An Ideal Sperm Selection Method for the Intrauterine Insemination Treatment of Normozoospermic Infertile Patients*

Yıl 2020, Cilt: 25 Sayı: 1, 19 - 26, 15.01.2020

Sevilay Erimşah , Süheyla Gonca , Aysel Kükner Bülent Duran Yusufhan Yazır

Öz

Aim: In this study, we evaluated the DNA integrity of sperms selected by using
the conventional swim-up and
density gradient centrifugation techniques and the new nanotechnology-based microfluidic
chip method in order to determine the ideal
sperm selection method for the intrauterine
insemination treatment of
normozoospermic infertile patients (NIPs).

Materials and Methods: Semen samples obtained from 20 patients were divided into four equal fractions. Control, density gradient centrifugation, swim-up, and microfluidic chip (MC) groups were created, and the untreated (control) and treated (other) sperm samples from the four groups were examined for DNA integrity. Acidic aniline blue staining and the TUNEL method were used respectively for evaluating sperm chromatin condensation defects and DNA fragmentation. Dichlorofluorescein diacetate and flow cytometry were used to determine the reactive oxygen species levels. 

Results: We measured significantly lower values of chromatin
condensation defects, DNA fragmentation and reactive oxygen species for the sperms
selected with the MC method, compared to the sperms selected with the conventional methods (p<0.0001).

Discussion and Conclusion: We found that the new MC method was more
effective in selecting sperms with high
DNA integrity, compared to the conventional methods. Accordingly, the MC method can be an ideal sperm selection method for
use in the intrauterine insemination treatment of NIPs with high DNA fragmentation,
apoptosis, and reactive oxygen species levels.

Anahtar Kelimeler

DNA integrity, intrauterine insemination, microfluidic chip, sperm selection

Normozoospermik İnfertil Hastaların İntrauterin İnseminasyon Tedavisi İçin İdeal Bir Sperm Seçim Yöntemi

Öz

Amaç: Bu çalışmada normozoospermik infertil hastaların (NİH) intrauterin
inseminasyon tedavisi için ideal sperm seçim yöntemini belirlemek amacıyla geleneksel
yüzdürme ve yoğunluk gradyanlı santrifüjleme teknikleri ve yeni nanoteknoloji
bazlı mikroakışkan çip yöntemi ile seçilen spermlerin DNA bütünlüğü değerlendirilmiştir.

Gereç ve Yöntemler: Yirmi hastadan toplanan semen
örnekleri dört eşit parçaya bölündü. Kontrol, yoğunluk gradyanlı santrifüjleme,
yüzdürme ve mikroakışkan çip (MÇ) grupları oluşturuldu ve bu dört gruba ait
işlem görmemiş (kontrol) ve görmüş (diğer) sperm örnekleri DNA bütünlüğü
açısından incelendi. Sperm kromatin yoğunlaşma kusurlarını ve DNA kırıklarını değerlendirmek
için asidik anilin mavisi ve TUNEL boyaması kullanıldı. Reaktif oksijen türevi seviyelerini tespit etmek için akış
sitometrisi ile diklorofloresan diasetat kullanıldı.

Bulgular: MÇ yöntemiyle seçilen spermlerde kromatin kondensasyon
defekti, DNA fragmantasyonu ve reaktif oksijen türevi değerleri, geleneksel yöntemlerle
seçilen spermlere göre anlamlı şekilde düşük bulundu (p<0,0001).

Tartışma ve Sonuç: Yeni MÇ yönteminin geleneksel yöntemlere göre yüksek DNA bütünlüğüne sahip sperm
seçiminde daha etkili olduğu görüldü. Buna göre MÇ yöntemi yüksek DNA kırığı,
apopitoz ve reaktif oksijen türevi görülen NİH’lerin intrauterin inseminasyon tedavisinde
ideal bir sperm seçim yöntemi olabilir. 

Anahtar Kelimeler

DNA bütünlüğü, intrauterin inseminasyon, mikroakışkan çip, sperm seçimi

Kaynakça

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