Relationship between the v2PO4/amide III ratio assessed by Raman spectroscopy and the calcium content measured by quantitative backscattered electron microscopy in healthy human osteonal bone

Roschger, Andreas; Gamsjaeger, Sonja; Hofstetter, B.; Mašić, Admir; Blouin, Stéphane; Messmer, Phaedra; Berzlanovich, Andrea; Paschalis, Eleftherios P.; Roschger, Paul; Klaushofer, Klaus; Fratzl, Peter

doi:10.1117/1.jbo.19.6.065002

Cited by 60 publications

(40 citation statements)

References 38 publications

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“…In the calibrated qBEI images, each gray value is assigned to a certain Ca content (weight percent Ca, wt%Ca), where a difference of one gray value corresponds to 0.17 wt%Ca (27) . qBEI results had been previously validated using energy dispersive x-ray (EDX) analysis and are shown to be consistent with the mineral/matrix ratio obtained with Raman spectroscopy (27,33) .…”

Section: Quantitative Backscattered Electron Imaging (Qbei)supporting

confidence: 58%

The contribution of the pericanalicular matrix to mineral content in human osteonal bone

Roschger

Wagermaier

et al. 2019

Bone

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The osteocyte lacunar-canalicular network (LCN) penetrates bone and houses the osteocytes and their processes. Despite its rather low volume fraction, the LCN represents an outstanding large surface that is possibly used by the osteocytes to interact with the surrounding mineralized bone matrix thereby contributing to mineral homeostasis. The aim of this study was to quantitatively describe such contributions by spatially correlating the local density of the LCN with the mineral content at the same location in micrometer-sized volume elements in human osteons. For this purpose, 65 osteons from the femur midshaft from healthy adults (n=4) and children (n=2) were structurally characterized with two different techniques. The 3D structure of the LCN in the osteons was imaged with confocal laser scanning microscopy after staining the bone samples with rhodamine. Subsequent image analysis provided the canalicular length density, i.e. the total length of the canaliculi per unit volume (µm/µm 3). Quantitative information on the mineral content (wt%Ca) from the identical regions was obtained using quantitative backscattered electron imaging. As the LCN-porosity lowers the mineral content, a negative correlation between Ca content and network density was expected. Calculations predict a reduction of around-0.97 fmol Ca per µm of network. However, the experiment revealed for 62 out of 65 osteons a positive correlation resulting in an average additional Ca loading of +1.15 fmol per µm of canalicular network, i.e. an accumulation of mineral has occurred at dense network regions. We hypothesize that this accumulation happens in the close vicinity of canaliculi forming mineral reservoirs that can be utilized by osteocytes. Significant differences found between individuals indicate that the extent of mineral loading of the reservoir zone reflects an important parameter for mineral homeostasis.

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Section: Quantitative Backscattered Electron Imaging (Qbei)supporting

confidence: 58%

The contribution of the pericanalicular matrix to mineral content in human osteonal bone

Roschger

Wagermaier

et al. 2019

Bone

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“…The Raman metrics investigated included mineral crystallinity, taken as the reciprocal of the full‐width at half‐maximum (1/FWHM) of the ν 1

{PO}_{4}^{3 -}

peak, the carbonate‐to‐phosphate ratio ( ν 1

{CO}_{3}^{2 -}

/ ν 1

{PO}_{4}^{3 -}

; ν 1

{CO}_{3}^{2 -}

/ ν 2

{PO}_{4}^{3 -}

), the apatite‐to‐collagen ratio, also referred to as the mineral‐to‐matrix ratio ( ν 2

{PO}_{4}^{3 -}

/Amide III; ν 1

{PO}_{4}^{3 -}

/Amide I), the carbonate‐to‐matrix ( ν 1

{CO}_{3}^{2 -}

/Amide I) ratio, and the Amide I/Amide III ratios . The amount of amino acids proline/hydroxyproline (Pro + Hyp), and the relative amounts of proline (Pro/ ν 1

{PO}_{4}^{3 -}

), phenylalanine (Phe/ ν 1

{PO}_{4}^{3 -}

), and tyrosine (Tyr/ ν 1

{PO}_{4}^{3 -}

) in relation to the main phosphate peak at 959 cm −1 were also investigated (Supporting Information Table S1).…”

Section: Methodsmentioning

confidence: 99%

“…the apatite-to-collagen ratio, also referred to as the mineralto-matrix ratio (m 2 PO 3-4 /Amide III 25 ; m 1 PO 3-4 /Amide I 26 ), the carbonate-to-matrix (m 1 CO 2-3 /Amide I 27 ) ratio, and the Amide I/Amide III ratios. 28 The amount of amino acids proline/hydroxyproline (Pro 1 Hyp), and the relative amounts of proline (Pro/m 1 PO 3-4 ), phenylalanine (Phe/m 1 PO 3-4 ), and tyrosine (Tyr/m 1 PO [3][4] ) in relation to the main phosphate peak at 959 cm 21 were also investigated (Supporting Information Table S1).…”

Section: Extracellular Matrix Compositionmentioning

confidence: 99%

Multiscale characterization of cortical bone composition, microstructure, and nanomechanical properties in experimentally induced osteoporosis

Shah

Stoica

Cardemil

et al. 2017

J Biomedical Materials Res

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Cortical bone plays a vital role in determining overall bone strength. We investigate the structural, compositional, and nanomechanical properties of cortical bone following ovariectomy (OVX) of 12-week-old Sprague Dawley rats, since this animal model is frequently employed to evaluate the performance of implantable biomaterials in compromised bone healing conditions. Morphological parameters and material properties of bone in the geometrical center of the femoral cortex were investigated four and eight weeks post-OVX and in unoperated controls (Ctrl), using X-ray micro-computed tomography, backscattered electron scanning electron microscopy, Raman spectroscopy, and nanoindentation. The OVX animals showed increase in body weight, diminished bone mineral density, increased intracortical porosity, but increased bone mass through periosteal apposition (e.g., increases in periosteal perimeter, cortical cross-sectional thickness, and cross-sectional area). However, osteocyte densities, osteocyte lacunar dimensions, and the nanomechanical behavior on the single mineralized collagen fibril level remained unaffected. Our correlative multiscale investigation provides structural, chemical, and nanomechanical evidence substantiating earlier reports suggesting that rats ovariectomized at 12 weeks undergo simultaneous bone loss and growth, resulting in the effects of OVX being less obvious. Periosteal apposition contradicts the conventional view of bone loss in osteoporosis but appears advantageous for the greater functional demand imposed on the skeleton by increased body weight and fragility induced by increased intracortical porosity. Through a variety of morphological changes, it is likely that 12-week-old rats are able to adapt to OVX-related microstructural and compositional alterations. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 997-1007, 2018.

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“…The existing relationships from Raman data on bone (Table 2) were determined mostly for use in medical settings: mineral to matrix ratio (to assess bone quality), carbonate to phosphate index, mineral maturity to crystallinity and parameters regarding collagen quality (137)(138)(139). These ratios are valid for diagenetically altered bones, including burned samples (34,56).…”

Section: Raman Spectroscopymentioning

confidence: 99%

Burned bones tell their own stories: A review of methodological approaches to assess heat-induced diagenesis

Mamede

Gonçalves

Marques

et al. 2017

Applied Spectroscopy Reviews

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One of the biggest struggles of biological anthropology is to estimate the biological profile from burned human skeletal remains. Bioanthropological methods are seriously compromised due to bone heat-induced alterations in shape and size. Therefore, it is urgent to improve our ability to estimate sex, age at death, stature and ancestrality, to recognize peri mortem traumas and differentiate them from fractures due to fire, and to determine what was the intensity of burning, namely maximum temperature and heat exposure length. This review focuses on different methodologies to assess heat prompted changes in bone submicrostructure. Some of these are extensively used in burned bones research, namely infrared and Raman spectroscopy and X-ray diffraction, while others such as neutron spectroscopy and diffraction are rarely applied to bone samples although their contribution may be crucial for establishing new bioanthropological methods for a reliable examination of burned victims.

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Relationship between the v2PO4/amide III ratio assessed by Raman spectroscopy and the calcium content measured by quantitative backscattered electron microscopy in healthy human osteonal bone

Cited by 60 publications

References 38 publications

The contribution of the pericanalicular matrix to mineral content in human osteonal bone

The contribution of the pericanalicular matrix to mineral content in human osteonal bone

Multiscale characterization of cortical bone composition, microstructure, and nanomechanical properties in experimentally induced osteoporosis

Burned bones tell their own stories: A review of methodological approaches to assess heat-induced diagenesis

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