Vitamins and Minerals (Oral Session 19)

(OR19-04-25) Dietary Menaquinone-9 Supplementation Does Not Influence Bone Tissue Quality or Bone Mineral Density in Mice

Tuesday, June 3, 2025

9:06 AM – 9:18 AM ET

Location: S220 E-G

Presenting Author(s)

ML

Minying Liu, MS

PhD Candidate
Jean Mayer USDA Human Nutrition Research Center on Aging (HNRCA) at Tufts University
Brighton, Massachusetts, United States

Disclosure information not submitted.

Co-Author(s)

CL

Chongshan Liu

Cornell University, United States
NC

Nicolas Cevallos

University of California, San Francisco, United States
TS

Tristan Silfverskiold

Jean Mayer USDA Human Nutrition Research Center on Aging (HNRCA) at Tufts University, United States
BO

Benjamin N. Orbach, n/a

Research Assistant
Tufts University
Boston, Massachusetts, United States
CH

Christopher J. Hernandez

University of California, San Francisco, United States
XF

Xueyan Fu, PhD

Scientist II
Tufts University
Boston, Massachusetts, United States
Jennifer Lee, PhD (she/her/hers)

Scientist II
JM-USDA HNRCA at Tufts University
Boston, Massachusetts, United States
Sarah L. Booth, PhD

Director
Tufts University
Tufts University
Boston, Massachusetts, United States
MS

M.Kyla Shea

Jean Mayer USDA Human Nutrition Research Center on Aging (HNRCA) at Tufts University, United States

Objectives: Vitamin K has been implicated in skeletal health because vitamin K-dependent proteins are present in bone. While there are multiple forms of vitamin K, most research has focused on phylloquinone, which is found mainly in plant-based foods, and menaquinone-4 (MK4), which is a metabolite of phylloquinone. However, there are additional forms of vitamin K that are bacterially produced that may influence bone health but have not yet been studied extensively. Herein, we evaluated the effects of menaquinone-9 (MK9), a bacterially produced form of vitamin K on bone tissue quality and bone mineral density (BMD) in young mice.

Methods: Four-week-old male (n=32) and female (n=32) C57BL/6 mice were supplemented with 0.06 mg/kg diet or 2.1 mg/kg diet of MK9 for 12 weeks. During week 11, a sub-group of mice (n=7/sex/group) received daily deuterium-labeled MK9 to trace its metabolic fate in bone. Skeletal outcomes, including bone geometry, tissue quality, and total body BMD were assessed. Liver and bone vitamin K concentrations were quantified using mass spectrometry. Difference in outcome measures between groups within sex were analyzed using an independent t-test.

Results: Liver MK4 and MK9 concentrations were significantly higher in mice fed 2.1 mg MK9/kg diet compared to those receiving 0.06 mg MK9/kg diet, regardless of sex (all p ≤ 0.017). MK4 was the only vitamin K form detected in bone, with 63-67% of skeletal MK4 in mice fed 2.1 mg MK9/kg diet derived from deuterium-labeled MK9. Femoral tissue strength, maximum bending moment, section modulus, and BMD did not differ significantly between diet groups in either sex (all p≥0.083). Cross-sectional area (p=0.003) and moment of inertia (p=0.001) were lower in female mice receiving 2.1 mg MK9/kg diet compared to those receiving 0.06 mg MK9/kg diet, but no differences were found in male mice.

Conclusions: Despite dietary MK9 being a dietary precursor to MK4 in bone, dietary MK9 supplementation did not affect bone tissue quality or bone mineral density.

Funding Sources: NIAMS R01AG067997 and US Department of Agriculture Agricultural Research Service Cooperative Agreement No. 58-8050-3-003