Intrauterine administration of autologous peripheral blood mononuclear cells regulates the endometrium estrogen and progesterone receptor expression: An RCT.

Background: Repeated implantation failure (RIF) affects 15% of women of reproductive age. There is a high endometrial expression of both estrogen receptors and progesterone receptors (PRs) during the window of implantation in women with RIF. Objective: To evaluate the effects of intrauterine administration of human peripheral blood mononuclear cells (PBMC) on estrogen receptor α (ERα) and PRs expression in the endometrium of women with RIF during the implantation window. Materials and Methods: This randomized clinical trial study was conducted on 22 women with RIF history from January 2018 to August 2019 in Erfan hospital, Tehran, Iran. Participantswere divided into 2 groups (PBMC-treated group [n = 11] and control group [n = 11]). Endometrial tissue samples were collected at the implantation window time, during the mid-secretory phase (luteinizing hormone surge +7 days) of each menstrual cycle. The quantitative real-time polymerase chain reaction technique was used to measure the mRNA levels of ERα and PRs isoforms (PR-A and PR-B) in endometrial tissues. Furthermore, the protein expression of ERα and PRs was investigated using immunohistochemical staining. Results: PBMC treatment significantly decreased the mRNA expression of endometrial ERα and PRs isoforms at the time of the implantation window (p < 0.001). Moreover, the endometrial ERα and PRs protein localization were significantly lower in PBMC-treated women compared with controls (p = 0.01, and p < 0.001 respectively). Conclusion: The intrauterine administration of PBMC decreased the endometrial ERα and PRs expression during the window of implantation in women with RIF. This local response to PBMC therapy could promote endometrial receptivity and embryo implantation.

Impact of sperm DNA fragmentation on ICSI outcome and incidence of apoptosis of human pre-implantation embryos obtained from in vitro matured MII oocytes.

BACKGROUND: Sperm DNA integrity and oocyte quality significantly affect embryo development and survival. The current study evaluated embryo development and quality, as well as the expression level of apoptosis-related genes and microRNAs in embryo derived from in vitro matured MII oocytes according to sperm DNA fragmentation (SDF) level. METHODS: The semen and immature oocytes were collected from 50 ICSI cycles with any recognizable female factor infertility. After ovarian stimulation, germinal vesicle stage (GV) oocytes were collected and incubated in in vitro maturation (IVM) medium for 24 h. Next, reactive oxygen species (ROS) level of media culture was determined. Using by sperm chromatin dispersion (SCD) test, the SDF levels of processed semen were assessed and categorized into SDF ≤ 30% and SDF>30%. Seventy two hours after intracytoplasmic injection, the embryo development and quality score were recorded in the groups I (GV-MII + SDF≤ 30%) and II (GV-MII + SDF> 30%). Also, the apoptosis incidence of embryos at morula stage was evaluated at molecular and cellular levels by quantitative real time PCR and TUNEL staining, respectively. RESULTS: Cleavage rate did not differ between two groups. The quality score of embryos obtained from IVM matured oocytes and high level of SDF was significantly lower than that of low level of SDF (P < 0.05). The embryos from group II had a significant reduction of the expression of BCL-2 compared to group I (P < 0.05). Also, they showed an increase in relative transcription of pro-apoptotic microRNAs; miR 15a and miR 16-1 versus group I (P < 0.05). A rise of TUNEL positive blastomers of embryo was observed at group II versus group I, but it did not reach to significantly level. CONCLUSION: The IVM oocytes, probably, did not suffice to recover the high level of paternal genomic damage and inhibition of apoptosis pathway beginning.

Granulocyte-colony stimulating factor may improve pregnancy outcome in patients with history of unexplained recurrent implantation failure: An RCT.

BACKGROUND: Family of colony-stimulating factors (CSF) have an essential role on early cross talk between embryo and uterine endometrium. OBJECTIVE: The aim of this study was to evaluate the effects of the single dose of Granulocyte-CSF (G-CSF) injection on clinical outcome of assisted reproductive technology cycle in patients with repeated implantation failures. MATERIALS AND METHODS: This randomized control trial study was performed on 52 infertile women who referred to the clinic with the history of more than three previous In vitro fertilization/Intracytoplasmic sperm injection-embryo transfer failures. All patients were stimulated with standard long protocol. All embryos were transferred on day five in blastocyst stage in both groups. The treated group received 300 µg (0.5 ml) recombinant human G-CSF subcutaneously which was injected 30 min before blastocyst embryo transfer. RESULTS: There was not statistically significant differences in abortion rate in G-CSF and control group (p=0.09). G-CSF treated group showed higher clinical pregnancy rate in comparison with control group (56.2% vs. 40.0%) but it was not statistically significant (p=0.09). Although live birth rate in G-CSF group was higher than control group (53.1% vs. 35.0%) but there wasn't statistically significant difference in the overall live birth rate between the two groups (p=0.10). G-CSF group had a twin pregnancies while in control group there was no twin pregnancy. CONCLUSION: Our result demonstrates the possibility that pregnancy outcome is better in women with repeated unexplained In vitro fertilization failure who are treated with G-CSF.

Is the ovarian reserve influenced by vitamin D deficiency and the dress code in an infertile Iranian population?

BACKGROUND: In the recent years, vitamin D has become a topical subject and a focus of research not only in reproductive medicine but across many medical disciplines. In reproductive medicine, studies have identified an association between vitamin D status in women and ovarian reserve. In humans, exposure of the skin to sunlight is the main important source of vitamin D. A dress code of wearing concealing clothing is a risk factor for vitamin D deficiency. The objective of this prospective observational study was to evaluate the correlation between vitamin D deficiency and ovarian reserve in a population of infertile women in Iran. As part of the basic fertility assessment of study participants, blood tests were taken to measure vitamin D concentration and transvaginal ultrasound scans were performed on day 2-5 of the cycle to determine antral follicle count (AFC). All study participants were assessed by a reproductive medicine specialist and consultant dermatologist to classify their skin types according to the Fitzpatrick classification. In addition, the dress code of each study participant was recorded noting the percentage of exposed skin not covered by concealing clothing. RESULTS: 189 infertility patients were included in this study. The mean concentration of vitamin D in this study population was 15.46 ng/ml, indicating severe vitamin D deficiency. A statistically significant negative correlation between age and vitamin D (p = 0.008) and age and AFC (p = 0.001) was identified. This study revealed a highly significant correlation between vitamin D concentrations and AFC (p < 0.001). CONCLUSIONS: A concealing dress code is an independent risk factor for vitamin D deficiency due to a lack of skin exposure to sunlight. Our study suggests that the so caused severe vitamin D deficiency may play a crucial role in reduced ovarian reserve in the herein described group of an infertile female Iranian population.

Modified natural cycle frozen-thawed embryo transfer in patients with repeated implantation failure: An observational study.

BACKGROUND: Natural endometrium in Frozen-thawed Embryo Transfer (FET) may have some benefits upon implantation in patients with Repeated Implantation Failure (RIF). It might be due to possible differences between natural and stimulated endometrial growth factors and cytokins secretions. OBJECTIVE: The objective of this study was to compare the pregnancy rate of FET on modified natural cycle versus hormone replacement therapy (HRT) cycle endometrium in patients with RIF. MATERIALS AND METHODS: In this observational study the pregnancy rate of patients with RIF undergoing day 3 FET in natural cycle endometrium (group 1, n=56), were compared with another group of patients with RIF in whom frozen-thawed day 3 embryos were transferred on HRT cycle (group 2, n=52). RESULTS: The pregnancy rate in group 1 was 41.07%, compared with the pregnancy rate of group 2; 36.5% (p=0.63). The abortion rate was not significantly different among the groups. CONCLUSION: It can be concluded that FET in a modified natural cycle is comparable with HRT cycle in patients with RIF.

Histone modification of embryonic stem cells produced by somatic cell nuclear transfer and fertilized blastocysts.

OBJECTIVE: Nuclear transfer-embryonic stem cells (NT-ESCs) are genetically identical to the donor's cells; provide a renewable source of tissue for replacement, and therefore, decrease the risk of immune rejection. Trichostatin A (TSA) as a histone deacetylase in- hibitor (HDACi) plays an important role in the reorganization of the genome and epigenetic changes. In this study, we examined whether TSA treatment after somatic cell nuclear transfer (SCNT) can improve the developmental rate of embryos and establishment rate of NT-ESCs line, as well as whether TSA treatment can improve histone modification in NT-ESCs lines. MATERIALS AND METHODS: In this experimental study, mature oocytes were recovered from BDF1 [C57BL/6×DBA/2) F 1 mice] mice and enucleated by micromanipulator. Cumulus cells were injected into enucleated oocytes as donor. Reconstructed embryos were ac- tivated in the presence or absence of TSA and cultured for 5 days. Blastocysts were transferred on inactive mouse embryonic fibroblasts (MEF), so ESCs lines were estab- lished. ESCs markers were evaluated by reverse transcription-polymerase chain reaction (RT-PCR). Histone modifications were analyzed by enzyme linked immunosorbent assay (ELISA). RESULTS: Result of this study showed that TSA treatment after SCNT can improve devel- opmental rate of embryos (21.12 ± 3.56 vs. 8.08 ± 7.92), as well as establishment rate of NT-ESCs line (25 vs. 12.5). We established 6 NT-ESCs in two experimental groups, and three embryonic stem cells (ESCs) lines as control group. TSA treatment has no effect in H3K4 acetylation and H3K9 tri-methylation in ESCs. CONCLUSION: TSA plays a key role in the developmental rate of embryos, establishment rate of ESC lines after SCNT, and regulation of histone modification in NT-ESCs, in a man- ner similar to that of ESCs established from normal blastocysts.

Melatonin modulates the expression of BCL-xl and improve the development of vitrified embryos obtained by IVF in mice.

PURPOSE: Antioxidant and anti-apoptotic effects of melatonin on development of in vitro fertilization (IVF)/vitrified two-cell mouse embryos were evaluated in this study. METHODS: The IVF two-cell embryos were vitrified by cryotop, and were cultured in KSOM medium in different concentrations of melatonin (10(-6), 10(-9), 10(-12) M) and without melatonin. The blastocyst cell number, apoptotic cells and glutathione (GSH) level were evaluated by differential, TUNEL and cell tracker blue staining, respectively. The expression of Bax and Bcl-xl genes was evaluated by qPCR. The expression of melatonin receptors (Mtnr1a and Mtnr1b) in mouse 2-cell embryos and blastocysts was evaluated by RT-PCR. RESULTS: Melatonin increased the rate of cleavage and blastulation at 10(-12) M concentration (p < 0.05). The number of trophectoderm and inner cell mass showed a significant increase (p < 0.05) in 10(-9) M melatonin. The 10(-9) M and 10(-12) M melatonin treatments significantly reduced (p < 0.05) the apoptotic index. The significant increase in the expression of Bcl-xl observed at 10(-9) M concentration however, reduced expression of Bax was not statistically significant. The levels of GSH in 10(-9) and 10(-12) M groups were significantly improved relative to the control group (p < 0.05). The Mtnr1a was expressed in 2-cell embryos and blastocysts in all groups, but the expression of Mntr1b was not detected. CONCLUSION: Melatonin may have a special role against oxidative stress in protection of IVF/vitrified embryos.