Abstract
Bone may be similar to geological formulations in many ways. Therefore, it may be logical to apply laser-based geological techniques in bone research. The mineral and element oxide composition of bioapatite can be estimated by mathematical models. Laser-induced plasma spectrometry (LIPS) has long been used in geology. This method may provide a possibility to determine the composition and concentration of element oxides forming the inorganic part of bones. In this study, we wished to standardize the LIPS technique and use mathematical calculations and models in order to determine CaO distribution and bone homogeneity using bovine shin bone samples. We used polished slices of five bovine shin bones. A portable LIPS instrument using high-power Nd++YAG laser pulses has been developed (OpLab, Budapest). Analysis of CaO distribution was carried out in a 10 × 10 sampling matrix applying 300-μm sampling intervals. We assessed both cortical and trabecular bone areas. Regions of interest (ROI) were determined under microscope. CaO peaks were identified in the 200–500 nm wavelength range. A mathematical formula was used to calculate the element oxide composition (wt%) of inorganic bone. We also applied two accepted mathematical approaches, the Bartlett’s test and frequency distribution curve-based analysis, to determine the homogeneity of CaO distribution in bones. We were able to standardize the LIPS technique for bone research. CaO concentrations in the cortical and trabecular regions of B1–5 bones were 33.11 ± 3.99% (range 24.02–40.43%) and 27.60 ± 7.44% (range 3.58–39.51%), respectively. CaO concentrations highly corresponded to those routinely determined by ICP-OES. We were able to graphically demonstrate CaO distribution in both 2D and 3D. We also determined possible interrelations between laser-induced craters and bone structure units, which may reflect the bone structure and may influence the heterogeneity of CaO distributions. By using two different statistical methods, we could confirm if bone samples were homogeneous or not with respect to CaO concentration distribution. LIPS, a technique previously used in geology, may be included in bone research. Assessment of element oxide concentrations in the inorganic part of bone, as well as mathematical calculations may be useful to determine the content of CaO and other element oxides in bone, further analyze bone structure and homogeneity and possibly apply this research to normal, as well as diseased bones.
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Acknowledgements
The authors would like to thank the Department of Laboratory Medicine at Faculty of Medicine of the University of Debrecen for carrying out the qCT tests and the staff of the Laboratory Department of the Geological and Geophysical Institute of Hungary (GGIH; namely István Kovács and Ferenc Fenesi) for the preparation of animal bone samples (thin sections) as well as for performing the ICP-OES assessment.
Funding
This research was supported by grant OTKA K105073 from the National Scientific Research Fund of Hungary (HPB and ZS) and by the European Union and the State of Hungary co-financed by the European Social Fund in the framework of TÁMOP-4.2.4.A/2-11/1-2012-0001 ‘National Excellence Program’ (ZS).
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Andrássy, L., Gomez, I., Horváth, Á. et al. Laser-induced plasma spectroscopy (LIPS): use of a geological tool in assessing bone mineral content. Lasers Med Sci 33, 1225–1236 (2018). https://doi.org/10.1007/s10103-018-2462-4
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DOI: https://doi.org/10.1007/s10103-018-2462-4