The Effect of Cholesterol‐Loaded Cyclodextrin and Resveratrol Compounds on Post‐Thawing Quality of Ram Semen

ABSTRACT Ram sperm are more vulnerable to freezing than those of most other farm animals. During sperm freezing, the cell membrane loses some of its cholesterol, which regulates signalling mechanisms and prevents premature capacitation. Resveratrol (RES) increases the fluidity of the cell membrane,...

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Main Authors: Eser Ahmet, Arıcı Ramazan, Yağcıoğlu Selin, Sandal Asiye İzem, Ersoy Nur, Demir Kamber, Evecen Mithat, Ak Kemal
Format: Article
Language:English
Published: Wiley 2025-01-01
Series:Veterinary Medicine and Science
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Online Access:https://doi.org/10.1002/vms3.70172
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author Eser Ahmet
Arıcı Ramazan
Yağcıoğlu Selin
Sandal Asiye İzem
Ersoy Nur
Demir Kamber
Evecen Mithat
Ak Kemal
author_facet Eser Ahmet
Arıcı Ramazan
Yağcıoğlu Selin
Sandal Asiye İzem
Ersoy Nur
Demir Kamber
Evecen Mithat
Ak Kemal
author_sort Eser Ahmet
collection DOAJ
description ABSTRACT Ram sperm are more vulnerable to freezing than those of most other farm animals. During sperm freezing, the cell membrane loses some of its cholesterol, which regulates signalling mechanisms and prevents premature capacitation. Resveratrol (RES) increases the fluidity of the cell membrane, which becomes peroxidized during freezing and reduces free radicals. In this study, the effectiveness of RES, cholesterol‐loaded cyclodextrin (CLC) and their combinations in ram sperm cryopreservation were investigated. The collected semen was divided into two equal volumes: One was diluted with tris‐citric acid‐glucose medium (TCG) containing CLC, whereas the other was diluted with a CLC‐free TCG solution. After examining motility, both groups were further divided into two equal volumes, forming the following working groups: control (no RES, no CLC); RES (20 µg/mL); CLC (2 mg CLC/120 × 106 sperm); and RES + CLC (RES 20 µg/mL + 2 mg CLC/120 × 106 sperm). These groups were diluted with media containing their respective additives. Post‐thawing, the samples were analysed for motility, acrosome and membrane integrity, membrane functionality, mitochondrial activity, capacitation status, oxidative stress and DNA integrity. CLC preserved sperm total motility, acrosome and plasma membrane integrity and decreased the rate of early capacitation (p < 0.05). RES had no significant effect on sperm quality before freezing and post‐thawing (p > 0.05). However, RES + CLC increased mitochondrial activity post‐thawing (p < 0.05). In conclusion, CLC minimized sperm membrane damage caused by cryopreservation in ram sperm. RES alone was ineffective, and the combination of RES and CLC did not yield a positive synergistic effect on ram spermatological parameters.
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spelling doaj-art-12fc62a9dea6409192ce4b90618f4e712025-08-20T03:40:17ZengWileyVeterinary Medicine and Science2053-10952025-01-01111n/an/a10.1002/vms3.70172The Effect of Cholesterol‐Loaded Cyclodextrin and Resveratrol Compounds on Post‐Thawing Quality of Ram SemenEser Ahmet0Arıcı Ramazan1Yağcıoğlu Selin2Sandal Asiye İzem3Ersoy Nur4Demir Kamber5Evecen Mithat6Ak Kemal7Department of Reproduction and Artificial Insemination, Faculty of Veterinary Medicine Siirt University Siirt TurkeyDepartment of Reproduction and Artificial Insemination, Faculty of Veterinary Medicine İstanbul University—Cerrahpasa Avcilar İstanbul TurkeyDepartment of Reproduction and Artificial Insemination, Faculty of Veterinary Medicine İstanbul University—Cerrahpasa Avcilar İstanbul TurkeyDepartment of Reproduction and Artificial Insemination, Faculty of Veterinary Medicine İstanbul University—Cerrahpasa Avcilar İstanbul TurkeyGraduate Education Institute Istanbul University—Cerrahpasa Avcılar İstanbul TurkeyDepartment of Reproduction and Artificial Insemination, Faculty of Veterinary Medicine İstanbul University—Cerrahpasa Avcilar İstanbul TurkeyDepartment of Reproduction and Artificial Insemination, Faculty of Veterinary Medicine İstanbul University—Cerrahpasa Avcilar İstanbul TurkeyDepartment of Reproduction and Artificial Insemination, Faculty of Veterinary Medicine İstanbul University—Cerrahpasa Avcilar İstanbul TurkeyABSTRACT Ram sperm are more vulnerable to freezing than those of most other farm animals. During sperm freezing, the cell membrane loses some of its cholesterol, which regulates signalling mechanisms and prevents premature capacitation. Resveratrol (RES) increases the fluidity of the cell membrane, which becomes peroxidized during freezing and reduces free radicals. In this study, the effectiveness of RES, cholesterol‐loaded cyclodextrin (CLC) and their combinations in ram sperm cryopreservation were investigated. The collected semen was divided into two equal volumes: One was diluted with tris‐citric acid‐glucose medium (TCG) containing CLC, whereas the other was diluted with a CLC‐free TCG solution. After examining motility, both groups were further divided into two equal volumes, forming the following working groups: control (no RES, no CLC); RES (20 µg/mL); CLC (2 mg CLC/120 × 106 sperm); and RES + CLC (RES 20 µg/mL + 2 mg CLC/120 × 106 sperm). These groups were diluted with media containing their respective additives. Post‐thawing, the samples were analysed for motility, acrosome and membrane integrity, membrane functionality, mitochondrial activity, capacitation status, oxidative stress and DNA integrity. CLC preserved sperm total motility, acrosome and plasma membrane integrity and decreased the rate of early capacitation (p < 0.05). RES had no significant effect on sperm quality before freezing and post‐thawing (p > 0.05). However, RES + CLC increased mitochondrial activity post‐thawing (p < 0.05). In conclusion, CLC minimized sperm membrane damage caused by cryopreservation in ram sperm. RES alone was ineffective, and the combination of RES and CLC did not yield a positive synergistic effect on ram spermatological parameters.https://doi.org/10.1002/vms3.70172cholesterolcryopreservationram spermresveratrolspermatological examination
spellingShingle Eser Ahmet
Arıcı Ramazan
Yağcıoğlu Selin
Sandal Asiye İzem
Ersoy Nur
Demir Kamber
Evecen Mithat
Ak Kemal
The Effect of Cholesterol‐Loaded Cyclodextrin and Resveratrol Compounds on Post‐Thawing Quality of Ram Semen
Veterinary Medicine and Science
cholesterol
cryopreservation
ram sperm
resveratrol
spermatological examination
title The Effect of Cholesterol‐Loaded Cyclodextrin and Resveratrol Compounds on Post‐Thawing Quality of Ram Semen
title_full The Effect of Cholesterol‐Loaded Cyclodextrin and Resveratrol Compounds on Post‐Thawing Quality of Ram Semen
title_fullStr The Effect of Cholesterol‐Loaded Cyclodextrin and Resveratrol Compounds on Post‐Thawing Quality of Ram Semen
title_full_unstemmed The Effect of Cholesterol‐Loaded Cyclodextrin and Resveratrol Compounds on Post‐Thawing Quality of Ram Semen
title_short The Effect of Cholesterol‐Loaded Cyclodextrin and Resveratrol Compounds on Post‐Thawing Quality of Ram Semen
title_sort effect of cholesterol loaded cyclodextrin and resveratrol compounds on post thawing quality of ram semen
topic cholesterol
cryopreservation
ram sperm
resveratrol
spermatological examination
url https://doi.org/10.1002/vms3.70172
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