Nanoscale structure of type I collagen fibrils: Quantitative measurement of D-spacing

Authors

  • Blake Erickson,

    1. Program in Biophysics, University of Michigan, Ann Arbor, MI, USA
    2. Michigan Nanotechnology Institute for Medicine and Biological Sciences, Ann Arbor, MI, USA
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  • Ming Fang,

    1. Michigan Nanotechnology Institute for Medicine and Biological Sciences, Ann Arbor, MI, USA
    2. Department of Chemistry, University of Michigan, Ann Arbor, MI, USA
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  • Prof. Joseph M. Wallace,

    1. Biomedical Engineering, Indiana University-Purdue University, Indianapolis, IN, USA
    2. Department of Biomedical Engineering, 723 W. Michigan SL220D, Indiana University-Purdue University Indianapolis, Indianapolis, IN 46202, USA
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  • Prof. Bradford G. Orr,

    1. Michigan Nanotechnology Institute for Medicine and Biological Sciences, Ann Arbor, MI, USA
    2. Program in Applied Physics, University of Michigan, Ann Arbor, MI, USA
    3. Department of Physics, University of Michigan, Ann Arbor, MI, USA
    4. Department of Physics, 450 Church Street, University of Michigan, Ann Arbor, MI 48109, USA
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  • Clifford M. Les,

    1. Bone and Joint Center, Henry Ford Hospital, Detroit, MI, USA
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  • Dr. Mark M. Banaszak Holl

    Corresponding author
    1. Program in Biophysics, University of Michigan, Ann Arbor, MI, USA
    2. Michigan Nanotechnology Institute for Medicine and Biological Sciences, Ann Arbor, MI, USA
    3. Department of Chemistry, University of Michigan, Ann Arbor, MI, USA
    4. Program in Applied Physics, University of Michigan, Ann Arbor, MI, USA
    • Department of Chemistry, University of Michigan, 930 N. University Ave., Ann Arbor, MI 48109-1055, USA
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Abstract

This article details a quantitative method to measure the D-periodic spacing of type I collagen fibrils using atomic force microscopy coupled with analysis using a two-dimensional fast fourier transform approach. Instrument calibration, data sampling and data analysis are discussed and comparisons of the data to the complementary methods of electron microscopy and X-ray scattering are made. Examples of the application of this new approach to the analysis of type I collagen morphology in disease models of estrogen depletion and osteogenesis imperfecta (OI) are provided. We demonstrate that it is the D-spacing distribution, not the D-spacing mean, that showed statistically significant differences in estrogen depletion associated with early stage osteoporosis and OI. The ability to quantitatively characterize nanoscale morphological features of type I collagen fibrils will provide important structural information regarding type I collagen in many research areas, including tissue aging and disease, tissue engineering, and gene knockout studies. Furthermore, we also envision potential clinical applications including evaluation of tissue collagen integrity under the impact of diseases or drug treatments.

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