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Proceedings Paper

Polarization Raman spectroscopy to explain rodent models of brittle bone
Author(s): Alexander J. Makowski; Jeffry S. Nyman; Anita Mahadevan-Jansen
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Paper Abstract

Activation Transcription Factor 4 (Atf-4) is essential for osteoblast maturation and proper collagen synthesis. We recently found that these bones demonstrate a rare brittleness phenotype, which is independent of bone strength. We utilized a confocal Renishaw Raman microscope (50x objective; NA=.75) to evaluate embedded, polished cross-sections of mouse tibia from both wild-type and knockout mice at 8 weeks of age (24 mice, n<=8 per group). Analysis of peak ratios indicated statistically significant changes in both mineral and collagen; however, compositional changes did not fully encompass biomechanical differences. To investigate the impact of material organization, we acquired colocalized spectra aligning the polarization angle parallel and perpendicular to the long bone axis from wet intact femurs. To validate our results, we used MMP9-/- mice, which have a brittleness phenotype that is not explained by compositional Raman measures. Polarization angle difference spectra show marked significant changes in orientation of these compositional differences when comparing wild type to knockout bones. Relative to wild-type, Atf4 -/- and MMP9 -/- bones show significant differences (t-test; p<0.05) in prominent collagen peaks. Further investigation of known peak ratios illustrates that this physical anisotropy of molecular organization is tightly clustered in brittle knockout bones. Such findings could have alternate interpretations about net collagen orientation or the angular distribution of collagen molecules. Use of polarization specific Raman measurements has implicated a structural profile that furthers our understanding of models of bone brittleness. Polarization content of Raman spectra may prove significant in future studies of brittle fracture and human fracture risk.

Paper Details

Date Published: 8 March 2013
PDF: 8 pages
Proc. SPIE 8565, Photonic Therapeutics and Diagnostics IX, 85656L (8 March 2013); doi: 10.1117/12.2006487
Show Author Affiliations
Alexander J. Makowski, Vanderbilt Univ. (United States)
Tennessee Valley Healthcare System (United States)
Jeffry S. Nyman, Vanderbilt Univ. (United States)
Tennessee Valley Healthcare System (United States)
Anita Mahadevan-Jansen, Vanderbilt Univ. (United States)

Published in SPIE Proceedings Vol. 8565:
Photonic Therapeutics and Diagnostics IX
Andreas Mandelis; Brian Jet-Fei Wong; Anita Mahadevan-Jansen; Henry Hirschberg M.D.; Hyun Wook Kang; Nikiforos Kollias; Melissa J. Suter; Kenton W. Gregory M.D.; Guillermo J. Tearney M.D.; Stephen Lam; Bernard Choi; Steen J. Madsen; Bodo E. Knudsen M.D.; E. Duco Jansen; Justus F. Ilgner M.D.; Haishan Zeng; Matthew Brenner; Laura Marcu, Editor(s)

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