The Effects of Serum and Follicular Fluid Vitamin D Levels on Assisted Reproductive Techniques: A Prospective Cohort Study

Document Type : Original Article


1 Department of Midwifery and Reproductive Health, Isfahan University of Medical Sciences, Isfahan, Iran

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

3 Isfahan Fertility and Infertility Center, Isfahan, Iran

4 School of Nursing and Midwifery, Faculty of Medicine, Nursing and Health Sciences, Monash University, Melbourne, Australia


Background: Based on studies on animal models, vitamin D plays an essential role in reproduction by controlling
Ca and Mg levels. Despite these findings, the effects of vitamin D deficiency and supplementation on the outcome of
assisted reproductive techniques (ART) remain controversial. Therefore, the aim of the present study was to assess the
relationship between serum and follicular fluid 25-OH vitamin D levels on reproductive outcomes of infertile women.
Materials and Methods: This prospective cohort study included 150 infertile women who underwent in vitro fertilization
(IVF) or intracytoplasmic sperm injection (ICSI). The participants were allocated to one of the three groups according
to their serum and follicular fluid 25-OH vitamin D concentrations (less than 10 ng/ml, between 10 and 30 ng/ml and more than 30 ng/ml), and fertilization, cleavage and biochemical and clinical pregnancy rates were compared among the groups. Data was analyzed by SPSS software and using Chi-square and Spearman correlation coefficient.
Results: Serum and follicular fluid vitamin D levels significantly correlated with biochemical (P=0.008), (P=0.003)
and clinical pregnancy (P=0.017), (P=0.001) rates respectively . However, the quality of embryos (P=0.125), (P=0.106)
and fertilization rate (P=0.082), (P=0.059) were not associated with the level of serum and follicular fluid vitamin D.
Conclusion: This study found that women with higher levels of vitamin D in their serum and follicular fluid are significantly more likely to achieve pregnancy but without affecting the quality of embryo and fertility rate.


  1. Hansen KR, He ALW, Styer AK, Wild RA, Butts S, Engmann L, et al. Predictors of pregnancy and live-birth in couples with unexplained infertility after ovarian stimulation–intrauterine insemination. Fertil Steril. 2016; 105(6): 1575-1583. e2.
  2. Bushnik T, Cook JL, Yuzpe AA, Tough S, and Collins J. Estimating the prevalence of infertility in Canada. Hum Reprod. 2012; 27(3): 738-746.
  3. Anderson K, Nisenblat V, Norman R. Lifestyle factors in people seeking infertility treatment-a review. Aust N Z J Obstet Gynaecol. 2010; 50(1): 8-20.
  4. Paffoni A, Ferrari S, Viganò P, Pagliardini L, Papaleo E, Candiani M, et al. Vitamin D deficiency and infertility: insights from in vitro fertilization cycles. J Clin Endocrinol Metab. 2014; 99(11): E2372-E2376.
  5. Holick MF. Vitamin D deficiency. N Engl J Med. 2007; 357(3): 266- 281.
  6. Haussler MR, Whitfield GK, Kaneko I, Haussler CA, Hsieh D, Hsieh JC, et al. Molecular mechanisms of vitamin D action. Calcif Tissue Int. 2013; 92(2): 77-98.
  7. Jensen MB, Nielsen JE, Jørgensen A, Rajpert-De Meyts E, Kristensen DM, Jørgensen N, et al. Vitamin D receptor and vitamin D metabolizing enzymes are expressed in the human male reproductive tract. Hum Reprod. 2010; 25(5): 1303-1311.
  8. Ozkan S, Jindal S, Greenseid K, Shu J, Zeitlian G, Hickmon C, et al. Replete vitamin D stores predict reproductive success following in vitro fertilization. Fertil Steril. 2010; 94(4): 1314-1319.
  9. Pludowski P, Holick MF, Pilz S, Wagner CL, Hollis BW, Grant WB, et al. Vitamin D effects on musculoskeletal health, immunity, autoimmunity, cardiovascular disease, cancer, fertility, pregnancy, dementia and mortality—a review of recent evidence. Autoimmun Rev. 2013; 12(10): 976-989.
  10. Barrera D, Avila E, Hernández G, Halhali A, Biruete B, Larrea F, et al. Estradiol and progesterone synthesis in human placenta is stimulated by calcitriol. J Steroid Biochem Mol Biol. 2007; 103(3-5): 529-532.
  11. Evans KN, Bulmer JN, Kilby MD, Hewison M. Vitamin D and placental-decidual function. J Soc Gynecol Investig. 2004; 11(5): 263-271.
  12. Asadi M, Matin N, Frootan M, Mohamadpour J, Qorbani M, Davari Tanha F. Vitamin D improves endometrial thickness in PCOS women who need intrauterine insemination: a randomized double-blind placebo-controlled trial. Arch Gynecol Obstet. 2014; 289(4): 865-870.
  13. Shin JS, Choi MY, Longtine MS, Nelson DM. Vitamin D effects on pregnancy and the placenta. Placenta. 2010; 31(12): 1027-1034.
  14. Aleyasin A, Agha Hosseini M, Mahdavi A, Safdarian L, Fallahi P, Mohajeri MR, et al. Predictive value of the level of vitamin D in follicular fluid on the outcome of assisted reproductive technology. Eur J Obstet Gynecol Reprod Biol. 2011; 159(1): 132-137.
  15. Anifandis GM, Dafopoulos K, Messini CI, Chalvatzas N, Liakos N, Pournaras S, et al. Prognostic value of follicular fluid 25-OH vitamin D and glucose levels in the IVF outcome. Reprod Biol Endocrinol. 2010; 8: 91.
  16. Rudick BJ, Ingles SA, Chung K, Stanczyk FZ, Paulson RJ, Bendikson KA. Influence of vitamin D levels on in vitro fertilization outcomes in donor-recipient cycles. Fertil Steril. 2014; 101(2): 447-452.
  17. Pelzer ES, Allan JA, Waterhouse MA, Ross T, Beagley KW, Knox CL. Microorganisms within human follicular fluid: effects on IVF. PLoS One. 2013; 8(3): e59062.
  18. Revelli A, Delle Piane L, Casano S, Molinari E, Massobrio M, Rinaudo P. Follicular fluid content and oocyte quality: from single biochemical markers to metabolomics. Reprod Biol Endocrinol. 2009; 7: 40.
  19. Mendoza C, Ruiz-Requena E, Ortega E, Cremades N, Martinez F, Bernabeu R, et al. Follicular fluid markers of oocyte developmental potential. Hum Reprod. 2002; 17(4): 1017-1022.
  20. Somigliana E, Panina-Bordignon P, Murone S, Di Lucia P, Vercellini P, Vigano P. Vitamin D reserve is higher in women with endometriosis. Hum Reprod. 2007; 22(8): 2273-2278.
  21. van de Vijver A, Drakopoulos P, Van Landuyt L, Vaiarelli A, Blockeel C, Santos-Ribeiro S, et al. Vitamin D deficiency and pregnancy rates following frozen–thawed embryo transfer: a prospective cohort study. Hum Reprod. 2016; 31(8): 1749-1754.
  22. Vanni VS, Vigano P, Somigliana E, Papaleo E, Paffoni A, Pagliardini L, et al. Vitamin D and assisted reproduction technologies: current concepts. Reprod Biol Endocrinol. 2014; 12: 47.
  23. Zhao J, Huang X, Xu B, Yan Y, Zhang Q, Li Y. Whether vitamin D was associated with clinical outcome after IVF/ICSI: a systematic review and meta-analysis. Reprod Biol Endocrinol. 2018; 16(1): 13.
  24. Nandi A, Sinha N, Ong E, Sonmez H, Poretsky L. Is there a role for vitamin D in human reproduction? Horm Mol Biol Clin Investig. 2016; 25(1): 15-28.
  25. Abadia L, Gaskins AJ, Chiu YH, Williams PL, Keller M, Diane L, et al. Serum 25-hydroxyvitamin D concentrations and treatment outcomes of women undergoing assisted reproduction. Am J Clin Nutr. 2016; 104(3): 729-735.
  26. Garbedian K, Boggild M, Moody J, Liu KE. Effect of vitamin D status on clinical pregnancy rates following in vitro fertilization. CMAJ Open. 2013;1(2): E77-E82.
  27. Polyzos NP, Anckaert E, Guzman L, Schiettecatte J, Van Landuyt L, Camus M, et al. Vitamin D deficiency and pregnancy rates in women undergoing single embryo, blastocyst stage, transfer (SET) for IVF/ICSI. Hum Reprod. 2014; 29(9): 2032-2040.
  28. Franasiak JM, Molinaro T, Dubell EK, Scott K, Ruiz AR, Forman EJ, et al. Vitamin D levels do not affect IVF outcomes following the transfer of euploid blastocysts. Am J Obstet Gynecol. 2015; 212(3): 315. e1-e6.
  29. Firouzabadi RD, Rahmani E, Rahsepar M, Firouzabadi MM. Value of follicular fluid vitamin D in predicting the pregnancy rate in an IVF program. Arch Gynecol Obstet. 2014; 289(1): 201-206.
  30. Estes SJ, Ye B, Qiu W, Cramer D, Hornstein MD, Missmer SA. A proteomic analysis of IVF follicular fluid in women. Fertil Steril. 2009; 92(5): 1569-1578.
  31. Aleyasin A, Hosseini MA, Mahdavi A, Safdarian L, Fallahi P, Mohajeri MR, et al. Predictive value of the level of vitamin D in follicular fluid on the outcome of assisted reproductive technology. Eur J Obstet Gynecol Reprod Biol. 2011; 159(1): 132-137.
  32. Zanatta A, Rocha AM, Carvalho FM, Pereira RMA, Taylor HS, Motta ELA, et al. The role of the Hoxa10/HOXA10 gene in the etiology of endometriosis and its related infertility: a review. J Assist Reprod Genet. 2010; 27(12): 701-710.
  33. Evans KN, Nguyen L, Chan J, Innes BA, Bulmer JN, Kilby MD, et al. Effects of 24-hydroxyvitamin D3 and 1,25-dihydroxyvitamin D3 on cytokine production by human decidual cells. Biol Reprod. 2006; 75(6): 816-822.
  34. Das SK. Regional development of uterine decidualization: Molecular signaling by Hoxa-10. Mol Reprod Dev. 2010; 77(5): 387-396.
  35. Bodnar LM, Catov JM, Simhan HN, Holick MF, Powers RW, Roberts JM. Maternal vitamin D deficiency increases the risk of preeclampsia. J Clin Endocrinol Metab. 2007; 92(9): 3517-3522.
  36. Hammoud AO, Meikle AW, Peterson CM, Stanford J, Gibson M, Carrell DT. Association of 25-hydroxy-vitamin D levels with semen and hormonal parameters. Asian J Androl. 2012; 14(6): 855-859.
  37. Bodnar LM, Catov JM, Zmuda JM, Cooper ME, Parrott MS, Roberts JM, et al. Maternal serum 25-hydroxyvitamin D concentrations are associated with small-for-gestational age births in white women. J Nutr. 2010; 140(5): 999-1006.
  38. Mannion CA, Gray-Donald K, Koski KG. Association of low intake of milk and vitamin D during pregnancy with decreased birth weight. CMAJ. 2006; 174(9): 1273-1277.
  39. Brooke OG, Brown IR, Bone CD, Carter ND, Cleeve HJ, Maxwell JD, et al. Vitamin-D supplements in pregnant Asian women:effects on calcium status and fetal growth. Br Med J. 1980; 280(6216): 751-754.
  40. Poel YHM, Hummel P, Lips P, Stam F, van der Ploeg T, Simsek S. Vitamin D and gestational diabetes: a systematic review and metaanalysis. Eur J Intern Med. 2012; 23(5): 465-469.