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Fourier Transform Infrared Spectroscopic Characterization of Mineralizing Type I Collagen Enzymatic Trivalent Cross-Links

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Abstract

The most abundant protein of bone’s organic matrix is collagen. One of its most important properties is its cross-linking pattern, which is responsible for the fibrillar matrices’ mechanical properties such as tensile strength and viscoelasticity. We have previously described a spectroscopic method based on the resolution of the Amide I and II Fourier transform Infrared (FTIR) bands to their underlying constituent peaks, which allows the determination of divalent and pyridinoline (PYD) collagen cross-links in mineralized thin bone tissue sections with a spatial resolution of ~6.3 μm. In the present study, we used FTIR analysis of a series of biochemically characterized collagen peptides, as well as skin, dentin, and predentin, to examine the potential reasons underlying discrepancies between two different analytical methodologies specifically related to spectral processing. The results identified a novel distinct FTIR underlying peak at ~1,680 cm−1, correlated with deoxypyridinoline (DPD) content. Furthermore, the two different methods of spectral resolution result in widely different results, while only the method employing well-established spectroscopic routines for spectral resolution provided biologically relevant results, confirming our earlier studies relating the area of the underlying 1,660 cm−1 with PYD content. The results of the present study describe a new peak that may be used to determine DPD content, confirm our earlier report relating spectroscopic parameters to PYD content, and highlight the importance of the selected spectral resolution methodology.

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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).

Conflict of Interest

E. P. Paschalis, S. Gamsjaeger, D. N. Tatakis, N. Hassler, S. P. Robins, and K. Klaushofer do not have any conflict of interest to declare.

Human and Animal Rights and Informed Consent

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional research committee and with the 1964 Helsinki declaration and its later amendments.

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Authors and Affiliations

  1. Ludwig Boltzmann Institute of Osteology at the Hanusch Hospital of WGKK and AUVA Trauma Centre Meidling, 1st Medical Department, Hanusch Hospital, Hanusch Krankenhaus, Heinrich Collin Str. 30, 1140, Vienna, Austria

    E. P. Paschalis, S. Gamsjaeger, N. Hassler & K. Klaushofer

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

    D. N. Tatakis

  3. Matrix Biochemistry, Rowett Institute of Nutrition and Health, University of Aberdeen, Bucksburn, Aberdeen, AB21 9SB, UK

    S. P. Robins

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  1. E. P. Paschalis
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  2. S. Gamsjaeger
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  4. N. Hassler
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Correspondence to E. P. Paschalis.

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Paschalis, E.P., Gamsjaeger, S., Tatakis, D.N. et al. Fourier Transform Infrared Spectroscopic Characterization of Mineralizing Type I Collagen Enzymatic Trivalent Cross-Links. Calcif Tissue Int 96, 18–29 (2015). https://doi.org/10.1007/s00223-014-9933-9

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  • DOI: https://doi.org/10.1007/s00223-014-9933-9

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