Stimulation by 1,25-dihydroxyvitamin D3 of in vitro mineralization induced by osteoblast-like MC3T3-E1 cells

Journal Article · Tue Jan 01 04:00:00 EST 1991 · Bone; (United States)

DOI:https://doi.org/10.1016/8756-3282(91)90051-J· OSTI ID:5464141

Matsumoto, T; Igarashi, C; Takeuchi, Y; Harada, S; Kikuchi, T; Yamato, H; Ogata, E ^[1]

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1. Fourth Department of Internal Medicine, University of Tokyo School of Medicine (Japan)

Although vitamin D is essential for mineralization of bone, it is as yet unclear whether vitamin D has a direct stimulatory effect on the bone mineralization process. In the present study, the effect of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) on in vitro mineralization mediated by osteoblast-like MC3T3-E1 cells was examined. MC3T3-E1 cells continued to grow after they reached confluency, and DNA content and alkaline phosphatase activity increased linearly until about 16 days of culture, whereas 45Ca accumulation into cell and matrix layer remained low. After this period, DNA content plateaued, and 45Ca accumulation increased sharply. Histological examination by von Kossa staining revealed that calcium was accumulated into extracellular matrix. In addition, needle-shaped mineral crystals similar to hydroxyapatite crystals could be demonstrated in between collagen fibrils by electron microscopy. Thus, MC3T3-E1 cells differentiate in vitro into cells with osteoblastic phenotype and exhibit mineralization. When MC3T3-E1 cells were treated with 1,25(OH)2D3 at this stage of culture, there was a dose-dependent stimulation of 45Ca accumulation by 1,25(OH)2D3, and a significant stimulation of 45Ca accumulation was observed with 3 x 10(-10) M 1,25(OH)2D3. Although 1,25(OH)2D3 enhanced alkaline phosphatase activity and collagen synthesis at the early phase of culture, it did not affect any of these parameters at the late phase when 1,25(OH)2D3 stimulated mineralization. Neither 24,25-dihydroxyvitamin D3 nor human PTH(1-34) affected mineralization in the presence or absence of 1,25(OH)2D3. These results demonstrate that 1,25(OH)2D3 stimulates matrix mineralization induced by osteoblastic MC3T3-E1 cells, and are consistent with the possibility that 1,25(OH)2D3 has a direct stimulatory effect on bone mineralization process.

• OSTI ID: 5464141
• Journal Information: Bone; (United States), Journal Name: Bone; (United States) Vol. 12:1; ISSN BONED; ISSN 8756-3282
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ALKALINE EARTH ISOTOPES
ALKALINE PHOSPHATASE
ANIMAL CELLS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOLOGICAL EFFECTS
BIOSYNTHESIS
BONE CELLS
CALCIUM 45
CALCIUM ISOTOPES
COLLAGEN
CONNECTIVE TISSUE CELLS
DAYS LIVING RADIOISOTOPES
DNA
DOSE-RESPONSE RELATIONSHIPS
ELECTRON MICROSCOPY
ENZYME ACTIVITY
ENZYMES
ESTERASES
EVEN-ODD NUCLEI
HYDROLASES
INTERMEDIATE MASS NUCLEI
ISOTOPE APPLICATIONS
ISOTOPES
MICROSCOPY
MINERALIZATION
NUCLEI
NUCLEIC ACIDS
ORGANIC COMPOUNDS
PHENOTYPE
PHOSPHATASES
PROTEINS
RADIOISOTOPES
SCLEROPROTEINS
SOMATIC CELLS
SYNTHESIS
TRACER TECHNIQUES
VITAMIN D
VITAMINS