Rapidly Forming Apatitic Mineral in an Osteoblastic Cell Line (UMR 106—01 BSP)

Clark M. Stanford; Paul A. Jacobson; E. David Eanes; Lois A. Lembke; Ronald J. Midura

doi:10.1074/jbc.270.16.9420

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Rapidly Forming Apatitic Mineral in an Osteoblastic Cell Line (UMR 106—01 BSP)

Journal article

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Rapidly Forming Apatitic Mineral in an Osteoblastic Cell Line (UMR 106—01 BSP)

Clark M. Stanford, Paul A. Jacobson, E. David Eanes, Lois A. Lembke and Ronald J. Midura

The Journal of biological chemistry, Vol.270(16), pp.9420-9428

04/1995

DOI: 10.1074/jbc.270.16.9420

PMID: 7721867

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Abstract

This study evaluated a rapid biomineralization phenomenon exhibited by an osteoblastic cell line, UMR 106-01 BSP, when treated with either organic phosphates [β-glycerophosphate (β-GP), Ser-P, or Thr-P], inorganic phosphate/P(i)), or calcium. In a dose-dependent manner, these agents (2-10 mM) stimulated confluent cultures to deposit mineral in the cell layer (ED50 of ~ 4.6 mM for β-GP (30 ± 2 nmol Ca2+/μg DNA) and ~3.8 mM (29 ± 2 nmol Ca2+/μg DNA) for P(i)) with a plateau in mineral formation by 20 h (ET50 ≃ 12-15 h). β-GP or P(i) treatment 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 (ED50 ≃ 250 μM), pyrophosphate (ED50 ≃ 1-10 μM), actinomycin C1 (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, ongoing RNA and protein synthesis, as well as Golgi-mediated processing and secretion.

Details

Title: Subtitle: Rapidly Forming Apatitic Mineral in an Osteoblastic Cell Line (UMR 106—01 BSP)
Creators: Clark M. Stanford - University of Iowa
Paul A. Jacobson - Cleveland Clinic
E. David Eanes - Cleveland Clinic
Lois A. Lembke - Cleveland Clinic
Ronald J. Midura - Cleveland Clinic
Resource Type: Journal article
Publication Details: The Journal of biological chemistry, Vol.270(16), pp.9420-9428
DOI: 10.1074/jbc.270.16.9420
PMID: 7721867
NLM abbreviation: J Biol Chem
ISSN: 0021-9258
eISSN: 1083-351X
Language: English
Date published: 04/1995
Academic Unit: Roy J. Carver Department of Biomedical Engineering; Dentistry Administration; Prosthodontics; Orthopedics and Rehabilitation; Craniofacial Anomalies Research Center
Record Identifier: 9984367754302771

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