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. 2014 Feb;12(2):103-10.

Effects of ascorbic acid on sperm motility, viability, acrosome reaction and DNA integrity in teratozoospermic samples

Hamed Fanaei  1 , Samira Khayat  2 , Iman Halvaei  3 , Vahid Ramezani  4 , Yaser Azizi  5 , Amir Kasaeian  6 , Jalal Mardaneh  7 , Mohammad Reza Parvizi  5 , Maryam Akrami  8
Affiliations

Effects of ascorbic acid on sperm motility, viability, acrosome reaction and DNA integrity in teratozoospermic samples

Hamed Fanaei et al. Iran J Reprod Med. 2014 Feb.

Abstract

Background: Oxidative stress in teratozoospermic semen samples caused poor assisted reproductive techniques (ART) outcomes. Among antioxidants, ascorbic acid is a naturally occurring free radical scavenger and as such its presence assists various other mechanisms in decreasing numerous disruptive free radical processes.

Objective: The main goal of this study was to evaluate potential protective effects of ascorbic acid supplementation during in vitro culture of teratozoospermic specimens.

Materials and methods: Teratozoospermic semen samples that collected from 15 volunteers were processed, centrifuged and incubated at 37(o)C until sperm swimmed-up. Supernatant was divided into four groups and incubated at 37(o)C for one hour under different experimental conditions: Control, 10 µm A23187, 600µm ascorbic acid and 10 µm A23187+600 µm ascorbic acid. After incubation sperm motility, viability, acrosome reaction, DNA damage and malondialdehyde levels were evaluated.

Results: Our results indicated that after one hour incubation, ascorbic acid significantly reduced malondialdehyde level in ascorbic acid group (1.4±0.11 nmol/ml) compared to control group (1.58±0.13 nmol/ml) (p<0.001). At the end of incubation, progressive motility and viability in ascorbic acid group (64.5±8.8% and 80.3±6.4%, respectively) were significantly (p<0.05 and p<0.001, respectively) higher than the control group (54.5±6.8% and 70.9±7.3%, respectively). A23187 significantly (p<0.0001) increased acrosome reaction in A23187 group (37.3±5.6%) compared to control group (8.5±3.2%) and this effect of A23187 attenuated by ascorbic acid in ascorbic acid+A23187 group (17.2±4.4%). DNA fragmentation in ascorbic acid group (20±4.1%) was significantly (p<0.001) lower than controls (28.9±4.6%).

Conclusion: In vitro ascorbic acid supplementation during teratozoospermic semen processing for ART could protect teratozoospermic specimens against oxidative stress, and it could improve ART outcome.

Keywords: Acrosome reaction; Ascorbic acid; DNA fragmentation; Oxidative stress; Teratozoospermic sperm.

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Figures

Figure 1
Figure 1
Effects of ascorbic acid on MDA levels one hour after incubation (mean±SD). *Significant difference (p<0.001) compared to before incubation group. Φ Significant difference (p<0.001) with control group
Figure 2
Figure 2
Detection of (a) acrosomal reaction by FITC-PSA staining method. The arrows show acrosome reacted sperm cells. (b) DNA damage by TUNEL staining method. The greenish sperm cells are DNA fragmented
Figure 3
Figure 3
Effects of A23187, ascorbic acid and A23187 in combination with ascorbic acid on acrosome reaction of teratozoospermic samples one hour after incubation (mean±SD). *Significant difference (p<0.0001) compared to other groups. ϕ Significant difference (p<0.001) compared to ascorbic acid group
Figure 4
Figure 4
Effects of ascorbic acid on DNA integrity of teratozoospermic samples one hour after incubation (mean±SD). *Significant difference (p<0.0001) compared to before incubation group. ϕ Significant difference (p<0.001) compared to control group
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