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Immunohistochemical Identification of Decorin and Biglycan in Human Dentin: A Correlative Field Emission Scanning Electron Microscopy/Transmission Electron Microscopy Study

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Abstract

Decorin and biglycan, two small leucine-rich proteoglycans, have been proposed to play important roles in matrix-mediated formation of mineralized tissues, and their three-dimensional arrangement in human dentin is still not completely understood. The aim of this study was to immunohistochemically analyze the distribution of decorin and biglycan in human predentin/dentin organic matrix under a high-resolution field emission in-lens scanning electron microscope (FEI-SEM) and a transmission electron microscope (TEM). Tooth dentin specimens were submitted to either a preembedding or a postembedding immunolabeling technique using primary antibodies antidecorin and antibiglycan and gold-conjugated secondary antibodies. Correlative FEI-SEM/TEM observations showed that the two antibodies yielded a similar labeling pattern over the processes of odontoblasts and the predentin. Decorin and biglycan were mainly associated with the collagen fibers within the predentin layer, revealing a moderate immunoreaction that was significantly higher compared to the one observed on dentin. Thus, a generally weak labeling for decorin was found in dentin, which, however, was significantly higher on odontoblast processes within dentinal tubules than in intertubular dentin. On the other hand, biglycan immunolocalization on dentin revealed few gold particles rather uniformly distributed, without showing significant differences between tubular and intertubular regions. In conclusion, this study reveals distinct distribution patterns of decorin and biglycan and their relation with collagen. Decorin’s and biglycan’s precise roles within prematrix and mineralized matrix in human teeth should be further clarified.

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Acknowledgements

The authors thank Mr. Marcello Piccirilli for extensive technical assistance, Mr. Aurelio Valmori for photographic work, and Prof. Lamberto Manzoli for the statistical analysis. We thank Dr. Larry Fisher (National Institute of Dental and Craniofacial Research, Bethesda, MD) for generously donating the anti-DCN (LF-122) and anti-BGN (LF-51) antibodies. The investigation was supported by grants (ex-60%) from Italian Ministry of Education, University and Research, Italy.

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Authors and Affiliations

  1. Department of Biomorphology, University of Chieti-Pescara, Via dei Vestini 31, 66100, Chieti, Italy

    G. Orsini

  2. Department of Anatomical Sciences, University of Bologna, Via Irnerio 48, 40100, Bologna, Italy

    A. Ruggeri Jr., A. Mazzoni, V. Papa & G. Mazzotti

  3. Department of Biomedicine, University of Trieste, Via Stuparich, 1, 34129, Trieste, Italy

    R. Di Lenarda & L. Breschi

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  1. G. Orsini
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  2. A. Ruggeri Jr.
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  3. A. Mazzoni
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  4. V. Papa
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  5. G. Mazzotti
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  6. R. Di Lenarda
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  7. L. Breschi
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Correspondence to L. Breschi.

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Orsini, G., Ruggeri, A., Mazzoni, A. et al. Immunohistochemical Identification of Decorin and Biglycan in Human Dentin: A Correlative Field Emission Scanning Electron Microscopy/Transmission Electron Microscopy Study. Calcif Tissue Int 81, 39–45 (2007). https://doi.org/10.1007/s00223-007-9027-z

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  • DOI: https://doi.org/10.1007/s00223-007-9027-z

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