The Effect of Recombinant Human Follicle-Stimulating Hormone on Sperm Quality, Chromatin Status and Clinical Outcomes of Infertile Oligozoospermic Men Candidate for Intracytoplasmic Sperm Injection: A Randomized Clinical Trial

Document Type : Original Article


1 Department of Biology, Fars Science and Research Branch, Islamic Azad University, Fars, Iran

2 Department of Biology, Shiraz Branch, Islamic Azad University, Shiraz, Iran

3 Department of Reproductive Biotechnology, Reproductive Biomedicine Research Center, Royan Institute for Biotechnology, ACECR,Isfahan, Iran

4 Isfahan Fertility and Infertility Center, Isfahan, Iran

5 Department of Biology, Marvdasht Branch, Islamic Azad University, Marvdasht, Iran

6 Department of Reproductive Biotechnology, Reproductive Biomedicine Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran


Background: Follicle-stimulating hormone (FSH) plays a crucial role in spermatogenesis; in this study, we assessed the effect of recombinant human FSH (rhFSH) on sperm parameters, chromatin status and clinical outcomes of infertile oligozoospermic men candidates for intracytoplasmic sperm injection (ICSI).

Materials and Methods: This interventional randomized clinical trials (IRCT) included 40 infertile oligozoospermic men undergoing ICSI. These individuals were randomized into two groups: 20 men received rhFSH drug for three months and the other 20 men who did not receive rhFSH drug were considered the control group. Before and 3 months after treatment initiation, sperm parameters (using computer-assisted semen analysis) and chromatin status [using chromomycin A3, aniline blue, and sperm chromatin dispersion (SCD) tests] were assessed in these individuals. Furthermore, hormonal profile was assessed using enzyme-linked immunosorbent assay (ELISA). Clinical outcomes of ICSI were also compared between the two groups.

Results: The rhFSH treated group showed a significant increase in the level of FSH, luteinizing hormone (LH), testosterone (T) and prolactin (PRL), as well as significant improvements in sperm parameters compared to the control group. Also, after administration of rhFSH, there was asignificant reduction in the percentage of sperm DNA damage, protamine deficiency and chromatin immaturity, while such a reduction in these parameters was not observed in the control group. Moreover, the percentage of embryos with grade Aquality, was significantly higher in the rhFSH group compared to the control group. The pregnancy rate in the rhFSH group was higher than the control group but the difference was insignificant.

Conclusion: Administration of rhFSH improves sperm quality in infertile oligozoospermic men and results in higher rates of good quality embryos post-ICSI (Registration number: IRCT20170923036334N2).


1. Mehta RH, Makwana S, Ranga GM, Srinivasan RJ, Virk SS. Prev­alences of oligozoospermia and azoospermia in male partners of infertile couples from different parts of India. Asian J Androl. 2006; 8(1): 89-93.
2. Zhang W, Song X, Ni F, Cheng J, Wu BL, Jiang H. Association analysis between HFM1 variations and idiopathic azoospermia or severe oligozoospermia in Chinese Men. Sci China Life Sci. 2017; 60(3): 315-318.
3. McLachlan RI. Approach to the patient with oligozoospermia. J Clin Endocrinol Metab. 2013; 98(3): 873-80.
4. Niederberger C. WHO manual for the standardized investigation, diagnosis and management of the infertile male. Urology. 2001; 57(1): 208.
5. Rowe P, Comhaire F, Hargreave T. WHO manual for the standard­ized investigation, diagnosis and management of the infertile male. Cambridge: Press Syndicate of the University of Cambridge; 2000.
6. Ben-Rafael Z, Farhi J, Feldberg D, Bartoov B, Kovo M, Eltes F, et al. Follicle-stimulating hormone treatment for men with idiopathic oligoteratoasthenozoospermia before in vitro fertilization: the im­pact on sperm microstructure and fertilization potential. Fertil Steril. 2000; 73(1): 24-30.
7. Farmakiotis D, Farmakis C, Rousso D, Kourtis A, Katsikis I, Panidis D. The beneficial effects of toremifene administration on the hy­pothalamic-pituitary-testicular axis and sperm parameters in men with idiopathic oligozoospermia. Fertil Steril. 2007; 88(4): 847-853.
8. Ratnayake GM, Weerathunga PN, Ruwanpura LP, Wickramasin­ghe A, Katulanda P. Isolated follicle stimulated hormone deficiency in male: case report. BMC Res Notes. 2018; 11(1): 24.
9. Yin W, Gore AC. The hypothalamic median eminence and its role in reproductive aging. Ann N Y Acad Sci. 2010; 1204: 113-122.
10. Sharpe RM. Environment, lifestyle and male infertility. Baillieres Best Pract Res Clin Endocrinol Metab. 2000; 14(3): 489-503.
11. Santi D, Granata AR, Simoni M. FSH treatment of male idiopathic infertility improves pregnancy rate: a meta-analysis. Endocr Con­nect. 2015; 4(3): R46-R58.
12. Nieschlag E, Simoni M, Gromoll J, Weinbauer GF. Role of FSH in the regulation of spermatogenesis: clinical aspects. Clin Endocrinol (Oxf). 1999; 51(2): 139-146.
13. Ding Ym, Zhang XJ, Li JP, Chen SS, Zhang RT, Tan Wl, et al. Treatment of idiopathic oligozoospermia with recombinant human follicle-stimulating hormone: a prospective, randomized, double-blind, placebo-controlled clinical study in Chinese population. Clin Endocrinol (Oxf). 2015; 83(6): 866-871.
14. Lenzi A, Balercia G, Bellastella A, Colao A, Fabbri A, Foresta C, et al. Epidemiology, diagnosis, and treatment of male hypogonado­tropic hypogonadism. J Endocrinol Invest. 2009; 32(11): 934-938.
15. Colacurci N, De Leo V, Ruvolo G, Piomboni P, Caprio F, Pivonello R, et al. Recombinant FSH improves sperm DNA damage in Male Infertility: a phase II clinical trial. Front Endocrinol (Lausanne). 2018; 9: 383.
16. Colacurci N, Monti MG, Fornaro F, Izzo G, Izzo P, Trotta C, et al. Recombinant human FSH reduces sperm DNA fragmentation in men with idiopathic oligoasthenoteratozoospermia. J Androl. 2012; 33(4): 588-593.
17. Ruvolo G, Roccheri MC, Brucculeri AM, Longobardi S, Cittadini E, Bosco L. Lower sperm DNA fragmentation after r-FSH adminis­tration in functional hypogonadotropic hypogonadism. J Assist Re­prod Genet. 2013; 30(4): 497-503.
18. Simoni M, Santi D, Negri L, Hoffmann I, Muratori M, Baldi E, et al. Treatment with human, recombinant FSH improves sperm DNA fragmentation in idiopathic infertile men depending on the FSH receptor polymorphism p.N680S: a pharmacogenetic study. Hum Reprod. 2016; 31(9): 1960-1969.
19. Kamischke A, Behre HM, Bergmann M, Simoni M, Schäfer T, Ni­eschlag E. Recombinant human follicle stimulating hormone for treatment of male idiopathic infertility: a randomized, double-blind, placebo-controlled, clinical trial. Hum Reprod. 1998; 13(3): 596-603.
20. Paradisi R, Busacchi P, Seracchioli R, Porcu E, Venturoli S. Effects of high doses of recombinant human follicle-stimulating hormone in the treatment of male factor infertility: results of a pilot study. Fertil Steril. 2006; 86(3): 728-731.
21. Selice R, Garolla A, Pengo M, Caretta N, Ferlin A, Foresta C. The response to FSH treatment in oligozoospermic men depends on FSH receptor gene polymorphisms. Int J Androl. 2011; 34(4pt1): 306-312.
22. Casamonti E, Vinci S, Serra E, Fino MG, Brilli S, Lotti F, et al. Short-term FSH treatment and sperm maturation: a prospective study in idiopathic infertile men. Andrology. 2017; 5(3): 414-422.
23. Menkveld R. Clinical significance of the low normal sperm morphol­ogy value as proposed in the fifth edition of the WHO Laboratory Manual for the Examination and Processing of Human Semen. Asian J Androl. 2010; 12(1): 47-58.
24. Barbonetti A, Calogero AE, Balercia G, Garolla A, Krausz C, La Vi­gnera S, et al. The use of follicle stimulating hormone (FSH) for the treatment of the infertile man: position statement from the Italian Society of Andrology and Sexual Medicine (SIAMS). J Endocrinol Invest. 2018; 41(9): 1107-1122.
25. Muriel L, Meseguer M, Fernández JL, Alvarez J, Remohí J, Pellicer A, et al. Value of the sperm chromatin dispersion test in predicting pregnancy outcome in intrauterine insemination: a blind prospec­tive study. Hum Reprod. 2006; 21(3): 738-744.
26. Nasr-Esfahani MH, Razavi S, Mardani M. Andrology: relation be­tween different human sperm nuclear maturity tests and in vitro fertilization. J Assist Reprod Genet. 2001; 18(4): 219-225.
27. Terquem A, Dadoune JP. Aniline blue staining of human spermato­zoa chromatin: evaluation of nuclear maturation. In: Andre J, editor. The sperm cell. London: Martinus Nijhoff; 1983; 696-701.
28. Giorgetti C, Terriou P, Auquier P, Hans E, Spach JL, Salzmann J, et al. Embryo score to predict implantation after in-vitro fertilization: based on 957 single embryo transfers. Hum Reprod. 1995; 10(9): 2427-2431.
29. Terriou P, Giorgetti C, Hans E, Salzmann J, Charles O, Cignetti L, et al. Relationship between even early cleavage and day 2 embryo score and assessment of their predictive value for pregnancy. Re­ prod Biomed Online. 2007; 14(3): 294-299.
30.Zegers-Hochschild F, Adamson GD, Dyer S, Racowsky C, de Mou­zon J, Sokol R, et al. The international glossary on infertility and fertility care, 2017. Hum Reprod. 2017; 32(9): 1786-1801.
31.Simon L, Murphy K, Shamsi MB, Liu L, Emery B, Aston KI, et al.Paternal influence of sperm DNA integrity on early embryonic de­velopment. Hum Reprod. 2014; 29(11): 2402-2412.
32.Deng C, Li T, Xie Y, Guo Y, Yang QY, Liang X, et al. SpermDNA fragmentation index influences assisted reproductivetechnology outcome: a systematic review and meta-analysiscombined with a retrospective cohort study. Andrologia. 2019;51(6): e13263.
33.Collins JA, Barnhart KT, Schlegel PN. Do sperm DNA integritytests predict pregnancy with in vitro fertilization? Fertil Steril. 2008;89(4): 823-831.
34.Lin MH, Kuo-Kuang Lee R, Li SH, Lu CH, Sun FJ, Hwu YM.Sperm chromatin structure assay parameters are not related tofertilization rates, embryo quality, and pregnancy rates in in vitrofertilization and intracytoplasmic sperm injection, but might berelated to spontaneous abortion rates. Fertil Steril. 2008; 90(2):352-359.
35.Twigg JP, Irvine DS, Aitken RJ.Oxidative damage to DNA in humanspermatozoa does not preclude pronucleus formation at intracyto­plasmic sperm injection. Hum Reprod. 1998; 13(7): 1864-1871.
36.González-Marín C, Gosálvez J, Roy R. Types, causes, detectionand repair of DNA fragmentation in animal and human sperm cells.Int J Mol Sci. 2012; 13(11): 14026-1452.
37.Alvarez Sedó C, Bilinski M, Lorenzi D, Uriondo H, Noblía F, Lon­gobucco V, et al. Effect of sperm DNA fragmentation on embryodevelopment: clinical and biological aspects. JBRA Assist Reprod.2017; 21(4): 343-350.
38.Ahmadi A, Ng SC. Fertilizing ability of DNA-damaged spermato­zoa. J Exp Zool. 1999; 284(6): 696-704.
39.Seli E, Gardner DK, Schoolcraft WB, Moffatt O, Sakkas D. Extentof nuclear DNA damage in ejaculated spermatozoa impacts onblastocyst development after in vitro fertilization. Fertil Steril. 2004;82(2): 378-383.
40.Santi D, Crépieux P, Reiter E, Spaggiari G, Brigante G, Casarini L,et al. Follicle-stimulating hormone (FSH) action on spermatogen­esis: a focus on physiological and therapeutic roles. J Clin Med.2020; 9(4): 1014.