Tehran University Medical Journal

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Background: Hyaluronan has an important role on the permeability and motility of sperm and the interaction of gametes and these can play a considerable role on the fertility rate. Therefore, in this study, we assessed the effect of different doses of hyaluronan on the morphology, motility, vitality and fertility rate of mice.

Methods: We used 40 mice (6-8 week) in this study which twenty of them were male and the rest were female. The sperm of each male mouse were divided into four groups. The group 1 (control): They were maintained in RPMI media without any hyaluronan supplementation for 2 hour. Hyaluronan with the doses of 750, 1000 and 1250 µg/ml were added into RPMI media in groups 2, 3 and 4, respectively. After 2 hour. incubation, the numbers of sperms were assessed, using haemocytometer. Also, their morphology with papanicolaeu staining and their vitality with Eosin B dye were assessed. As well as sperms motility measured under inverted microscope by observation and fertility rate evaluated after routine IVF by counting two-cell stage embryos.

Results: Our results demonstrated that, the dose of 750 µ g/ml has the greatest effect on the motility, vitality and fertility rate of sperms. The effect of dose of 1000 µ g/ml also was positive on them. On the other hand, none of these doses had any effect on sperm morphology.

Conclusion: Hyaluronan may have an influence on motility, vitality and fertility rate of sperms and the dose of 750µ g/ml had a significant effect on these factors.

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Background: Leukemia inhibitory factor (LIF) is a group of secreted glycoproteins with molecular weights ranging from 38-67 kD, resulting from differential protein glycosylation. LIF is constitutively expressed at high levels in the human fallopian tube epithelium and has an important role in the motility and vitality of sperm. In the present study, the effect of human recombinant LIF on human sperm motility and survival in vitro was investigated.

Methods: Normal spermatozoa of 30 fertile men were collected and after preparation were incubated in Ham's F10+FCS 10% medium, containing various concentrations (0, 3, 5, 10, and 50 ng/ml) of LIF at 37 ºC under 5% CO2 for 6, 24 and 48 hours. Sperm motion characteristics were measured using a Makler chamber. Sperm survival was determined using the hypoosmotic swelling test. Collected data were analyzed by one-way ANOVA and LSD test using SPSS version 11. The difference in values were considered significant when p<0.05.

Results: Sperm motility was significantly higher after 24 h exposure to 5-10 ng/ml LIF (p<0.05). The survival rate of sperm was significantly prolonged when exposed to 50 ng/ml LIF (p<0.05). Nonprogressive motility and survival rate of sperm were significantly higher after 48 h exposure to 50 ng/ml and 10-50 ng/ml LIF, respectively. (p<0.05). There was no significant difference in progressive sperm motility during the 48 h exposure of sperm to the various concentrations of LIF.

Conclusion: According to our results, the effect of LIF on sperm motility and survival were dependent on the dose of LIF supplementation and the length of incubation.

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Background: Pi-GST and Mu-GST are subclasses of glutathione S-transferase that present on human sperm surface and play an important role against oxidative stress. Therefore, any defects in the enzyme activity may be associated with male infertility.In this study the polymorphisms of GSTM1 and GSTP1 in association with enzyme activity and sperm parameters were studied.

Methods: This case-control study involved 95 men with oligoastenoteratozoospermia and 26 controls with normozoospermia. Semen analyses were carried out according to WHO guidelines. Blood DNA was extracted using salting out procedures. GSTM1 and GSTP1 polymorphisms gene were determined through PCR-RFLP and multiplex PCR, respectively. Finally, Glutathione S-transferase activity was measured.

Results: Frequencies of GSTM1 null genotype in oligoastenoteratospermic and normospermic groups were 52.1% and 53.8% respectively. There were no statistically significant differences in sperm parameters and enzyme activity between GSTM1 null and positive genotypes in two groups. There were no statistically significant differences in glutathione S-transferase activity between oligoastenoteratospermia and normospermic groups (p>0.05). All the 121 men in this study had Ile/Ile genotypes at 105 codon of GSTP1. Frequency of normal homozygote (114Ala/Ala), heterozygote (114Ala/Val) and mutant homozygote (114Val/Val) genotypes in oligoastenoteratospermic group were 81.1%, 17.9% and 1.1% respectively but in the control group they were 88.5%, 11.5% and null.

Conclusions: Total glutathione S-transferase activity and sperm parameters were not affected by deficient Glutathione S-transferase activity in GSTM1 null genotype. Compensate activity of other sperm surface glutathione S-transferase isozymes, like GSTP1, may justify the cause.

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Normal 0 false false false EN-US X-NONE AR-SA MicrosoftInternetExplorer4 Background: Defective sperm function is now recognized as one of the most important causes of male infertility. Seminal plasma possesses a rich source of different enzymatic and non-enzymatic antioxidants such as vitamin C (ascorbic acid) that protect spermatozoa against oxidative stress as one of the mediators of infertility causing sperm dysfunction and low sperm quality. The aim of this study was investigation of seminal total antioxidant capacity and determination of vitamin C effects on sperm motility.

Methods: We designed a case-control study with a total subject of 62 males. Sperm parameters were analyzed according to World Health Organization guidelines (WHO, 1999). Total antioxidant capacity and vitamin C level of seminal plasma were measured in the 32 normozoospermic as the control group and 32 asthenospermic men as the case group using FRAP (Ferric Reducing of Antioxidants Powers) and RP-HPLC (Reverse Phase High Performance Liquid Chromatography) methods, respectively.

Results: Our results indicated that total antioxidant capacity levels in the seminal plasma of asthenospermic men were significantly lower than healthy men (p=0.002). In addition, we found a positive correlation between reduced total antioxidant capacity levels and low sperm motility. Vitamin C levels of seminal plasma in asthenospermic men were statistically lower than control men (p=0.01).

Conclusions: It is suggested that asthenospermia could be related to an antioxidant deficiency or it&aposs reduction.

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Background: Use of certain antipsychotic drugs has severe effects on fertility in males. Hypothalamus and hypophysial impressions and changes in plasma hormones concentration like prolactin, LH and FSH can affect sperm production. In this study, we investigated the effects of sulpiride on sperm quality, maturation and DNA damage.

Methods: Twenty for adult male mice (age: 6-8 weeks) were divided into three groups. The treatment group received 40 mg/kg sulpiride solution and the control sham group was given carrier of the drug intraperitoneally (IP) daily for 45 days but the control group received nothing. Finally, all the mice were sacrificed by cervical dislocation and their cauda epididymis were removed surgically. The excised specimens were placed in 1 ml HTF medium and incubated for 30 min in CO2 incubator to allow the spermatozoa to swim out. Later, sperm count, motility and viability were analyzed. Additionally, sperm chromatin quality and DNA integrity were assessed by aniline blue and acridine orange staining.

Results: Significant decrease in sperm motility and count were observed in the treatment group while the number of abnormal sperm increased as compared with the other two groups. Sperm viability and DNA maturation showed significant reduction and the rate of DNA damage increased in comparison with the control sham and the control groups (P<0.05).

Conclusion: The study showed that sulpiride has negative effects on sperm parameters in treated animals and in some cases it could cause secondary infertility.

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Background: Achillea millefolium or yarrow is a native plant in Europe and Iran. Yarrow has been used as a medicine historically, mainly because of its astringent effects. It is reported to be associated with the treatment of several ailments. Nowadays use of plants for medical purpose has become very common. Achillea millefolium L, Yarrow, is being used in traditional and modern medicine due to various chemical compounds. Considering the importance of birth control, finding a drug with less side effects inhibiting spermatogenesis seems to be necessary. The aim of the present study was to investigate the effect of ethanol extract of Achillea millefolium L. on spermato-genesis of male wistar rats.
Methods: In this study, 32 adult male wistar rats were used. The animals were divided to four groups of eight rats. The first group, received 200 mg/kg Achillea millefolium L. interaperitoneally, the second and third groups received 400, 800 mg/kg Achillea millefolium L. interaperitoneally, respectively. In the fourth group (control) distilled water was administered. After 20 days, the rats were sacrificed and testis tissues were histologically evaluated.
Results: Comparing to control group, in the experimental groups received the high doses of the extract, thickening of seminiferous tubules basement membrane, loss of germinal epithelium and testicular hyperemia were demonstrated (P<0.001).
Conclusion: Based on the results, high concentrations of Achillea millefolium L. leaded to structural and spermatogenesis changes in testis tissue.

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Background : One of the important problems in fertilization in vitro (IVF) is failure of implantation. This could be the result of estrogen and progesterone effects in endometrial acceptance during ovulation stimulation. Although progesterone has a vital role in primary phase of pregnancy, but the estradiol role in luteal phase is unknown. The aim of this study is assessment of the ratio of estradiol to progesterone in embryo transfer day on Intracytoplasmic sperm injection (ICSI) outcomes.

Methods : This is a cohort study. The subjects were 311 infertile women referred to Urmia Kosar infertility clinic & Urmia reproductive health research center who treated with ICSI method between August-Jan 2011 . Five cc blood was drawn for determine of estradiol and progesterone in transfer day. Transfer occurred after a variable in vitro culture period ranging from 48 to 72 hours after ovulation induction. Chemiluminescent ELICA the level of mention hormones was used to determine the ratio of the progesterone level to the estradiol serum level and was compared based on treatment outcomes.

Results : A total of 311 patients, 115 (37%) were pregnant and happened abortion were 18 (5.8%) . The mean ratio of estradiol to progesterone in transfer day in two groups of miscarriage/ non miscarriage and pregnant/ non pregnant was 32.26±23.86 , 28.17±26.5 and 28.58±2.4 , 36.09±4.39 respectively. There is no significant difference between two groups in regard of estradiol on progesterone ratio (P=0.5, P= 0.2) .

Conclusion: The results of this prospective cohort study show that there is no effect of estradiol to progesterone ratio on day of embryo transfer, successful pregnancy in ICSI cycles and abortion rate following of ICSI .

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Background: Infertility is one of the most serious social problems. Illicit drug use can be an important cause of male factor infertility. Kerack which its use is rising up in Iran refers to a high purity street-level heroin (heroin Kerack). Heroin Kerack used in Iran is an opioid and has harmful effects on body organs. The aim of this study is to investigate the effects of Kerack used in Iran on fertility adult mice.
Methods: In this study, 25 male mice were divided into five groups (control, sham and three experimental). Experimental groups of Kerack-dependent mice (received ascend-ing dose of Kerack for seven days) were divided into three categories, experimental I, II and III. Experimental I was given Kerack at a dose of 5 mg/kg, experimental II 35 mg/kg and experimental III 70 mg/kg, intraperitoneally twice a day for a period of 35 days. The sham group received normal saline and lemon juice (2.6 µl/ml) whilst the control group just received water and food. Mice were then scarified and sperm removed from cauda epididymis were analyzed for sperm count, motility, morphology (normal/abnormal) and viability. Testes were also removed, weighed and processed for light microscopic studies.
Results: The results showed that fertility were significantly decreased in addicted mice compared with control groups (P≤0.05). Epididymal sperm parameters and thickness of seminiferous epithelium were significantly decreased in experimental groups (dose-dependent) compared with sham and control groups (P≤0.05). Gonadosomatic index was significantly reduced with high dose Kerack injected (70 mg/kg) in comparison with control testes (P≤0.05).
Conclusion: This study has shown the deleterious effects of Kerack used in addicted Iranian people on fertility for the first time. This effect is especially on epididymal sperm parameters in adult mice.

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Background: Colorectal carcinoma is a common malignancy, in treatment of which pelvic radiotherapy plays an important role. But this may lead to azospermia. We designed a study to determine the delivered dose to the testis with thermoluminescence dosimetry (TLD) and compare it to the dose calculated by the Three-dimensional planning software. Methods: We measured the testicular doses by TLD the TLDs were fixed to the scrotum in six points anteriorly and posteriorly in two random fractions of the radiation course. All patients received a 50-50.4 Gy radiation dose to the pelvis in a prone position with standard fractionation and 3-dimensional planning, through three or four fields. The average dose of the TLD measurements was compared to the average of 6 relevant point doses calculated by the planning software. Results: In 33 patients with a mean age of 56 years, the mean testis dose of radiation measured by TLD was 3.77 Gy, equal to 7.5% of the total prescribed dose. The mean of point doses calculated by the 3-dimensional planning software was 4.11 Gy, equal to 8.1% of the total prescribed dose. A significant relationship was seen between the position of the inferior edge of the fields and the mean testis dose (P= 0.04). Also body mass index (BMI) was inversely related with the testicular dose (P= 0.049). Conclusion: In this study, the mean testis dose of radiation was 3.77 Gy, similar to the dose calculated by the planning software (4.11 Gy). This dose could be significantly harmful for spermatogenesis, though low doses of scattered radiation to the testis in fractionated radiotherapy might be followed with better recovery. Based on above findings, careful attention to testicular dose in radiotherapy of rectal cancer for the males desiring continued fertility seems to be required.

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Background: With introduction of intracytoplasmic sperm injection with testicular sperm extraction or precutaneouse epididymal sperm aspiration, effective treatment was provided for azoospermic men. The aim of present study was to compare clinical outcome following intracytoplasmic sperm injection using extracted testicular/epididymal sperm or ejaculated severe oligoasthenoteratozoospermic sperm. Methods: After retrospective evaluation of more than four hundred medical records of patients undergoing intracytoplasmic sperm injection Mehr medical institute (between 2011-2012), 45 cycles with severe eligoasthenoteratozoospermia and 34 cycles with azoospermia were included. Patients were treated with gonadotropin releasing hormone agonist. The clinical characteristics and intracytoplasmic sperm injection outcome such as the rate of fertilization, implantation and clinical pregnancy were compared between the two groups. Results were presented as mean±standard deviation and number (percent). Differences between variables were analyzed using student's t test and the chi-square test was used to examine differences between categorical variables. P value less than 0.05 were considered as statistically significant. Results: Mean of female age (29±4.9 vs. 30.2±5.8), body mass index (26.9±5.3 vs. 26.9±3.8), estradiol level on human chorionic gonadotropin administration day (1375.6±843.9 vs. 1181.8±673.1), total number of retrieved oocytes (9.7±5.3 vs. 9.2±5.9) and metaphase II oocytes (7.7±5.1 vs. 7.5±5.4) were similar between the two groups. Of 436 and 313 retrieved oocytes, respectively 232 and 163 oocytes were ferti-lized in oligoasthenoteratozoospermic and azoospermic groups (53.2% vs. 52.1%, P=0.214). There were not statistical differences between groups in number of trans-ferred top quality embryos (1.5±1.2 vs. 1±1.2, P=0.09), implantation rate (22.7% vs. 16.9%, P=0.238) and clinical pregnancy rate (21 (47.7%) vs. 11 (35.4%), P=0.199). Conclusion: Intracytoplasmic sperm injection with precutaneouse epididymal sperm aspiration and testicular sperm extraction are effective methods to treat azoospermic men and its clinical outcome were comparable to ejaculated sever oligoasthenoterato-zoospermic cycles. It can be concluded that the influence of sperm quality and origin on intracytoplasmic sperm injection outcome are the same.

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Background: Last decades, we have observed major improvements in treating infertility by using microinjection. However, reduction in abortion or increase in fertility has not been significant. It seems use of corticosteroids improves clinical outcomes during assisted reproductive technology (ART) techniques. Therefore, this study tried to show how corticosteroids therapy improves the results of intracytoplasmic sperm injection (ICSI). Methods: This semi clinical trial study without control group was included infertile women with more than one year’s infertility and were candidate to ICSI which were referred to Kosar Infertility Center, Urmia University of Medical Sciences from April 2011 to September 2013. Patients received prednisolone which was started 20 mg/day from one day before embryo transfer to 7 days, then for 2 days more 1 tablet and stopped. Pregnancy outcomes were chemical and clinical pregnancy, ectopic pregnancy, multiple pregnancy and rate of abortion before 20 weeks. Beta human chorionic gonadotropin (HCG) test was performed. Patients with positive pregnancy test were followed by sonography in 6, 12 weeks and 20 weeks of pregnancy. Results: One hundred and eighty one patients entered to the study. The mean± SD of age and fertility duration were 30.42± 6.07 and 7.69± 5.54 years. The mean± SD of transferred embryo was 4.60± 1.10 and embryo grading was 138 (44.7%) grade A, 124 (40.1%) grade B, 47 (15.2%) grade C, respectively. There were no significant difference between fresh/frozen embryo transfer in pregnancy outcomes (P> 0.05 in all of outcomes comparison). The incidence rate of biochemical pregnancy was 48.1% (87), clinical pregnancy rate with appearance of fetal heart was 44.2% (80), incidence rate of abortion before appearance of fetal heart (6 weeks) in women with intra-uterine pregnancy was 5.9% (5), incidence rate of abortion before 20 weeks was 12.9% (11). Incidence rate of ectopic pregnancy was 2.3% (2) and rate of multiple pregnancies was 32.5% (26). Conclusion: Our study suggests that the infertile patients who receive prednisolone in ICSI cycle, had improved pregnancy outcomes. Additional confirmatory studies are needed.

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Background: Infertility is defined as failure to achieve pregnancy after one year of unprotected sexual intercourse. Infertility can be related to male or female factors. Varicocele is the most common cause of infertility in men that is correctable with surgery. The purpose of this study was to determine the effects of recombinant follicle-stimulating hormone (rFSH) on semen parameters in infertile men.

Methods: This randomized clinical trial was done on 96 infertile men admitted to the Women's General Hospital Mohebe-Yas from September 2014 to September 2015. Inclusion criteria were to include varicocelectomy for unilateral idiopathic varicoceles and consent to participate in the study. Allergy to the drug combination and patient dissatisfaction were exclusion criteria. Patients participating in the study were divided into two groups randomly, one group received recombinant FSH three times a week and the other group received a placebo (normal saline) in the same way. After three months, the improvement of semen parameters, including motility, morphology and sperm count as well as the complications were determined in both groups. The data were analyzed with statistical software SPSS version 13 (Chicago, IL, USA).

Results: A total of 96 patients were enrolled in two groups of 48 men and women both groups were matched in terms of underlying factors. The rate of improvement in the morphology and motility of sperm in the treated group was significantly more than the placebo group (P= 0.0001) but the changes in sperm count were not significantly different between the groups (P= 0.495).

Conclusion: In summary, based on the results obtained in this study, it can be concluded that recombinant FSH is effective on improving semen parameters in infertile men after varicocelectomy compared with a placebo group and its major impact is on the morphology and motility of sperm.

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Background: Receptivity of endometrium has a critical role in the establishment of pregnancy after embryo transfer in the treatment process of infertile couples. As the glycoprotein CA-125 is a product of human endometrium and is measurable in the peripheral circulation, it is investigated whether it might serve as an indicator of endometrial receptivity and predictor of pregnancy following Intracytoplasmic sperm injection (ICSI).

Methods: In an observational diagnostic study, over a twelve-month period (from August 2013 to July 2014), all couples with male-factor infertility who attended to infertility clinic of Moheb Yas Hospital, Tehran and were candidate of performing ICSI, were invited to participate in the study. Based on the inclusion criteria of study, 64 women were eligible to take part in the study. They were assessed for serum CA-125 levels on the day of human chorionic gonadotropin (HCG) administration and also on the day of oocyte retrieval. After ICSI, the possibility of pregnancy was assessed by measuring serum concentration of &beta-HCG on 14 days after embryo transfer and also by visualizing the gestational sac by trans-vaginal ultrasound examination on four to five weeks after transfer. The pregnancy rate was compared between those with normal and high CA-125 levels.

Results: Among the subjects, 15 patients (23.4%) had high CA-125 levels, and totally 19 patients (29.7%) experienced pregnancy. Among those with normal and high CA-125 levels, 16 patients (32.7%) and 3 subjects (20%) experienced pregnancy, respectively, that showed no statistically significant difference according to Chi-square test (P=0.348). Also, according to the Fisher’s exact test, there was no correlation between CA-125 levels and the rate of pregnancy on the basis of body mass index (BMI).

Conclusion: Totally, according to the obtained results in current study, it may be concluded that serum CA-125 levels has no prognostic value in prediction of the outcomes of ICSI among infertile couples with male-factor infertility.

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Background: Spermatogenesis is a complex and highly organized process of proliferation and differentiation of spermatogonial stem cells. Spermatogonial stem cells (SSCs) as a unique stem cell have the potential to self-renewal, differentiation and transmit genetic information to the next generation and play a vital role in maintaining fertility. Sertoli cells as the only somatic cells within the seminiferous epithelium play central roles in the formation of niche and balance between self-renewal and differentiation by secrete many growth factors. Given the importance and widespread use of SSCs, particularly in the treatment of infertility, the aim of this study was to create an optimal environment for the proliferation of SSCs. So we decided to study of undifferentiated (ID4) and differentiated (c-Kit) gene expression in SSCs followed by co-culture with Sertoli cells for a one-month.

Methods: This experimental study was conducted from November 2013 to December 2014 in Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, on immature NMRI mouse (6-3 days old). Initially, Sertoli cells and SSCs were isolated from neonates mouse testes during the two-step enzymatic digestion characteristics Sertoli cells with vimentin marker and SSCs with promyelocytic leukemia zinc-finger (PLZF) marker were confirmed. Then SSCs were cultured in two groups: co-culture with Sertoli and without co-culture (control). Undifferentiated (ID4) and differentiation (c-Kit) gene expression were evaluated by Real-time PCR technique.

Results: Spermatogonial stem cells purity was obtained 66.91% by flow cytometry. The relative expression levels of gene ID4 in co-culture group at the end of each week, compared to the control group showed a significant increase (P<0.05). While the expression of this gene significantly decreased in each group over time (P<0.05). The results of the comparison of the relative expression of c-Kit gene in co-culture group are indicated significant decrease than the control group at the end of each week (P<0.05). In addition, this gene expression was showed significant increase in each group individually over time (P<0.05) ID4 gene expression showed a significant (P<0.05) increase toward the control group, while in the expression of c-Kit was observed a significant (P<0.05) decrease compared with the control group at the end of each week.

Conclusion: According to the results of this study, co-culture with Sertoli cells maintains SSCs in the prolifration stage for long-term, so can be used to optimize the culture medium at the clinic.

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Background: Lots of vitamin D functions are mediated by its steroid family receptor (VDR). Vitamin D role in infertility is reported by significant fertility reduction in many male laboratory animals with vitamin D deficiency. The reason for reduced fertility in male VDR-null mouse model has been reported to be reduced sperm count and sperm motility. Vitamin D has effects on sperm motility, sperm-ovum coupling, and acrosome reaction stimulation. As VDR is expressed in human male reproductive system, the aim of the current study was to investigate the role of rs2228570 polymorphism of VDR gene in male infertility.

Methods: Investigation was done as a case-control study on infertile azoospermic or oligospermic men referring to Avicenna Research Institute from March 2014 to April 2015. Rs2228570 single nucleotide polymorphism (SNP) located in exon 1 of VDR gene was chosen according to its role on protein function. Blood sampling was done on cases and control groups and after DNA extraction the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) reaction was designed and performed on 100 normal cases, 100 azoospermic and 100 oigospermic control samples. Distribution of quantitative age variable was done using Student’s t-test and qualitative variables (genotype and allelic frequencies) was done using SPSS, ver. 22 (Chicago, IL, USA).

Results: Chi-square test didn’t show significant difference between two case groups and controls (Azoosperm and control P=0.5 and oligosperm and control P=0.09). Comparing CC genotype frequency with TT and CT genotypes (azoosperm and control P=0.48 OR=0.77, oligosperm and control, P=0.17 OR=0.77) and in comparing between TT genotype with CT and CC genotypes (azoosperm and control P=0.49 OR=3.03, oligosperm and control P=0.19 OR=7.21) the difference between these groups was not significant and didn’t increase the probability of disease and didn’t show protective role against it.

Conclusion: According to the findings, the association between rs2228570 polymorphism of vitamin D receptor gene and infertility was not significant and investigation of other polymorphisms might show a relationship with male infertility.

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Results: Out of 253 couples, eighty-five (33.6%) became pregnant by IVF. The mean age of women with successful pregnancies was 33.96±5.2 years and in the unsuccessful pregnancies was 35.84±5.07 years. The results of this study showed that women who had a successful pregnancy after IVF were significantly younger than women who did not have a successful pregnancy (P=0.006). In the classification of patients into two groups of positive and negative pregnancies, younger age, adequate vitamin D levels, and higher sperm motility were the determinants of pregnancy in the first period of fertilization (Fresh protocol). Also, the existence of a failed history of infertility treatment in couples has been introduced as a negative factor for IVF fertility. 13(15.9%) abortions occurred and 11(12.9%) infants who were born with this method needed intensive care. Conclusion: Lower age at the time of fertilization, higher vitamin D levels, and more active sperms were found to have important and prognostic roles in the success of pregnancy by in vitro fertilization technique. The results of the present study showed that, Increased unsuccessful treatment has been associated with the history of failed treatment. Therefore, to respond to the relationship between previous infertility treatments and success in subsequent pregnancies in the IVF process, It is recommended to conduct studies with larger sample size and at a higher time efficiency.

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