Infertility affects around 20% of couples of reproductive age; however, in some societies, as many as one-third of couples are unable to conceive. Different factors contribute to the decline of male fertility, such us environmental and professional exposure to endocrine disruptors, oxidative stress, and life habits with the risk of de novo epigenetics dysregulation. Since the fantastic development of new “omes and omics” technologies, the contribution of inherited or de novo genomes and epigenome disorders to male infertility have been further elucidated. Many other techniques have become available to andrology laboratories for the investigation of genome and epigenome integrity and the maturation and the competency of spermatozoa. All these new methods of assessment are highlighting the importance of genetics and epigenetics investigation for assisted reproduction pathology and for supporting professionals in counselling patients and proposing different management strategies for male infertility. This aims to improve clinical outcomes while minimizing the risk of genetics or health problems at birth.

Male infertility is a multifactorial condition often associated with disruptions in sperm metabolism and mitochondrial function, yet traditional semen analysis provides limited insight into these molecular mechanisms. Understanding sperm bioenergetics and metabolic dysfunctions is crucial for improving the diagnosis and treatment of conditions such as asthenozoospermia and azoospermia. This systematic review synthesizes recent literature, focusing on advanced tools and techniques—including omics technologies, advanced imaging, spectroscopy, and functional assays—that enable comprehensive molecular assessment of sperm metabolism and development. The reviewed studies highlight the effectiveness of metabolomics, proteomics, and transcriptomics in identifying metabolic biomarkers linked to male infertility. Non-invasive imaging modalities such as Raman and magnetic resonance spectroscopy offer real-time metabolic profiling, while the seminal microbiome is increasingly recognized for its role in modulating sperm metabolic health. Despite these advances, challenges remain in clinical validation and implementation of these techniques in routine infertility diagnostics. Integrating molecular metabolic assessments with conventional semen analysis promises enhanced diagnostic precision and personalized therapeutic approaches, ultimately improving reproductive outcomes. Continued research is needed to standardize biomarkers and validate clinical utility. Furthermore, these metabolic tools hold significant potential to elucidate the underlying causes of previously misunderstood and unexplained infertility cases, offering new avenues for diagnosis and treatment.

Endometriosis is a chronic, estrogen-dependent disorder associated with the presence of endometrial cells mainly in the pelvic cavity, causing systemic immune inflammation, infertility, epigenetic dysregulation of differential DNA methylation, coelomic metaplasia, and pain. It affects approximately 10–12% of women. Despite decades of research, full pathophysiology, a diagnostic roadmap, and clinical management strategies for endometriosis are not yet fully elucidated. Cell-free DNA (Cf-DNA) in the peripheral blood of diseased and healthy individuals was discovered in the 1950s. Quantifying peripheral Cf-DNA and the specific differential methylation of a group of genes have been proposed as potential non-invasive diagnostic biomarkers for somatic and constitutional genetics and for various other pathological disorders. In this study, we investigated the Cf-DNA levels of 78 young women, 38 of whom had endometriosis confirmed via laparoscopy and 40 of whom were healthy. We found a significant difference between the two groups when Cf-DNA was quantified, with 3.9 times more Cf-DNA in the serum of women with endometriosis. We also identified nine target genes potentially involved in the pathogenesis of endometriosis, with a different methylation profile between the two groups. Our data suggest that the combination of cell-free DNA quantification and the assessment of the epigenetic signature of differential methylation of nine genes can be proposed as a non-invasive predictive and diagnostic test for endometriosis.

Cadmium (Cd) is a toxic heavy metal that disrupts spermatogenesis and steroidogenesis due to its long half-life. This study evaluated the impact of recovery alone or with zinc (Zn) supplementation on Cd-induced testicular toxicity. A total of 42 pubertal male Wistar rats were divided into seven groups of six rats each. The control group (1) received NaCl (0.9%). Groups 2, 3, and 4 were treated with Cd 10 μg/kg/d by intraperitoneal injection for 1, 2, and 3 months respectively. Group 5 received Cd for 3 months with a recovery period of 1 month; Group 6 was exposed to Cd for 3 months, followed by a 1-month recovery period combined with Zn supplementation. (0.5 mg/kg/d). The last group was treated with zinc at a dose of 0.5 mg/kg/day for one month. The results showed decreased body weight, testicular and epididymal weight, testicular dimensions, and sperm parameters, along with Cd accumulation in the testes. Cd caused testicular damage and reduced serum testosterone levels, with more pronounced effects in the 3-month treatment group. Recovery alone did not significantly reverse Cd’s toxic effects, whereas Zn supplementation mitigated most of the damage. Recovery combined with Zn supplementation was more effective in correcting Cd-induced testicular toxicity than recovery alone.

Background/Objectives: Interleukin-6 (IL-6), a pleiotropic cytokine involved in immune regulation, is consistently detected in human semen, even in the absence of overt infection. Its contribution to sperm dysfunction, oxidative stress, and inflammation remains incompletely understood. This study evaluated the associations between seminal IL-6 concentrations and markers of semen quality, oxidative stress, nuclear integrity, and genital tract inflammation in infertile men. Methods: A cohort of 204 infertile men was assessed. Seminal IL-6 was quantified by electrochemiluminescence immunoassay. Semen parameters, malondialdehyde (MDA), catalase (CAT) activity, sperm DNA fragmentation index (DFI), sperm chromatin decondensation index (SDI), leukocytospermia, and bacteriospermia were measured. Analyses included correlation testing, IL-6 threshold stratification (<30, 30–60, 60–100, ≥100 pg/mL), and multivariate regression. Results: IL-6 was detectable in all samples (median: 31.52 pg/mL; range: 1.5–5000 pg/mL). Higher IL-6 levels were significantly associated with reduced sperm concentration, progressive motility, and vitality, and with increased DFI, SDI, MDA, leukocyte counts, and bacteriospermia (p < 0.001). In multivariate models, IL-6 independently predicted reduced progressive motility (β = −0.005; p = 0.032) and elevated leukocyte count (β = 0.0018; p < 0.0001). Logistic regression further showed that IL-6 increased the odds of DFI ≥ 30%, SDI ≥ 30%, and bacteriospermia (p < 0.05). Conclusions: Seminal IL-6 emerges as a sensitive biomarker of immuno-oxidative stress and sperm dysfunction in infertile men. Its integration into clinical evaluation may improve the assessment of inflammatory and oxidative contributors to male infertility.

Sperm cryopreservation provides patients undergoing oncological, surgical, or infertility treatments the opportunity to conceive their own children, using assisted reproductive technologies. However, the freezing-thawing process can negatively influence both the quantity and the quality of spermatozoa, mainly due to an excessive production of reactive oxygen species and/or an impaired antioxidant defense system in sperm. Aromatic and medicinal plants synthesize essential oils with antioxidant proprieties as a part of their ecological adaptation to environmental stress, thanks to their rich bioactive phytochemical components. This study aimed to assess sperm progressive motility, viability, plasma membrane functionality, and lipid peroxidation levels of human cryopreserved normozoospermic (n = 51) and asthenozoospermic (n = 51) samples without or with the addition of Thymus satureoides (TSEO) (20 µg/mL), Artemisia vulgaris (AVEO) (48 µg/mL), and Rosmarinus officinalis (ROEO) (13 µg/mL) essential oils. Sperm parameters were significantly better preserved with ROEO in both normozoospermic (p < 0.05) and asthenozoospermic samples (p < 0.01). In contrast, TSEO had a negative impact for both groups (p < 0.05). Meanwhile, no significant effects were observed with AVEO. In summary, the study revealed that in vitro addition of essential oils as antioxidant agents during cryopreservation can be either beneficial, which helps preserve sperm parameters and fertilizing potential, or detrimental as spermicidal agents.