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Bone‐specific heparan sulfates induce osteoblast growth arrest and downregulation of retinoblastoma protein
Author(s) -
Manton Kerry J.,
Sadasivam Murali,
Cool Simon M.,
Nurcombe Victor
Publication year - 2006
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.20727
Subject(s) - osteoblast , cell growth , extracellular matrix , chemistry , microbiology and biotechnology , cell cycle , retinoblastoma protein , fibroblast growth factor , heparan sulfate , biochemistry , cell , receptor , biology , in vitro
The heparan sulfate (HSs) sugars of the extracellular matrix (ECM) play a key role during both development and wound repair in regulating the flow of growth and adhesive factors across their cell surface receptors. The aim of this study was to assess the structural and functional differences of HS chains extracted from the conditioned media (soluble), cell surface, and ECM of primary human osteoblast cultures, and to analyze their effects on osteoblast cell growth. HS chains from these compartments were characterized through a combination of enzymatic degradation, anion exchange chromatography, and molecular sieving. Although the chains were all approximately the same size, they varied systematically in their sulfate content, suggesting differences in their protein‐binding domains. When added to pre‐confluent hFOB1.19 osteoblast cultures, HS doses exceeding 500 ng/ml inhibited proliferation, without affecting viability, irrespective of their origin. Furthermore, HS doses of 500 ng/ml also downregulated retinoblastoma, Cyclin A and CDK1 protein expression, indicating that high doses of osteoblast HS negatively regulate cell cycle, resulting in growth arrest; when high doses of HS were withdrawn after a prolonged period, linear cell growth was reestablished. Thus, despite differences in sulfation, HS from either the soluble, cell surface, or matrix compartments of primary human osteoblast cultures are functionally similar with respect to their effects on growth. Binding assays revealed that the HS chains bound TGFβ1, a known inhibitor of osteoprogenitor growth, at higher affinity than a suite of other bone‐related, heparin‐binding growth factors. Overcoming such sugar‐mediated inhibition may prove important for wound repair. J. Cell. Physiol. 209: 219–229, 2006. © 2006 Wiley‐Liss, Inc.