Mitochondria, fundamental organelles in cell metabolism, and ATP synthesis are responsible for generating reactive oxygen species (ROS), calcium homeostasis, and cell death. Mitochondria produce most ROS, and when levels exceed the antioxidant defenses, oxidative stress (OS) is generated. These changes may eventually impair the electron transport chain, resulting in decreased ATP synthesis, increased ROS production, altered mitochondrial membrane permeability, and disruption of calcium homeostasis. Mitochondria play a key role in the gamete competence to facilitate normal embryo development. However, iatrogenic factors in assisted reproductive technologies (ART) may affect their functional competence, leading to an abnormal reproductive outcome. Cryopreservation, a fundamental technology in ART, may compromise mitochondrial function leading to elevated intracellular OS that decreases sperm and oocytes’ competence and the dynamics of fertilization and embryo development. This article aims to review the role played by mitochondria and ROS in sperm and oocyte function and the close, biunivocal relationships between mitochondrial damage and ROS generation during cryopreservation of gametes and gonadal tissues in different species. Based on current literature, we propose tentative hypothesis of mechanisms involved in cryopreservation-associated mitochondrial dysfunction in gametes, and discuss the role played by antioxidants and other agents to retain the competence of cryopreserved reproductive cells and tissues.

Mitochondrial Dysfunction and Oxidative Stress Caused by Cryopreservation in Reproductive Cells.

Lipid metabolic disorders due to poor eating habits are on the rise in both developed and developing countries, with a negative impact of the "Western diet" on sperm count and quality. Dietary lipid imbalance can involve cholesterol, fatty acids, or both, under different pathophysiological conditions grouped under the term dyslipidemia. The general feature of dyslipidemia is the development of systemic oxidative stress, a well-known deleterious factor for the quality of male gametes and associated with infertility. Sperm are particularly rich in polyunsaturated fatty acids (PUFA), an important characteristic associated with normal sperm physiology and reproductive outcomes, but also targets of choice for oxidative thrust. This review focuses on the effects of dietary cholesterol or different fatty acid overload on sperm composition and function in both animals and humans. The links between oxidative stress induced by dyslipidemia and sperm dysfunction are then discussed, including possible preventive or therapeutic strategies to preserve gamete quality, longevity when stored in cryobanking, and male fertility.

/pdf/dietary-cholesterol-and-lipid-overload-impact-on-male-363e97gy08.pdf

Dietary Cholesterol and Lipid Overload: Impact on Male Fertility

Simple Summary Here we present a comprehensive review of the current literature describing mechanisms of damage incurred by cryopreservation, and we highlight several classical and novel strategies to mitigate this damage and increase sperm survival during the cryopreservation process. Abstract Cryopreservation is a way to preserve germplasm with applications in agriculture, biotechnology, and conservation of endangered animals. Cryopreservation has been available for over a century, yet, using current methods, only around 50% of spermatozoa retain their viability after cryopreservation. This loss is associated with damage to different sperm components including the plasma membrane, nucleus, mitochondria, proteins, mRNAs, and microRNAs. To mitigate this damage, conventional strategies use chemical additives that include classical cryoprotectants such as glycerol, as well as antioxidants, fatty acids, sugars, amino acids, and membrane stabilizers. However, clearly current protocols do not prevent all damage. This may be due to the imperfect function of antioxidants and the probable conversion of media components to more toxic forms during cryopreservation.

/pdf/cryopreservation-of-semen-in-domestic-animals-a-review-of-1y5bbtvd.pdf

Cryopreservation of Semen in Domestic Animals: A Review of Current Challenges, Applications, and Prospective Strategies

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9968670

Tenofovir antiviral drug solubility enhancement with β-cyclodextrin inclusion complex and in silico study of potential inhibitor against SARS-CoV-2 main protease (Mpro)



Alpha-tocopherol is a potent antioxidant involved in sperm protection particularly during cryopreservation. However, its poor solubility limits the optimal protection in aqueous solutions. 




The aim of this study was to enhance the solubility of α-tocopherol by the use of liposomes. 




 The experimental approach consisted to load vitamin E in liposomes prepared by ethanol injection method and the optimization carried out by an experimental design. The optimum solution was characterized by high performance liquid chromatography and scanning electron microscope. Finely, the impact on sperm motility protection was studied by the freezing technic of bovine sperm. 




 The optimum solution was obtained when using 10.9 mg/ml of phospholipids, 1.7 mg/ml of cholesterol and 2 mg/ml of vitamin E. The liposome size was 99.86 nm, providing 78.47% of loaded efficiency. The results showed also a significant positive impact on sperm motility after hours of preservation. 




 In conclusion, the current results showed the interest of liposome preparation as an alternative to enhance vitamin E solubility and to protect spermatozoa during cryopreservation.



Alpha Tocopherol Loaded in Liposome: Preparation, Optimization, Characterization and Sperm Motility Protection

Background During cryopreservation sperm cells suffer from two major deleterious impacts: oxidative stress and cold shock. Objective To investigate in bovine species the benefit of cholesterol and vitamin E, both loaded in cyclodextrins, as a double protection against oxidative stress and cold shock. Materials and methods Semen was collected from nine mature bulls using an artificial vagina and each ejaculate was split into four equal aliquots. The control aliquot was diluted with Fraction A (Tris+citric acid+fructose+penicillin) without further supplementation; the treated samples were diluted in Fraction A supplemented with cholesterol-loaded cyclodextrins (CD-CHL), vitamin E-loaded cyclodextrins (CD-Vit E) or CD-CHL in association with CD-Vit E (CD-CHL-VitE). After incubation at 22°C for 15 min and addition of Fraction B (Fraction A+egg yolk+glycerol), all aliquots were frozen in 0.25 ml straws. Straws were then thawed individually at 37C for 30 seconds in a water bath and immediately analyzed for motility, using Computer Aided Semen Analysis (CASA), membrane integrity and oxidative stress status. Results The results showed that samples treated with CD-CHL and CD-Vit E were protected against the deleterious impact of freezing thawing process. However, the optimal protection was observed when the two complexes CD-CHL and CD-Vit E were simultaneously used. All analysed semen parameters including motility, membrane integrity and oxidative stress status were significantly improved in CD-CHL-Vit E compared to all other treatments. Conclusion Cholesterol and vitamin E, both preloaded in cyclodextrins to increase their solubility, appeared as a powerful protection in cryopreserved bovine semen to fight simultaneously cold shock and oxidative stress.

The Complementary Effect of Cholesterol and Vitamin E Preloaded in Cyclodextrins on Frozen Bovine Semen: Motility Parameters, Membrane Integrity and Lipid Peroxidation.

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The Complementary Effect of Cholesterol and Vitamin E Preloaded in Cyclodextrins on Frozen Bovine Semen: Motility Parameters, Membrane Integrity and Lipid Peroxidation.