Curcumin-loaded niosomal nanocarriers offer a promising approach to improve quality characteristics, apoptotic gene expression, and flow cytometry assessments of stallion spermatozoa after thawing.
Abstract: The optimization of cryopreservation media to reduce oxidative damage on post-thaw spermatozoa is crucial. This research aimed to assess the antioxidant properties of curcumin-loaded niosomal nanocarriers (CurLNN) on the functional characteristics, the relative expression of apoptotic genes, and flow cytometry assessments of apoptotic-like changes, reactive oxygen species production (ROS), mitochondrial membrane potential, and chromatin integrity in stallion spermatozoa following thawing. Twenty-five ejaculates were diluted in INRA96 freezing media supplemented with 20 μM of either curcumin (Cur) or CurLNN and then cryopreserved. Results demonstrated that spermatozoa treated with Cur, particularly CurLNN, exhibited higher percentages of total and progressive motility, as well as VAP, VSL, and STR kinematics. Additionally, the functionality of the plasma membrane was enhanced, and there was a decrease in spermatozoa abnormality (P < 0.05). The incorporation of cryo-diluent medium with Cur and CurLNN led to increased viability (P < 0.05), while simultaneously reducing the levels of MDA. Flow cytometry analysis revealed a significant enhancement in mitochondrial potential activity, a reduction (P < 0.05) in ROS production, and an increase (P < 0.05) in the proportion of live and a decrease in late apoptotic stallion post-thawed spermatozoa treated with both Cur and CurLNN. Moreover, the relative expression of the Bcl anti-apoptotic gene increased (P < 0.05) by the addition of cur and CurLNN in cryo-diluent extender, while inclusion of CurLNN in cryo-diluent medium resulted in a significant reduction (P < 0.05) in the relative expression of the Bax pro-apoptotic gene in stallion post-thawed spermatozoa. In summary, the findings of this study demonstrated that CurLNN exhibits enhanced antioxidant properties, which contribute to the improved functional quality of spermatozoa by alleviating oxidative stress during the cryopreservation process.
Copyright © 2024 Society for Cryobiology. Published by Elsevier Inc. All rights reserved.
Publication Date: 2024-12-29 PubMed ID: 39706284DOI: 10.1016/j.cryobiol.2024.105188Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
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Overview
- This study investigates the use of curcumin-loaded niosomal nanocarriers (CurLNN) to improve the quality and function of stallion spermatozoa after thawing from cryopreservation.
- The research focuses on how CurLNN affects sperm motility, apoptosis-related gene expression, oxidative stress indicators, and cellular integrity measures.
Background
- Cryopreservation is a widely used technique for preserving spermatozoa, but the freezing and thawing process often leads to oxidative damage that reduces sperm quality and fertility potential.
- Oxidative stress during cryopreservation can cause membrane damage, DNA fragmentation, impaired mitochondrial function, and apoptosis-like changes in sperm cells.
- Antioxidants are explored as additives to cryopreservation media to protect sperm from oxidative damage and improve post-thaw viability and functionality.
- Curcumin is a natural antioxidant with potential to reduce oxidative stress, but its use is limited by poor bioavailability; loading curcumin into niosomal nanocarriers (CurLNN) can enhance delivery and effectiveness.
Objectives
- To evaluate the antioxidant effects of curcumin alone (Cur) and curcumin-loaded niosomal nanocarriers (CurLNN) on stallion sperm post-thaw characteristics.
- To analyze the impact on sperm motility parameters, membrane functionality, oxidative stress markers, and apoptosis-related gene expression in cryopreserved stallion spermatozoa.
- To assess the efficacy of CurLNN compared to free curcumin in improving sperm quality after thawing.
Methodology
- Twenty-five stallion ejaculates were collected and diluted in INRA96 freezing medium supplemented either with 20 μM curcumin or 20 μM CurLNN.
- The samples were cryopreserved and later thawed for analyses.
- Sperm motility was assessed using parameters such as total motility, progressive motility, average path velocity (VAP), straight line velocity (VSL), and straightness (STR).
- Plasma membrane functionality was evaluated to determine membrane integrity.
- The level of sperm abnormalities was recorded.
- Viability was measured alongside malondialdehyde (MDA) levels, a marker of lipid peroxidation and oxidative damage.
- Flow cytometry was employed to analyze mitochondrial membrane potential, reactive oxygen species (ROS) production, apoptotic-like changes, and chromatin integrity.
- Expression of apoptotic regulatory genes was quantified, focusing on Bcl-2 (anti-apoptotic) and Bax (pro-apoptotic) genes.
Key Findings
- Sperm treated with both Cur and CurLNN showed improved total and progressive motility and superior kinematic parameters (VAP, VSL, STR), with CurLNN showing greater enhancement.
- Plasma membrane functionality improved significantly, indicating better membrane integrity after thawing.
- The rate of sperm abnormality decreased significantly with both treatments.
- Viability of sperm cells increased, especially with CurLNN.
- MDA levels were reduced, indicating lowered lipid peroxidation and oxidative stress.
- Flow cytometry results demonstrated enhanced mitochondrial membrane potential and reduced ROS production, suggesting improved mitochondrial function and reduced oxidative damage.
- The proportion of live sperm increased, whereas late apoptotic sperm decreased post-thaw when treated with Cur and CurLNN.
- Bcl-2 (anti-apoptotic) gene expression increased with both treatments, supporting enhanced cell survival.
- CurLNN caused a significant reduction in Bax (pro-apoptotic) gene expression, indicating decreased apoptosis.
Conclusions and Implications
- The encapsulation of curcumin into niosomal nanocarriers (CurLNN) enhanced its antioxidant properties, resulting in better protection of stallion spermatozoa during cryopreservation.
- CurLNN improved multiple aspects of sperm quality post-thaw, including motility, membrane integrity, oxidative stress reduction, and gene expression related to apoptosis.
- This approach shows promise for improving the outcomes of sperm cryopreservation by minimizing oxidative damage and cellular apoptosis.
- Such improvements in sperm quality are crucial for assisted reproduction and breeding programs in equine species.
- Future work may further optimize these nanocarrier systems and explore their application across other species and cryopreservation protocols.
Cite This Article
APA
Nasiri-Foomani N, Hassani S, Najafi M, Samadi F.
(2024).
Curcumin-loaded niosomal nanocarriers offer a promising approach to improve quality characteristics, apoptotic gene expression, and flow cytometry assessments of stallion spermatozoa after thawing.
Cryobiology, 118, 105188.
https://doi.org/10.1016/j.cryobiol.2024.105188 Publication
Researcher Affiliations
- Department of Animal and Poultry Physiology, Faculty of Animal Science, Gorgan University of Agricultural Science and Natural Resources, Golestan, Gorgan, Iran.
- Department of Animal and Poultry Physiology, Faculty of Animal Science, Gorgan University of Agricultural Science and Natural Resources, Golestan, Gorgan, Iran.
- Department of Animal and Poultry Physiology, Faculty of Animal Science, Gorgan University of Agricultural Science and Natural Resources, Golestan, Gorgan, Iran.
- Department of Animal and Poultry Physiology, Faculty of Animal Science, Gorgan University of Agricultural Science and Natural Resources, Golestan, Gorgan, Iran. Electronic address: f.samadi@gau.ac.ir.
MeSH Terms
- Male
- Animals
- Curcumin / pharmacology
- Horses
- Spermatozoa / drug effects
- Spermatozoa / cytology
- Spermatozoa / physiology
- Spermatozoa / metabolism
- Cryopreservation / veterinary
- Cryopreservation / methods
- Semen Preservation / veterinary
- Semen Preservation / methods
- Flow Cytometry
- Apoptosis / drug effects
- Reactive Oxygen Species / metabolism
- Membrane Potential, Mitochondrial / drug effects
- Cryoprotective Agents / pharmacology
- Sperm Motility / drug effects
- Antioxidants / pharmacology
- Oxidative Stress / drug effects
- Liposomes
Conflict of Interest Statement
Declarations of competing interest The authors declare that they possess no recognized financial conflicts of interest or personal affiliations that might have seemingly impacted the research presented in this paper.
Citations
This article has been cited 2 times.- Khalil WA, Elkhamy SA, Hegazy MM, Hassan MAE, Tafish AM, Abdelnour SA, El-Harairy MA. Beneficial effects of carvacrol loaded phytosomes on enhancing cryotolerance of Buffalo semen following cryopreservation.. Sci Rep 2025 Jul 14;15(1):25366.
- Khalil WA, Elkhamy SA, Hegazy MM, Hassan MAE, Abdelnour SA, El-Harairy MA. The cryoprotective effects of celastrol nanoemulsion on post-thawed attributes and fertilizing ability of cryopreserved buffalo semen.. Vet Res Commun 2025 Jun 5;49(4):214.
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