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. 2023 Aug 26;12(9):1673.
doi: 10.3390/antiox12091673.

Antioxidant Effects of Myo-Inositol Improve the Function and Fertility of Cryopreserved Boar Semen

Rana Osman  1 , Seongju Lee  1 , Areeg Almubarak  1   2 , Jae-Ik Han  3 , Il-Jeoung Yu  1 , Yubyeol Jeon  1
Affiliations

Antioxidant Effects of Myo-Inositol Improve the Function and Fertility of Cryopreserved Boar Semen

Rana Osman et al. Antioxidants (Basel). .

Abstract

During cryopreservation, sperm undergoes structural and molecular changes such as ice crystal formation, DNA fragmentation, and reactive oxygen species (ROS) production, leading to decreased sperm quality after thawing. Antioxidants play a crucial role in preventing these damages, both in vivo and in vitro. One potent antioxidant is myo-inositol, known for its protective effects on sperm against ROS. This study aimed to investigate the protective effect of myo-inositol on cryopreserved boar semen. The semen was diluted, cooled, and cryopreserved using a BF5 extender. It was then divided into five groups: control and different concentrations of myo-inositol (0.5, 1, 1.5, and 2 mg/mL). The post-thaw evaluation included assessments of motility, viability, acrosome integrity, mitochondrial membrane potential (MMP), caspase activity, gene expression, ROS levels, apoptosis, and IVF with treated semen. Results showed that myo-inositol at 0.5 mg/mL improved motility, acrosome integrity, and fertilization ability. It also reduced the expression of pro-apoptotic genes and increased SMCP expression. Lower concentrations also demonstrated improved viability and reduced apoptosis and ROS levels. In conclusion, myo-inositol treatment during cryopreservation improved sperm quality, reduced apoptosis and ROS levels, and enhanced fertility rates in boar semen.

Keywords: ROS; apoptosis; boar semen; cryopreservation; in vitro fertilization; myo-inositol.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Effects of different concentrations of myo-inositol on sperm motility parameters after the freezing–thawing process. Control: 0 mg/mL, 0.5 mg/mL, 1 mg/mL, 1.5 mg/mL, and 2 mg/mL myo-inositol-treated groups; TM: total motility; PM: progressive motility; RPM: rapid progressive motility; MPM: medium progressive motility. Letters a, b, and c represent significant differences between groups (p < 0.05). Error bars show the SEM.
Figure 2
Figure 2
Effect of myo-inositol on sperm survival after the freezing–thawing process. (A) Semen viability; (B) acrosome integrity; (C) sperm caspase activity; and (D) sperm mitochondrial membrane potential. Control and 0.5 mg/mL, 1 mg/mL, 1.5 mg/mL, and 2 mg/mL myo-inositol-treated groups. Letters a, b, and c represent significant differences between the groups (p < 0.05). Error bars show the SEM.
Figure 3
Figure 3
Effect of myo-inositol on gene expression after the freezing–thawing process. Control: 0 mg/mL myo-inositol; myo-inositol: 0.5 mg/mL myo-inositol. (*) Indicates a significant difference between the groups (p < 0.05). Error bars show the SEM. BAX: Bcl-2-associated X protein; Bcl-2: Bcl-2 apoptosis regulator; ROMO1: reactive oxygen species modulator 1; SMOX: spermine oxidase; NOX5: NADPH oxidase 5; SMCP: sperm mitochondria associated cysteine rich protein.
Figure 4
Figure 4
(A) Effect of myo-inositol-treated cryopreserved semen on the cleavage rate. (B) Effect of myo-inositol-treated cryopreserved semen on the blastocyst rate. Control: 0 mg/mL myo-inositol; myo-inositol: 0.5 mg/mL myo-inositol. (*) Indicates a significant difference between the groups (p < 0.05). Error bars show the SEM.
Figure 5
Figure 5
Effect of myo-inositol on sperm damage after the freezing–thawing process. (A) Sperm MDA levels; (B) sperm ROS; (C) sperm apoptosis; (D) control ROS; and (E) control apoptosis. Control: 0 mg/mL myo-inositol; myo-inositol: 0.5 mg/mL myo-inositol. (*) Indicates a significant difference between the groups (p < 0.05). Error bars show the SEM.
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