Effect of magnesium oxide nanoparticles on varicocele-induced testicular damage in male Wistar rats
Subject Areas :
Veterinary Clinical Pathology
Masoumeh Tolu Ghamari
1
,
Akram Eidi
2
,
Pejman Mortazavi
3
,
Ahmad Asghari
4
1 - Ph.D. Graduate, Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.
2 - Professor, Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.
3 - Associate Professor, Department of Pathology, Faculty of Specialized Veterinary Sciences, Science and Research Branch, Islamic Azad University, Tehran, Iran.
4 - Associate Professor, Department of Clinical Sciences, Faculty of Specialized Veterinary Sciences, Science and Research Branch, Islamic Azad University, Tehran, Iran.
Received: 2022-12-02
Accepted : 2023-04-09
Published : 2023-04-21
Keywords:
sperm,
Rat,
Varicocele,
Magnesium oxide nanoparticles,
Testicular damage,
Abstract :
Varicocele is a pathological dilation of the venous network of the spermatic cord and considering that magnesium oxide nanoparticles play a key role in various physiological functions, the aim of this study was to evaluate the performance of nanoparticles on sperm characteristics affected by experimental varicocele. A total of 54 male Wistar rats were randomly divided into 9 equal groups including healthy control group (untouched animals), sham-operated group (underwent sham surgery), three healthy experimental groups (animals in these groups received magnesium oxide nanoparticles at doses of 1.25, 2.5 and 5 mg/kg respectively by gavage for 6 weeks), varicocele control group (varicocele was induced by renal vein ligation) and three experimental varicocele groups (in addition to varicocele induction, magnesium oxide nanoparticles were given at doses of 1.25, 2.5 and 5 mg/kg respectively by gavage for 6 weeks). At the end of the 6th week, the abdomen was opened and semen samples were collected from the tail of the epididymis to determine the indices of concentration, survival and motility of sperm and the data were statistically analyzed (p<0.05). The results showed that magnesium oxide nanoparticles were able to significantly increase the concentration, viability, progressive, moderate and non-progressive movements of sperm compared to the varicocele group (p<0.001) and also caused a significant decrease (p<0.001) in the number of non-moving sperms in the varicocele experimental groups. Therefore, magnesium oxide nanoparticles may possibly reduce the destructive effects of varicocele due to their antioxidant activity and be effective in its treatment by improving sperm properties during varicocele.
References:
Abinaya, S., Kavitha, H.P., Prakash, M. and Muthukrishnaraj, A. (2021). Green synthesis of magnesium oxide nanoparticles and its applications: A review. Sustainable Chemistry and Pharmacy, 19(7) 1-9.
Agarwal, A., Durairajanayagam, D., Halabi, J., Peng, J. and Vazquez-Levin, M. (2014). Proteomics, oxidative stress and male infertility. Reproductive BioMedicine Online, 29(1): 32-58.
Ahmadi Asr Badr, Y., Hasanzadeh, K. and Jahantabi, E. (2016). Evaluation of the effect of the number and size of veins ligated at left sided microsurgical inguinal varicocelectomy on the semen analysis outcomes. Medical Journal of Tabriz University of Medical Sciences, 38(3): 6-11.
Aleksandra, B., Zorica, B., Danijela, D., Vesna, M. and Buha, A. (2012). Magnesium effects against cd-induced stress in rats. Archives of Industrial Hygiene and Toxicology, 63(1): 247-254.
Amouoghli Tabrizi, B. and Khakpour, M. (2013). Protective effects of ginger (Zingiber officinale) rhizome extract on heatinduced testicular damage in the mouse. Journal of Veterinary Clinical Pathology, 27(7): 183-261. [In Persian]
Bahmanzadeh, M., Abolhassani, F., Amidi, F., Ejtemaiemehr, Sh., Salehi, M. and Abbasi M. (2008). The effects of nitric oxide synthase inhibitor (L-NAME) on epididymal sperm count, motility, and morphology in varicocelized rat. DARU Journal of Pharmaceutical Sciences, 16(1): 23-28.
Bisht, S., Faiq, M., Tolahunase, M. and Dada, R. (2017). Oxidative stress and male infertility. Nature Reviews Urology, 14(8): 470-485.
Brown, D.G. and Burk, R.F. (1973). Selenium retention in tissues and sperm of rats fed a torula yeast diet. The Journal of Nutrition, 102(1):102-108.
Cai, L., Chen, J., Liu, Z., Wang, H., Yang, H. and Ding, W. (2018). Magnesium Oxide Nanoparticles: Effective agricultural antibacterial agent against ralstonia solanacearum. Frontiers in Microbiology, 790(9): 1-19.
Celik-Ozenci, C., Bayram, Z., Akkoyunlu, G., Türkay Korgun, E.,Erdogru, T., Seval, Y., et al. (2006). Localization of NGF and nNOS in varicocele-induced rat testis. Acta Histochemica, 107(6): 435-442.
Chandra, A.K., Sengupta, P., Goswam, H. and Sarkar, M. (2013). Effects of dietary magnesium on testicular histology, steroidogenesis, spermatogenesis and oxidative stress markers in adult rat. Indian Journal of Experimental Biology, 51(1): 37-47.
Comhaire, F. and Vermeulen, A. (1974). Varicocele sterility: Cortisol and catecholamines. Fertility and Sterility, 25(1): 88-95.
Goodson, S.G., Zhang, Z., Tsuruta, J.K., Wang, W. and O'Brien, D.A. (2011). Classification of mouse sperm motility patterns using an automated multiclass support vector machines model. Biology of Reproduction, 84(6):1207-15.
Girardi, S.K. and Goldestein, M. (1997). Varicocele. Current Therapy in Endocrinology and Metabolism, 6(1): 355-358.
Giri, A., Khynriam, D. and Prasad, S.B. (1998).Vitamin C mediated protection on cisplatin induced mutagenicity in mice. Mutation Research, 421(2):139-148.
Gual-Frau, J., Abad, C., Amengual, M.J., Hannaoui, N., Checa, M.A., Ribas-Maynou, J. and et al. (2015). Oral antioxidant treatment partly improves integrity of human sperm DNA in infertile grade I varicocele patients. Human Fertility, 18(3): 225-9.
Hamada, A., Esteves, S.C. and Agarwal, A. (2013). Insight into oxidative stress in varicocele-associated male infertility. Part 2. Nature Reviews Urology, 10(1): 26-37.
Han, J.W., Jeong, J.K., Gurunathan, S., Choi, Y.J., Das, J., Kwon, D.N., et al. (2016). Male and female-derived somatic and germ cell-specific toxicity of silver nanoparticles in mouse. Nanotoxicology, 10(1): 361-373.
Ishikawa, T., Kondo, Y., Goda, K. and Fujisawa, M. (2005). Overexpression of endothelial nitric oxide synthase in transgenic mice accelerates testicular germ cell apoptosis induced by experimental cryptorchidism. Journal of Andrology, 26(2): 281-288.
Jintakosol, T. and Singjai, P. (2009). Effect of annealing treatment on luminescence property of MgO nanowires. Current Applied Physics, 9(6): 1288-1292.
Kang, S., Herzberg, M., Rodrigues, D.F. and Elimelech, M. (2008). Antibacterial effects of carbon nanotubes: size does matter. Langmuir, 24(13): 6409-6413.
Kaur, R. and Kaur, K. (2000). Effects of dietary selenium on morphology of testis and cauda epididymis in rats. Indian Journal of Pharmacology, 44(3): 265-272.
Kavak, E.C., Bulmus, F.G., Bulmus, O., Burcin Kavak, S. and Kocaman, N. (2018). Magnesium: Does it reduce ischemia/reperfusion injury in an adnexal torsion rat model? Drug Design. Development and Therapy, 12(1): 409-415.
Katz, D.F., Yanagimach, R. (1980). Movement characteristics of hamster spermatozoa within the oviduct. Biology Reproduction, 22(4): 759-764.
Kesmati, M., Konani, M., Torabi, M. and Khajehpour, L. (2016). Magnesium oxide nanoparticles reduce anxiety induced by morphine withdrawal in adult male mice. Physiology and Pharmacology, 20(3): 197-205.
Koksal, T., Erdogru, T., Toptas, B., Gulkesen, K.H., Usta, M.,Baykal, A., et al. (2002). Effect of experimental varicocele in rats on testicular oxidative stress status. Andrologia, 34(1): 242-247.
Mann, S.L., Patton, W.C., King, A. and Chan, P.J. (2002). Comparative genomic hybridization analysis of sperm DNA apoptosis after exposure to heat shock. Journal of Assisted Reproduction and Genetics, 19(4): 195-200.
Marmar, J.L. (2001). The pathophysiology of varicoceles in the light of current molecular and genetic information. Human Reproduction Update, 7(5): 461-472.
Martinez-Boubeta, C., Balcells, L., Cristofol, R., Sanfeliu, C., Rodriguez, E. and Weissleder, R. (2010). Self-assembled multifunctional Fe/MgO nanospheres for magnetic resonance imaging and hyperthermia. Nanomedicine: Nanotechnology, Biology and Medicine Journal, 6(2): 362-370.
Matovic, V., Buha, A., Buha, Z., Dukic-Cosic, D., Miljkovic, M., Ivanisevic, J. and et al. (2012). Route-dependent effects of cadmium/cadmium and magnesium acute treatment on parameters of oxidative stress in rat liver. Food Chemistry Toxicology, 50(1): 552-557.
Mohajeri, D., Mousavi, G.H. and Mansouri M.B. (2012). Histopathological study on the effects of turmeric (Curcuma longa linn.) powder on renal ischemia-reperfusion injury in rats. Journal of Veterinary Clinical Pathology, 6(1): 1493-1503. [In Persian]
Mohammad nejad, D., Soleimani rad, J. and Mohammadi-roshandeh, A. (2013). Preventive effects of Cetrorelix on the changes induced by Cisplatin on spermatogenic and myoid cells and basal lamina of seminiferous ducts in the testis of Balb/C mouse. Journal of Veterinary Clinical Pathology, 6(4): 1665-1674. [In Persian]
Mostafa, T., Anis, T., Imam, H. and El-Nashar, A.R. (2009). Seminal reactive oxygen species-antioxidant relationship in fertile males with and without varicocele. Andrologia, 41(2): 125-129.
NavaeianKalat, E., Tavalaee, M., Abasi, H. and Nasr Esfahani, M.H. (2012). Comparison of sperm parameters and DNA integrity between fertile and varicocele individuals. Journal of Cell, 3(2): 171-177.
Park, J.Y., Lee, Y.J., Jun, K.W., Baeg, J.O. and Yim, D.J. (2006). Chemical synthesis and characterization of highly oil dispersed MgO nanoparticles. Journal of Industrial and Engineering Chemistry, 12(6): 882-887.
Park, E.J., Bae, E., Yi, J., Kim, Y., Choi, K., Lee, S.H., et al., (2010). Repeated-dose toxicity and inflammatory responses in mice by oral administration of silver nanoparticles. Environmental Toxicology and Pharmacology, 30(2): 162-168.
Pasqualotto, F.F., Sundaram, A., Sharma, R.K. and Borges, E. (2008). Semen quality and oxidative stress scores in fertile and infertile patients with varicocele. Fertility and Sterility, 89(3): 602-607.
Razavi, S., Nasr-Esfahani, M., Mardani, M. and Javanmardi, S. (2006). Relation between protamine deficiency and sperm parameters, pronuclear morphology, cleavage and embryo quality. Cell Journal (Yakhteh), 2(1): 80-87.
Ren, C., Hu, X. and Zhou, Q. (2018). Graphene oxide quantum dots reduce oxidative stress and inhibit neurotoxicity in vitro and in vivo through catalase-like activity and metabolic regulation. Advanced Science, 5(5): 1-13.
Rudramurthy, G., Swamy, M., Sinniah, U. and Ghasemzadeh, A. (2016). Nanoparticles: alternatives against drug-resistant pathogenic microbes. Molecules, 21(7): 836-866.
Shittu, O.K., Aaron, S.Y., Oladuntoye, M.D. and Lawal, B. (2018). Diminazene aceturate modified nanocomposite for improved efficacy in acute trypanosome infection. Journal of Acute Disease, 7(1): 36-42.
Twigg, J., Fulton, N., Gomez, E., Irvine, D.S. and Aitken, R.J. (1998). Analysis of the impact of intracellular reactive oxygen species generation on the structural and functional integrity of human spermatozoa: lipid peroxidation, DNA fragmentation and effectiveness of antioxidants. Human Reproduction, 13(6): 1429-1436.
Valsa, J., Padmanabhan Skandhan, K., Sahab Khan, P., Sumangala, B. and Gondalia, M. (2012). Split ejaculation study: semen parameters and calcium and magnesium in seminal plasma. Central European Journal of Urology, 65(4): 216-218.
Villanueva-Diaz, C.A., Vega-Hernandez, E.A., Diaz-Perez, M.A., Echavarria-Sanchez, M., et al. (1999). Sperm dysfunction in subfertile patients with varicocele and marginal semen analysis. Andrologia, 31(5): 263-267.
Visiki, S., Szollosi, L., Kiss, A.S. and Csikkel-Szolnoki, A. (1997). Effects of magnesium on spermiogenesis. Magnesium: Current Status and New Developments, 1(1): 335-339.
(2010). World Health Organization laboratory manual for the examination and processing of human semen, Geneva: World Health Organization. 5th edition.
Wu, G.J., Chang, F.W., Lee, S.S. and Cheng, Y.Y. (2009). Apoptosis-related phenotype of ejaculated spermatozoa in patients with varicocele. Fertility and Sterility, 91(3): 831-837.
Yang, Y., Wu, Z., Chen, Y., Qiao, M., Yuan, J., Nie, W., et al. (2006). Magnesium deficiency enhances hydrogen peroxide production and oxidative damage in chick embryo hepatocyte in vitro. Bio Metals, 19(1): 71-81.
Zhang, H., Zhou, Q.M., Li, X.D., Yi, X., Duan, X., Liu, B., et al. (2006). Ginsenoside Re increases fertile and asthenozoospermic infertile human sperm motility by induction of nitric oxide synthase. Archives of Pharmacal Research, 29(2): 145-151.
_||_
Abinaya, S., Kavitha, H.P., Prakash, M. and Muthukrishnaraj, A. (2021). Green synthesis of magnesium oxide nanoparticles and its applications: A review. Sustainable Chemistry and Pharmacy, 19(7) 1-9.
Agarwal, A., Durairajanayagam, D., Halabi, J., Peng, J. and Vazquez-Levin, M. (2014). Proteomics, oxidative stress and male infertility. Reproductive BioMedicine Online, 29(1): 32-58.
Ahmadi Asr Badr, Y., Hasanzadeh, K. and Jahantabi, E. (2016). Evaluation of the effect of the number and size of veins ligated at left sided microsurgical inguinal varicocelectomy on the semen analysis outcomes. Medical Journal of Tabriz University of Medical Sciences, 38(3): 6-11.
Aleksandra, B., Zorica, B., Danijela, D., Vesna, M. and Buha, A. (2012). Magnesium effects against cd-induced stress in rats. Archives of Industrial Hygiene and Toxicology, 63(1): 247-254.
Amouoghli Tabrizi, B. and Khakpour, M. (2013). Protective effects of ginger (Zingiber officinale) rhizome extract on heatinduced testicular damage in the mouse. Journal of Veterinary Clinical Pathology, 27(7): 183-261. [In Persian]
Bahmanzadeh, M., Abolhassani, F., Amidi, F., Ejtemaiemehr, Sh., Salehi, M. and Abbasi M. (2008). The effects of nitric oxide synthase inhibitor (L-NAME) on epididymal sperm count, motility, and morphology in varicocelized rat. DARU Journal of Pharmaceutical Sciences, 16(1): 23-28.
Bisht, S., Faiq, M., Tolahunase, M. and Dada, R. (2017). Oxidative stress and male infertility. Nature Reviews Urology, 14(8): 470-485.
Brown, D.G. and Burk, R.F. (1973). Selenium retention in tissues and sperm of rats fed a torula yeast diet. The Journal of Nutrition, 102(1):102-108.
Cai, L., Chen, J., Liu, Z., Wang, H., Yang, H. and Ding, W. (2018). Magnesium Oxide Nanoparticles: Effective agricultural antibacterial agent against ralstonia solanacearum. Frontiers in Microbiology, 790(9): 1-19.
Celik-Ozenci, C., Bayram, Z., Akkoyunlu, G., Türkay Korgun, E.,Erdogru, T., Seval, Y., et al. (2006). Localization of NGF and nNOS in varicocele-induced rat testis. Acta Histochemica, 107(6): 435-442.
Chandra, A.K., Sengupta, P., Goswam, H. and Sarkar, M. (2013). Effects of dietary magnesium on testicular histology, steroidogenesis, spermatogenesis and oxidative stress markers in adult rat. Indian Journal of Experimental Biology, 51(1): 37-47.
Comhaire, F. and Vermeulen, A. (1974). Varicocele sterility: Cortisol and catecholamines. Fertility and Sterility, 25(1): 88-95.
Goodson, S.G., Zhang, Z., Tsuruta, J.K., Wang, W. and O'Brien, D.A. (2011). Classification of mouse sperm motility patterns using an automated multiclass support vector machines model. Biology of Reproduction, 84(6):1207-15.
Girardi, S.K. and Goldestein, M. (1997). Varicocele. Current Therapy in Endocrinology and Metabolism, 6(1): 355-358.
Giri, A., Khynriam, D. and Prasad, S.B. (1998).Vitamin C mediated protection on cisplatin induced mutagenicity in mice. Mutation Research, 421(2):139-148.
Gual-Frau, J., Abad, C., Amengual, M.J., Hannaoui, N., Checa, M.A., Ribas-Maynou, J. and et al. (2015). Oral antioxidant treatment partly improves integrity of human sperm DNA in infertile grade I varicocele patients. Human Fertility, 18(3): 225-9.
Hamada, A., Esteves, S.C. and Agarwal, A. (2013). Insight into oxidative stress in varicocele-associated male infertility. Part 2. Nature Reviews Urology, 10(1): 26-37.
Han, J.W., Jeong, J.K., Gurunathan, S., Choi, Y.J., Das, J., Kwon, D.N., et al. (2016). Male and female-derived somatic and germ cell-specific toxicity of silver nanoparticles in mouse. Nanotoxicology, 10(1): 361-373.
Ishikawa, T., Kondo, Y., Goda, K. and Fujisawa, M. (2005). Overexpression of endothelial nitric oxide synthase in transgenic mice accelerates testicular germ cell apoptosis induced by experimental cryptorchidism. Journal of Andrology, 26(2): 281-288.
Jintakosol, T. and Singjai, P. (2009). Effect of annealing treatment on luminescence property of MgO nanowires. Current Applied Physics, 9(6): 1288-1292.
Kang, S., Herzberg, M., Rodrigues, D.F. and Elimelech, M. (2008). Antibacterial effects of carbon nanotubes: size does matter. Langmuir, 24(13): 6409-6413.
Kaur, R. and Kaur, K. (2000). Effects of dietary selenium on morphology of testis and cauda epididymis in rats. Indian Journal of Pharmacology, 44(3): 265-272.
Kavak, E.C., Bulmus, F.G., Bulmus, O., Burcin Kavak, S. and Kocaman, N. (2018). Magnesium: Does it reduce ischemia/reperfusion injury in an adnexal torsion rat model? Drug Design. Development and Therapy, 12(1): 409-415.
Katz, D.F., Yanagimach, R. (1980). Movement characteristics of hamster spermatozoa within the oviduct. Biology Reproduction, 22(4): 759-764.
Kesmati, M., Konani, M., Torabi, M. and Khajehpour, L. (2016). Magnesium oxide nanoparticles reduce anxiety induced by morphine withdrawal in adult male mice. Physiology and Pharmacology, 20(3): 197-205.
Koksal, T., Erdogru, T., Toptas, B., Gulkesen, K.H., Usta, M.,Baykal, A., et al. (2002). Effect of experimental varicocele in rats on testicular oxidative stress status. Andrologia, 34(1): 242-247.
Mann, S.L., Patton, W.C., King, A. and Chan, P.J. (2002). Comparative genomic hybridization analysis of sperm DNA apoptosis after exposure to heat shock. Journal of Assisted Reproduction and Genetics, 19(4): 195-200.
Marmar, J.L. (2001). The pathophysiology of varicoceles in the light of current molecular and genetic information. Human Reproduction Update, 7(5): 461-472.
Martinez-Boubeta, C., Balcells, L., Cristofol, R., Sanfeliu, C., Rodriguez, E. and Weissleder, R. (2010). Self-assembled multifunctional Fe/MgO nanospheres for magnetic resonance imaging and hyperthermia. Nanomedicine: Nanotechnology, Biology and Medicine Journal, 6(2): 362-370.
Matovic, V., Buha, A., Buha, Z., Dukic-Cosic, D., Miljkovic, M., Ivanisevic, J. and et al. (2012). Route-dependent effects of cadmium/cadmium and magnesium acute treatment on parameters of oxidative stress in rat liver. Food Chemistry Toxicology, 50(1): 552-557.
Mohajeri, D., Mousavi, G.H. and Mansouri M.B. (2012). Histopathological study on the effects of turmeric (Curcuma longa linn.) powder on renal ischemia-reperfusion injury in rats. Journal of Veterinary Clinical Pathology, 6(1): 1493-1503. [In Persian]
Mohammad nejad, D., Soleimani rad, J. and Mohammadi-roshandeh, A. (2013). Preventive effects of Cetrorelix on the changes induced by Cisplatin on spermatogenic and myoid cells and basal lamina of seminiferous ducts in the testis of Balb/C mouse. Journal of Veterinary Clinical Pathology, 6(4): 1665-1674. [In Persian]
Mostafa, T., Anis, T., Imam, H. and El-Nashar, A.R. (2009). Seminal reactive oxygen species-antioxidant relationship in fertile males with and without varicocele. Andrologia, 41(2): 125-129.
NavaeianKalat, E., Tavalaee, M., Abasi, H. and Nasr Esfahani, M.H. (2012). Comparison of sperm parameters and DNA integrity between fertile and varicocele individuals. Journal of Cell, 3(2): 171-177.
Park, J.Y., Lee, Y.J., Jun, K.W., Baeg, J.O. and Yim, D.J. (2006). Chemical synthesis and characterization of highly oil dispersed MgO nanoparticles. Journal of Industrial and Engineering Chemistry, 12(6): 882-887.
Park, E.J., Bae, E., Yi, J., Kim, Y., Choi, K., Lee, S.H., et al., (2010). Repeated-dose toxicity and inflammatory responses in mice by oral administration of silver nanoparticles. Environmental Toxicology and Pharmacology, 30(2): 162-168.
Pasqualotto, F.F., Sundaram, A., Sharma, R.K. and Borges, E. (2008). Semen quality and oxidative stress scores in fertile and infertile patients with varicocele. Fertility and Sterility, 89(3): 602-607.
Razavi, S., Nasr-Esfahani, M., Mardani, M. and Javanmardi, S. (2006). Relation between protamine deficiency and sperm parameters, pronuclear morphology, cleavage and embryo quality. Cell Journal (Yakhteh), 2(1): 80-87.
Ren, C., Hu, X. and Zhou, Q. (2018). Graphene oxide quantum dots reduce oxidative stress and inhibit neurotoxicity in vitro and in vivo through catalase-like activity and metabolic regulation. Advanced Science, 5(5): 1-13.
Rudramurthy, G., Swamy, M., Sinniah, U. and Ghasemzadeh, A. (2016). Nanoparticles: alternatives against drug-resistant pathogenic microbes. Molecules, 21(7): 836-866.
Shittu, O.K., Aaron, S.Y., Oladuntoye, M.D. and Lawal, B. (2018). Diminazene aceturate modified nanocomposite for improved efficacy in acute trypanosome infection. Journal of Acute Disease, 7(1): 36-42.
Twigg, J., Fulton, N., Gomez, E., Irvine, D.S. and Aitken, R.J. (1998). Analysis of the impact of intracellular reactive oxygen species generation on the structural and functional integrity of human spermatozoa: lipid peroxidation, DNA fragmentation and effectiveness of antioxidants. Human Reproduction, 13(6): 1429-1436.
Valsa, J., Padmanabhan Skandhan, K., Sahab Khan, P., Sumangala, B. and Gondalia, M. (2012). Split ejaculation study: semen parameters and calcium and magnesium in seminal plasma. Central European Journal of Urology, 65(4): 216-218.
Villanueva-Diaz, C.A., Vega-Hernandez, E.A., Diaz-Perez, M.A., Echavarria-Sanchez, M., et al. (1999). Sperm dysfunction in subfertile patients with varicocele and marginal semen analysis. Andrologia, 31(5): 263-267.
Visiki, S., Szollosi, L., Kiss, A.S. and Csikkel-Szolnoki, A. (1997). Effects of magnesium on spermiogenesis. Magnesium: Current Status and New Developments, 1(1): 335-339.
(2010). World Health Organization laboratory manual for the examination and processing of human semen, Geneva: World Health Organization. 5th edition.
Wu, G.J., Chang, F.W., Lee, S.S. and Cheng, Y.Y. (2009). Apoptosis-related phenotype of ejaculated spermatozoa in patients with varicocele. Fertility and Sterility, 91(3): 831-837.
Yang, Y., Wu, Z., Chen, Y., Qiao, M., Yuan, J., Nie, W., et al. (2006). Magnesium deficiency enhances hydrogen peroxide production and oxidative damage in chick embryo hepatocyte in vitro. Bio Metals, 19(1): 71-81.
Zhang, H., Zhou, Q.M., Li, X.D., Yi, X., Duan, X., Liu, B., et al. (2006). Ginsenoside Re increases fertile and asthenozoospermic infertile human sperm motility by induction of nitric oxide synthase. Archives of Pharmacal Research, 29(2): 145-151.
