Comparative study of three delivery systems of menaquinone-7

In an earlier study, beneficial effects of a nutrient-enriched dairy product on
vitamin K-status were observed. The study product contained menaquinone-7
(MK-7) and omega-3 fatty acids for vascular health and extra minerals and
antioxidants to support general health. It was the first time that such low
dose of MK-7 (50 µg) could significantly improve vascular and general vitamin
K-status already after short-term treatment. From the results of this study, it
was assumed that the matrix of the dairy product and/or other components in the
product, in which the vitamin was offered, contributed to optimal absorption
and availability of vitamin K2. In this new study, the uptake and efficacy of
MK-7 from the nutrient enriched product is investigated and compared to the
uptake and efficacy from a MK-7-containing supplement and a basic dairy product
enriched with MK-7.

To investigate the absorption of MK-7 from a nutrient-enriched dairy product
compared to a general dairy product enriched with MK-7 and a MK-7 supplement.

An open intervention study; 114 healthy men and postmenopausal women between 45
and 65 years will be recruited in the southern region of Limburg. Eligible
participants will be randomly divided over three parallel study groups:
• Treatment group I: 38 subjects will receive daily a supplement consisting of
50 µg MK-7
• Treatment group II: 38 subjects will receive daily two basic dairy products
enriched with MK-7 (50 µg)
• Treatment group III: 38 subjects will receive daily two nutrient-enriched
dairy products (smart-mix) containing extra nutrients (calcium, magnesium,
vitamin D3, vitamin C, MK-7 and omega-3 FA (fish-oil); all in a concentration
of 15% of the recommended allowed daily intake (RDI)
The first blood sample will be taken after an overnight fast (t = 0 h) on the
first test day. After the first blood sampling, participants start consuming
the study products daily for an intervention period of 6 weeks followed by a
2-week wash-out period. During the intervention period, it is recommended that
the two yoghurt products are consumed during the first part of the day, namely
one product at breakfast and one product at lunch. Participants of the
treatment group I are not allowed to consume their study product (MK-7
containing supplement) together with any milk-or yoghurt product. Subsequent
blood samplings will take place at the following time-points: at t = 2, 4, and
6 weeks during intervention and t = 3 days, 1 and 2 weeks during the wash-out
period.

During the 6-wk intervention period, the subjects will be instructed to report
any signs of illness, medication used, and any deviations from the study
protocol. Subjects will be urged not to change their general dietary habits
(except for vitamin K-containing products), smoking habits, level of physical
exercise or use of alcohol during the study. Furthermore, the subjects will be
instructed to keep a stable body weight; therefore body weight will be measured
at the first and last study visit. In addition, two weeks (14 days) before the
start of the study until the final blood sampling, participants will be asked
to restrict their intake of vitamin K-containing foods, namely no curd
products, maximum of 1 slice of cheese (25 g) per day and maximum of 200 gram
of green vegetables. At each study visit, subjects will be questioned about
their compliance with these restrictions and their answers will be recorded on
a case report form (CRF). During the intervention period, also compliance of
the treatment will be checked by questions at each study visit.

The risks for the subjects are minimal. No adverse effects are to be expected
from the nutrient-enriched yogurt products. The subjects will visit the
BioPartner Center Maastricht 8 times for the information-visit and consequent
study visits and blood samplings at t=0, 2, 4 and 6 weeks, and at t=3 days, 1
and 2 weeks during the wash-out period. The major burden for the subjects
consists of 7 venipunctures in 6 weeks. The venipunctures will be performed by
experienced researchers.