Effects of antioxidants on motility and DNA integrity in frozen-thawed sperm

Year 2020, Volume: 12 Issue: 2, 41 - 48, 30.08.2020

Seren Can Çelik , Ümit Özekici Mehmet Cıncık Belgin Selam Yaprak Dönmez Çakıl

https://doi.org/10.35514/mtd.2020.28

Abstract

Aims: This study aims to investigate whether the addition of the antioxidant complex including taurine, ascorbic acid, and glutathione into the cryopreservation medium affects the damage to sperm during the freezing process. Methods: Ejaculate samples of patients who applied for semen analysis to the Assisted Reproduction Unit of Private Adatip Hospital were used. Fresh samples were analyzed for standard semen quality parameters according to the World Health Organization guidelines. Samples within the normal range were evaluated for sperm DNA fragmentation using the Halosperm technique. Remaining ejaculates were washed with the gradient method before cryopreservation. Sperm samples of each patient were divided equally for freezing in a cryopreservation medium with or without the antioxidant supplementation. One month later, the samples were thawed. Post-thaw total motility and DNA fragmentation were determined for each sample. Results: Semen samples of 40 patients were analyzed. We observed decreased total motility (34.8 ±5.32 % vs. 65.5 ±6.42 %, P= 0.002) and increased sperm DNA fragmentation (52.3±5.42 % vs. 26.4±3.12 %, P= 0.002) in post-thaw semen samples following cryopreservation in comparison to fresh samples. The addition of antioxidants to the freezing medium did not have a statistically significant effect on sperm motility (38.3± 6.22 % vs. 34.8 ±5.32 %, P =0.07) and DNA damage (47.5±4.7 % vs. 52.3±5.42, P =0.08) when compared to control samples following the freezing process. Conclusions: We observed increased sperm DNA fragmentation and decreased total motility following cryopreservation. No significant improvement in sperm motility or DNA integrity was obtained after the addition of 5 µM of the antioxidants taurine, ascorbic acid, and glutathione to the freezing media.

Keywords

Halosperm, DNA fragmentation, antioxidants, sperm freezing, motility, cryopreservation

Thanks

The authors greatly acknowledge Ender Yalçınkaya Kalyan, MSc., IVF Lab Director of Private Adatip Hospital, for her contributions and support during laboratory work.

Antioksidanların donmuş çözülmüş spermde motilite ve DNA bütünlüğü üzerindeki etkileri

Abstract

Amaç: Bu çalışma, taurin, askorbik asit ve glutatyon içeren antioksidan kompleksin kriyoprezervasyon ortamına eklenmesinin dondurma işlemi sırasında spermdeki hasarı etkileyip etkilemediğini araştırmayı amaçlamaktadır. Yöntemler: Özel Adatip Hastanesi Yardımcı Üreme Ünitesine meni analizi için başvuran hastalardan ejakülat örnekleri kullanıldı. Taze numuneler, Dünya Sağlık Örgütü yönergelerine göre standart semen kalite parametreleri için analiz edildi. Normal aralıktaki numuneler, Halosperm tekniği kullanılarak sperm DNA fragmantasyonu açısından değerlendirildi. Geri kalan ejakülatlar kriyoprezervasyon öncesi gradyan yöntemi ile yıkandı. Her hastanın sperm örnekleri, antioksidan takviyesi olan veya olmayan bir kriyoprezervasyon ortamında dondurulmak üzere eşit olarak bölünmüştür. Bir ay sonra numuneler çözüldü. Her numune için çözülme sonrası toplam hareketlilik ve DNA fragmantasyonu belirlendi. Bulgular: 40 hastanın semen örnekleri analiz edildi. Dondurarak saklamayı takiben eritme sonrası semen örneklerinde toplam motilitede azalma (%34,8 ±5,32 %65.5 ±6.42, P= 0.002) ve sperm DNA fragmantasyonunda artış (%52.3±5.42'ye karşı %26.4±3.12, P= 0.002) gözlemledik. taze örneklerle karşılaştırıldığında. Dondurma ortamına antioksidanların eklenmesi, sperm motilitesi (%38,3±6,22'ye karşı %34,8 ±5,32, P=0,07) ve DNA hasarı (47,5±4,7'ye karşı 52,3±5,42, P =) üzerinde istatistiksel olarak anlamlı bir etkiye sahip değildi. 0.08), dondurma işlemini takip eden kontrol numuneleriyle karşılaştırıldığında. Sonuçlar: Dondurarak saklamayı takiben artan sperm DNA fragmantasyonu ve toplam motilitede azalma gözlemledik. Dondurma ortamına 5 uM antioksidan taurin, askorbik asit ve glutatyon ilave edildikten sonra sperm motilitesinde veya DNA bütünlüğünde önemli bir gelişme elde edilmedi.

Keywords

cryopreservation, Halosperm, DNA fragmantasyon, antioksidanlar, sperm dondurma, hareketlilik

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