International Journal Of Fertility and Sterility
Volume:18 Issue: 2, Apr-Jun 2024

Some failures in ovary function, like folliculogenesis and oogenesis, can give rise to various infertility-associated problems, including polycystic ovary syndrome (PCOS) and premature ovarian insufficiency (POI). PCOS influences 8 to 20% of women; while POI occurs in at least 1% of all women. Regrettably, the current therapies for these diseaseshave not sufficiently been effective, and finding a suitable strategy is still a puzzle. One of the helpful strategies for managing and treating these disorders is understanding the contributing pathogenesis and mechanisms. Recently, it has been declared that abnormal expression of microRNAs (miRNAs), as a subset of non-coding RNAs, is involved in thepathogenesis of reproductive diseases. Among the miRNAs, the roles of miRNA-21 in the pathogenesis of PCOS and POI have been highlighted in some documents; hence, the purpose of this mini-review was to summarize the evidence in conjunction with the functions of this miRNA and other effective microRNAs in the normal or abnormal functions of the ovary (i.e., PCOS and POI) with a mechanistic insight.

Premature menopause (PM) is the cessation of ovarian function before age 40. PM women are more likelyto have cardiovascular diseases (CVDs), diabetes, and mental disorders. This is the first study that assessed the associationof single nucleotide polymorphisms (SNPs) with anti-heat shock protein 27 (Hsp27), High-sensitivity C-reactive protein(hs-CRP), and PM and serum pro-oxidant-antioxidant balance (PAB), as putative risk factors for CVDs. We aimed toexplore the association of oxidative stress markers with eight different SNPs shown to be related to premature menopause. In this cross-sectional research, we included 183 healthy women and 117 premature menopausalwomen. We determined baseline characteristics for all participants and measured serum hs-CRP, anti-HSP-27 antibody titer, and PAB levels using the established methods. Genotyping for eight SNPs was done usingthe tetra amplification refractory mutation system polymerase chain reaction (Tetra-ARMS PCR) and allele-specificoligonucleotide PCR (ASO-PCR) methods. We found a significant difference between mean serum PAB levels and the genetic variant of rs16991615(P=0.03). ANCOVA showed a significant effect of the genotypes rs4806660 and rs10183486 on hs-CRP serum levelsin the case and control groups, respectively (P=0.04 and P=0.007). ANCOVA also showed an association betweenrs244715 genotypes and anti-hsp27 serum levels in the case group (P=0.02). There was a significant effect of thegenotypes of rs451417 on the serum hs-CRP level in the control group (P=0.03). There was a significant association of the genetic variants related to PM with oxidative stress and inflammatorymarkers (serum PAB, anti-hsp27 antibody, and hs-CRP). Accordingly, this seems to be an effective approach topredicting susceptible subjects for cardiovascular and mental disorders as well as various cancers.

Oxidative/nitrosative stress in the oocyte microenvironment could have an impact on intracytoplasmicsperm injection (ICSI) outcomes. The presence of reactive oxygen species (ROS) and reactive nitrogen species (RNS) canstimulate the secretion of pro-inflammatory cytokines, leading to chronic inflammation and potentially affecting embryo aswell as oocyte quality. This study aimed to examine the relationship of lipid peroxidation [measured by the malondialdehyde(MDA) assay] with protein carbonyl [measured by the 2,4 dinitrophenylhydrazine (DNPH) assay] levels in cumulus cells(CCs), as well as nitric oxide (NO), peroxynitrite (ONOO-), and C-reactive protein (CRP) levels in follicular fluid (FF).The potential relationship of these levels with ICSI outcomes was also evaluated. In this prospective study, 63 FF samples and their corresponding CCs were collected forICSI procedures. Spectrophotometry was used to assess levels of DNPH, MDA, NO, and ONOO-. CRP levels wereevaluated using an immunoturbidimetric assay. The patients under 37 years with normal ovarian reserve had significantly lower levels of MDA, DNPH,NO, ONOO-, and CRP compared to those over 37 years. Additionally, we observed higher levels of MDA, DNPH,NO, ONOO-, and CRP in the group with an oocyte maturity rate of less than 60%. No significant difference was observedbetween the DNPH levels and factors such as infertility duration, embryo quality, pregnancy, or the number ofretrieved oocytes. A higher level of MDA, NO, ONOO-, and CRP was found to be significantly related to the lowernumber of retrieved oocytes, longer periods of infertility, poor embryo quality, and negative pregnancy outcomes. Oxidative/nitrosative stress, linking to inflammation in the oocyte microenvironment, can be consideredas a potentially useful biomarker for assessing the development and competence of oocytes and embryos and predictingICSI outcomes.

This research was aimed at assessing the relationship between the follicular fluid (FF) antioxidantsactivity, aging and in vitro fertilization (IVF) outcome. The present cross-sectional study was carried out on 65 women undergoing IVF/intracytoplasmicsperm injection (IVF/ICSI) cycles due to unexplained infertility. Ovarian stimulation was performed using thelong gonadotropin-releasing hormone (GnRH) agonist protocol. After ovum pickup, FF was collected and processedto measure the level of superoxide dismutase (SOD), catalase (CAT) activity, total antioxidant capacity (TAC) andglutathione (GSH). Day 3 after ICSI, fresh embryos were transferred and later, possible pregnancy was assessed.Patients participating in this study were divided into four groups on the basis of age and pregnancy outcome. SOD activity was not significantly different between the groups (P=0.218). GSH in the group whose participantswere aged ≤35 years and were pregnant was higher than that in other groups. CAT activity in groups withyounger participants was higher compared to the other groups. The mean TAC was higher in groups with pregnantparticipants compared to the non-pregnant women. Correlation analysis showed that: GSH level had a significantnegative correlation with age (P<0.001, R -0.55) and a significant positive correlation with pregnancy (P=0.015,R=0.30). CAT level also had a significant negative correlation with age (P<0.001, R=-0.42) and the level of TAC hada significant positive correlation with pregnancy (P<0.001, R=0.59). According to our results, the levels of TAC, GSH and CAT in younger and pregnant women were highercompared with those undergoing ICSI cycles. Given the correlation of FF antioxidant activity with age and pregnancy,it is necessary to carry out more research on these compounds and the maintenance of pregnancy.

Myometrial thickness has been expected to be a prognosticator for lower uterine segment function. Anabnormal function of the uterine muscle layer can cause common and important reproductive problems. This studyaimed to evaluate the relationship between the baseline myometrial thickness and assisted reproductive technologies(ART) outcomes. In this prospective cohort study, 453 infertile women undergoing ART cycles without anyobvious uterine pathology, participated in this prospective cohort study from February 2013 to May 2015. In order tomeasure the myometrial thickness in the anterior and posterior of the uterine, trans-vaginal ultrasounds were conducted ondays 2-4 of the cycle (menstrual phase) preceding ovarian stimulation and the day of human chorionic gonadotropin(hCG) injection. We defined three groups based on the baseline myometrial thickness in the anterior and posterior, including(A) <25 mm, (B) 25-29.9 mm and (C) ≥30 mm. Ovarian stimulation, oocyte retrieval and luteal phase support wereperformed in accordance with the standard long protocol. Two weeks after embryo transfer, the patients underwent apregnancy test by checking their serum β-hCG levels. The primary outcome measure was the clinical pregnancy rate. Secondaryoutcome measures were implantation rate, abortion rate and live birth rate. The clinical pregnancy (P=0.013) and implantation (P=0.003) rates were significantly lower in group A thanin two other groups. Although the live birth rate was lower in group A than two other groups, this decrease was not statisticallysignificant (P=0.058). The findings may be a way for clinicians to draw focus on providing therapeutic strategies and a specificsupportive care for women with a baseline myometrial thickness <25 mm in order to improve the reproductive outcomeof in vitro fertilization/intracytoplasmic sperm injection (IVF-ICSI).

The aim of this study was to evaluate the effects of thiamine (vitamin B1) on general health and infertilitytreatment outcomes in women with polycystic ovary syndrome (PCOS). The study is a triple-blinded, randomized, placebo-controlled clinical trial performed on 64 infertilewomen with PCOS referred to Sarem Hospital in Tehran, Iran. The primary outcomes of the study were general healthand infertility treatment outcomes. Eligible women were randomly assigned to the vitamin B1 group (n=32, vitamin B1tablet at a dose of 300 mg/day for 4 weeks) or the placebo group (n=32, placebo tablet daily for 4 weeks). A general healthquestionnaire was completed before and after the intervention by both groups and treatment success was evaluated at theend of the study. Data were analyzed using SPSS software ver.16 P<0.05 was considered statistically significant. The mean age of participants in the vitamin B1 (VB1) group was 30.4 ± 3.27 years and in the placebo (Pl)group was 29.1 ± 2.66 years with the mean duration of marriage 12.7 ± 3.01 and 13.2 ± 2.97 years respectively. Ourresults showed that there were significant differences between the two groups in overall score (P<0.001) and scoresfor all domains of the general health questionnaire including somatic symptoms (P<0.001), anxiety and insomnia(P<0.001), social dysfunction (P=0.028), and severe depression (P<0.001) after the intervention. Four weeks of consumptionof vitamin B1 also resulted in higher numbers of positive pregnancy tests (P=0.006), although the numberof fetuses was not significantly different between the two groups after the intervention. The results of the current study support a possible favourable effect of vitamin B1 on improving generalhealth, infertility treatment outcome, and retrieved follicle count without changing the number of fetuses in womenwith polycystic ovary syndrome (registration number: IRCT201510266917N3).

Various protocols have been approved to improve the response rate leading to successful fertilizationin poor ovarian responders (PORs). The application of double ovarian stimulation (DuoStim) in the follicular andluteal phases of the same ovarian cycle has been shown as an intriguing option to achieve more oocyte retrievals inthe shortest time. The aim of the current study, is to compare the outcomes of different protocols, minimal stimulation(MS) and Duostim. This randomized clinical trial was performed on 42 in vitro fertilization (IVF) candidateswith POR diagnosis. Patients were classified into two equal groups and treated with the DuoStim protocol and MSprotocol. The IVF outcomes, including retrieved follicles, oocytes, metaphase II (MII) oocytes and embryos, werecompared between these groups. The patients’ characteristics including age, anti-mullerian hormone (AMH), follicle-stimulating hormone(FSH), luteinizing hormone (LH), and antral follicle count (AFC) were collected and compared. It showed there wasno significant difference between the two groups' baseline characteristics (P>0.05). We observed that the DuoStimprotocol resulted in a significantly higher score in comparison with the MS protocols, including the number of follicles(6.23 ± 2.93 vs. 1.77 ± 1.66, P<0.001), retrieved oocytes (3.86 ± 2.57 vs. 1.68 ± 1.58, P=0.002), MII oocytes (3.36 ±2.42 vs. 1.27 ± 1.27, P=0.001) and obtained embryos (2.04 ± 1.64 vs. 0.77 ± 0.86, P=0.003). The DuoStim protocol is a favourable and time saving plan that is associated with more oocytes in a singlestimulation cycle. The DuoStim protocol significantly can result in more frequent MII oocytes and embryos. We figuredthat the higher number of oocytes and embryos might have led to a higher rate of pregnancy (registration number:IRCT20200804048303N1).

It is difficult to obtain healthy oocytes in poor ovarian responders with conventional treatment methods.Thus, the need to investigate new methods is essential. This study aims to investigate ovulation induction outcomesin patients with decreased ovarian reserve (DOR) in two groups treated with double stimulation (DuoStim) during thefollicular and luteal phases in comparison with the antagonist cycle. This was a randomised clinical trial that enrolled the patients with reduced ovarian reserve. Thepatients referred for in vitro fertilisation (IVF) at Molud Infertility Clinic, Ali Ebn Abitalib (AS) Hospital, Zahedan, Iranfrom 2020 to 2021. Participants were randomly divided into two groups, those who underwent treatment with DuoStimduring the follicular and luteal phase (case group) and those who received the conventional antagonist cycle (control group). The mean number of metaphase II (MII) eggs was 7.7 ± 3.1 in the case group and 6.1 ± 3.9 in the controlgroup (P=0.063). The mean total number of retrieved eggs in the case group was 9.2 ± 3.7 and in the control group, itwas 6.9 ± 4.4 (P=0.023). The mean number of embryos obtained in the case group was 6.5 ± 3.9; in the control group,it was 4.7 ± 2.8 (P=0.016). The DuoStim method can effectively play a role in increasing the total number of retrieved eggs and embryos(registration number: IRCT20120817010617N8).

There is a definite shift in assisted reproductive centres from cleavage-stage embryo transfer (ET) toblastocyst transfer that is attributed to improvements in laboratory environments and advances in the development ofembryo culture media. The aim of the study was to investigate the reproductive outcomes of thawed cleavage-stage ET versusblastocysts derived from an extended culture of these embryos. This open-label, randomised, parallel group clinical trial study enrolled 182 women aged ≤37years who underwent frozen-thawed ET from November 2015 to June 2020 at Royan Institute Research Centre, Tehran,Iran. The women were randomly assigned to either the thawed cleavage ET group (n=110) or the post-thaw extended cultureblastocysts group (n=72). The primary outcome measure was the clinical pregnancy rate. Secondary outcome measureswere implantation rate, live birth rate (LBR), and miscarriage rate. A P<0.05 indicated statistical significance. There were no significant differences between the two groups in terms of demographic characteristics. Boththe mean numbers of embryos transferred and good quality embryos transferred were significantly lower in the post-thawextended culture blastocysts group compared to thawed cleavage-stage ET cycles. However, the post-thaw extendedculture blastocysts group had higher clinical pregnancy (56.94 vs. 40.91%, P=0.034), implantation (34.43 vs.19.84%, P=0.001) and live birth (49.3 vs. 33.63%, P=0.036) rates compared to the thawed cleavage-stage ET group.Miscarriage and multiple gestations rates were comparable between the groups. These results allow us to take a position in favour of post-thaw extended culture blastocysts; thus, it isimportant to improve the post-thawing extended culture technique (registration number: NCT02681029).

The parallel and continued improvements in both infertility treatment and the management of malignancycases have brought to the forefront the potential for fertility preservation. Using ovarian follicular resourcescan effectively improve reproductive capacity and prevent infertility. The primary aim of this research was to try togenerate an appropriate in vivo environment for the growth of the mouse follicles. Hence, the possible effects of theovarian parenchyma cell suspension were explored on the growth and maturation of preantral follicles in vitro. In this experimental study, ovarian parenchymal cells were mechanically dissociated frompreantral follicles of 12-14 days-old NMRI mice and then divided into 5 experimental groups (G1: Control, G2: Freshfollicle with fresh parenchyma cell suspension, G3: Vitrified-warmed follicle with fresh parenchyma cell suspension,G4: Fresh follicle with frozen-thawed parenchyma cell suspension, and G5: Vitrified-warmed follicle with frozen-thawedparenchyma cell suspension). The diameter of the follicles and immature oocytes, viability, antrum formation,resumption of meiosis, in vitro fertilization (IVF), and Gdf9, Bmp6, and Bmp15 gene expression were examined ondifferent periods. The diameter of the follicles and the oocytes on days 4 and 8, as well as the survival rate of the follicles upto day 12, were significantly higher in G2 and G4 compared to the Ctrl group (G1: 73.66%, G2:87.99%, G3: 82.70%,G4: 94.37%, and G5: 78.59%). Expression of growth marker genes for G3, and G5 groups was significantly higherthan other groups, which indicated the protective effects of parenchyma cell suspension on follicles damaged by vitrificationsolutions. The growth, survival, and maturation of preantral follicles could be enhanced by co-culturing them withovarian parenchyma cells. Further studies are needed to optimize the conditions for a successful parenchyma cellsuspension-induced in vitro maturation (IVM) to occur in infertility clinics.

Platelet-rich plasma (PRP) therapy has been shown to enhance tissue regeneration by expressing severalcytokines and growth factors (GFs). This study investigated the effect of intrauterine infusion of PRP as a noninvasiveautologous GF on pregnancy outcomes in women with repeated implantation failure. This randomized clinical trial was conducted to compare the pregnancy rates between twogroups of women who were candidates for the frozen-thawed embryo transfer with a history of two or more implantationfailures. The PRP group (n=33) was treated with hormone replacement therapy+0.5 cc to 1 cc PRP infusedinto the uterine cavity two days before the embryo transfer. The control group (n=33) was only treated with hormonereplacement therapy. The endometrial preparation process was done similarly in both groups. The chemical, clinical,and ongoing pregnancy, and implantation rates were compared between the two groups. Our results showed that the chemical pregnancy rate was not statistically higher in the PRP group in comparisonwith the control group (36.4 vs. 24.2%). In addition, the clinical pregnancy, ongoing pregnancy, and implantationrates were higher in the PRP group than the control group; however, the difference between the two groups was notstatistically significant. Administration of intrauterine PRP before embryo transfer in women with repeated implantation failure(RIF) does not affect assisted reproductive technology (ART) outcomes (registration number: IRCT2016090728950N3).

Varicocele is one of the most common treatable causes of male infertility, and its treatment may bebeneficial for fertility. This study aimed to evaluate fertility rate and DNA fragmentation index (DFI) following varicocelectomyin primary infertile men with clinical varicocele. This prospective longitudinal study was conducted on primary infertility men, in a tertiarycenter from December 2018 to December 2019 with one-year follow-up. Data of the semen parameters, DFI (%), andfertility rate were gathered before, as well as 4 and 12 months after undergoing varicocelectomy. For data analysis,SPSS software and analytical test were used. Out of 76 patients who were analyzed, 22 (29%) became fertile and 54 (71%) remained infertile. Semenparameters and DFI (%) were improved significantly following varicocelectomy (P<0.001). Smoking history, occupationalheated exposure, body mass index (BMI), and infertility duration were determined as predictors associatedwith fertility status (P<0.05). Although varicocele repair improved the DFI, the fertility rate was achieved in less than one-third ofpatients; it seems that the other parameters, such as the history of smoking, occupational heated exposure, overweight,and duration of infertility should be considered as predictors of fertility status, in primary infertile men who are a candidatefor varicocelectomy.

Cryopreservation of sperm is essential for patients with low sperm counts and couples undergoing infertilitytreatment. The aim of this study was to compare the effects of Taurine (T) and Sucrose (S) in individual spermcryopreservation utilizing cryotop and petri dish and thawing at 37 and 42°C. In this experimental study, 17 normospermic semen samples were processed using the"Swim-up" procedure and progressively motile sperm were then isolated from these samples using an inverted microscope.Sperm were added to droplets of "sucrose medium" with 25 mM Taurine antioxidant (S+T) and the commercialcryoprotectant "Sperm Freeze" (CPA), loaded on a petri dish and cryotop. After rapid freezing of the samples, theywere thawed at two different temperatures (37°C and 42°C), and the sperm classical parameters, viability, and DNAfragmentation were assessed. Statistical analysis displayed a significant increase in total and progressive motility in individual spermfreezing on cryotop with CPA and thawing at 42°C (P<0.05). Other parameters did not show any differences betweenthe CPA and S+T groups and two thawing temperatures in either of the cryopreservation methods. Although, both cryoprotectants (CPA and S+T) may preserve individual sperm effectively using cryotop,the CPA and thawing at 42°C showed a better effect on the motility percentage of the small number of sperm.

Infertile men with multiple morphological abnormalities of the sperm flagella (MMAF) phenotypeexhibit mosaic sperm flagella abnormalities such as short, bent, coiled, and irregular flagella or absent flagella. Spermflagellum has an ultrastructurally axonemal structure that contains a large number of proteins. A-Kinase AnchoringProtein 3 (AKAP3) is expressed in spermatozoa. It may function as a regulator of motility and the acrosome reaction.This study aimed to compare genetic changes in infertile men suffering MMAF phenotype with the control group.

In this case-control study, genetic variants of the AKAP3 gene were evaluated in 60 infertilemen with MMAF phenotype and 40 fertile men, as control. As exon five of the AKAP3 gene encodes the functionaldomain of this protein, its genetic variants were studied. Therefore, polymerase chain reaction (PCR)-sequencing wasundertaken on the DNA extracted from control and patients’ blood samples.

Sixty infertile men with MMAF phenotype and 40 normozoospermic men, as control, were enrolled inthis study. Four haplotype variants 1378T>C (rs10774251), 1391C>G (rs11063266), 1437T>C (rs11063265), and1573G>A (rs1990312) were detected in all patients and controls. On the other hand, a missense mutation 1499T>C(rs12366671) was observed in four patients with the homozygous form while seven patients carried the heterozygousform. No mutation was identified in the controls (P=0.04). The difference between the variation allele frequencies wasassessed in the patient and control groups by the Fisher Exact Test.

In the homozygous form, this mutation changed Isoleucine to Threonine. This alternation occurred insidethe AKAP4 binding domain of the AKAP3 protein. The observed variants caused no significant deviation in thesecondary structure of AKAP3 protein and probably its function in spermatozoa flagella. So, these variants cannot beconsidered as the causes of MMAF phenotype in the studied patients.