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Identification of Pyridinoline Trivalent Collagen Cross-Links by Raman Microspectroscopy

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Abstract

Intermolecular cross-linking of bone collagen is intimately related to the way collagen molecules are arranged in a fibril, imparts certain mechanical properties to the fibril, and may be involved in the initiation of mineralization. Raman microspectroscopy allows the analysis of minimally processed bone blocks and provides simultaneous information on both the mineral and organic matrix (mainly type I collagen) components, with a spatial resolution of ~1 μm. The aim of the present study was to validate Raman spectroscopic parameters describing one of the major mineralizing type I trivalent cross-links, namely pyridinoline (PYD). To achieve this, a series of collagen cross-linked peptides with known PYD content (as determined by HPLC analysis), human bone, porcine skin, predentin and dentin animal model tissues were analyzed by Raman microspectroscopy. The results of the present study confirm that it is feasible to monitor PYD trivalent collagen cross-links by Raman spectroscopic analysis in mineralized tissues, exclusively through a Raman band ~1660 wavenumbers. This allows determination of the relative PYD content in undecalcified bone tissues with a spatial resolution of ~1 μm, thus enabling correlations with histologic and histomorphometric parameters.

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Acknowledgements

This study was supported by the AUVA (Research funds of the Austrian workers compensation board), and the WGKK (Viennese sickness insurance funds).

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Authors and Affiliations

  1. Ludwig Boltzmann Institute of Osteology at Hanusch Hospital of WGKK and AUVA Trauma Centre Meidling, 1st Med. Dept. Hanusch Hospital, Vienna, Austria

    Sonja Gamsjaeger, Klaus Klaushofer & Eleftherios P. Paschalis

  2. Rowett Institute of Nutrition and Health, Bucksburn, Aberdeen, AB21 9SB, Scotland, UK

    Simon P. Robins

  3. Division of Periodontology, College of Dentistry, The Ohio State University, Columbus, OH, USA

    Dimitris N. Tatakis

  4. Ludwig Boltzmann Institute of Osteolgy, Hanusch Kh., Heinrich Collin Str. 30, 1140, Vienna, Austria

    Eleftherios P. Paschalis

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  1. Sonja Gamsjaeger
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  5. Eleftherios P. Paschalis
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Correspondence to Eleftherios P. Paschalis.

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Human and Animal Rights and Informed Consent

All procedures followed were in accordance with the ethical standards of the institutional committee on human experimentation and with the Helsinki Declaration of 1975, as revised in 2008. Informed consent was obtained from all patients.

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Gamsjaeger, S., Robins, S.P., Tatakis, D.N. et al. Identification of Pyridinoline Trivalent Collagen Cross-Links by Raman Microspectroscopy. Calcif Tissue Int 100, 565–574 (2017). https://doi.org/10.1007/s00223-016-0232-5

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  • DOI: https://doi.org/10.1007/s00223-016-0232-5

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