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Antioxidants
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Oxidative Stress and Sperm: Technical, Biological and Clinical Aspects—2nd Edition
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Oxidative Stress and Sperm: Technical, Biological and Clinical Aspects—2nd Edition

A special issue of Antioxidants (ISSN 2076-3921). This special issue belongs to the section "Health Outcomes of Antioxidants and Oxidative Stress".

Deadline for manuscript submissions: closed (30 April 2026) | Viewed by 4635

Special Issue Editor

Dr. Monica Muratori

E-Mail Website
Guest Editor
Department of Experimental and Clinical Medicine, University of Florence, Viale Pieraccini, 6, 50139 Florence, Italy
Interests: male infertility; sperm biology; oxidative stress; sperm DNA fragmentation; semen analysis; flow cytometry; nutrition; environmental pollution
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

Oxidative stress appears to be an underlying cause for many cases of male infertility, as it could be the converging step for several environmental, biological and lifestyle factors impacting sperm formation and function. Induction of reactive oxygen species production is detrimental during in vitro sperm manipulation, including selection for oocyte insemination and semen cryopreservation, posing a potential risk to couples treated with assisted reproductive techniques. Oxidative stress is one of the main mechanisms responsible for sperm DNA damage, including DNA fragmentation, a genome anomaly negatively affecting both natural and assisted reproduction. In addition, emerging data suggest that oxidative stress could alter the sperm epigenome. Both genetic and epigenetic damage has the potential to impact not only male reproductive function but also embryo development and the health of offspring.

In this context, further research in this field appears to be of the upmost importance. However, despite a huge number of published studies, there are many areas that remain little explored, including, but not limited to, the following: reliable techniques for revealing oxidative stress; the biological mechanisms of inducing genetic and epigenetic sperm damages; and the clinical meaning and use of the knowledge in this field. The poor clinical results obtained up to now for in vitro and/or in vitro treatment with antioxidant compounds further underline the need for deeper knowledge on this topic.

Dr. Monica Muratori
Guest Editor

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Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Antioxidants is an international peer-reviewed open access monthly journal published by MDPI.

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Keywords

  • male infertility
  • ROS
  • oxidative sperm DNA damage
  • sperm DNA fragmentation
  • sperm epigenetic damage
  • assisted reproductive techniques
  • in vitro sperm manipulation
  • antioxidants
  • embryo development
  • pregnancy

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Published Papers (4 papers)

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16 pages, 2923 KB  
Article
Functional and Molecular Characterization of Melamine-Induced Disruption of Human Spermatozoa via Oxidative Stress and Apoptotic Pathways: An In Vitro Study
by Francesca Paola Luongo, Eugenia Annunzi, Rosetta Ponchia, Francesca Girolamo, Giuseppe Morgante, Paola Piomboni and Alice Luddi
Antioxidants 2026, 15(1), 122; https://doi.org/10.3390/antiox15010122 - 17 Jan 2026
Viewed by 680
Abstract
Melamine, a nitrogen-rich industrial chemical, has raised increasing concern as an emerging environmental contaminant with potential reproductive toxicity. While its nephrotoxic effects are well established, the direct impact of melamine on human sperm remains poorly defined. In this study, we investigated the in [...] Read more.
Melamine, a nitrogen-rich industrial chemical, has raised increasing concern as an emerging environmental contaminant with potential reproductive toxicity. While its nephrotoxic effects are well established, the direct impact of melamine on human sperm remains poorly defined. In this study, we investigated the in vitro effects of melamine on human sperm, under both capacitating and non-capacitating conditions. Functional analyses revealed that the exposure to 0.8 mM melamine, the highest non-cytotoxic concentration in vitro, significantly compromised sperm motility and disrupted key capacitation processes, including tyrosine phosphorylation patterns, cholesterol efflux, and the acrosome reaction. Molecular assessments demonstrated melamine-induced mitochondrial dysfunction, characterized by COX4I1 downregulation, reduced mitochondrial membrane potential, and altered reactive oxygen species production. In parallel, gene expression analyses revealed the activation of apoptotic pathways, with the upregulation of BAX and downregulation of BCL2, changes that were more pronounced during capacitation. Furthermore, melamine exposure significantly increased sperm DNA fragmentation and denaturation, indicating genotoxic stress. Collectively, these findings demonstrate that even low, non-cytotoxic concentrations of melamine compromise sperm function by disrupting capacitation, mitochondrial activity, and genomic integrity. This study identifies capacitation as a critical window of vulnerability and underscores the need to consider melamine as a potential environmental risk factor for male reproductive health. Full article
(This article belongs to the Special Issue Oxidative Stress and Sperm: Technical, Biological and Clinical Aspects—2nd Edition)
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21 pages, 2012 KB  
Article
Protective Effect of Raphanus sativus Seed Extract on Damage Induced by In Vitro Incubation and Cryopreservation of Human Spermatozoa
by Oumaima Ammar, Costanza Calamai, Mariachiara Marino, Elisabetta Baldi, Mario Maggi, Linda Vignozzi, Meriem Mehdi, Nadia Mulinacci and Monica Muratori
Antioxidants 2026, 15(1), 74; https://doi.org/10.3390/antiox15010074 - 6 Jan 2026
Viewed by 798
Abstract
In vitro manipulation of human spermatozoa during Assisted Reproductive Technology (ART) can induce several damages to sperm structure and functions. This study investigated the protective effects of Raphanus sativus seed extract and its active compounds on several sperm parameters during in vitro incubation [...] Read more.
In vitro manipulation of human spermatozoa during Assisted Reproductive Technology (ART) can induce several damages to sperm structure and functions. This study investigated the protective effects of Raphanus sativus seed extract and its active compounds on several sperm parameters during in vitro incubation and cryopreservation. Extracts from five seed-batches were characterized by HPLC-DAD-MS and 1H-NMR, identifying sinapine and sinipic glycosides as the main characteristic compounds. Sperm DNA fragmentation (sDF) was detected by the Sperm Chromatin Dispersion test and LiveTUNEL. Excessive reactive oxygen species (ROS) production was detected by MitoSOX Red in viable spermatozoa. Caspase activity was detected by FLICA. Cryopreservation was conducted with two alternative freezing media. In vitro incubation with the extract protected against the loss of motility and reduced the induction of sDF, sperm ROS production, and caspase activity. Similarly, during cryopreservation, it allowed much better recoveries of sperm viability, motility, and DNA integrity by decreasing sperm ROS production with both freezing media. Sinapine and sinapic acid completely mimicked the protective effects of the whole extract during both in vitro incubation and cryopreservation, suggesting that they are included among the active principles. These findings support Raphanus sativus seed extract and its active compounds as candidates for inclusion in handling and freezing media for human spermatozoa in ART. Full article
(This article belongs to the Special Issue Oxidative Stress and Sperm: Technical, Biological and Clinical Aspects—2nd Edition)
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14 pages, 2079 KB  
Article
Determination of the Antioxidant Capacity of Human Seminal Fluid Using a Fast and Accurate Electrochemical Approach
by Youssef Hibaoui, Slim Khedhri, Dorothea Wunder, Josefina Vargas, Alexandre Vallée, Jean-Marc Ayoubi and Anis Feki
Antioxidants 2026, 15(1), 35; https://doi.org/10.3390/antiox15010035 - 26 Dec 2025
Cited by 2 | Viewed by 1047
Abstract
Infertility affects around 10–15% of couples worldwide, out of which male factor contributes to 30–50% of cases of infertility. Oxidative stress, which corresponds to an imbalance between antioxidant capacities and reactive oxygen species, is considered a leading cause of male infertility. Therefore, the [...] Read more.
Infertility affects around 10–15% of couples worldwide, out of which male factor contributes to 30–50% of cases of infertility. Oxidative stress, which corresponds to an imbalance between antioxidant capacities and reactive oxygen species, is considered a leading cause of male infertility. Therefore, the ability to monitor antioxidant capacity in seminal fluid is critical as it sustains free radical balance in the sperm. Most currently available methods to assess antioxidant capacity in seminal fluid are time-consuming, require specialized equipment, or are not easily implemented in clinical routine practice. Here, we evaluate the applicability of an electrochemical approach to determine the antioxidant capacity of human seminal fluid. We show that the results of this electrochemical approach are comparable to those of two reference methods for evaluating free radical scavenging activity, namely 2,20-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH), when measuring the antioxidant capacity of seminal plasma or antioxidant molecules such as 6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid (Trolox), ascorbic acid, and uric acid. Furthermore, we demonstrate the applicability of the method for the assessment of the antioxidant capacity of seminal fluid isolated from 30 normozoospermic patients (528.2 ± 142 nW). Further analysis demonstrates a positive correlation between the antioxidant capacity measured through the electrochemical approach and sperm concentration. Overall, this electrochemical approach provides a fast and accurate assessment of total antioxidant capacity in human seminal fluid. It may be implemented as a complementary tool in the routine evaluation of male infertility. Full article
(This article belongs to the Special Issue Oxidative Stress and Sperm: Technical, Biological and Clinical Aspects—2nd Edition)
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12 pages, 1176 KB  
Article
Effect of Different Extenders on the Oxidative Status and Fertility of Sarda Ram Liquid Semen Stored at 15 °C
by Valeria Pasciu, Charbel Nassif, Maria Dattena, Sara Succu, Francesca Daniela Sotgiu, Antonello Cannas, Ignazio Cossu, Elena Baralla, Fabrizio Chessa, Fiammetta Berlinguer and Laura Mara
Antioxidants 2025, 14(8), 932; https://doi.org/10.3390/antiox14080932 - 30 Jul 2025
Cited by 1 | Viewed by 1302
Abstract
Liquid storage is an important tool used to prolong fresh semen shelf-life while protecting spermatozoa from damage, conserving their overall functionality, and ensuring better fertility than frozen semen from sheep. The increased production of reactive oxygen species (ROS) during sperm storage leads to [...] Read more.
Liquid storage is an important tool used to prolong fresh semen shelf-life while protecting spermatozoa from damage, conserving their overall functionality, and ensuring better fertility than frozen semen from sheep. The increased production of reactive oxygen species (ROS) during sperm storage leads to a decline in sperm quality, particularly with regard to sperm nuclear DNA damage and mitochondrial membrane potential (MMP). This study evaluated the effect of storing Sarda ram semen at 15 °C for 7 h on its redox status, motility, morphology, acrosome integrity, ATP content, mitochondrial potential membrane, and in vivo fertility after artificial insemination. Two different extenders were compared: a lab-made skimmed milk (SM)-based extender and a commercial extender (OviXcell®, IMV-Technologies, France). Lower ROS levels in the SM (p < 0.001) indicated that its oxidative status was better maintained compared to the commercial extender (CE). Antioxidant defenses (total antioxidant capacity, TEAC; superoxide dismutase, SOD; total thiols) were higher in the SM (p < 0.01) than in the CE. SM also had higher MMP (p < 0.05), acrosome integrity (p < 0.05), ATP content (p < 0.01), and in vivo fertilizing capacity (p < 0.05) compared to the CE, which indicated higher semen quality. In conclusion, the SM extender, while maintaining a better oxidative/antioxidant balance, ensured higher semen quality after 7 h of storage at 15 °C in vitro compared to the CE. Full article
(This article belongs to the Special Issue Oxidative Stress and Sperm: Technical, Biological and Clinical Aspects—2nd Edition)
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