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This study evaluated a rapid biomineralization phenomenon
exhibited by an osteoblastic cell line, UMR 106-01 BSP, when
treated with either organic phosphates [
Volume 270,
Number 16,
Issue of April 21, pp. 9420-9428, 1995
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
01 BSP)
-glycerophosphate
(-GP), Ser-P, or Thr-P], inorganic phosphate (P
),
or calcium. In a dose-dependent manner, these agents (2-10
m
M) stimulated confluent cultures to deposit mineral in the
cell layer (EDof
4.6 m
M for
-GP (30
± 2 nmol Ca/µg DNA) and
3.8
m
M (29 ± 2 nmol Ca
/µg DNA) for
P
) with a plateau in mineral formation by 20 h (ET
12-15 h). -GP or Ptreatment yielded
mineral crystals having an x-ray diffraction pattern similar to normal
human bone. Alizarin red-S histology demonstrated calcium mineral
deposition in the extracellular matrix and what appeared to be
intracellular paranuclear staining. Electron microscopy revealed small,
needle-like crystals associated with fibrillar, extracellular matrix
deposits and intracellular spherical structures. Mineral formation was
inhibited by levamisole (ED 250 µ
M),
pyrophosphate (ED 1-10 µ
M),
actinomycin C
(500 ng/ml), cycloheximide (50 µg/ml), or
brefeldin A (1 µg/ml). These results indicate that UMR 106-01
BSP cells form a bio-apatitic mineralized matrix upon addition of
supplemental phosphate. This process involves alkaline phosphatase
activity, on-going RNA and protein synthesis, as well as Golgi-mediated
processing and secretion.
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