Liens entre la morphologie et les marques épigénétiques, la qualité de l'ADN, le contenu chromosomique et les capacités fécondantes du spermatozoïde humain

Intrinsic sperm quality criteria (chromatin condensation, DNA fragmentation, chromosome content, fecundity, etc.) vary from one spermatozoon to another. It is critical to be able to link morphological aspects to these quality criteria, in order to improve the selection of high-quality, live spermatozoa and thus improve pregnancy and delivery rates. Since 2002, motile sperm organelle morphology examination has been used to screen for defects under high magnification and thus select vacuole-free spermatozoa (which are known to be associated with higher pregnancy rates in intracytoplasmic morphologically selected sperm injection (IMSI) programmes). Here, we demonstrate that sperm vacuoles are nuclear concavities associated with chromatin condensation failure (due to a lack of histone replacement during spermiogenesis). A number of other morphological abnormalities were found to be linked to chromatin condensation failure. In some cases, vacuolated spermatozoa were seen to be more DNA-fragmented than vacuole-free ones. In contrast, sperm morphology was not related to chromosomal content or fecundity characteristics. Our observations enabled us to identify new indications for IMSI and refine the criteria for selecting the best spermatozoon: the spermatozoon with the lowest risk of implantation failure or abnormalities in the offspringLes qualités intrinsèques spermatiques (état de condensation de la chromatine, intégrité de l’ADN, contenu chromosomique, capacités fécondantes..) sont très variables d’un spermatozoïde à l’autre. Le pari aujourd’hui en assistance médicale à la procréation est de relier un aspect morphologique spermatique à ces qualités intrinsèques dans le but de mieux choisir le spermatozoïde vivant à injecter et d’améliorer les taux de grossesse. Depuis 2002, le MSOME (Motile Sperm Organelle Morphology Examination) permet d’observer les spermatozoïdes à fort grossissement, en contraste interférentiel de Nomarski et d’observer des structures qu’on appelle les « vacuoles ». L’injection intra-ovocytaire de spermatozoïdes sélectionnés en MSOME (IMSI) comme non porteurs de vacuoles céphaliques permettrait d’améliorer les taux de grossesses évolutives. Ici, nous avons montré que les vacuoles spermatiques correspondaient à des cratères nucléaires en lien avec une non-condensation chromatinienne (non remplacement des histones au cours de la spermiogenèse). Certaines atypies morphologiques spermatiques sont aussi en lien avec une non-condensation de la chromatine. De plus, spermatozoïdes vacuolés présentent parfois des taux de fragmentation élevés. Aucun lien n’a cependant été retrouvé entre un aspect morphologique spermatique d’une part et un contenu chromosomique ou des capacités fécondantes particulières d’autre part. De ces liens, nous avons pu dégager de nouvelles indications d’IMSI. Ainsi, nous avons avancé dans la description des critères de sélection du spermatozoïde humain vivant donnant le moins de risque d’échec d’implantation embryonnaire et/ou d’anomalies pour la descendanc

Reply to Pallotti et al. comment on “Boitrelle et al. The sixth edition of the who manual for human semen analysis: A critical review and swot analysis. Life 2021, 11, 1368"

We would like to thank F. Pallotti and his colleagues for their positive comments [1] on our SWOT analysis of the strengths, weaknesses, threats and opportunities of the sixth edition of the WHO manual on semen analysis [2]. They have raised two concerns: the re-introduction of the category of rapidly moving sperm, and the dropping of the reference range. With regard to the re-introduction of the distinction between rapidly progressive (type a) motility and sluggish progressive (type b) motility, Pallotti et al. raised the point that this distinction is difficult to make visually and will, therefore, lead to “approximation” and subjective reporting with “reduced standardization”. This is a valid concern and was the reason why the editors of the fifth edition of the WHO manual removed this distinction (which was present in the fourth edition) and created the combined category of “progressive motility”

Impact of antioxidant therapy on natural pregnancy outcomes and semen parameters in infertile men: A systematic review and meta-analysis of randomized controlled trials

Purpose: Seminal oxidative stress (OS) is a recognized factor potentially associated with male infertility, but the efficacy of antioxidant (AOX) therapy is controversial and there is no consensus on its utility. Primary outcomes of this study were to investigate the effect of AOX on spontaneous clinical pregnancy, live birth and miscarriage rates in male infertile patients. Secondary outcomes were conventional semen parameters, sperm DNA fragmentation (SDF) and seminal OS.Materials and Methods: Literature search was performed using Scopus, PubMed, Ovid, Embase, and Cochrane databases. Only randomized controlled trials (RCTs) were included and the meta-analysis was conducted according to PRISMA guidelines.Results: We assessed for eligibility 1,307 abstracts, and 45 RCTs were finally included, for a total of 4,332 infertile patients. We found a significantly higher pregnancy rate in patients treated with AOX compared to placebo-treated or untreated controls, without significant inter-study heterogeneity. No effects on live-birth or miscarriage rates were observed in four studies. A significantly higher sperm concentration, sperm progressive motility, sperm total motility, and normal sperm morphology was found in patients compared to controls. We found no effect on SDF in analysis of three eligible studies. Seminal levels of total antioxidant capacity were significantly higher, while seminal malondialdehyde acid was significantly lower in patients than controls. These results did not change after exclusion of studies performed following varicocele repair.Conclusions: The present analysis upgrades the level of evidence favoring a recommendation for using AOX in male infertility to improve the spontaneous pregnancy rate and the conventional sperm parameters. The failure to demonstrate an increase in live-birth rate, despite an increase in pregnancy rates, is due to the very few RCTs specifically assessing the impact of AOX on live-birth rate. Therefore, further RCTs assessing the impact of AOX on live-birth rate and miscarriage rate, and SDF will be helpful

Technical aspects and clinical limitations of sperm DNA fragmentation testing in male infertility: A global survey, current guidelines, and expert recommendations

Purpose: Sperm DNA fragmentation (SDF) is a functional sperm abnormality that can impact reproductive potential, for which four assays have been described in the recently published sixth edition of the WHO laboratory manual for the examination and processing of human semen. The purpose of this study was to examine the global practices related to the use of SDF assays and investigate the barriers and limitations that clinicians face in incorporating these tests into their practice. Materials and Methods: Clinicians managing male infertility were invited to complete an online survey on practices related to SDF diagnostic and treatment approaches. Their responses related to the technical aspects of SDF testing, current professional society guidelines, and the literature were used to generate expert recommendations via the Delphi method. Finally, challenges related to SDF that the clinicians encounter in their daily practice were captured. Results: The survey was completed by 436 reproductive clinicians. Overall, terminal deoxynucleotidyl transferase deoxyuridine triphosphate Nick-End Labeling (TUNEL) is the most commonly used assay chosen by 28.6%, followed by the sperm chromatin structure assay (24.1%), and the sperm chromatin dispersion (19.1%). The choice of the assay was largely influenced by availability (70% of respondents). A threshold of 30% was the most selected cut-off value for elevated SDF by 33.7% of clinicians. Of respondents, 53.6% recommend SDF testing after 3 to 5 days of abstinence. Although 75.3% believe SDF testing can provide an explanation for many unknown causes of infertility, the main limiting factors selected by respondents are a lack of professional society guideline recommendations (62.7%) and an absence of globally accepted references for SDF interpretation (50.3%). Conclusions: This study represents the largest global survey on the technical aspects of SDF testing as well as the barriers encountered by clinicians. Unified global recommendations regarding clinician implementation and standard laboratory interpretation of SDF testing are crucial

A Web-Based Global Educational Model for Training in Semen Analysis during the COVID-19 Pandemic

Infertility affects between 2.5% and 12% of couples worldwide, with male factor infertility solely accounting for 20% to 30% and contributing to 50% of the overall infertility cases [1]. In the United States alone, infertility affects 9.5% of men [2]. The clinical evaluation of male infertility is based on the semen analysis where the results can significantly influence the diagnostic interpretation and management. While many clinicians rely on semen parameters as a surrogate marker of a man’s ability to father a child, the results of semen analysis should, however, be interpreted with caution considering its inherent limitations [3,4]. A properly performed semen analysis and an adequate clinical examination of the male along with questions regarding current medical conditions and lifestyle circumstances that could affect sample quality, can provide valuable information related to a man’s fertility potential. This information facilitates a better understanding of the physiology of the reproductive organs and the underlying causes of dysfunction [5- 7]. However, manual semen analysis has its inherent challenges associated with high subjectivity, lack of standardization, inadequate quality control and quality assurance, as well as inadequate assessment of competency, and training of laboratory personnel performing the test [7,8]. Unlike sperm concentration and motility, sperm morphology has even more subjectivity in reporting the results, with increased intra- and intervariability [8-10]. Therefore, quality control is imminent in preventing such variations and retaining uniformity in all assessments by all operators. This includes preanalytical (test requisition, correct sample collection, delivery of sample), analytical (mixing and loading of sample, correct preparation of smears or calculation of results), and post-analytical (correct reporting of results to the clinician) indicators. To minimize errors, daily, weekly, or monthly quality control of reagents and equipment is imperative

Impact of varicocele repair on semen parameters in infertile men: A systematic review and meta-analysis

Purpose: Despite the significant role of varicocele in the pathogenesis of male infertility, the impact of varicocele repair (VR) on conventional semen parameters remains controversial. Only a few systematic reviews and meta-analyses (SRMAs) have evaluated the impact of VR on sperm concentration, total motility, and progressive motility, mostly using a before-after analytic approach. No SRMA to date has evaluated the change in conventional semen parameters after VR compared to untreated controls. This study aimed to evaluate the effect of VR on conventional semen parameters in infertile patients with clinical varicocele compared to untreated controls. Materials and Methods: A literature search was performed using Scopus, PubMed, Embase, and Cochrane databases following the Population Intervention Comparison Outcome (PICOS) model (Population: infertile patients with clinical varicocele; Intervention: VR [any technique]; Comparison: infertile patients with clinical varicocele that were untreated; Outcome: sperm concentration, sperm total count, progressive sperm motility, total sperm motility, sperm morphology, and semen volume; Study type: randomized controlled trials and observational studies). Results: A total of 1,632 abstracts were initially assessed for eligibility. Sixteen studies were finally included with a total of 2,420 infertile men with clinical varicocele (1,424 patients treated with VR vs. 996 untreated controls). The analysis showed significantly improved post-operative semen parameters in patients compared to controls with regards to sperm concentration (standardized mean difference [SMD] 1.739; 95% CI 1.129 to 2.349; p<0.001; I2=97.6%), total sperm count (SMD 1.894; 95% CI 0.566 to 3.222; p<0.05; I2=97.8%), progressive sperm motility (SMD 3.301; 95% CI 2.164 to 4.437; p<0.01; I2=98.5%), total sperm motility (SMD 0.887; 95% CI 0.036 to 1.738; p=0.04; I2=97.3%) and normal sperm morphology (SMD 1.673; 95% CI 0.876 to 2.470; p<0.05; I2=98.5%). All the outcomes showed a high inter-study heterogeneity, but the sensitivity analysis showed that no study was sensitive enough to change these results. Publication bias was present only in the analysis of the sperm concentration and progressive motility. No significant difference was found for the semen volume (SMD 0.313; 95% CI -0.242 to 0.868; I2=89.7%). Conclusions: This study provides a high level of evidence in favor of a positive effect of VR to improve conventional semen parameters in infertile men with clinical varicocele. To the best of our knowledge, this is the first SRMA to compare changes in conventional semen parameters after VR with changes in parameters of a control group over the same period. This is in contrast to other SRMAs which have compared semen parameters before and after VR, without reference to a control group. Our findings strengthen the available evidence and have a potential to upgrade professional societies’ practice recommendations favoring VR to improve conventional semen parameters in infertile men

Post-vasectomy semen analysis: Optimizing laboratory procedures and test interpretation through a clinical audit and global survey of practices

Purpose: The success of vasectomy is determined by the outcome of a post-vasectomy semen analysis (PVSA). This article describes a step-by-step procedure to perform PVSA accurately, report data from patients who underwent post vasectomy semen analysis between 2015 and 2021 experience, along with results from an international online survey on clinical practice. Materials and Methods: We present a detailed step-by-step protocol for performing and interpretating PVSA testing, along with recommendations for proficiency testing, competency assessment for performing PVSA, and clinical and laboratory scenarios. Moreover, we conducted an analysis of 1,114 PVSA performed at the Cleveland Clinic’s Andrology Laboratory and an online survey to understand clinician responses to the PVSA results in various countries. Results: Results from our clinical experience showed that 92.1% of patients passed PVSA, with 7.9% being further tested. A total of 78 experts from 19 countries participated in the survey, and the majority reported to use time from vasectomy rather than the number of ejaculations as criterion to request PVSA. A high percentage of responders reported permitting unprotected intercourse only if PVSA samples show azoospermia while, in the presence of few non-motile sperm, the majority of responders suggested using alternative contraception, followed by another PVSA. In the presence of motile sperm, the majority of participants asked for further PVSA testing. Repeat vasectomy was mainly recommended if motile sperm were observed after multiple PVSA’s. A large percentage reported to recommend a second PVSA due to the possibility of legal actions. Conclusions: Our results highlighted varying clinical practices around the globe, with controversy over the significance of non-motile sperm in the PVSA sample. Our data suggest that less stringent AUA guidelines would help improve test compliance. A large longitudinal multi-center study would clarify various doubts related to timing and interpretation of PVSA and would also help us to understand, and perhaps predict, recanalization and the potential for future failure of a vasectomy.American Center for Reproductive Medicin

Standards in semen examination:publishing reproducible and reliable data based on high-quality methodology

Biomedical science is rapidly developing in terms of more transparency, openness and reproducibility of scientific publications. This is even more important for all studies that are based on results from basic semen examination. Recently two concordant documents have been published: the 6th edition of the WHO Laboratory Manual for the Examination and Processing of Human Semen, and the International Standard ISO 23162:2021. With these tools, we propose that authors should be instructed to follow these laboratory methods in order to publish studies in peer-reviewed journals, preferable by using a checklist as suggested in an Appendix to this article.Peer reviewe

Technical Aspects and Clinical Limitations of Sperm DNA Fragmentation Testing in Male Infertility: A Global Survey, Current Guidelines, and Expert Recommendations

PURPOSE: Sperm DNA fragmentation (SDF) is a functional sperm abnormality that can impact reproductive potential, for which four assays have been described in the recently published sixth edition of the WHO laboratory manual for the examination and processing of human semen. The purpose of this study was to examine the global practices related to the use of SDF assays and investigate the barriers and limitations that clinicians face in incorporating these tests into their practice. MATERIALS AND METHODS: Clinicians managing male infertility were invited to complete an online survey on practices related to SDF diagnostic and treatment approaches. Their responses related to the technical aspects of SDF testing, current professional society guidelines, and the literature were used to generate expert recommendations via the Delphi method. Finally, challenges related to SDF that the clinicians encounter in their daily practice were captured. RESULTS: The survey was completed by 436 reproductive clinicians. Overall, terminal deoxynucleotidyl transferase deoxyuridine triphosphate Nick-End Labeling (TUNEL) is the most commonly used assay chosen by 28.6%, followed by the sperm chromatin structure assay (24.1%), and the sperm chromatin dispersion (19.1%). The choice of the assay was largely influenced by availability (70% of respondents). A threshold of 30% was the most selected cut-off value for elevated SDF by 33.7% of clinicians. Of respondents, 53.6% recommend SDF testing after 3 to 5 days of abstinence. Although 75.3% believe SDF testing can provide an explanation for many unknown causes of infertility, the main limiting factors selected by respondents are a lack of professional society guideline recommendations (62.7%) and an absence of globally accepted references for SDF interpretation (50.3%). CONCLUSIONS: This study represents the largest global survey on the technical aspects of SDF testing as well as the barriers encountered by clinicians. Unified global recommendations regarding clinician implementation and standard laboratory interpretation of SDF testing are crucial