Increased serum osteocalcin levels and vitamin K status by daily cheese intake


  • Helge E. Lundberg Skjetten Medical Centre and Primary Medicine, Skjetten, Norway
  • Trond Holand Centre for Epidemiology and Biostatistics, Faculty of Veterinary Medicine, Norwegian University of Life Sciences, Oslo, Norway
  • Helge Holo Faculty of Biotechnology and Food Science, Norwegian University of Life Sciences, Aas, Norway
  • Stig Larsen Centre for Epidemiology and Biostatistics, Faculty of Veterinary Medicine, Norwegian University of Life Sciences, Oslo, Norway



Dose-finding study of cheese, Jarlsberg® cheese, Osteocalcin, Osteoporosis, Lipid pattern, Vitamin K2, Vital signs, Women


Background: Cheese is a major source of long-chained vitamin K2 variants. How intake of vitamin K2 rich cheese affects vitamin K and osteocalcin has not been studied. The aim was to establish a maximum efficacy dose (MED) after daily intake of vitamin K2-rich cheese (Jarlsberg®) based on increase in ratio between carboxylated and undercarboxylated osteocalcin during a five-week diet.

Methods: 20 healthy healthy volunteers (HV) were recruited. The daily intake of Jarlsberg® cheese in the study varied from 20 to 152 g. Clinical investigation was performed initially and after three, four and five weeks with measurement of vital signs, hematological and biochemical variables, carboxylated and undercarboxylated osteocalcin and vitamin K. The ratio OR= carboxylated/undercarboxylated osteocalcin was the main variable.

Results: The MED decreased with treatment duration and was estimated to 57 g/day (95% CI: 47-67) after five weeks diet, resulting in a mean OR increase of 30% (95% CI: 23.8-36.8). Both OR and serum osteocalcin followed a quadratic dose response curve. For osteocalcin, a maximal increase of 46% was estimated at 59 g/day for five weeks. The serum content of long-chained vitamin K2 increased significantly with increasing cheese dose. The increase were mainly obtained the first three weeks and kept unchanged the following two weeks. The cheese doses close to the MED caused nearly significant reductions in total cholesterol, LDL-cholesterol, the LDL/HDL ratio and significant reduction in the blood pressures after five weeks diet (p≤0.05).

Conclusions: MED of Jarlsberg® cheese was estimated to 57 g/day. Daily intake of Jarlsberg® cheese increased the osteocalcin level, vitamin K2 and positively affected the lipid patterns and blood pressure.


van de Peppel J, van Leeuwen JPTM. Vitamin D and gene networks in human osteoblasts. Front Physiol. 2014;5:137.

Levinger I, Scott D, Nicholson GC, Stuart AL, Duque G, McCorquodale T, et al. Undercarboxylated osteocalcin, muscle strength and indices of bone health in older women. Bone. 2014;64:8-12.

Kang J-H. Association of serum osteocalcin with insulin resistance and coronary atherosclerosis. J Bone Metabol. 2016;23(4):183-90.

Mera P, Ferron M, Mosialou I. Regulation of Energy Metabolism by Bone-Derived Hormones. Cold Spring Harb Perspect Med. 2018;8(6).

Mera P, Laue K, Ferron M, Confavreux C, Wei J, Galán-Díez M, Lacampagne A, Mitchell SJ, Mattison JA, Chen Y, et al. Osteocalcin Signaling in Myofibers Is Necessary and Sufficient for Optimum Adaptation to Exercise. Cell Metab. 2016;23(6):1078-92.

Mera P, Laue K, Wei J, Berger JM, Karsenty G. Osteocalcin is necessary and sufficient to maintain muscle mass in older mice. Mol Metab. 2016;5(10):1042-7.

Zoch ML, Clemens TL, Riddle RC. New insights into the biology of osteocalcin. Bone. 2016;82:42-9.

Lees JS, Chapman FA, Witham MD, Jardine AG, Mark PB. Vitamin K status, supplementation and vascular disease: a systematic review and meta-analysis. Heart. 2019;105(12):938-45.

Theuwissen E, Cranenburg EC, Knapen MH, Magdeleyns EJ, Teunissen KJ, Schurgers LJ, et al. Low-dose menaquinone-7 supplementation improved extra-hepatic vitamin K status, but had no effect on thrombin generation in healthy subjects. Brit J Nutr. 2012;108(9):1652-7.

Szulc P, Chapuy M, Meunier P, Delmas P. Serum undercarboxylated osteocalcin is a marker of the risk of hip fracture in elderly women. J Clin Invest. 1993;91(4):1769-74.

Vermeer C, Raes J, van ’t Hoofd C, Knapen M, Xanthoulea S. Menaquinone Content of Cheese. Nutrients. 2018;10(4):446.

Sato T, Schurgers LJ, Uenishi K. Comparison of menaquinone-4 and menaquinone-7 bioavailability in healthy women. Nutr J. 2012;11(1):93.

Schurgers LJ, Teunissen KJ, Hamulyak K, Knapen MH, Vik H, Vermeer C. Vitamin K-containing dietary supplements: comparison of synthetic vitamin K1 and natto-derived menaquinone-7. Blood. 2007;109(8):3279-83.

Geleijnse JM, Vermeer C, Grobbee DE, Schurgers LJ, Knapen MH, Van Der Meer IM, Hofman A, Witteman JC. Dietary intake of menaquinone is associated with a reduced risk of coronary heart disease: the Rotterdam Study. J Nutr. 2004;134(11):3100-5.

Manoury E, Jourdon K, Boyaval P, Fourcassie P. Quantitative measurement of vitamin K2 (menaquinones) in various fermented dairy products using a reliable high-performance liquid chromatography method. J Dairy Sci. 2013;96(3):1335-46.

Hojo K, Watanabe R, Mori T, Taketomo NJJods. Quantitative measurement of tetrahy dromenaquinone-9 in cheese fermented by propionibacteria. J Dairy Sci. 2007;90(9):4078-83.

Dewi S, Kristiansen V, Lindkær-Jensen S, Larsen S. Between-and within-patient n-level response surface pathway design in dose-finding studies. Open Access J Clin Trials. 2014;6:63.

Holand T, Ellingsen K, Dewi S, Larsen S. Randomized response surface pathway design with odd response outcomes in a Latin Square designed study. Open Access J Clin Trials. 2017;9:1-10.

Larsen S, Holand T, Bjornaes K, Glomsrod E, Kaufmann J, Garberg TH, et al. Randomized Two-Dimensional between-Patient Response Surface Pathway Design with Two Interventional-and One Response Variable in Estimating Minimum Efficacy Dose. Int J Clin Trials. 2018;8(4):1-9.

Altman DG. Practical statistics for medical research: London: Chapman & Hall; 1991.

Agresti A. Categorical Data Analysis. New Jersey: John Wiley & Sons, 2002.

Kleinbaum DG, Kupper LL, Muller KE, Nizam A. Applied regression analysis and other multivariable methods. CA: Duxbury Press Belmont; 1988.

Stylianou M, Flournoy N. Dose finding using the biased coin up‐and‐down design and isotonic regression. Biometrics. 2002;58(1):171-7.

Holand T, Evensen Ø, Dewi S, Larsen S. Randomized response surface pathway design with skewed starting point and stochastic dose window. Int J Trials. 2020;7(1):18-27.

Bugel S. Vitamin K and bone health in adult humans. Vitam Horm. 2008;78:393-416.

Theuwissen E, Magdeleyns EJ, Braam LA, Teunissen KJ, Knapen MH, Binnekamp IA, et al. Vitamin K status in healthy volunteers. Food Funct. 2014;5(2):229-34.

Knapen MH, Braam LA, Teunissen KJ, Zwijsen RM, Theuwissen E, Vermeer C. Yogurt drink fortified with menaquinone-7 improves vitamin K status in a healthy population. J Nutr Sci. 2015;4:35.

Koshihara Y, Hoshi K. Vitamin K2 Enhances Osteocalcin Accumulation in the Extracellular Matrix of Human Osteoblasts In Vitro. J Bone Miner Res. 1997;12(3):431-8.

Kanbur NO, Derman O, Sen TA, Kinik E. Osteocalcin. A biochemical marker of bone turnover during puberty. Int J Adolesc Med Health. 2002;14(3):235-44.

Choi HJ, Yu J, Choi H, An JH, Kim SW, Park KS, et al. Vitamin K2 supplementation improves insulin sensitivity via osteocalcin metabolism: a placebo-controlled trial. Diabetes Care. 2011;34(9):e147

Hill HS, Grams J, Walton RG, Liu J, Moellering DR, Garvey WT. Carboxylated and uncarboxylated forms of osteocalcin directly modulate the glucose transport system and inflammation in adipocytes. Horm Metab Res. 2014;46(5):341-7.

Kuźniewski M, Fedak D, Dumnicka P, Kapusta M, Stępień E, Chowaniec E, et al. Carboxylated and intact osteocalcin predict adiponectin concentration in hemodialyzed patients. Renal Failure. 2016;38(3):451-7.

Shea MK, Gundberg CM, Meigs JB, Dallal GE, Saltzman E, Yoshida M, et al. Gamma-carboxylation of osteocalcin and insulin resistance in older men and women. Am J Clin Nutr. 2009;90(5):1230-5.

Oosterwerff MM, van Schoor NM, Lips P, Eekhoff EM. Osteocalcin as a predictor of the metabolic syndrome in older persons: a population-based study. Clin Endocrinol (Oxf). 2013;78(2):242-7.

Puig J, Blasco G, Daunis-i-Estadella J, Moreno M, Molina X, Alberich-Bayarri A, Xifra G, Pedraza S, Ricart W, Fernandez-Aranda F, et al. Lower serum osteocalcin concentrations are associated with brain microstructural changes and worse cognitive performance. Clin Endocrinol (Oxf). 2016;84(5):756-63.

Millar SA, Patel H, Anderson SI, England TJ, O'Sullivan SE. Osteocalcin, Vascular Calcification, and Atherosclerosis: A Systematic Review and Meta-analysis. Front Endocrinol (Lausanne). 2017;8:183.

Yun S-H, Kim MJ, Choi B-h, Park K-C, Park K-S, Kim Y-S. Low level of osteocalcin is related with arterial stiffness in Korean adults: an inverse J-shaped relationship. J Clin Endocrinol. 2016;101(1):96-102.

Nilsen R, Pripp AH, Hostmark AT, Haug A, Skeie S. Effect of a cheese rich in angiotensin-converting enzyme-inhibiting peptides (Gamalost®) and a Gouda-type cheese on blood pressure: results of a randomised trial. Food Nutr Res. 2016;60:32017.

Biong AS, Rebnord HM, Fimreite RL, Trygg KU, Ringstad J, Thelle DS, et al. Intake of dairy fat and dairy products, and risk of myocardial infarction: a case-control study. Int J Food Sci Nutr. 2008;59(2):155-65.

Feeney EL, Barron R, Dible V, Hamilton Z, Power Y, Tanner L, et al. Dairy matrix effects: response to consumption of dairy fat differs when eaten within the cheese matrix—a randomized controlled trial. Am J Clin Nutr. 2018;108(4):667-74.






Original Research Articles