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Isolation of a native osteoblast matrix with a specific affinity for BMP2

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Abstract

During their commitment and differentiation toward the osteoblast lineage, mesenchymal stem cells secrete a unique extracellular matrix (ECM) that contains large quantities of glycosaminoglycans (GAGs). Proteoglycans (PGs) are major structural and functional components of the ECM and are composed of a core protein to which one or more glycosaminoglycan sugar chains (GAGs) attach. The association of BMP2, a member of the TGF-β super-family of growth factors, and a known heparin-binding protein, with GAGs has been implicated as playing a significant role in modulating the growth factor’s in vitro bioactivity. Here we have characterised an osteoblast-derived matrix (MX) obtained from decellularised MC3T3-E1 cell monolayers for its structural attributes, using SEM and histology, and for its functional ability to maintain cell growth and viability. Using a combination of histology and anion exchange chromatography, we first confirmed the retention of GAGs within MX following the decellularisation process. Then the binding specificity of the retained GAG species within the MX for BMP2 was examined using a BMP2-HBP/EGFP (BMP2 Heparin-Binding Peptide/Enhanced Green Fluorescent Protein) fusion protein. The results of this study provide further evidence for a central role of the ECM in the regulation of BMP2 bioactivity, hence on mesenchymal stem cell commitment to the osteoblast lineage.

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Acknowledgements

The authors would like to acknowledge the grant support from Singapore’s Agency for Science Technology and Research (A-STAR), the Biomedical Research Council (BMRC) of Singapore and the Institute of Molecular and Cell Biology (IMCB), Singapore.

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Authors and Affiliations

  1. Stem Cells and Tissue Repair Group, Institute of Molecular and Cell Biology, Proteos, 61 Biopolis Drive, Singapore, 138673, Singapore

    Martin Grünert, Christian Dombrowski, Murali Sadasivam, Kerry Manton, Simon M. Cool & Victor Nurcombe

  2. School of Biomedical Sciences, University of Queensland, Brisbane, QLD, 4072, Australia

    Martin Grünert

  3. Department of Orthopaedic Surgery, Yong Loo Lin School of Medicine, National University of Singapore, 119074, Singapore, Singapore

    Simon M. Cool & Victor Nurcombe

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  1. Martin Grünert
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  2. Christian Dombrowski
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  4. Kerry Manton
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  5. Simon M. Cool
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  6. Victor Nurcombe
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Correspondence to Victor Nurcombe.

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Grünert, M., Dombrowski, C., Sadasivam, M. et al. Isolation of a native osteoblast matrix with a specific affinity for BMP2. J Mol Hist 38, 393–404 (2007). https://doi.org/10.1007/s10735-007-9119-0

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  • DOI: https://doi.org/10.1007/s10735-007-9119-0

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