Differential accumulation of lead and zinc in double-tidemarks of articular cartilage A. Roschger, J.G. Hofstaetter, B. Pemmer, N. Zoeger, P. Wobrauschek,

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Differential accumulation of lead and zinc in double-tidemarks of articular cartilage A. Roschger, J.G. Hofstaetter, B. Pemmer, N. Zoeger, P. Wobrauschek, G. Falkenberg, R. Simon, A. Berzlanovich, H.W. Thaler, P. Roschger, K. Klaushofer, C. Streli Osteoarthritis and Cartilage Volume 21, Issue 11, Pages 1707-1715 (November 2013) DOI: 10.1016/j.joca.2013.06.029 Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Fig. 1 Spectrum of a single measurement point in a TM obtained after 8 s acquisition time. Bracketed numbers are the maximum counts of the corresponding peaks. Osteoarthritis and Cartilage 2013 21, 1707-1715DOI: (10.1016/j.joca.2013.06.029) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 Giemsa-stained histological undecalcified PMMA embedded section of sample #5, exhibiting a double TM region with non-mineralized cartilage (non-mdAC), superficial tidemark (supTM), deep tidemark (deepTM), mineralized cartilage (mdAC), subchondral bone (BONE). Osteoarthritis and Cartilage 2013 21, 1707-1715DOI: (10.1016/j.joca.2013.06.029) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Fig. 3 Quantitative backscattered electron image (qBEI), Pb fluorescence map (Pb) and Zn map (Zn) of a double TM region from sample #2. Note that the color-coding scale is changed between the elemental maps. In the illustration the histological regions are abbreviated. Osteoarthritis and Cartilage 2013 21, 1707-1715DOI: (10.1016/j.joca.2013.06.029) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Fig. 4 Whisker plots of Zn and Pb levels of the supTM and the deepTM compared to the subchondral bone matrix of the eight analyzed individuals (average age: 46 years, range: 33–85 years, Table I). The ‘+’ indicates a significant difference from a hypothetical equal distribution (dashed line; Wilcoxon-test). (a) Zn concentration in the supTM and the deepTM normalized to the Zn-BONE level. (b) Pb concentration in the supTM and the deepTM normalized to the Pb-BONE level. Osteoarthritis and Cartilage 2013 21, 1707-1715DOI: (10.1016/j.joca.2013.06.029) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Fig. 5 Concentration ratios between different histological regions. A ‘+’ indicates a significant difference from a hypothetical equal distribution (dashed line; Wilcoxon-test): (a) Ratios of Ca, Zn, and Pb between deepTM and supTM of the eight analyzed samples (average individual age: 46 years, range: 33–85 years, Table I). The circles represent a single sample (#5) with outlying low Pb content. (b) Ratios of Ca, Zn, and Pb between mdAC and BONE (n = 7, in sample #5 no mdAC could be evaluated). Osteoarthritis and Cartilage 2013 21, 1707-1715DOI: (10.1016/j.joca.2013.06.029) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Fig. 6 (a) Pb/Ca countrate ratios in regions representative for BONE and for the deepTM. (b) Zn/Ca countrate ratios in regions representative for BONE and for the deepTM. The empty circles represent a single sample (#5) with outlying low Pb content. Osteoarthritis and Cartilage 2013 21, 1707-1715DOI: (10.1016/j.joca.2013.06.029) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Fig. 7 (a) Comparison of single TMs (STs) with double TMs (supTM, deepTM) in two of the samples (sample #4: filled circles, sample #5: empty circles). Each datapoint represents the average Pb intensity ratio of the TM to BONE of three measurement fields. (b) Quantitative backscattered electron image (qBEI) of an osteoarthritic bone (sample #9) sample with a superimposed lead X-ray fluorescence map. The cartilage including the TM is totally worn-out. Osteoarthritis and Cartilage 2013 21, 1707-1715DOI: (10.1016/j.joca.2013.06.029) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions