P–062 Annexin-V MACS method could be a good choice for sperm selection with high PLCZ1 expression and high blastocyst rate in high DFI male factor

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
M Saleh. Novin ◽  
Z Zandie ◽  
M Bakhtiari ◽  
R Aflatoonian
Keyword(s):  
Dna Fragmentation ◽  
Sperm Motility ◽  
Annexin V ◽  
Fertilization Rate ◽  
Good Choice ◽  
Semen Parameters ◽  
Sperm Selection ◽  
Male Factor ◽  
Significant Difference ◽  
Blastocyst Rate

Abstract Study question What effect does Annexin-V MACS method have on sperm selection by pLCZ1 expression and blastocyst rate in male DNA fragmentation? Summary answer AnnexinV-MACS method could be a good choice for sperm selection with high pLCZ1 expression and high blastocyst rate in high DNA fragmentation male factor. What is known already Sperm selection based on morphology and motility in ART techniques, is not enough for choosing the best sperm especially in male factor patients. In Annexin-V magnetic activated cell sorting (MACS) technique, apoptotic sperm are separated from non-apoptotic one by negative selection. So, this method can improved quality of compaction rate in embryo. PLCζ is oocyte activating factors that it starts oscillations of calcium ca2+ in oocyte and it has a significant effect on fertilization and implantation. Study design, size, duration Semen samples from 30 male factor infertile couples with high DFI (DFI>30%) were selected and divided into two group (control and experimental) in each patient. Participants/materials, setting, methods Control was washed with DGC and experimental one was selected by MACS-DGC. Retrieved eggs in each patient, were divided in 2. Both group were injected by DGC and MACS respectively. Semen parameters and DFI (SCD test) were analyzed before and after processing. After ICSI, rate of fertilization, embryo development and blastocyst formation were evaluated. Real time PCR evaluate expression of PLCζ. Comparison between results of two groups was assessed by SPSS analysis. Main results and the role of chance Results showed that, sperm motility and morphology after MACS method (45%, 1.7%) was significantly higher than DGC method (40%, 1.1%) and before washing (35%, 0.9%). Percent of DFI in MACS group (36%) was significantly decreased compared to DGC (45%) and primitive group (55%). The number of oocytes were injected in DGC group was 93 and in MACS group was 111. Fertilization rate in both groups was almost the same (72.07% in MACS vs 73.11 in DGC). Rate of day 3 embryo with good grade in MACS group (72. 5%) was significantly higher than DGC (51.47%) (P < 0.05). The blastocyst rate in MACS-DGC group (69.69%) was significantly increased compared to DGC group (48%). PLCζ gene expression in MACS-DGC was significantly higher than DGC group (p-value=0.046). Limitations, reasons for caution This experiment was performed invivo. Wider implications of the findings: Sperm selection by MACS-DGC method can improve sperm motility, morphology and reduce sperm DFI. No significant difference was observed in fertilization rate, but percent of high-quality embryo on days 3 and 5 was significantly higher by this method,also it can be suggested as a good choice for patients with highDF. Trial registration number Not applicable

P–235 Are morphokinetics parameters and profitability of an assisted reproduction cycle related to sperm selection technique? Microfluidic sperm sorter chip versus magnetic activated cell sorting

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
D Agud. Garcillan ◽  
E Santamarí. López ◽  
C Gonzále. Ravina ◽  
A Pachec. Castro ◽  
M Cru. Palomino ◽  
...  
Keyword(s):  
Assisted Reproduction ◽  
Cell Sorting ◽  
Statistical Significance ◽  
Fertilization Rate ◽  
Sperm Selection ◽  
Fragmented Dna ◽  
Significant Difference ◽  
Blastocyst Rate ◽  
Sperm Sorting ◽  
Magnetic Activated Cell Sorting

Abstract Study question Does microfluidic sperm sorter offer any biological improvement over magnetic activated cell sorting (MACS)? Summary answer Microfluidic approach for selecting high profile spermatozoa is as good as magnetic activated cell sorting in terms of morphokinetic, fertilization and good quality blastocyst rate. What is known already Microfluidic sperm sorter chip is a method for select non-fragmented DNA sperm. We know that the use of these non-fragmented DNA sperms can improve the clinical results and the useful blastocyst rate. As several studies have shown previously, embryos morphokinetics parameters are affected by culture medium, ovarian stimulation, oxygen tension, origin of the oocytes, or the age of the patient, this is why we wanted to compare if the cleavage times using time lapse technology, are different depending on the sperm selection method used to select sperm with the non-fragmented DNA. Study design, size, duration Prospective and observational study performed between May 2019 to January 2021 in IVI Madrid and IVI Sevilla. Seminal samples from couples participating in the study were divided into two aliquots; each of them was processed according to one of the study methods. 53 couples were included in the study. Half of the oocyte from each donor were microinjected with sperm selected through MACS (n = 281) and the other half through a microfluidic device (n = 275). Participants/materials, setting, methods These oocytes were microinjected with both types of sperm samples and incubated in EmbryoScope. Cellular events studied in this study included cellular divisions until blastocyst stage, appearance and fading of some cellular structures and the duration of the first, second and third cellular cycle (cc1, cc2 and cc3) as well as their synchrony (S1, S2, S3). Data were exported from the EmbryoViewer data base. We perform an ANOVA statistical analysis to analyze the data. Main results and the role of chance No significant differences between both sperm selection methods were found regarding the time of cell division (from T2 to Tblstocyst), the cellular cycles duration (cc1, cc2 and cc3) or the synchrony of the cellular cycles divisions (s1, s2 and s3). However, a clear trend towards statistical significance has been found in both duration of cc2 (p = 0.052) being longer in MACS embryos than in microfluidic sperm sorting embryos, and in the expansion of the blastocyst, which occurs earlier in embryos that come from MACS than in those that come from microfluidic sperm sorting (p = 0.097). These two events could indicate a better embryo cleavage dynamic in the case of MACS embryos, with a better blastocyst expandability and the necessary time to carry out all the biological events that must occur in the cc2. However, significant difference was found in the direct cleavage from 1 to 3 cells embryo stage, which is one of the adverse events that more affects embryo implantation, being higher in microfluidic sperm sorting group (p = 0,037). Finally, the fertilization rate (73.1% vs 76.9%) and the good quality blastocyst rate (53.7% vs 56.5%) were higher in MACS embryos than in microfluidic sperm sorting embryos, although no significant differences were found. Limitations, reasons for caution This study has been performed in donated oocytes, so these results may not be extrapolated to other groups of assisted reproduction patients. However, more data are needed to draw firm conclusions. Furthermore, it´s crucial to increase the sample size to check if the trends founded reach statistical significance. Wider implications of the findings: Microfluidic sorting of unprocessed semen in un unselected population is as efficacious as magnetic activated cell sorting according embryo morphokinetic, fertilization rate and useful blastocyst rate. Microfluidic sperm sorting does not show clinical advantage over MACS considering this data collection. Trial registration number NCT04061484

scholarly journals Impact of percutaneous embolization versus subinguinal microsurgical ligation on semen parameters in primary varicocele patients: comparative study

2020 ◽  
Vol 51 (1) ◽  
Author(s):  
Haytham M. Nasser ◽  
Ahmed Hussein ◽  
Gad M. Behairy ◽  
Mostafa Abdo
Keyword(s):  
Sperm Count ◽  
Semen Analysis ◽  
Statistical Significance ◽  
Semen Parameters ◽  
Male Factor ◽  
Percutaneous Embolization ◽  
Significant Difference ◽  
Group 2 ◽  
Group 1

Abstract Background Varicocele is an abnormally dilated pampiniform plexus of the veins within the spermatic cord and is considered the most common correctable cause of male factor infertility. Many approaches are described for treatment either surgical (tradition inguinal, subinguinal, and laparoscopic) or non-surgical percutaneous embolization. During the period from August 2017 to December 2018, we prospectively analyzed the preoperative and post-operative alteration of semen parameters (at 3 and 9 months) of the data collected from 63 patients with clinically evident varicocele referred to our tertiary hospital. Patients were divided into two groups: group 1, thirty-three patients who underwent subinguinal microsurgical ligation, and group 2, thirty patients who underwent percutaneous embolization. Results Sixty-three patients enrolled in this study were divided in two groups: group 1, patients who underwent surgery, and group 2, patients who underwent embolization; the mean age is 24.6 ± 1.27 years in group 1 and 23.7 ± 2 years in group 2; there was no statistically significant difference between the two groups as regards BMI, diabetes, hypertension, and smoking. Bilaterality was present in 15.2% of group 1 patients and 10% in group 2 patients (P value 0.06). Most of the patients were classified as grades 2 and 3 with no statistical significance regarding severity of the disease. Preoperative semen parameters for patients including sperm count, motility, and abnormal forms showed no statistically significant difference between the two groups. Post-intervention semen analysis was done twice during follow-up after 3 months and 9 months from the date of intervention. After 3 months, the semen parameters were improved in both groups in spite of the higher sperm count in group 2 but with no statistical significance. After 9 months follow-up, semen analysis showed persistent increase in sperm mobility in group 1 patients in comparison to group 2 patients. Both groups had better improvement in count of normal form with no statistical significant change. Conclusion Improvement of semen parameters while treating primary varicocele by either subinguinal microsurgery approach or percutaneous embolization shows equivalent outcomes.

P–025 Sperm selection using a modified “swim up” technique in absence of sperm centrifugation improve sperm DNA fragmentation and decreases miscarriage rate

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
Y Cabell. Vives ◽  
P Belchin ◽  
C Lopez-Fernandez ◽  
M Fernandez-Rubio ◽  
J Guerrero-Sanchez ◽  
...  
Keyword(s):  
Dna Fragmentation ◽  
Sperm Quality ◽  
P Value ◽  
Sperm Dna Fragmentation ◽  
Sperm Selection ◽  
Male Factor ◽  
Miscarriage Rate ◽  
Sperm Dna ◽  
Group 2

Abstract Study question Is it useful to avoid sperm centrifugation in laboratory routine work to improve sperm quality and reproductive outcome in Assisted Reproduction Techniques (ART)? Summary answer Exclusion of sperm centrifugation for sperm selection using neat sperm samples (IO-lix), increases sperm quality in the collected subpopulation decreasing miscarriage rate after using ICSI. What is known already Inclusion of sperm centrifugation in ART is an aggressive intervention for sperm selection with ineludible production iatrogenic damage affecting sperm integrity. The application of IMSI, PICSI or microfluidic devices avoid sperm centrifugation and may improve the quality of the subsample obtained. However, these methodologies may result time consuming, expensive or producing poor results when the quality of the sperm is limited. We have already shown that a modified swim-up avoiding centrifugation (called IO-lix) is a low-cost and efficient alternative to microfluidic devices, recovers 100 times more concentration and reduces sperm DNA fragmentation with no significant differences to other methodologies. Study design, size, duration This is a retrospective study from 2018 to 2020 which includes patients with an average of age of 38.2 years using their own oocytes with ICSI as fertilization technique. Two aleatory groups of patients were made: Group 1: 88 cycles with 503 fertilized oocytes and 206 blastocysts were obtained with sperm samples processed by IO-lix and Group 2: 303 cycles, 1451 fertilized oocytes and 591 blastocysts using a standard “swim up” technique to process sperm. Participants/materials, setting, methods A total of 391 ICSI cycles were included in this retrospective study. The male factor was similar in both groups and they showed altered SDF previously to the cycle. We compared data of the motility and SDF of sperm samples before and after applying IO-lix and we analyzed by X2 contingence test differences on miscarriage rates between groups 1 and 2. Main results and the role of chance General sperm parameter changes after IO-lix showed that averaged sperm concentration observed in neat ejaculated samples was 62M/SD=46.4. Values obtained after IO-lix in the same samples were 12.3M/SD8.0. Averaged sperm motility in neat samples was 54%/SD=9.3 and 70.9%/SD=13.2 after IO-lix. Finally, sperm DNA fragmentation in neat samples was 35.8%/SD17.3, while these values decreased to 9.2%/SD=3.9 after IO-lix. About reproductive outcome results, significant differences were not obtained on the development to blastocyst stage rate comparing both groups (X2=0.003; p value = 0.954; Alpha 0.05). In the case of IO-lix processed samples, the pregnancy rate was 59.42% in Group 1 and 44.72% in Group 2 (X2=0.651; p value =0.419; Alpha 0.05). A total of 9 miscarriages of 41 clinical pregnancies (21.95%) were observed after IO-lix, while this number increases to 59 out of 123 clinical pregnancies, which means the 47.96% of the embryo transfers, when “swim-up” was used. In this case significant differences were obtained (X2=3.935; p value = 0.0.047; Alpha 0.05). Limitations, reasons for caution Being a pilot study aimed to understand the results of IO-lix in ART, correlations have not been stablished between the levels of sperm improvement after IO-lix and paired results of ART. This study would be necessary, specially to identify the possible origin of miscarriage associated to the male factor. Wider implications of the findings: Elimination of sperm centrifugation using a combined strategy of gradients and “swim-up” for sperm isolation, reduce miscarriage rate and produce equivalent results of blastocyst development to those obtained with “swim-up”. Being a cost-effective and improving laboratory workload, its use for sperm selection is recommended. Trial registration number Not applicable

scholarly journals Randomized Clinical Trial: Effect of Walnuts on Semen Parameters and Male Fertility (P18-042-19)

2019 ◽  
Vol 3 (Supplement_1) ◽  
Author(s):  
Wendie Robbins ◽  
Howard Kim ◽  
Justin Houman ◽  
Geng-Wei Lee
Keyword(s):  
Clinical Trial ◽  
Los Angeles ◽  
Randomized Clinical Trial ◽  
Sperm Motility ◽  
Clinical Care ◽  
Sperm Concentration ◽  
Nutritional Supplement ◽  
Semen Parameters ◽  
Male Factor Infertility ◽  
Male Factor

Abstract Objectives Infertility affects ∼8–12% of couples worldwide with ∼40% attributed to male factors. Recent studies suggest a role for paternal diet in fertility. Walnuts contain a variety of nutrients essential in the development of spermatozoa. We conducted a randomized clinical trial (RCT) to determine if consumption of walnuts improves semen parameters and fertility in men seeking clinical care for male factor infertility. Methods This was a two arm, single blind, RCT. The comparison groups both received usual care for male factor infertility. One group added 42 gm/d walnuts to their diet, and the other group added a daily nutritional supplement recommended for male reproductive health. Participants (n = 75) were enrolled at an infertility clinic located in a large metropolitan medical center. Eligibility was determined by history, physical exam, and lab tests collected as part of clinical care. Research measures included semen analysis and blood sample at baseline and 3 months; ASA24 dietary recall at baseline, 2 and 3 months; and fertility report at 3 months and 1 year. Results Age range was 27 to 61 years (39.7 ± 7.0); BMI range 19.6 to 46.9 (26.8 ± 4.5); participant race was Asian 26.2%, White 44.3%, Hispanic White 16.4%, Black 8.2%, other 4.9%. Baseline sperm concentration was 39.4 ± 30 million per ml; sperm motility 31.1 ± 23.4%; and progressive motility 21.2 ± 15.8%. At 3 months, the walnut group demonstrated increased sperm motility and concentration, P = .04 and P = .07, respectively, whereas no significant changes from baseline were found in the nutritional supplement group. Both groups showed improved sperm morphology, P < .03. Preliminary data from the subset of men with 1-year follow-up data shows higher frequency of pregnancy in the walnut group compared to nutritional supplement, although not statistically significant, P = .09. We continue to follow the remaining participants until their 1-year fertility report. Conclusions This RCT demonstrated a beneficial effect of adding walnuts to the diet on sperm motility and morphology in men seeking care for infertility. Preliminary fertility data suggests walnuts may enhance the probability of pregnancy for men with male factor infertility. Funding Sources Center for Occupational and Environmental Health, University of California, Los Angeles; California Walnut Commission.

scholarly journals DNA fragmentation index and its correlation with other sperm parameters in 4833 infertile males

2018 ◽  
Vol 7 (9) ◽  
pp. 3718
Author(s):  
Priya S. Patil ◽  
Satish M. Patki ◽  
Shweta S. Patki ◽  
Ujjwala S. Patki
Keyword(s):  
Dna Fragmentation ◽  
Sperm Motility ◽  
Sperm Count ◽  
Semen Analysis ◽  
Pearson Correlation ◽  
Significant Negative Correlation ◽  
Male Factor ◽  
Fragmentation Index ◽  
Success Of Treatment ◽  
Dna Fragmentation Index

Background: Semen analysis forms the basic investigation for male factor of infertility. According to WHO Manual sperm values of normalcy are decreasing. Recent studies have highlighted newer parameters of sperm playing their role in the outcome of pregnancy. The aim was to study the DNA fragmentation index (DFI) and to know its correlation with other sperm parameters.Methods: A total of 4833 cases underwent routine semen analysis along with the DFI. The statistical analysis was done using SPSS using t test and ANOVA test.Results: It was observed that 52% infertile males presented late for treatment. All parameters showed a decline with advancing age. There was definite correlation between sperm count and sperm motility as well as morphology. The DFI showed significant negative correlation with sperm count. Pearson correlation showed positive correlation between DFI and sperm motility and morphology.Conclusions: The study highlights the need for increasing awareness about male factor of infertility. A thorough semen analysis along with DFI can help the clinician to explain the couple in a better way about the choice of ART and success of treatment.

P–091 Magnetic Activated Cell Sorting (MACS) improves euploid blastocysts rate in pre-implantation genetic testing cycles with high levels of sperm DNA fragmentation and advanced paternal age

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
F Scarselli ◽  
E Cursio ◽  
A Colasante ◽  
V Zazzaro ◽  
P Andrea ◽  
...  
Keyword(s):  
Genetic Testing ◽  
Dna Fragmentation ◽  
Sperm Quality ◽  
Annexin V ◽  
Paternal Age ◽  
Control Group ◽  
Sperm Dna Fragmentation ◽  
Sperm Dna ◽  
Number Of Patients ◽  
Blastocyst Rate

Abstract Study question Can MACS increase euploid blastocyst rate in Pre-implantation Genetic Testing (PGT) cycles for AMA-APA (Advanced Maternal-Paternal Age) in patients with high sperm DNA fragmentation (SDF)? Summary answer A slight increase in euploid blastocyst rate was found using MACS in infertile patients with high SDF undergoing PGT cycles compared to the control group. What is known already Many authors have shown a close correlation between the presence of apoptotic markers on spermatozoa and the failure of assisted reproduction treatments. In normal physiological conditions, apoptotic spermatozoa with phosphatidylserine (PS) residues externalized on the plasma membrane, are eliminated along female genital tract, preventing oocyte fertilization. MACS eliminates apoptotic sperm whit PS residues using superparamagnetic microbeads conjugated with annexin V. This technique reduces the proportion of sperm with high rates of SDF and can be used to maximize ART procedures results. MACS application improves sperm quality, fertilization, cleavage and pregnancy rates reducing miscarriage rate. Study design, size, duration From June to November 2020, 10 couples in which MACS was applied to select non-apoptotic spermatozoa, were randomly enrolled in our study (MACS group) and 8 couples without MACS were considered as controls (No-MACS Group). All couples in both groups underwent a PGT cycle and had high sperm DNA Fragmentation (&gt; 20%). A higher rate of euploid and diploid-euploid mosaic blastocysts were obtained in the MACS group compared to the control group. Participants/materials, setting, methods Patients with severe oligoastenoteratozoospermia were excluded. MACS protocol was performed as follows: semen sample was analyzed (WHO 2010) and washed with buffered medium; pellet was removed and a swim-up was performed. Retrieved spermatozoa were washed with a binding buffer (Miltenyi Biotec), centrifuged (400 g x 4 minutes) and supernatant discarded. Pellet was covered with Annexin-V and re-suspended. After 15 minutes incubation at room temperature, the sample was eluted through the column and collected for ICSI. Main results and the role of chance In MACS group, female and male mean age ± SD were 41.6 ± 2.1 and 43.5 ± 7.3, respectively. Female and male mean age ± SD were 41.7 ± 2.8 and 44.6 ± 8.1 in the No-MACS group, respectively. In MACS and No-MACS groups, injected oocytes were 44 and 35, fertilized oocytes were 32 (72.3%) and 27 (77.1%) (NS), blastocyst formation rates were 71.8% (23/32) and 48.1% (13/27) (NS), respectively. In No-MACS group, only 1 euploid and 1 diploid-euploid mosaic blastocysts were obtained (1/13 = 8%) (NS). In MACS group, 4 euploid blastocysts were formed (4/23 = 17.4%) whereas mosaic diploid-euploid blastocysts were 3/23 (13.0%) (NS). Aneuploid blastocysts were 16/23 (69.6%) in MACS group and 11/13 (84.6%) in No-MACS group (NS). Limitations, reasons for caution AMA and APA of couples enrolled should be considered as a limit of the study. A larger number of patients and biopsied blastocysts are needed to analyze clinical results and perform a robust statistical analysis establishing if MACS is useful to improve transferable blastocyst rate in patients with high SDF. Wider implications of the findings: MACS is useful to select non apoptotic sperms; although fertilization, cleavage and blastocyst rates are not improved, aneuploid blastocysts rate slightly decreases using MACS. It I possible that, selecting spermatozoa free from PS residues, MACS allows to choose spermatozoa with a better DNA packaging, thus affecting the embryo ploidy. Trial registration number non applicable

P–122 Benefit of use of theophilline in ICSI with testicular sperm

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
M Be. Khelif. Jerbi ◽  
I Chabchoub ◽  
S Sfaxi ◽  
M H Be. aribia ◽  
S Mnallah ◽  
...  
Keyword(s):  
Pregnancy Rate ◽  
Sperm Motility ◽  
Time Interval ◽  
Sperm Selection ◽  
Testicular Sperm ◽  
Cumulative Pregnancy Rate ◽  
Significant Difference ◽  
Group A ◽  
Icsi Outcomes ◽  
Group B

Abstract Study question Would the use of theophylline have an effect on ICSI outcomes? Summary answer The cumulative pregnancy rate after the transfer of fresh and frozen embryos (FET) becomes more important justifying the addition of theophylline as an efficacy variable. What is known already Absolute immotile spermatozoa is one of the most important causes of reduced fertilization and pregnancy rates after ICSI, immotility of testicular spermatozoa is a physiological event resulting from metabolic sperm immaturity. Over the years, there have been numerous attempts to resolve this problem by identification of pharmacological agents that might improve sperm motility. In particular, theophylline turned out to be an effective tool in stimulating motility in human semen and identifying viable sperm in testicular sperm extraction sperm (TESE) patients. aim of this study is to evaluate ICSI outcomes after the use of theophylline to select viable spermatozoa Study design, size, duration This prospective, comparative randomised study was conducted in Fertillia ART center in Tunisia, between november 2017 to november 2020. All patients underwent ICSI cycles with testicular sperm were included. The exclusion criteria consist of cycles with female partner age &gt;42 years and/or cycles with no information about pregnancy outcomes. In our study, the cycles were categorized into two groups according to sperm selection method used in ICSI procdure. Participants/materials, setting, methods The present study include 678 cycles. This cohort was randomly divided into two groups according to method of spermatozoa selection. Treatements arms were performed by theophylline to improve sperm motility (Group A) and the Hyposmotic swelling (HOS) test to indicate sperm vitality (Group B). Clinical and biological parameters, the duration of sperm selection and ICSI outcomes were compared between the two groups. A statistical significant difference was accepted when the p value was &lt;0.05. Main results and the role of chance Baseline clinical parameters were found to be comparable in the two groups. No differences regarding number of oocytes retrieved or MII oocytes were reported . he sperm selection was easier in the group A (time interval : 12±7,2 minutes) than group B (time interval : 22±10 minutes) (p &lt; 0,05). No significative difference was observed between goups in the Fertilization rate (group A : 66% vs group B : 68%) ; Cleavage rate (group A : 76.20% vs. Group B : 75.56%) and Blasulation rate (group A : 63.60% vs. Group B 60.61%) p &gt; 0 ,05. The Cumulative Pregnancy rate for group A (31. 5%) was higher than group B (30%, p &gt; 0 ,05). Indeed, The rate of frozen cycles is significantly higher for group A compared to group B (30% vs. 4.5% p &lt; 0,05). Limitations, reasons for caution Need large sample size. Wider implications of the findings: Theophylline reduces significantly the time needed for sperm selection, Which positively affects the ICSI result. Trial registration number Not applicable

scholarly journals Antioxidant pretreatment for male partner before ART for male factor subfertility: a randomized controlled trial

2020 ◽  
Vol 2020 (4) ◽  
Author(s):  
Treasa Joseph ◽  
Mariano Mascarenhas ◽  
Reka Karuppusami ◽  
Muthukumar Karthikeyan ◽  
Aleyamma T Kunjummen ◽  
...  
Keyword(s):  
Pregnancy Rate ◽  
Embryo Transfer ◽  
Controlled Trial ◽  
Male Partner ◽  
Clinical Pregnancy ◽  
Clinical Pregnancy Rate ◽  
Semen Parameters ◽  
Male Factor ◽  
Sperm Dna ◽  
Significant Difference

Abstract STUDY QUESTION Does oral antioxidant pretreatment for the male partner improve clinical pregnancy rate in couples undergoing ART for male factor subfertility? SUMMARY ANSWER There was no significant difference in clinical pregnancy rate following oral antioxidant pretreatment for male partner in couples undergoing ART for male factor subfertility compared to no pretreatment. WHAT IS KNOWN ALREADY Damage to sperm mediated by reactive oxygen species (ROS) contributes significantly to male factor infertility. The ROS-related injury reduces fertilization potential and adversely affects the sperm DNA integrity. Antioxidants act as free radical scavengers to protect spermatozoa against ROS induced damage. During ART, use of sperms which have been exposed to ROS-mediated damage may affect the treatment outcome. Pretreatment with antioxidants may reduce the ROS-mediated sperm DNA damage. Currently, antioxidants are commonly prescribed to men who require ART for male factor subfertility but there is ambiguity regarding their role. STUDY DESIGN, SIZE, DURATION This was an open label, randomized controlled trial conducted at a tertiary level infertility clinic between February 2013 and October 2019. The trial included 200 subfertile couples who were undergoing ART treatment for male factor subfertility. PARTICIPANTS/MATERIALS, SETTING, METHODS Couples were randomized into treatment arm (n = 100) and control arm (n = 100). In the treatment arm, the male partner received oral antioxidants (Vitamin C, Vitamin E and Zinc) for 3 months just prior to the ART cycle. In the control arm, no antioxidant was given to the male partner. The primary outcome was clinical pregnancy rate, while live birth rate (LBR), miscarriage rate and changes in semen parameters were the secondary outcomes. MAIN RESULTS AND THE ROLE OF CHANCE Out of 200 women randomized, 135 underwent embryo transfer as per protocol. Following intention to treat analysis, no significant difference was noted in clinical pregnancy (36/100, 36% vs 26/100, 26%; odds ratio (OR) 1.60, 95% CI 0.87 to 2.93) and LBR (25/100, 25% vs 22/100, 22%; OR 1.18, 95% CI 0.61 to 2.27) between antioxidant and no pretreatment arms. The clinical pregnancy rate per embryo transfer was significantly higher following antioxidant pretreatment (35/64, 54.7% vs 26/71, 36.6%; OR 2.09, 95% CI 1.05 to 4.16) compared to no pretreatment. There was no significant difference in LBR per embryo transfer (25/64, 39.1%, vs 22/71, 31.0%; OR 1.43, 95% CI 0.70 to 2.91) after antioxidant pretreatment versus no pretreatment. The semen parameters of sperm concentration (median, interquartile range, IQR) (18.2, 8.6 to 37.5 vs 20.5, 8.0 to 52.5, million/ml; P = 0.97), motility (median, IQR) (34, 20 to 45 vs 31, 18 to 45%; P = 0.38) and morphology (mean ± SD) (2.0 ± 1.4 vs 2.2 ± 1.5%; P = 0.69) did not show any significant improvement after intake of antioxidant compared to no treatment, respectively. LIMITATIONS, REASONS FOR CAUTION The objective assessment of sperm DNA damage was not carried out before and after the antioxidant pretreatment. Since the clinicians were aware of the group allotment, performance bias cannot be ruled out. WIDER IMPLICATIONS OF THE FINDINGS The current study did not show any significant difference in clinical pregnancy and LBR following antioxidant pretreatment for the male partner in couples undergoing ART for male subfertility. The findings need further validation in a larger placebo-controlled randomized trial. STUDY FUNDING/COMPETING INTEREST(S) This trial has been funded by Fluid Research grant of Christian Medical College, Vellore (internal funding). The authors have no conflicts of interest to declare. TRIAL REGISTRATION NUMBER CTRI/2013/02/003431 TRIAL REGISTRATION DATE 26 February 2013 DATE OF FIRST PATIENT’S ENROLMENT 11 February 2013

P–120 Selecting spermatozoa with the highest genomic integrity in order to enhance clinical outcomes in men with high DNA fragmentation levels

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
D Tavares ◽  
P Xie ◽  
M Haddad ◽  
Z Rosenwaks ◽  
G D Palermo
Keyword(s):  
Embryonic Development ◽  
Semen Analysis ◽  
Implantation Rate ◽  
Testicular Biopsy ◽  
Fertilization Rate ◽  
Semen Parameters ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Sperm Chromatin ◽  
Genomic Integrity ◽  
Male Factor

Abstract Study question What are the best methods of selecting spermatozoa with the highest genomic integrity in order to improve embryo implantation and term pregnancy rates with ICSI? Summary answer Testicular or ejaculate spermatozoa isolated by microfluidic sperm selection (MFSS) were characterized by superior genomic integrity with improved clinical pregnancy and delivery rates. What is known already In couples with unexplained infertility, a subtle male factor can often be identified. Both single-strand (ss) and double-strand (ds) DNA nicks and breaks hinder the ability of the male gamete to support embryonic development. Surgical retrieval of spermatozoa from the proximal male genital tract can prevent their exposure to oxidative stress. Moreover, use of membrane-based microfluidics chips has been shown to allow for selection of the most progressively motile spermatozoa with higher genomic integrity. Study design, size, duration Over the course of 48 months, 86 consenting men presenting with high sperm chromatin fragmentation (SCF) in their ejaculate with prior ART failure underwent a subsequent cycle with specimens retrieved by testicular biopsy or ejaculate processed by MFSS. A concurrent TUNEL assay was performed on samples collected or selected by each method. Sperm specimens of both origins were utilized for ICSI cycles. Semen parameters, chromatin integrity, and pregnancy outcomes were compared between the two methods. Participants/materials, setting, methods Fresh ejaculates from consenting men were collected for standard semen analysis (WHO 2010). Testicular biopsy and MFSS were used to isolate spermatozoa with a higher genomic integrity after previous ART failure. SCF was assessed by terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) on at least 500 spermatozoa under a fluorescent microscope with a threshold of ≥ 15%. MFSS was carried out by Zymot® chips. ICSI was performed in the standard fashion. Main results and the role of chance A total of 86 men (36.5±5 years) had the following semen parameters: volume of 2.6 ±1mL, concentration of 27.0±33 x 106/mL, 35.6±15% motility, and high SCF (24.1±10%). They underwent 146 ICSI cycles with their partners (maternal age, 33.7±3) resulting in a high incidence of pregnancy loss (100%; 13/13). Of those who failed to conceive, 22 couples used surgically retrieved spermatozoa (SRS) with a concentration of 1.8 ± 4 x 106/mL (P &lt; 0.01), 5.0±11% motility (P &lt; 0.01), and an SCF of 12.6 ± 6% (P &lt; 0.0001). SRS was used in 37 ICSI cycles, yielding a fertilization rate of 61.6% (204/331, P &lt; 0.01), an implantation rate of 10.6% (9/85, P &lt; 0.01), a CPR of 23.5% (8/34, P &lt; 0.01), and a delivery rate of 17.6% (6/34, P &lt; 0.01). Another 24 couples underwent ICSI cycles with ejaculated spermatozoa processed by MFSS with a concentration of 1.8±3 x 106/mL (P &lt; 0.01), but an increased motility of 99±1% (P &lt; 0.01) and an SCF of 1.2 ±1%, lower than both the raw and testicular specimens (P &lt; 0.0001). MFSS-processed specimens resulted in a fertilization rate of 76% (335/441, P &lt; 0.01), an implantation rate of 26.3% (15/57, P &lt; 0.05), and a CPR of 67.9% (19/28, P &lt; 0.01), of which 15 patients delivered and 2 pregnancies are ongoing (89.5%; P &lt; 0.01). Limitations, reasons for caution This is a preliminary study on a small number of subjects. A randomized prospective study conducted on a larger cohort would be required to confirm our findings. Wider implications of the findings: SCF severely affects pregnancy by impairing embryonic development, consequently promoting implantation failure. While retrieving spermatozoa from the germinal epithelium is a viable option, MFSS provides an alternative. Although MFSS requires an adequate number of sperm with good kinetic characteristics, it provides a more palatable option, reducing surgical risk and costs. Trial registration number Not applicable

scholarly journals Successful Pregnancy in a Couple with Severe Male Factor Infertility after Selection of Sperm with Cytoplasmic Droplets

2015 ◽  
Vol 2015 ◽  
pp. 1-4 ◽  
Author(s):  
Jenna Bellish ◽  
David H. McCulloh ◽  
Khaliq Ahmad ◽  
Peter G. McGovern
Keyword(s):  
Dna Fragmentation ◽  
Embryo Development ◽  
Male Partner ◽  
Male Factor Infertility ◽  
Sperm Selection ◽  
Selection Methods ◽  
Male Factor ◽  
Successful Pregnancy ◽  
Live Births ◽  
Selection Of

We present live births resulting from two separate IVF cycles in a couple in which ICSI was performed with sperm specifically selected for presence of small cytoplasmic droplets. These cycles followed previous cycles using standard sperm selection methods in which very poor embryo development and no pregnancies ensued. The male partner was diagnosed with severe male factor infertility including elevated DNA fragmentation.

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