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Bone increased : Stiffness (Vitamin D), Flexibility (Vitamin K2) – Sept 2020

Vitamin K2 Modulates Vitamin D-Induced Mechanical Properties of Human 3D Bone Spheroids In Vitro

JBMR Plus, . 2020 Aug 3;4(9):e10394. doi: 10.1002/jbm4.10394
Maria Schröder 1, Elisabeth Aurstad Riksen 1, Jianying He 2, Bjørn Helge Skallerud 2, Mona Elisabeth Møller 3, Aina-Mari Lian 4, Unni Syversen 4 5 6, Janne Elin Reseland 1



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Rotational culture promotes primary human osteoblasts (hOBs) to form three-dimensional (3D) multicellular spheroids with bone tissue-like structure without any scaffolding material. Cell-based bone models enable us to investigate the effect of different agents on the mechanical strength of bone. Given that low dietary intake of both vitamin D and K is negatively associated with fracture risk, we aimed to assess the effect of these vitamins in this system. Osteospheres of hOBs were generated with menaquinone-4 (MK-4; 10µM) and 25-hydroxyvitamin D3 [25(OH)D3; 0.01µM], alone and in combination, or without vitamins. The mechanical properties were tested by nanoindentation using a flat-punch compression method, and the mineralized extracellular bone matrix was characterized by microscopy. The in vitro response of hOBs to MK-4 and 25(OH)D3 was further evaluated in two-dimensional (2D) cultures and in the 3D bone constructs applying gene expression analysis and multiplex immunoassays.
Mechanical testing revealed that 25(OH)D3 induced a stiffer and MK-4 a softer or more flexible osteosphere compared with control. Combined vitamin conditions induced the same flexibility as MK-4 alone. Enhanced levels of periostin (p < 0.001) and altered distribution of collagen type I (COL-1) were found in osteospheres supplemented with MK-4. In contrast, 25(OH)D3 reduced COL-1, both at the mRNA and protein levels, increased alkaline phosphatase, and stimulated mineral deposition in the osteospheres. With the two vitamins in combination, enhanced gene expression of periostin and COL-1 was seen, as well as extended osteoid formation into the central region and increased mineral deposition all over the area. Moreover, we observed enhanced levels of osteocalcin in 2D and osteopontin in 3D cultures exposed to 25(OH)D3 alone and combined with MK-4.
In conclusion, the two vitamins seem to affect bone mechanical properties differently: vitamin D enhancing stiffness and K2 conveying flexibility to bone. These effects may translate to increased fracture resistance in vivo.

Created by admin. Last Modification: Thursday October 1, 2020 15:53:10 GMT-0000 by admin. (Version 5)

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14382 Bone MK-4.jpg admin 01 Oct, 2020 43.09 Kb 486
14381 Bone flexibility.jpg admin 01 Oct, 2020 41.96 Kb 516
14380 Vitamin K2 bone.pdf admin 01 Oct, 2020 1.90 Mb 343