Supplementation of sperm freezing medium with myoinositol improve human sperm parameters and protects it against DNA fragmentation and apoptosis
- 42 Downloads
The aim of this study is to evaluate the beneficial effect of Myoinositol (MYO) supplement in freezing media on the post thaw sperm quality. Semen samples from 40 normozoospermic men were divided into two aliquots and frozen with simple or 2 mg/mL MYO supplemented freezing medium. Post thaw process including, computer-assissted sperm analysis was used to analyze sperm motility and morphology. Reactive oxygen species was evaluated by the fluorometry of DCFH-DA, as well as total antioxidant capacity and lipid peroxidation were measured based on colorimetric assay by ELISA reader. Eventually, DNA fragmentation was assessed using TUNEL staining. MYO significantly improved progressive motility and normal morphology in treated samples (p < 0.05). Lipid peroxidation (malondialdehyde level) can be diminished in samples were frozen by MYO supplemented freezing media (p < 0.05). While MYO did not affect the amount of ROS (p > 0.05), it was associated with high values of total antioxidant capacity (p < 0.05). DNA integrity was significantly affected by MYO, as in MYO treated samples, DNA fragmentation was decreased compared to control ones (p < 0.001). The findings support the use of 2 mg/mL myoinositol supplemented freezing media in sperm cryopreservation to increase sperm quality after freezing–thawing procedures.
KeywordsMyoinositol Human sperm cryopreservation Sperm parameters Apoptosis DNA fragmentation
We wish to thank Dr.Joghataei for his kind courtesy and attempt to improve the article.
Compliance with ethical standards
Conflicts of interest
The authors declare that they have no conflicts of interest.
All procedures performed in this study were in accordance with the ethical standards of the national research committee.
This research was supported by Shahid Akbarabadi Clinical Research Unit, Iran University Of Medical Sciences.
- Agarwal A, Varghese AC, Sharma RK (2009) Markers of oxidative stress and sperm chromatin integrity Mol Endocrinol (pp 377–402): SpringerGoogle Scholar
- Asadpour R, Jafari R, Tayefi-Nasrabadi H (2012) The effect of antioxidant supplementation in semen extenders on semen quality and lipid peroxidation of chilled bull spermatozoa. Iran J Vet Res 13(3):246–249Google Scholar
- Brugnon F, Ouchchane L, Pons-Rejraji H, Artonne C, Farigoule M, Janny L (2013) Density gradient centrifugation prior to cryopreservation and hypotaurine supplementation improve post-thaw quality of sperm from infertile men with oligoasthenoteratozoospermia. Hum Reprod 28(8):2045–2057CrossRefGoogle Scholar
- Colagar AH, Karimi F, Jorsaraei SGA (2013) Correlation of sperm parameters with semen lipid peroxidation and total antioxidants levels in astheno-and oligoasheno-teratospermic men. Iran Red Crescent Med J 15(9):780Google Scholar
- Das P, Choudhari A, Singh A, Singh R (2009) Correlation among routine semen parameters, sperm viabilty and malondialdehyde levels in human subjects with different fertility potentialGoogle Scholar
- Gupta S, AA, Sharma R, Ahmady A (2014) Sperm banking via cryopreservation Rizk Botros RMB, Aziz N, Agarwal A, Sabanegh E Medical and Surgical Management of Male Infertility. New Delhi Jaypee Brothers Medical Publishers LtdGoogle Scholar
- Hamada A, Wasik M, Gupta S, Agarwal A (2012) Sperm banking: indications and regulations. Infertility: diagnosis, management and IVF. Jaypee Brothers Medical Publishing, New Delhi, pp 409–434Google Scholar
- Korosi T, Barta C, Rokob K, Torok T (2017) Physiological Intra-Cytoplasmic Sperm Injection (PICSI) outcomes after oral pretreatment and semen incubation with myo-inositol in oligoasthenoteratozoospermic men: results from a prospective, randomized controlled trial. Eur Rev Med Pharmacol Sci 21(2 Suppl):66–72PubMedGoogle Scholar
- Marat AL, Haucke V (2016) Phosphatidylinositol 3‐phosphates—at the interface between cell signalling and membrane traffic. EMBO J e201593564Google Scholar
- Montanino Oliva M, Minutolo E, Lippa A, Iaconianni P, Vaiarelli A (2016) Effect of myoinositol and antioxidants on sperm quality in men with metabolic syndrome. Int J Endocrinol 2016. https://doi.org/10.1155/2016/1674950
- Nagy ZP, Varghese AC, Agarwal A (2017) Cryopreservation of Mammalian Gametes and Embryos. Methods Mol Biol 1568Google Scholar
- Organization WH (2010) WHO laboratory manual for the examination and processing of human semenGoogle Scholar
- Palmieri M, Papale P, Della Ragione A, Quaranta G, Russo G, Russo S (2016) In vitro antioxidant treatment of semen samples in assisted reproductive technology: effects of myo-inositol on nemaspermic parameters. Int J Endocrinol 2016. https://doi.org/10.1155/2016/2839041
- Rubino P, Palini S, Chigioni S, Carlomagno G, Quagliariello A, De Stefani S, Bulletti C (2015) Improving fertilization rate in ICSI cycles by adding myoinositol to the semen preparation procedures: a prospective, bicentric, randomized trial on sibling oocytes. J Assist Reprod Genet 32(3):387–394CrossRefGoogle Scholar
- Santamaria A, Di Benedetto A, Petrella E, Pintaudi B, Corrado F, D’Anna R, Facchinetti F (2016) Myo-inositol may prevent gestational diabetes onset in overweight women: a randomized, controlled trial. J Maternal Fetal Neonatal Med 29(19):3234–3237Google Scholar
- Varghese AC, Nandi P, Mahfouz R, Athayde KS, Agarwal A (2010) Human sperm cryopreservation Rao AR, Agarwal A, Srinivas MS. Andrology laboratory manual. Jaypee Brothers Medical Publishers Ltd., New Delhi Google Scholar
- Zandieh Z, Vatannejad A, Doosti M, Zabihzadeh S, Haddadi M, Bajelan L, Amanpour S (2017) Comparing reactive oxygen species and DNA fragmentation in semen samples of unexplained infertile and healthy fertile men. Irish J Med Sci 187:1–6Google Scholar