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Measuring Differences in Compositional Properties of Bone Tissue by Confocal Raman Spectroscopy

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Abstract

The full range of fracture risk determinants arise from each hierarchical level comprising the organization of bone. Raman spectroscopy is one tool capable of characterizing the collagen and mineral phases at a near submicron-length scale, but the ability of Raman spectra to distinguish compositional differences of bone is not well defined. Therefore, we analyzed multiple Raman peak intensities and peak ratios to characterize their ability to distinguish between the typically less mineralized osteonal tissue and the more mineralized interstitial tissue in intracortical human bone. To further assess origins of variance, we collected Raman spectra from embedded specimens and for two orientations of cut. Per specimen, Raman peak intensities or ratios were averaged among multiple sites within five osteons and five neighboring interstitial tissue. The peak ratios of ν1 phosphate (PO4) to proline or amide III detected the highest increases of 15.4 or 12.5%, respectively, in composition from osteonal to interstitial tissue. The coefficient of variance was less than 5% for each as opposed to a value of ~8% for the traditional ν1PO4/amide I, a peak ratio that varied the most between transverse and longitudinal cuts for each tissue type. Although embedding affected Raman peaks, it did not obscure differences in most peak ratios related to mineralization between the two tissue types. In studies with limited sample size but sufficient number of Raman spectra per specimen for spatial averaging, ν1PO4/amide III or ν1PO4/proline is the Raman property that is most likely to detect a compositional difference between experimental groups.

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Acknowledgments

Supported in part by NIH grants NCI U54 CA-126505 and NIAMS R21 AG-029413.

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Authors and Affiliations

  1. Department of Veterans Affairs, Tennessee Valley Healthcare System, Nashville, TN, USA

    Jeffry S. Nyman & Alexander J. Makowski

  2. Department of Orthopaedics and Rehabilitation, Vanderbilt University Medical Center, Nashville, TN, USA

    Jeffry S. Nyman & Elizabeth C. O’Quinn

  3. Center for Bone Biology, Vanderbilt University Medical Center, Nashville, TN, USA

    Jeffry S. Nyman

  4. Department of Biomedical Engineering, Vanderbilt University, Nashville, TN, USA

    Jeffry S. Nyman, Alexander J. Makowski, Chetan A. Patil, Xiaohong Bi & Anita Mahadevan-Jansen

  5. Department of Chemical and Biomolecular Engineering, Vanderbilt University, Nashville, TN, USA

    T. Philip Masui & Scott A. Guelcher

  6. Materials Engineering Department, Southwest Research Institute, San Antonio, TX, USA

    Daniel P. Nicollela

  7. Vanderbilt Orthopaedic Institute, Medical Center East, South Tower, Suite 4200, Nashville, TN, 37232, USA

    Jeffry S. Nyman

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  1. Jeffry S. Nyman
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Correspondence to Jeffry S. Nyman.

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Nyman, J.S., Makowski, A.J., Patil, C.A. et al. Measuring Differences in Compositional Properties of Bone Tissue by Confocal Raman Spectroscopy. Calcif Tissue Int 89, 111–122 (2011). https://doi.org/10.1007/s00223-011-9497-x

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