Skip to main content

Associations of total and free 25OHD and 1,25(OH)2D with serum markers of inflammation in older men



Vitamin D is hypothesized to suppress inflammation. We tested total and free vitamin D metabolites and their association with inflammatory markers. Interleukin-6 levels were lower with higher 25-hydroxyvitamin D. 1,25-dihydroxyvitamin D and free 25OHD associations mirrored those of 25OHD. However, associations for the two metabolites diverged for tumor necrosis factor alpha (TNF-α) soluble receptors.


Vitamin D is hypothesized to suppress inflammation, and circulating 25-hydroxyvitamin D (25OHD) and inflammatory markers are inversely correlated. However, total serum 25OHD may not be the best indicator of biologically active vitamin D.


We tested serum total 25OHD, total 1,25(OH)2D, vitamin D binding protein (DBP), and estimated free 25OHD and free 1,25(OH)2D associations with inflammatory markers serum interleukin-6 (IL-6), TNF-α and their soluble receptors, interleukin-10 (IL-10), and C-reactive protein (CRP) as continuous outcomes and the presence of ≥2 inflammatory markers in the highest quartile as a dichotomous outcome, in a random subcohort of 679 men in the Osteoporotic Fractures in Men (MrOS) study.


IL-6 was lower in men with higher 25OHD (−0.23 μg/mL per standard deviation (SD) increase in 25OHD, 95 % confidence intervals (CI) −0.07 to −0.38 μg/mL) and with higher 1,25(OH)2D (−0.20 μg/mL, 95 % CI −0.0004 to −0.39 μg/mL); free D associations were slightly stronger. 25OHD and DBP, but not 1,25(OH)2D, were independently associated with IL-6. TNF-α soluble receptors were inversely associated with 1,25(OH)2D but positively associated with 25OHD, and each had independent effects. The strongest association with ≥2 inflammatory markers in the highest quartile was for free 1,25(OH)2D (odds ratios (OR) 0.70, 95 % CI 0.54 to 0.89 per SD increase in free 1,25(OH)2D).


Associations of 1,25(OH)2D and free 25OHD with IL-6 mirrored those of 25OHD, suggesting that 1,25(OH)2D and free D do not improve upon 25OHD in population-based IL-6 studies. However, associations for the two metabolites diverged for TNF-α soluble receptor, warranting examination of both metabolites in studies of TNF-α and its antagonists.

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2
Fig. 3


  1. 1.

    Franceschi C (2007) Inflammaging as a major characteristic of old people: can it be prevented or cured? Nutr Rev 65:S173–176

    Article  PubMed  Google Scholar 

  2. 2.

    Collin-Osdoby P, Rothe L, Anderson F, Nelson M, Maloney W, Osdoby P (2001) Receptor activator of NF-kappa B and osteoprotegerin expression by human microvascular endothelial cells, regulation by inflammatory cytokines, and role in human osteoclastogenesis. J Biol Chem 276:20659–20672

    CAS  Article  PubMed  Google Scholar 

  3. 3.

    Doherty TM, Asotra K, Fitzpatrick LA, Qiao JH, Wilkin DJ, Detrano RC, Dunstan CR, Shah PK, Rajavashisth TB (2003) Calcification in atherosclerosis: bone biology and chronic inflammation at the arterial crossroads. Proc Natl Acad Sci U S A 100:11201–11206

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  4. 4.

    Cauley JA, Danielson ME, Boudreau RM, Forrest KY, Zmuda JM, Pahor M, Tylavsky FA, Cummings SR, Harris TB, Newman AB (2007) Inflammatory markers and incident fracture risk in older men and women: the Health Aging and Body Composition Study. J Bone Miner Res 22:1088–1095

    CAS  Article  PubMed  Google Scholar 

  5. 5.

    Ding C, Parameswaran V, Udayan R, Burgess J, Jones G (2008) Circulating levels of inflammatory markers predict change in bone mineral density and resorption in older adults: a longitudinal study. J Clin Endocrinol Metab 93:1952–1958

    CAS  Article  PubMed  Google Scholar 

  6. 6.

    Arnson Y, Itzhaky D, Mosseri M, Barak V, Tzur B, Agmon-Levin N, Amital H (2013) Vitamin D inflammatory cytokines and coronary events: a comprehensive review. Clin Rev Allergy Immunol 45:236–247

    CAS  Article  PubMed  Google Scholar 

  7. 7.

    Inanir A, Ozoran K, Tutkak H, Mermerci B (2004) The effects of calcitriol therapy on serum interleukin-1, interleukin-6 and tumour necrosis factor-alpha concentrations in post-menopausal patients with osteoporosis. J Int Med Res 32:570–582

    CAS  Article  PubMed  Google Scholar 

  8. 8.

    Swanson CM, Srikanth P, Lee CG, Cummings SR, Jans I, Cauley JA, Bouillon R, Vanderschueren D, Orwoll ES, Nielson CM (2015) Associations of 25-hydroxyvitamin D and 1,25-dihydroxyvitamin D with bone mineral density, bone mineral density change, and incident nonvertebral fracture. J Bone Miner Res 30:1403–1413

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  9. 9.

    Lange U, Jung O, Teichmann J, Neeck G (2001) Relationship between disease activity and serum levels of vitamin D metabolites and parathyroid hormone in ankylosing spondylitis. Osteoporos Int 12:1031–1035

    CAS  Article  PubMed  Google Scholar 

  10. 10.

    Zhang Y, Leung DY, Richers BN, Liu Y, Remigio LK, Riches DW, Goleva E (2012) Vitamin D inhibits monocyte/macrophage proinflammatory cytokine production by targeting MAPK phosphatase-1. J Immunol 188:2127–2135

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  11. 11.

    Rostkowska-Nadolska B, Sliupkas-Dyrda E, Potyka J, Kusmierz D, Fraczek M, Krecicki T, Kubik P, Zatonski M, Latocha M (2010) Vitamin D derivatives: calcitriol and tacalcitol inhibits interleukin-6 and interleukin-8 expression in human nasal polyp fibroblast cultures. Adv Med Sci 55:86–92

    CAS  Article  PubMed  Google Scholar 

  12. 12.

    Querfeld U (2013) Vitamin D and inflammation. Pediatr Nephrol 28:605–610

    Article  PubMed  Google Scholar 

  13. 13.

    Ding C, Wilding JP, Bing C (2013) 1,25-dihydroxyvitamin D3 protects against macrophage-induced activation of NFkappaB and MAPK signalling and chemokine release in human adipocytes. PLoS One 8:e61707

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  14. 14.

    Amital H, Szekanecz Z, Szucs G et al (2010) Serum concentrations of 25-OH vitamin D in patients with systemic lupus erythematosus (SLE) are inversely related to disease activity: is it time to routinely supplement patients with SLE with vitamin D? Ann Rheum Dis 69:1155–1157

    CAS  Article  PubMed  Google Scholar 

  15. 15.

    Rossini M, Maddali Bongi S, La Montagna G, Minisola G, Malavolta N, Bernini L, Cacace E, Sinigaglia L, Di Munno O, Adami S (2010) Vitamin D deficiency in rheumatoid arthritis: prevalence, determinants and associations with disease activity and disability. Arthritis Res Ther 12:R216

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  16. 16.

    Merlino LA, Curtis J, Mikuls TR, Cerhan JR, Criswell LA, Saag KG (2004) Vitamin D intake is inversely associated with rheumatoid arthritis: results from the Iowa Women’s Health Study. Arthritis Rheum 50:72–77

    CAS  Article  PubMed  Google Scholar 

  17. 17.

    Munger KL, Zhang SM, O’Reilly E, Hernan MA, Olek MJ, Willett WC, Ascherio A (2004) Vitamin D intake and incidence of multiple sclerosis. Neurology 62:60–65

    CAS  Article  PubMed  Google Scholar 

  18. 18.

    Blank JB, Cawthon PM, Carrion-Petersen ML, Harper L, Johnson JP, Mitson E, Delay RR (2005) Overview of recruitment for the osteoporotic fractures in men study (MrOS). Contemp Clin Trials 26:557–568

    Article  PubMed  Google Scholar 

  19. 19.

    Orwoll E, Blank JB, Barrett-Connor E et al (2005) Design and baseline characteristics of the osteoporotic fractures in men (MrOS) study—a large observational study of the determinants of fracture in older men. Contemp Clin Trials 26:569–585

    Article  PubMed  Google Scholar 

  20. 20.

    Orwoll E, Nielson CM, Marshall LM, Lambert L, Holton KF, Hoffman AR, Barrett-Connor E, Shikany JM, Dam T, Cauley JA (2009) Vitamin D deficiency in older men. J Clin Endocrinol Metab 94:1214–1222

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  21. 21.

    Singh RJ, Taylor RL, Reddy GS, Grebe SK (2006) C-3 epimers can account for a significant proportion of total circulating 25-hydroxyvitamin D in infants, complicating accurate measurement and interpretation of vitamin D status. J Clin Endocrinol Metab 91:3055–3061

    CAS  Article  PubMed  Google Scholar 

  22. 22.

    Vanderschueren D, Pye SR, O’Neill TW et al (2013) Active vitamin D (1,25-dihydroxyvitamin D) and bone health in middle-aged and elderly men: the European Male Aging Study (EMAS). J Clin Endocrinol Metab 98:995–1005

    CAS  Article  PubMed  Google Scholar 

  23. 23.

    Bouillon R, van Baelen H, de Moor P (1977) The measurement of the vitamin D-binding protein in human serum. J Clin Endocrinol Metab 45:225–231

    CAS  Article  PubMed  Google Scholar 

  24. 24.

    Chun RF, Peercy BE, Adams JS, Hewison M (2012) Vitamin D binding protein and monocyte response to 25-hydroxyvitamin D and 1,25-dihydroxyvitamin D: analysis by mathematical modeling. PLoS One 7:e30773

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  25. 25.

    Jones SA, Horiuchi S, Topley N, Yamamoto N, Fuller GM (2001) The soluble interleukin 6 receptor: mechanisms of production and implications in disease. FASEB J 15:43–58

    CAS  Article  PubMed  Google Scholar 

  26. 26.

    Washburn RA, Smith KW, Jette AM, Janney CA (1993) The physical Activity Scale for the Elderly (PASE): development and evaluation. J Clin Epidemiol 46:153–162

    CAS  Article  PubMed  Google Scholar 

  27. 27.

    Pahor M, Chrischilles EA, Guralnik JM, Brown SL, Wallace RB, Carbonin P (1994) Drug data coding and analysis in epidemiologic studies. Eur J Epidemiol 10:405–411

    CAS  Article  PubMed  Google Scholar 

  28. 28.

    Levey AS, Coresh J, Greene T, Marsh J, Stevens LA, Kusek JW, Van Lente F (2007) Expressing the Modification of Diet in Renal Disease Study equation for estimating glomerular filtration rate with standardized serum creatinine values. Clin Chem 53:766–772

    CAS  Article  PubMed  Google Scholar 

  29. 29.

    Lumley T, Diehr P, Emerson S, Chen L (2002) The importance of the normality assumption in large public health data sets. Annu Rev Public Health 23:151–169

    Article  PubMed  Google Scholar 

  30. 30.

    Laird E, McNulty H, Ward M et al (2014) Vitamin d deficiency is associated with inflammation in older Irish adults. J Clin Endocrinol Metab 99:1807–1815

    CAS  Article  PubMed  Google Scholar 

  31. 31.

    Wobke TK, Sorg BL, Steinhilber D (2014) Vitamin D in inflammatory diseases. Front Physiol 5:244

    PubMed  PubMed Central  Google Scholar 

  32. 32.

    Nonn L, Peng L, Feldman D, Peehl DM (2006) Inhibition of p38 by vitamin D reduces interleukin-6 production in normal prostate cells via mitogen-activated protein kinase phosphatase 5: implications for prostate cancer prevention by vitamin D. Cancer Res 66:4516–4524

    CAS  Article  PubMed  Google Scholar 

  33. 33.

    Shea MK, Booth SL, Massaro JM et al (2008) Vitamin K and vitamin D status: associations with inflammatory markers in the Framingham Offspring Study. Am J Epidemiol 167:313–320

    Article  PubMed  Google Scholar 

  34. 34.

    Bratke K, Wendt A, Garbe K, Kuepper M, Julius P, Lommatzsch M, Virchow JC (2014) Vitamin D binding protein and vitamin D in human allergen-induced endobronchial inflammation. Clin Exp Immunol 177:366–372

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  35. 35.

    Toss G, Sorbo B (1986) Serum concentrations of 25-hydroxyvitamin D and vitamin D-binding protein in elderly people. Effects of institutionalization, protein-energy malnutrition and inflammation. Acta Med Scand 220:273–277

    CAS  Article  PubMed  Google Scholar 

  36. 36.

    Zehnder D, Bland R, Chana RS, Wheeler DC, Howie AJ, Williams MC, Stewart PM, Hewison M (2002) Synthesis of 1,25-dihydroxyvitamin D(3) by human endothelial cells is regulated by inflammatory cytokines: a novel autocrine determinant of vascular cell adhesion. J Am Soc Nephrol 13:621–629

    CAS  PubMed  Google Scholar 

  37. 37.

    Edfeldt K, Liu PT, Chun R et al (2010) T-cell cytokines differentially control human monocyte antimicrobial responses by regulating vitamin D metabolism. Proc Natl Acad Sci U S A 107:22593–22598

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  38. 38.

    Hewison M, Burke F, Evans KN, Lammas DA, Sansom DM, Liu P, Modlin RL, Adams JS (2007) Extra-renal 25-hydroxyvitamin D3-1alpha-hydroxylase in human health and disease. J Steroid Biochem Mol Biol 103:316–321

    CAS  Article  PubMed  Google Scholar 

  39. 39.

    Hummel DM, Fetahu IS, Groschel C, Manhardt T, Kallay E (2014) Role of proinflammatory cytokines on expression of vitamin D metabolism and target genes in colon cancer cells. J Steroid Biochem Mol Biol 144PA:91–95

    Article  Google Scholar 

  40. 40.

    Welsh P, Peters MJ, McInnes IB et al (2011) Vitamin D deficiency is common in patients with RA and linked to disease activity, but circulating levels are unaffected by TNFalpha blockade: results from a prospective cohort study. Ann Rheum Dis 70:1165–1167

    Article  PubMed  Google Scholar 

  41. 41.

    Waldron JL, Ashby HL, Cornes MP, Bechervaise J, Razavi C, Thomas OL, Chugh S, Deshpande S, Ford C, Gama R (2013) Vitamin D: a negative acute phase reactant. J Clin Pathol 66:620–622

    CAS  Article  PubMed  Google Scholar 

  42. 42.

    Reid D, Toole BJ, Knox S, Talwar D, Harten J, O’Reilly DS, Blackwell S, Kinsella J, McMillan DC, Wallace AM (2011) The relation between acute changes in the systemic inflammatory response and plasma 25-hydroxyvitamin D concentrations after elective knee arthroplasty. Am J Clin Nutr 93:1006–1011

    CAS  Article  PubMed  Google Scholar 

  43. 43.

    Hopkins MH, Owen J, Ahearn T, Fedirko V, Flanders WD, Jones DP, Bostick RM (2011) Effects of supplemental vitamin D and calcium on biomarkers of inflammation in colorectal adenoma patients: a randomized, controlled clinical trial. Cancer Prev Res (Phila) 4:1645–1654

    CAS  Article  Google Scholar 

  44. 44.

    Peake JM, Kukuljan S, Nowson CA, Sanders K, Daly RM (2011) Inflammatory cytokine responses to progressive resistance training and supplementation with fortified milk in men aged 50+ years: an 18-month randomized controlled trial. Eur J Appl Physiol 111:3079–3088

    CAS  Article  PubMed  Google Scholar 

  45. 45.

    Pittas AG, Harris SS, Stark PC, Dawson-Hughes B (2007) The effects of calcium and vitamin D supplementation on blood glucose and markers of inflammation in nondiabetic adults. Diabetes Care 30:980–986

    CAS  Article  PubMed  Google Scholar 

  46. 46.

    Shab-Bidar S, Neyestani TR, Djazayery A, Eshraghian MR, Houshiarrad A, Kalayi A, Shariatzadeh N, Khalaji N, Gharavi A (2012) Improvement of vitamin D status resulted in amelioration of biomarkers of systemic inflammation in the subjects with type 2 diabetes. Diabetes Metab Res Rev 28:424–430

    CAS  Article  PubMed  Google Scholar 

  47. 47.

    Verstuyf A, Carmeliet G, Bouillon R, Mathieu C (2010) Vitamin D: a pleiotropic hormone. Kidney Int 78:140–145

    CAS  Article  PubMed  Google Scholar 

  48. 48.

    Hopkins MH, Flanders WD, Bostick RM (2012) Associations of circulating inflammatory biomarkers with risk factors for colorectal cancer in colorectal adenoma patients. Biomark Insights 7:143–150

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  49. 49.

    Barbour KE, Boudreau R, Danielson ME et al (2012) Inflammatory markers and the risk of hip fracture: the Women’s Health Initiative. J Bone Miner Res 27:1167–1176

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  50. 50.

    Barbour KE, Lui LY, Ensrud KE, Hillier TA, LeBlanc ES, Ing SW, Hochberg MC, Cauley JA (2014) Inflammatory markers and risk of hip fracture in older white women: the study of osteoporotic fractures. J Bone Miner Res 29:2057–2064

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  51. 51.

    Penninx BW, Kritchevsky SB, Newman AB, Nicklas BJ, Simonsick EM, Rubin S, Nevitt M, Visser M, Harris T, Pahor M (2004) Inflammatory markers and incident mobility limitation in the elderly. J Am Geriatr Soc 52:1105–1113

    Article  PubMed  Google Scholar 

  52. 52.

    Karim Y, Turner C, Dalton N, Roplekar R, Sankaralingam A, Ewang M, Fogelman I, Hampson G (2013) The relationship between pro-resorptive inflammatory cytokines and the effect of high dose vitamin D supplementation on their circulating concentrations. Int Immunopharmacol 17:693–697

    CAS  Article  PubMed  Google Scholar 

  53. 53.

    Willis KS, Smith DT, Broughton KS, Larson-Meyer DE (2012) Vitamin D status and biomarkers of inflammation in runners. Open Access J Sports Med 3:35–42

    PubMed  PubMed Central  Google Scholar 

Download references


The Osteoporotic Fractures in Men (MrOS) Study is supported by National Institutes of Health funding. The following institutes provide support: the National Institute on Aging (NIA), the National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS), the National Center for Advancing Translational Sciences (NCATS), and NIH Roadmap for Medical Research under the following grant numbers: U01 AG027810, U01 AG042124, U01 AG042139, U01 AG042140, U01 AG042143, U01 AG042145, U01 AG042168, U01 AR066160, and UL1 TR000128.

Funding for this study was supported in part by the following NIH grants: NIAMS R01 AR063910 (PIs Martin Hewison and John Adams), P60 AR054731 (PI Jane Cauley), and NIAMS K01 AR062655 (PI Carrie Nielson). Supported in part by an independent investigator grant (SRA-12-009) from Merck &Co, Inc.

Author information




Corresponding author

Correspondence to E. S. Orwoll.

Ethics declarations

Conflicts of interest

Roger Bouillon received lecture fees from Amgen, Novartis, Novo Nordisk, Chugai, and Teijin and gave a license to a university patent on vitamin D analogs to Hybrigenix (France). Eric S. Orwoll consults for and has received research support from Merck, Lilly, and Amgen. Carrie Nielson, Priya Srikanth, Rene F Chun, Martin Hewison, John S Adams, Dirk Vanderschueren, Nancy E Lane, Peggy Cawthon, Tien Dam, Elizabeth Barrett-Connor, Lori B Daniels, James Shikany, Marcia L Stefanick, and Jane Cauley declare that they have no conflict of interest.

Electronic supplementary material

Below is the link to the electronic supplementary material.


(DOC 38 kb)


(DOC 36 kb)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Srikanth, P., Chun, R.F., Hewison, M. et al. Associations of total and free 25OHD and 1,25(OH)2D with serum markers of inflammation in older men. Osteoporos Int 27, 2291–2300 (2016).

Download citation


  • Elderly
  • Free 1,25(OH)2D
  • Free 25OHD
  • Inflammation
  • Men
  • Total 1,25(OH)2D
  • Total 25OHD