Search Results (262)

Infertility is a significant global health concern, and incorporating antioxidants into sperm preparation media is one strategy to enhance sperm quality and decrease infertility rates. This study aimed to investigate the phytochemical compounds of red cotton stamen extracts and their effects as antioxidants in improving the quality of bull frozen semen. Among the extracts, RCU contained the highest levels of total phenolics, total tannins, and total monomeric anthocyanins along with the strongest ABTS free radical scavenging activity and protein denaturation inhibition. Exposing sperm to FeSO₄-induced oxidative stress resulted in significantly reduced motility, viability, and normal morphology. However, treatment with RCD, RCU, and RCM improved these parameters. Additionally, the FeSO₄-induced group showed elevated levels of reactive oxygen species (ROS) and advanced glycation end products (AGEs) compared to the normal control, whereas all red cotton stamen extracts effectively reduced these levels. In conclusion, red cotton stamen extracts, rich in phenolic bioactive compounds, demonstrated strong free radical scavenging capacity and improved sperm motility, viability, and morphology by neutralizing free radicals and enhancing antioxidant defenses. These findings suggest that the red cotton stamen extracts, particularly RCD and RCU, offer benefits for sperm preservation. Full article

The conservation of small and genetically vulnerable brown bear populations, such as the Cantabrian subpopulation in Spain, depends on developing species-specific assisted reproductive technologies and genetic resource banks. However, the lack of standardized sperm collection and cryopreservation protocols hinders their application. This study provides the first comparative analysis of sperm quality and proteomic profiles from three different origins: epididymal, pre-ejaculated, and ejaculated. Sperm quality parameters —motility and kinetic, viability, apoptosis, and oxidative stress— and protein expression were assessed. Although yields were similar, marked differences were observed in sperm quality and protein profiles. Sixty-three proteins involved in metabolism, stress response, and oxidative balance were differentially expressed depending on sperm origin. Epididymal sperm showed the highest viability and motility, lowest apoptosis, and a proteomic profile indicative of active spermatogenesis and enhanced oxidative stress defense. In contrast, ejaculated sperm had increased oxidative stress and reduced expression of metabolic proteins, while pre-ejaculated sperm exhibited lower motility, likely due to urine contamination and mitochondrial protein alterations, despite comparable viability and apoptosis. These findings offer novel insights into brown bear sperm biology and highlight the importance of sperm origin in developing optimized assisted reproduction strategies, ultimately supporting ex situ conservation efforts for this species. Full article

Sperm membrane stability is a key factor in determining sperm viability and fertilization capability, with broad implications ranging from basic reproductive biology to livestock breeding practices. This comprehensive review examines the structural and functional mechanisms underlying sperm membrane integrity, including defensive barrier functions, potentiometric ion channel regulation, and motility modulation that collectively optimize sperm survival, motility, and fertilization potential. Environmental factors such as temperature fluctuations, abnormal pH levels (outside the optimal 7.2–8.2 range), pathological conditions, and hormonal imbalances can compromise membrane stability by inducing oxidative stress and protein denaturation. Key regulatory proteins, notably NPC2 for cholesterol homeostasis, Flotillin proteins for lipid raft organization, and Annexin V for membrane repair mechanisms, demonstrate essential roles in maintaining structural integrity. In livestock reproduction, membrane stability research facilitates the optimization of cryoprotectant formulations and freezing protocols, resulting in 15–25% improvements in post-thaw sperm survival rates and enhanced artificial insemination success. These findings provide valuable insights for advancing assisted reproductive technologies and improving reproductive efficiency in animal husbandry. Full article

Sexing procedures and subsequent freezing still impact sperm cells, leading to decreased fertility of gender-ablated semen. This study aimed to enhance cryo-survivability and reduce the capacitation-like change rate of gender-ablated semen by adding 2 mg of cholesterol-loaded cyclodextrin (CLC) per mL of extended semen containing 67 × 10⁶ sperm cells. This marks the first use of CLC with gender-ablated semen. Semen from four Jersey bulls was used for this study. Viability, motility, and mitochondrial activity were evaluated and adjusted to account for the inactivation of undesired sex sperm cells during processing. Binding ability to oviduct cells, fertilizing ability, and acrosome status were also evaluated. Adding CLC did not increase sperm motility. The population with intact membranes and acrosomes was significantly increased (p < 0.05) from 28.9 ± 1.2% to 34.1 ± 1.2% in the CLC-treated group. Mitochondrial potential, capacitation status at the membrane, calcium levels, and binding ability to oviduct cells were maintained. CLC treatment did not delay capacitation while significantly improving fertilization rates after 8 and 12 h of co-incubation (77 ± 3% vs. 67 ± 3% and 82 ± 3% vs. 74 ± 3%, respectively; p < 0.05). In conclusion, CLC addition significantly improved gender-ablated post-thaw sperm viability, acrosome integrity, and fertilizing ability while preserving motility, capacitation progress, and binding ability to oviduct cells. Full article

Background: Doxorubicin (DOX) is a potent anti-cancer agent that is widely described in cancer treatment. However, its administration is often limited by its adverse effects, particularly its testicular toxicity, which can induce infertility in male patients. DOX-induced testicular damage is due to oxidative stress, apoptosis, and inflammation. Nanocurcumin (NCR) is a nano-formulated edition of curcumin with a higher therapeutic potential. NCR has demonstrated antioxidant and anti-inflammatory properties. Methods: This study is designed to inspect the potential validity of NCR on DOX-induced testicular damage in male rats. We used thirty-two Wistar albino rats (150–200 g) and divided them into four groups. NCR (80 mg/kg/ dissolved in 1% CMC) was given orally by oral gavage for 14 days. A single dose of DOX (15 mg/kg) (i.p.) was injected on the 7th day of the experiment. Results: DOX treatment reduced the sperm viability and motility rate, cellular antioxidants, and gonadal hormones; it led to higher levels of inflammatory mediators, necrosis, and sloughing in seminiferous tubules. Conversely, NCR treatment significantly alleviated these side effects by improving sperm count/motility and reducing sperm abnormalities. The testicular function recovery was likely driven by stimulating the cytoprotective SIRT1/NF-κB pathway, depressing the testicular level of oxidative indicators such as MDA, TNF-α, iNOS, IL-1β, and NO, and increasing levels of antioxidants such as GSH and SOD. In addition, NCR contradicted the apoptotic changes by downregulating the pro-apoptotic signals Bax and caspase-3, while inducing Bcl-2 upregulation. Moreover, NCR increased levels of gonadal hormones, attenuated histological abnormalities, and preserved testicular structure when compared with the DOX group. Conclusions: NCR treatment can effectively ameliorate DOX-induced testicular toxicity. Full article

Artificial insemination in goats commonly relies on refrigerated semen doses, yet the optimal energetic substrate to support sperm metabolism remains unclear. This study aimed to evaluate the effects of different energetic substrates on goat buck sperm metabolism and motility when refrigerated at 17 °C. Semen from six Murciano-Granadina male goats were collected and diluted in PBS supplemented with 35 mM of either glucose, fructose, pyruvate, or lactate in the first experiment. In the second experiment, the effects of varying concentrations of pyruvate and/or glucose, NaCl supplementation, and the osmolarity on sperm quality parameters were assessed. Semen was stored at 17 °C for 48 h and evaluated for motility using the CASA system, as well as for viability, mitochondrial membrane potential, and mitochondrial ROS by flow cytometry. The results show that pyruvate and lactate extenders outperformed the others, preserving higher total motility, progressivity, and viability of spermatozoa over 48 h, even at a concentration lower than 35 mM, as in the case of pyruvate. In contrast, glucose had a detrimental effect on sperm quality, reducing viability and healthy population rates while increasing motility, especially at higher concentrations. NaCl supplementation and osmolarity had no significant effect on any of the sperm quality parameters. In conclusion, pyruvate maintains a higher quality and motility of sperm stored at 17 °C in PBS in comparison with a glucose-supplemented extender. Full article

The physiological functions of mammalian sperm, such as motility, hyperactivation, and capacitation, require substantial energy. This study investigates the effects of two classic cryopreservation extenders—TCG (tris-citrate-glucose) and LEY (lactose-egg yolk)—on the energy metabolism of frozen–thawed boar semen. By comparing the quality indicators, key metabolite levels, and the activities of critical enzymes involved in glycolysis and the tricarboxylic acid cycle, we aim to understand how these different semen extenders influence the spermatozoa vitality of frozen–thawed boar semen. Following thawing, the LEY-cryopreserved sperm demonstrated significantly elevated motility parameters (viability, VCL, VSL, and VAP) and enhanced plasma membrane and acrosomal integrity compared with the TCG group (p < 0.05), though both cryopreserved groups exhibited significantly reduced performance relative to fresh semen controls. Cryopreservation markedly reduced intracellular adenosine triphosphate (ATP), pyruvate, and acetyl coenzyme A (A-CoA) levels (fresh > LEY > TCG; p < 0.05). The LEY-preserved spermatozoa retained higher activities of glycolysis-related enzymes (phosphofructokinase, PFK; pyruvate kinase, PK) compared with the TCG group, which, in turn, showed elevated lactate dehydrogenase (LDH) activity. Critically, TCG-suppressed pyruvate dehydrogenase (PDH) activity (p < 0.05) coincided with diminished A-CoA, indicating impaired mitochondrial oxidative phosphorylation. These results demonstrate LEY’s superior preservation of motility and membrane stability but highlight cryodamage-induced energy metabolism dysregulation, particularly TCG’s disruption of the glycolysis–TCA cycle coordination essential for spermatozoa function. In conclusion, the choice of semen extender has a significant impact on the energy metabolism and overall quality of frozen–thawed semen, highlighting the importance of optimizing cryopreservation protocols for improved spermatozoa viability and functionality. Full article

Sperm cryopreservation is a fundamental reproductive biotechnology, enabling the long-term storage of genetic material and supporting assisted reproduction programs. Despite its widespread application, the process is associated with significant limitations due to the cryo-induced cellular damage that occurs during freezing and thawing. These injuries primarily affect the plasma membrane, nuclear DNA, and motility, thereby compromising the fertilizing potential of spermatozoa. Furthermore, interspecies variability in terms of cryo-sensitivity poses a major challenge to the development of standardized cryopreservation protocols. Recent advances have focused on mitigating cryodamage through the use of various strategies. The inclusion of antioxidants in cryopreservation media has proven effective in reducing oxidative stress, thereby enhancing cellular protection. Similarly, the addition of lipid-based supplements contributes to membrane stabilization, improving post-thaw sperm viability and functionality. Moreover, the application of omics technologies, such as transcriptomics and proteomics, has facilitated a deeper understanding of molecular damage and protective responses, paving the way for the development of tailored, species-specific protocols. These integrated approaches optimize cryopreservation conditions, maximizing post-thaw survival and the fertilizing capacity of sperm. Enhancing cryopreservation techniques not only improves the outcomes of assisted reproductive technologies, but also plays a crucial role in the conservation of genetically valuable livestock species. In conclusion, the integration of biotechnological and molecular tools holds significant promise for overcoming the current limitations and advancing the efficacy of sperm cryopreservation. Full article

The cryopreservation of boar sperm effectively extends its storage period but often leads to increased reactive oxygen species (ROS) production, compromising sperm quality. Plant extracts, rich in bioactive compounds such as polyphenols and flavonoids, have been shown to reduce ROS. Djulis (Chenopodium formosanum), also known as the “ruby of cereals”, is nutritionally rich and holds potential as a cryoprotective additive. This study aimed to determine the optimal concentration of extracts from different parts of djulis, including unhulled seeds and stems, for effective boar semen cryopreservation. Fresh semen from Taiwan indigenous boars was diluted with a modified GLT-cryoprotectant extender containing glycerol, low-density lipoprotein (LDL), and trehalose. The experimental groups included DSS25, DSS50, DS25, and DS50—representing djulis unshelled seed at 25 mg/mL and 50 mg/mL, and djulis stem at 25 mg/mL and 50 mg/mL in distilled water, respectively—alongside a control group without additives. Post-thaw assessments included sperm motility, kinetic parameters, viability, acrosome integrity, and the antioxidant properties of djulis extracts, such as DPPH radical scavenging activity and total phenolic acid content. Results showed that total motility (TM) was significantly higher in the DSS25 (48.8 ± 3.9), DSS50 (49.0 ± 6.7), and DS50 (49.0 ± 2.4) groups compared to the control group (31.3 ± 4.8). Similarly, progressive motility (PM) was significantly improved in DSS25 (27.5 ± 2.7) and DSS50 (26.8 ± 4.1) versus the control (12.8 ± 3.2). However, for straightness (STR), the control group (87.8 ± 1.3) exhibited significantly higher values than the DS50 group (83.5 ± 1.3) (p < 0.05). Viability and acrosome integrity showed no significant differences across groups. In conclusion, djulis extracts positively influence sperm motility and forward movement, with 1% djulis extract confirmed to enhance the quality of cryopreserved semen. Future research will focus on determining the optimal dosage of djulis extract for improved cryopreservation outcomes. Full article

Male contraception remains an unmet need. Na,K-ATPase α4 (NKA α4), a specific Na⁺/K⁺ transporter of the sperm flagellum, is an attractive target for male contraception. NKA α4 is critical for sperm motility and fertility, and its deletion in male mice causes complete infertility. Our previous structure–activity relationship (SAR) studies on a cardenolide scaffold identified a highly selective, safe NKAα4 inhibitor, but its complex, heavily hydroxylated structure posed challenges for modification and optimization. To address this, we employed a structural simplification strategy to synthesize novel steroidal and non-steroidal analogs and examined their effects on NKAα4 inhibition and sperm motility. Both series reduced sperm motility (up to ~50%), with IC₅₀ values in the picomolar range. Compounds 13 and 45 displayed specificities for NKAα4 over NKAα1, did not affect sperm viability, and showed no reversibility in vitro. Notably, 45, featuring a hexahydronaphthalene core and a benzyltriazole moiety at C5, exhibited potent, highly selective NKAα4 inhibition, reduced sperm motility in vitro and in vivo, and blocked fertilization in vitro. This highlights 45 as a promising lead for non-hormonal male contraception and indicates that the newly generated series of compounds possess the key characteristics needed for further development as potential non-hormonal male contraceptive agents. Full article

Oxidative stress plays a critical role in regulating sperm function, yet species-specific antioxidant mechanisms remain poorly understood. This study compared hydrogen peroxide (H₂O₂) tolerance in horse and human sperm and investigated the roles of catalase and glutathione peroxidase (GPx) in horses. A H₂O₂ dose–response assay (0–2000 µM) showed that horse sperm were significantly more resistant to oxidative damage, with an IC₅₀ for progressive motility over 14-fold higher than that of human sperm (391.6 µM vs. 27.3 µM). Horse sperm also accumulated more intracellular H₂O₂ without loss of motility or viability. DNA damage assays (Halo and SCSA) revealed H₂O₂-induced fragmentation in human but not horse sperm. Enzyme inhibition experiments in horse sperm using 3-amino-1,2,4-triazole (catalase inhibitor) and (1S,3R)-RSL3 (GPx inhibitor) at 250 µM H₂O₂ showed that catalase inhibition severely impaired motility and increased intracellular H₂O₂ > 100-fold, while GPx inhibition had a milder effect (~5-fold increase). Immunocytochemistry localized catalase to the sperm head, particularly the post-acrosomal region, challenging the notion that sperm lack peroxisomes. The dependence of horse sperm on oxidative phosphorylation may drive the need for enhanced antioxidant defenses. These findings reveal species-specific oxidative stress adaptations and highlight catalase as a key antioxidant in equine sperm. Full article

Background/Objectives: Infertility in both men and women can be significantly influenced by oxidative stress (OS), which occurs due to an imbalance between reactive oxygen species (ROS) and the body’s antioxidant defenses. In women, OS disrupts oocyte maturation, implantation, and the viability of the embryo; in men, it impairs sperm quality, reduces motility, and damages DNA integrity. This review explores existing research on how dietary and lifestyle interventions can reduce OS and enhance reproductive health outcomes. Methods: We conducted a comprehensive review of clinical, translational, and molecular studies exploring the mechanisms by which OS affects fertility, as well as the efficacy of nutritional and behavioral strategies. The interventions evaluated include weight management, regular exercise, micronutrient supplementation, antioxidant-rich diets, smoking and alcohol cessation, and stress-reduction techniques. Results: Specific dietary components such as zinc, selenium, vitamins C and E, and polyphenols have been found to neutralize reactive oxygen species (ROS) and enhance gamete function. OS is additionally reduced through lifestyle modifications, including minimizing harmful exposures, managing stress, and participating in moderate physical activity. Biomarkers such as ROS levels, total antioxidant capacity, 8-OHdG, and DNA fragmentation index are essential for assessing the effectiveness of interventions. Conclusions: Fertility in both sexes can be improved, and oxidative stress significantly reduced, through a multimodal approach incorporating dietary and lifestyle changes. There are encouraging opportunities to improve reproductive health through customized approaches that are informed by biomarker profiles. To incorporate these treatments into regular fertility care, future studies should concentrate on standardized procedures and long-term results. Full article

This study investigated the effects of dietary supplementation with the probiotic Pediococcus acidilactici on the reproductive performance and offspring viability of male Atlantic salmon (Salmo salar). A total of 48 mature males were divided into three groups—Group A (control, no probiotic), Group B (60-day supplementation), and Group C (120-day supplementation). The fish were kept in 10 m³ tanks at a constant temperature of 9 °C under a natural photoperiod and evaluated for multiple reproductive and physiological parameters. The results demonstrate that Group C exhibited significantly higher gonad weight, gonadosomatic index (GSI), and sperm concentration than Groups A and B. The blood biochemistry revealed reduced bilirubin and alanine aminotransferase (ALT) levels in Group C; however, values were within normal reference ranges for the species in all groups. While sperm quality parameters such as membrane integrity and motility showed no significant differences, fertility and embryo viability were significantly higher in Group C across all three developmental stages (180, 300, and 380 accumulated thermal units, ATUs). Additionally, Group C showed lower rates of embryonic malformation and mortality at later stages of development. Statistical analyses confirmed a significant interaction between treatment duration and embryo viability, with the 120-day probiotic treatment (Group C) producing the best outcomes. These findings support the hypothesis that P. acidilactici supplementation positively influences male reproductive parameters and improves embryo quality and survival in Atlantic salmon. Full article

Oxidative stress (OS) contributes to poor sperm parameters and increased sperm DNA fragmentation (sDF), yet effective therapeutic strategies remain limited. This study aimed to evaluate the in vitro efficacy of pentoxifylline (PTX) in improving sperm motility and reducing OS and sDF in men with isolated asthenozoospermia. Thirty semen samples from patients with asthenozoospermia were processed using density gradient centrifugation. Each sample was divided into two aliquots: one treated with PTX at a dose of 3.6 mM and the other without PTX treatment. The sperm viability and motility were assessed at 30 min, 1 h, 2 h, and 24 h post-treatment. OS was evaluated using nitro blue tetrazolium staining and a chemiluminescence assay. sDF was assessed using the alkaline Comet assay. The sperm samples treated with PTX, compared to the controls, exhibited a significant increase in total sperm motility (71.8 ± 23.03% versus 47.47 ± 4.88%, respectively; p < 0.0001). However, no significant difference was observed in the sperm viability. PTX treatment significantly reduced ROS production and sDF levels compared to controls (p < 0.01). These findings suggest that in vitro PTX supplementation enhances sperm motility and reduces the nuclear sperm injury associated with seminal ROS production. Therefore, PTX supplementation in vitro may be beneficial in assisted reproductive technology procedures involving men with asthenozoospermia. Full article

Previous studies have shown mouse antigenotoxic and chemopreventive potential with the administration of Citrus paradisi juice (GJ). To evaluate another activity, the aim of the present report was to determine the beneficial effect of GJ on male mouse reproductive damage induced by cadmium chloride (CC). Seven groups of mice were intragastrically (IG) administered for 11 days. A control group was administered purified water daily, three groups were administered GJ daily (4.1, 16.6, and 33.2 µL/g), plus a single administration of CC (3 mg/kg) on the fifth day of the assay, another group was treated daily with 33.2 µL/g GJ, and a positive control group was treated with 3 mg/kg of CC on day 5 of the experiment. The results with the high GJ dose on the CC-treated mice showed a mean reduction of 88% in sperm quality endpoints (viability, motility, malformations) and a 94% sperm concentration increase. With the same dose, we also determined an 81% reduction in the DNA breaking potential and in the number of micronuclei in the spermatids. We also found an 87% decrease in lipoperoxidation and a 68% decrease in protein oxidation with respect to the CC damage, and a strong DPPH scavenging ability. Our results suggest the potential involvement of the GJ antioxidant in the observed effect. Full article