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Antioxidants in Male Human and Animal Reproduction: In Vitro and In Vivo Studies

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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 June 2023) | Viewed by 16196

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Dr. Elena Moretti

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Guest Editor

Department of Molecular and Developmental Medicine, University of Siena, 53100 Siena, Italy
Interests: human spermatozoa; male infertility; oxidative stress in seminal plasma and in spermatozoa; human sperm morphology; spermatoza as a cell model in in vitro experiments

Dr. Cinzia Signorini

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Guest Editor

Department of Molecular and Developmental Medicine, University of Siena, 53100 Siena, Italy
Interests: isoprostanes; neuroprostanes; markers of oxidative stress; oxidative stress in human diseases
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Dear Colleagues,

Oxidative stress (OS) affects the quality of gametes and plays a central role in the pathogenesis of male infertility. OS is exacerbated during laboratory procedures for semen handling, such as centrifugation and cryopreservation, as well as during the protocols for assisted fertilization technologies, both in human and animal species. Treatment strategies for OS include the use of both synthetic and natural antioxidants administered in vivo and used in vitro, for example, cell supplement in culture and cryopreservation media. In the last few years, various interesting bioactive factors, such as ghrelin, obestatin, melatonin, and some adipocytokines, have demonstrated a protective antioxidant effect worthy of investigation in the field of animal and human reproduction. Studies on the effects of diets supplemented with different antioxidant molecules in animal models may represent a starting point for managing male infertility.

In vitro studies of antioxidants in human and animal sperm;
Relevance of antioxidants in semen cryopreservation and semen handling;
Antioxidants and spermatogenesis;
Antioxidant effect on human male infertility—mechanisms, molecular signaling, sperm quality;
Molecular effects of antioxidants on male reproduction—investigations in animal models;
Relevance of constitutive antioxidants in male infertility and animal models;
Antioxidant-based diet in animal model of male infertility.

Dr. Elena Moretti
Dr. Cinzia Signorini
Guest Editors

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Keywords

animal models
antioxidants
antioxidant signaling
male infertility
natural antioxidants
oxidative stress
spermatogenesis
spermatozoa
synthetic antioxidants

Published Papers (9 papers)

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Research

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22 pages, 3738 KiB

Open AccessArticle

Impact of Chromium Picolinate on Leydig Cell Steroidogenesis and Antioxidant Balance Using an In Vitro Insulin Resistance Model

by Rúben Moreira, Ana D. Martins, Rita Ferreira, Marco G. Alves, Maria de Lourdes Pereira and Pedro F. Oliveira

Antioxidants 2024, 13(1), 40; https://doi.org/10.3390/antiox13010040 - 23 Dec 2023

Cited by 1 | Viewed by 1113

Abstract

Leydig cells (LCs) play a pivotal role in male fertility, producing testosterone. Chromium (III) picolinate (CrPic₃), a contentious supplement with antidiabetic and antioxidant properties, raises concerns regarding male fertility. Using a rodent LC line, we investigated the cytotoxicity of increasing CrPic₃ doses. An insulin resistance (IR) model was established using palmitate (PA), and LCs were further exposed to CrPic₃ to assess its antioxidant/antidiabetic activities. An exometabolome analysis was performed using ¹H-NMR. Mitochondrial function and oxidative stress were evaluated via immunoblot. Steroidogenesis was assessed by quantifying androstenedione through ELISA. Our results uncover the toxic effects of CrPic₃ on LCs even at low doses under IR conditions. Furthermore, even under these IR conditions, CrPic₃ fails to enhance glucose consumption but restores the expression of mitochondrial complexes CII and CIII, alleviating oxidative stress in LCs. While baseline androgen production remained unaffected, CrPic₃ promoted androstenedione production in LCs in the presence of PA, suggesting that it promotes cholesterol conversion into androgenic intermediates in this context. This study highlights the need for caution with CrPic₃ even at lower doses. It provides valuable insights into the intricate factors influencing LCs metabolism and antioxidant defenses, shedding light on potential benefits and risks of CrPic₃, particularly in IR conditions. Full article

(This article belongs to the Special Issue Antioxidants in Male Human and Animal Reproduction: In Vitro and In Vivo Studies)

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19 pages, 2644 KiB

Open AccessArticle

Goji Berry in the Diet of the Rabbit Buck: Effects on Semen Quality, Oxidative Status and Histological Features of the Reproductive Tract

by Gabriele Brecchia, Gerald Muça, Albana Munga, Laura Menchetti, Livio Galosi, Giacomo Rossi, Olimpia Barbato, Grazia Pastorelli, Stella Agradi, Valentina Serra, Majlind Sulçe, Enkeleda Ozuni, Luigj Turmalaj, Marta Castrica, Maria Rachele Ceccarini, Federica Riva, Bernard Fioretti, Alda Quattrone, Maria Laura Marongiu and Giulio Curone

Antioxidants 2023, 12(11), 1959; https://doi.org/10.3390/antiox12111959 - 02 Nov 2023

Viewed by 1517

Abstract

Goji berry (GB) shows beneficial effects on human health, although its effects on the male rabbit have been little investigated. This study examines the impact of GB dietary supplementation on the semen traits, antioxidant capacity of seminal plasma, and histological features of the reproductive tract of rabbit buck. Eighteen rabbits were distributed into two dietary groups: one receiving a commercial feed (Control), and the other a feed supplemented with 1% of GB (Goji). After a nutritional adaptation period of 60 days, the animals were subjected to semen collection every 15 days. The semen traits, libido, antioxidant, and inflammatory parameters were collected and analyzed. The rabbits were sacrificed after 60 days, and tissues of the genital tract were analyzed. Compared to the Control group, the Goji group showed higher spermatozoa concentration, motility, and vitality (p < 0.05), as well as fewer abnormal spermatozoa and a higher libido (p < 0.1). Histological features such as functional activity and hyperplasia were improved by GB and correlated with some semen traits (p < 0.05). Conversely, antioxidant and anti-inflammatory parameters were unaffected by the diet. These findings suggest that GB acts on the tissues of the reproductive tract positively influencing semen quality, although further studies are needed to understand the effect on oxidative stress. Full article

(This article belongs to the Special Issue Antioxidants in Male Human and Animal Reproduction: In Vitro and In Vivo Studies)

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16 pages, 1336 KiB

Open AccessArticle

Pro-Atherogenic and Pro-Oxidant Diets Influence Semen and Blood Traits of Rabbit Bucks

by Simona Mattioli, Elisa Angelucci, Alessandro Dal Bosco, Cinzia Signorini, Lakamy Sylla, Luigia Bosa, Giulia Collodel, Thierry Durand, Jean-Marie Galano, Camille Oger and Cesare Castellini

Antioxidants 2023, 12(10), 1880; https://doi.org/10.3390/antiox12101880 - 19 Oct 2023

Viewed by 857

Abstract

Many dietary factors can affect sperm traits. We compared the effect of diets rich in pro-oxidant (flaxseed oil) and pro-atherogenic (coconut oil) substances without added antioxidants on semen traits, using the rabbit as an animal model. Thirty rabbit bucks (8 months old) were fed three diets for 150 days: CNT (control) a standard diet; HA (high-atherogenic) standard diet + 3% coconut oil, and HO (high-oxidizing) standard diet + 3% flaxseed oil. Semen samples were collected weekly for the evaluation of qualitative traits (kinetics, viability) and the oxidative damage (MDA and cytokines). Blood was collected at the start (T0) and end (T8) of the experimental period for the assessment of the oxidative damage (MDA and isoprostanoids), lipid profile, and testosterone. A worsening of sperm kinetics and viability was recorded in the HA group. Lipid oxidation in seminal plasma, as well as isoprostanoids in blood (F₃-IsoPs and F₄-NeuroPs), increased in both the HO and HA groups. A high level of TNF-α, a marker of inflammatory status, was recorded in the seminal plasma of the HA group. The resulting outcomes were mainly attributable to the different fatty acid profiles (SFA vs. PUFA) of the diets, which modulated an inflammatory/oxidative response. Full article

(This article belongs to the Special Issue Antioxidants in Male Human and Animal Reproduction: In Vitro and In Vivo Studies)

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14 pages, 2164 KiB

Open AccessArticle

Loss of Nuclear/DNA Integrity in Mouse Epididymal Spermatozoa after Short-Term Exposure to Low Doses of Dibutyl Phthalate or Bisphenol AF and Its Mitigation by Oral Antioxidant Supplementation

by Elisa Hug, Pauline Villeneuve, Stephanie Bravard, Areski Chorfa, Christelle Damon-Soubeyrand, Stephen G. Somkuti, Aron Moazamian, R. John Aitken, Parviz Gharagozloo, Joël R. Drevet and Fabrice Saez

Antioxidants 2023, 12(5), 1046; https://doi.org/10.3390/antiox12051046 - 05 May 2023

Cited by 1 | Viewed by 2328

Abstract

Routine exposure to chemicals omnipresent in the environment, particularly the so-called endocrine-disrupting chemicals (EDCs), has been associated with decreased sperm quality and increased anomalies in testis. The decline in semen quality and testicular abnormalities have been attributed to the disruption of endocrine signaling as well as oxidative stress. The present study set out to examine the effect of short-term exposure of two common EDCs widely used in the plastic industry: Dibutyl Phthalate (DBP) and Bisphenol AF (BPAF). Our research objective was to focus on the post-testicular compartment of the epididymis, where spermatozoa acquire their functional capacity and are stored. The data obtained indicated no significant effect for either chemicals on sperm viability, motility or acrosome integrity. Neither of the EDCs had a noticeable effect on the structures of the testis and epididymis. However, substantial impact on the integrity of the sperm nucleus and DNA structure was evidenced by a significant increase in nuclear decondensation and DNA base oxidation. The damage observed was postulated to arise from the pro-oxidant properties of the EDCs generating excess of reactive oxygen species (ROS) and triggering a state of oxidative stress. This hypothesis was confirmed when the observed damage was largely blocked by co-administering EDCs with an evidenced-based antioxidant formulation. Full article

(This article belongs to the Special Issue Antioxidants in Male Human and Animal Reproduction: In Vitro and In Vivo Studies)

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28 pages, 18469 KiB

Open AccessArticle

Entomotherapeutic Role of Periplaneta americana Extract in Alleviating Aluminum Oxide Nanoparticles-Induced Testicular Oxidative Impairment in Migratory Locusts (Locusta migratoria) as an Ecotoxicological Model

by Esraa A. Arafat, Doaa S. El-Sayed, Hussein K. Hussein, Justin Flaven-Pouchon, Bernard Moussian, Lamia M. El-Samad, Abeer El Wakil and Mohamed A. Hassan

Antioxidants 2023, 12(3), 653; https://doi.org/10.3390/antiox12030653 - 06 Mar 2023

Cited by 15 | Viewed by 2052

Abstract

In this study, we shed light for the first time on the usage of migratory locusts (Locusta migratoria) as an insect model to investigate the nanotoxicological influence of aluminum oxide (Al₂O₃) nanoparticles at low doses on testes, and evaluate the capacity of a whole-body extract of American cockroaches (Periplaneta americana) (PAE) to attenuate Al₂O₃ NPs-induced toxicity. Energy dispersive X-ray microanalyzer (EDX) analysis verified the bioaccumulation of Al in testicular tissues due to its liberation from Al₂O₃ NPs, implying their penetration into the blood–testis barrier. Remarkably, toxicity with Al engendered disorders of antioxidant and stress biomarkers associated with substantial DNA damage and cell apoptosis. Furthermore, histopathological and ultrastructural analyses manifested significant aberrations in the testicular tissues from the group exposed to Al₂O₃ NPs, indicating the overproduction of reactive oxygen species (ROS). Molecular docking analysis emphasized the antioxidant capacity of some compounds derived from PAE. Thus, pretreatment with PAE counteracted the detrimental effects of Al in the testes, revealing antioxidant properties and thwarting DNA impairment and cell apoptosis. Moreover, histological and ultrastructural examinations revealed no anomalies in the testes. Overall, these findings substantiate the potential applications of PAE in preventing the testicular impairment of L. migratoria and the conceivable utilization of locusts for nanotoxicology studies. Full article

(This article belongs to the Special Issue Antioxidants in Male Human and Animal Reproduction: In Vitro and In Vivo Studies)

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13 pages, 1675 KiB

Open AccessArticle

Blood Leukocyte ROS Production Reflects Seminal Fluid Oxidative Stress and Spermatozoa Dysfunction in Idiopathic Infertile Men

by Matteo Becatti, Gianmartin Cito, Flavia Rita Argento, Eleonora Fini, Alessandra Bettiol, Serena Borghi, Amanda Mannucci, Rossella Fucci, Claudia Giachini, Rita Picone, Giacomo Emmi, Niccolò Taddei, Maria Elisabetta Coccia and Claudia Fiorillo

Antioxidants 2023, 12(2), 479; https://doi.org/10.3390/antiox12020479 - 14 Feb 2023

Cited by 4 | Viewed by 1633

Abstract

A large proportion of infertile men do not receive a clear diagnosis, being considered as idiopathic or unexplained cases due to infertility diagnosis based on standard semen parameters. Particularly in unexplained cases, the search for new indicators seems mandatory to provide specific information. In the etiopathogenesis of male infertility oxidative stress displays important roles by negatively affecting sperm quality and function. In this study, performed in a population of 34 idiopathic infertile men and in 52 age-matched controls, redox parameters were assessed in blood, leukocytes, spermatozoa, and seminal fluid and related to semen parameters. The main findings indicate that blood oxidative stress markers reflect seminal oxidative stress. Interestingly, blood leukocyte ROS production was significantly correlated to sperm ROS production and to semen parameters. Overall, these results suggest the potential employ of blood redox markers as a relevant and adjunctive tool for sperm quality evaluation aimed to preconception care. Full article

(This article belongs to the Special Issue Antioxidants in Male Human and Animal Reproduction: In Vitro and In Vivo Studies)

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16 pages, 1426 KiB

Open AccessArticle

Male Infertility Coexists with Decreased Sperm Genomic Integrity and Oxidative Stress in Semen Irrespective of Leukocytospermia

by Kamil Gill, Tomasz Machalowski, Patryk Harasny, Michal Kups, Marta Grabowska, Ewa Duchnik, Olimpia Sipak, Monika Fraczek, Maciej Kurpisz, Rafal Kurzawa and Malgorzata Piasecka

Antioxidants 2022, 11(10), 1987; https://doi.org/10.3390/antiox11101987 - 05 Oct 2022

Cited by 6 | Viewed by 2090

Abstract

Our research was designed to verify the relationship between male infertility, basic semen characteristics (with respect to detailed sperm morphology), sperm DNA fragmentation (SDF), oxidation-reduction potential in semen (ORP), and leukocytospermia. The obtained results showed that infertile groups (with or without leukocytospermia) had significantly lower basic semen characteristics and higher SDF, raw ORP, and static ORP (sORP) than fertile controls. The thresholds of 13% SDF (AUC = 0.733) and 1.40 sORP (AUC = 0.857) were predictive values for discriminating infertile from fertile men. In infertile groups, a higher prevalence and risk for >13% SDF and >1.40 sORP were revealed. Unexpectedly, leukocytospermic subjects had lower sORP, prevalence, and risk for >1.40 sORP than leukocytospermic-negative men. These groups did not differ in SDF and raw ORP. Both SDF and sORP negatively correlated with basic semen parameters but positively correlated with sperm head and midpiece defects. sORP positively correlated with sperm tail defects, immature sperm cells with excess residual cytoplasm, and SDF. In turn, raw ORP negatively correlated with sperm count but positively correlated with SDF and sORP. These findings indicate that (1) there is a relationship between male infertility, SDF, and OS in semen; (2) in infertile men, there is a clinically significant risk of SDF and OS irrespective of leukocytospermia; and (3) the assessment of SDF and oxidative stress should be independent of leukocytospermia. Full article

(This article belongs to the Special Issue Antioxidants in Male Human and Animal Reproduction: In Vitro and In Vivo Studies)

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Review

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17 pages, 1662 KiB

Open AccessReview

On Males, Antioxidants and Infertility (MOXI): Certitudes, Uncertainties and Trends

by Manuel Alfaro Gómez, María del Rocío Fernández-Santos, Alejandro Jurado-Campos, Pedro Javier Soria-Meneses, Vidal Montoro Angulo, Ana Josefa Soler, José Julián Garde and Virginia Rodríguez-Robledo

Antioxidants 2023, 12(8), 1626; https://doi.org/10.3390/antiox12081626 - 17 Aug 2023

Cited by 3 | Viewed by 1457

Abstract

Male infertility (MI) involves various endogenous and exogenous facts. These include oxidative stress (OS), which is known to alter several physiological pathways and it is estimated to be present at high levels in up to 80% of infertile men. That is why since the late 20th century, the relationship between OS and MI has been widely studied. New terms have emerged, such as Male Oxidative Stress Infertility (MOSI), which is proposed as a new category to define infertile men with high OS levels. Another important term is MOXI: Male, Antioxidants, and Infertility. This term refers to the hypothesis that antioxidants could improve male fertility without the use of assisted reproductive technology. However, there are no evidence-based antioxidant treatments that directly improve seminal parameters or birth ratio. In this regard, there is controversy about their use. While certain scientists argue against their use due to the lack of results, others support this use because of their safety profile and low price. Some uncertainties related to the use of antioxidants for treating MI are their questionable efficacy or the difficulties in knowing their correct dosage. In addition, the lack of quality methods for OS detection can lead to excessive antioxidant supplementation, resulting in “reductive stress”. Another important problem is that, although the inflammatory process is interdependent and closely linked to OS, it is usually ignored. To solve these uncertainties, new trends have recently emerged. These include the use of molecules with anti-inflammatory and antioxidant potential, which are also able to specifically target the reproductive tissue; as well as the use of new methods that allow for reliable quantification of OS and a quality diagnosis. This review aims to elucidate the main uncertainties about MOXI and to outline the latest trends in research to develop effective therapies with clinically relevant outcomes. Full article

(This article belongs to the Special Issue Antioxidants in Male Human and Animal Reproduction: In Vitro and In Vivo Studies)

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25 pages, 1522 KiB

Open AccessReview

Human Sperm as an In Vitro Model to Assess the Efficacy of Antioxidant Supplements during Sperm Handling: A Narrative Review

by Elena Moretti, Cinzia Signorini, Roberta Corsaro, Maria Giamalidi and Giulia Collodel

Antioxidants 2023, 12(5), 1098; https://doi.org/10.3390/antiox12051098 - 15 May 2023

Cited by 5 | Viewed by 2306

Abstract

Spermatozoa are highly differentiated cells that produce reactive oxygen species (ROS) due to aerobic metabolism. Below a certain threshold, ROS are important in signal transduction pathways and cellular physiological processes, whereas ROS overproduction damages spermatozoa. Sperm manipulation and preparation protocols during assisted reproductive procedures—for example, cryopreservation—can result in excessive ROS production, exposing these cells to oxidative damage. Thus, antioxidants are a relevant topic in sperm quality. This narrative review focuses on human spermatozoa as an in vitro model to study which antioxidants can be used to supplement media. The review comprises a brief presentation of the human sperm structure, a general overview of the main items of reduction–oxidation homeostasis and the ambivalent relationship between spermatozoa and ROS. The main body of the paper deals with studies in which human sperm have been used as an in vitro model to test antioxidant compounds, including natural extracts. The presence and the synergic effects of different antioxidant molecules could potentially lead to more effective products in vitro and, in the future, in vivo. Full article

(This article belongs to the Special Issue Antioxidants in Male Human and Animal Reproduction: In Vitro and In Vivo Studies)

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