changes in Iranian Patients Undergoing Female-to-Male Transsexual Surgery: A Prospective Study.

BACKGROUND: Gender identity disorder (GID) is associated with various adverse health outcomes as well as psychiatric problems. Quality of life (QOL) in patients after surgery is an important issue, as some cases report dissatisfaction and regret after surgery. The present study compared QOL in female-to-male (FTM) GID patients before and after gender reassignment surgery (GRS) in an Iranian population. In the present study, 42 natal female patients with Gender Disorder diagnosis, who were referred to the Fertility Research Center of Tehran University of Medical Sciences for GRS from December 2014 to December 2015, were prospectively recruited. METHODS: The demographic characteristics, including age, body mass index (BMI), occupational and marital status, educational level, and family support were recorded. Then, participants were asked to complete the Persian version of the 36-Item Short Form Health Survey (SF-36) questionnaire under supervision of the researcher, once at baseline and once 6 months after surgery. RESULTS: The changes in SF-36 scores and association with the demographic variables were then evaluated using SPSS 16 at significance level of 0.05. Mean age of patients was 34.17 ± 5.58 years with a mean BMI of 43.14 ± 6.91 kg/m2 . Most were single (54.8%), had primary school education (50.5%), and acceptable family support (59.5%). Total mean score of QOL significantly improved from 26.43±6.81 to 37.52 ± 8.67 (P < 0.001), 6 months after surgery and also in all domains (P < 0.001), although the increase in emotional problem was not statistically significant (P = 0.05). CONCLUSION: In conclusion, as the results of the present study highlight, FTM GD patients have a low QOL before surgery that is significantly improved after surgery.

Morphological, Ultrastructural, and Molecular Aspects of In Vitro Mouse Embryo Implantation on Human Endometrial Mesenchymal Stromal Cells in The Presence of Steroid Hormones as An Implantation Model.

OBJECTIVE: This experimental study aimed to evaluate the effects of 17ß-estradiol (E2) and progesterone (P4) on the interaction between mouse embryo and human endometrial mesenchymal stromal cells, and gene expressions related to implantation [αV and ß3 integrins, interleukin-1 receptor (IL-1R), and leukemia inhibitory factor receptor (LIFR)] using an in vitro twodimensional model. MATERIALS AND METHODS: In this experimental study, the endometrial stromal cells were isolated enzymatically and mechanically, and cultured to the fourth passage. Next, their immunophenotype was confirmed by flow cytometric analysis as mesenchymal stromal cells. The cells were cultured as either the experimental group in the presence of E2 (0.3 nmol) and P4 (63.5 nmol) or control group without any hormone treatment. Mouse blastocysts were co-cultured with endometrial mesenchymal stromal cells in both groups for 48 hours. Their interaction was assessed under an inverted microscope and scanning electron microscopy (SEM). Expressions of αV and ß3 integrins, LIFR, and IL-1R genes were analyzed by real-time reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: Similar observations were seen in both groups by light microscopy and SEM. We observed the presence of pinopode-like structures and cell secretions on the apical surfaces of endometrial mesenchymal stromal cells in both groups. The trophoblastic cells expanded and interacted with the mesenchymal monolayer cells. At the molecular level, expression of IL-1R significantly increased in the hormonal treated group compared to the control (P≤0.05). Expressions of the other genes did not differ. CONCLUSION: This study has shown that co-culture of endometrial mesenchymal stromal cells with mouse embryo in media that contained E2 (0.3 nmol) and P4 (63.5 nmol) could effectively increase the expression of IL-1R, which is involved in embryo implantation. However, there were no significant effects on expressions of αV and ß3 integrins, LIFR, and on the morphology and ultrastructure of endometrial mesenchymal stromal cells.

Trans dermal estrogen (oestrogel) for endometrial preparation in freeze embryo transfer cycle: An RCT.

BACKGROUND: Estrogen and progesterone are two crucial factors for endometrial preparation in frozen embryo transfer (FET) cycles. Studies assessing different forms of estradiol in FET have published already but literature lacks enough surveys on transdermal estrogen application in reproductive medicine. OBJECTIVE: To investigate the effects of trans dermal estrogen (Oestrogel) on pregnancy rates in patients that candidate for FET cycle. MATERIALS AND METHODS: In this randomized clinical trial, 100 women undergoing FET cycles referred to Imam Khomeeini Hospital were enrolled in two groups, randomly. Group I received 8 mg/day estradiol valerate (E2 tablet) orally and group II were treated with 6 mg/day transdermal oestrogel gel after suppression with gonadotropin releasing hormone agonist. In both groups medication were started in the first day of menstruation cycle and continued until endometrial thickness reached 8 mm. Pregnancy rates (chemical, clinical, and ongoing), abortion rate, live birth rate, and frequency of complications were compared between two groups. RESULTS: Chemical and clinical pregnancy rates were not significantly different between two groups (p=0.384). The abortion rate was significantly lower in group II than group I (p=0.035). Ongoing pregnancy and the live birth rates were significantly higher in group II (p=0.035). The rate of complication was not different in two groups. CONCLUSION: Oestrogel seems to enhance ongoing pregnancy and live birth rates in comparison to estradiol valerate tablet.

Vitrification and in vitro culture had no adverse effect on the follicular development and gene expression of stimulated human ovarian tissue.

AIM: The study assesses the effect of the vitrification procedure on the integrity, morphology, follicular development and gene expression of stimulated human ovarian tissue after warming and two weeks of in vitro culture. METHODS: Ovarian specimens were divided into non-vitrified and vitrified groups and were cultured for two weeks. Morphological analysis and immunohistochemistry were performed. The 17-ß estradiol and anti-Müllerian hormone levels in collected media were assessed. Gene expression was analyzed using real-time reverse transcription polymerase chain reaction. RESULTS: The morphology and immunohistochemistry of bcl-2-like protein 4 and B-cell lymphoma 2 of human stimulated ovarian tissue were similar in both groups. There was no significant difference in the percentage of normal follicles between the groups before and after in vitro culture. In spite of an increase in the percentage of growing follicles in cultured tissues compared to the non-cultured groups, the rate of normal follicles was significantly decreased in both cultured groups (P < 0.05). Gene expression was no different in vitrified tissues compared to the control; however, the expression of growth differentiation factor 9 and follicle stimulating hormone receptor genes were increased and factor in germ line alpha and kit ligand genes were decreased during in vitro culture (P < 0.05). In the two cultured groups, the level of 17-ß estradiol was increased (P < 0.05), but the anti-Müllerian hormone concentration was not statistically altered. CONCLUSIONS: These results showed that the integrity of stimulated human ovarian tissue after vitrification/warming was well preserved; however, the in vitro culture condition needs improvement.

Short Term Culture of Vitrified Human Ovarian Cortical Tissue to Assess the Cryopreservation Outcome: Molecular and Morphological Analysis.

BACKGROUND: The aim of the present study was to evaluate the effectiveness of human ovarian vitrification protocol followed with in vitro culture at the morphological and molecular levels. METHODS: Ovarian tissues were obtained from 10 normal transsexual women and cut into small pieces and were divided into non-vitrified and vitrified groups and some of the tissues fragments in both groups were randomly cultured for two weeks. The morphological study using hematoxylin and eosin and Masson's trichrome staining was done. The analysis of mean follicular density, 17-ß estradiol (E2) and anti mullerian hormone (AMH), and real-time RT-PCR was down for the evaluation of expression of genes related to folliculogenesis. Data were compared by paired-samples and independent-samples T test. Values of p<0.05 were considered statistically significant. RESULTS: The proportion of normal follicles did not show significant difference between vitrified and non-vitrified groups before and after culture but these rates and the mean follicle density significantly decreased in both cultured tissues (p<0.05). The expression of genes was similar in vitrified and non-vitrified groups but in cultured tissues the expression of GDF9 and FSHR genes increased and the expression of FIGLA and KIT-L genes decreased (p<0.05). An increase in E2 and AMH concentration was observed after 14 days of culture in both groups. CONCLUSION: In conclusion, the present study indicated that the follicular development and gene expression in vitrified ovarian tissue was not altered before and after in vitro culture, thus this method could be useful for fertility preservation; however, additional studies are needed to improve the culture condition.