Journal of Hematology and Oncology Research
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Research Article | Open Access
  • Available online freely | Peer Reviewed
  • | Provisional

    The Silver, Cobalt, Chromium, Iron, Mercury, Rubidium, Antimony, Selenium, and Zinc Contents in Human Bone Affected by Chondrosarcoma

    Vladimir Zaichick 1       Sofia Zaichick 2    

    1Radionuclide Diagnostics Department, Medical Radiological Research Centre, Obninsk 249036, Russia

    2Department of Medicine, University of Illinois, College of Medicine at Chicago, Chicago, IL 60612, USA

    Abstract

    Objectives:

    To clarify the role of trace elements in the etiology and the pathogenesis of the chondrosarcoma, a non-destructive neutron activation analysis with high resolution spectrometry of long-lived radionuclides were performed.

    Methodology:

    The silver (Ag), cobalt (Co), chromium (Cr), iron (Fe), mercury (Hg), rubidium (Rb), antimony (Sb), selenium (Se), and zinc (Zn)mass fractions and Co/Zn, Cr/Zn, Fe/Zn, Hg/Zn, Sb/Zn, Co/Rb, Cr/Rb, Fe/Rb, Hg/Rb, Sb/Rb, and Se/Rb mass fraction ratios were estimated in normal bone samples from 27 patients with intact bone (12 females and 15 males, aged from 16 to 49 years), who had died from various non bone related causes, mainly unexpected from trauma, and in tumor samples, obtained from open biopsies or after operation of 16 patients with chondrosarcoma ((3 females and 13 males, 8 to 65 years old). The reliability of difference in the results between intact bone and chondrosarcoma tissues was evaluated by Student’s t-test.

    Key Results:

    In the chondrosarcoma tissue the mass fractions of Co, Fe, and Se are significantly higher while the mass fraction of Rb is lower than in normal bone tissues. Moreover, significantly higher Co/Zn, Fe/Zn, Co/Rb, Cr/Rb, Fe/Rb, Sb/Rb, and Se/Rb mass fraction ratios are typical of the chondrosarcoma tissue compared to intact bone. In the chondrosarcoma tissue many correlations between trace elements found in the control group was no longer evident.

    Major Conclusions:

    In chondrosarcoma transformed bone tissues the trace element homeostasis is significantly disturbed.

    Received 15 Apr 2015; Accepted 22 Jun 2015; Published 08 Aug 2015;

    Academic Editor:Krzysztof Roszkowski, Department of Radiotherapy, the F. Lukaszczyk Oncology Center, Bydgoszcz, PL; Chair of Gynecology, Oncology and Gynecological Nursing, Collegium Medicum, Nicolaus Copernicus University, PL

    Checked for plagiarism: Yes

    Review by: Single-blind

    Copyright©  2015 Vladimir Zaichick, et al.

    License
    Creative Commons License    This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

    Competing interests

    The authors have declared that no competing interests exist.

    Citation:

    Vladimir Zaichick, Sofia Zaichick (2015) The Silver, Cobalt, Chromium, Iron, Mercury, Rubidium, Antimony, Selenium, and Zinc Contents in Human Bone Affected by Chondrosarcoma. Journal of Hematology and Oncology Research - 1(4):19-30.
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    DOI10.14302/issn.2372-6601.jhor-15-666

    Introduction

    The roles of trace elements in the development and inhibition of cancer have a complex character and have risen many questions because of their essential and toxic effects on human health. The effects of trace elements are related to content and recorded observations range from a deficiency state, to function as biologically essential components, to an unbalance when excess of one element interferes with the function of another, to pharmacologically active doses, and finally to toxic and even life-threatening levels.1, 2 Thus, in normal environmental and health conditions there is a trace element homeostasis in tissues and fluids of human body and an unbalance of trace element contents could be a causative factor for many diseases, including cancer.2

    It is well known that the tissues of human body differ greatly in their contents of trace elements. Our detailed previous studies have shown this using a chemical composition analysis of bone tissue.3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 Bone tumors form a heterogeneous group of benign or malignant neoplastic diseases since they can derive from all the tissue components of bone (cartilage, osteoid, fibrous tissue, and bone marrow elements). Each tissue can be subject to inflammation, benign or malignant tumors.

    Chondrosarcoma is the second most common primary bone cancer, whose tumor cells produce a pure hyaline cartilage that results in abnormal bone and/or cartilage growth. About one fourth of malignant bone cancers are chondrosarcoma. Although any bone can be affected, the long bones such as legs, arms, fingers, toes are most commonly involved. Chondrosarcoma is typically seen in adulthood between the ages of late 20 to 60 and occurs more commonly in men than women.30, 31

    No single morphologic or functional imaging method provides findings for a specific diagnosis of chondrosarcoma, but the results do contribute to tumor staging. Therefore, obtaining a histologic specimen of the lesion in all patients is essential in recognizing this tumor and planning therapy. However, histological examination of bone tumors is one of the most difficult subjects in pathology.32It is the reason, why molecular techniques to assist in distinguishing among the various types of bone tumors are being developed, but these techniques have not yielded any clinically significant contribution.30

    To our knowledge, no data are available for the trace element contents of chondrosarcoma, to permit conclusion about their role in malignant transformation or pathogenesis, and also about their usefulness as the tumor’s markers.

    The aim of the study was to compare and to correlate the trace element mass fractions and the mass fraction ratios of selected traceelement in two groups of samples (normal bone and chondrosarcoma tissue). For this purpose, the silver (Ag), cobalt (Co), chromium (Cr), iron(Fe), mercury (Hg), rubidium (Rb), antimony (Sb), selenium (Se), and zinc (Zn) contents were determined in the two groups of samples using nondestructive instrumental neutron activation analysis (INAA) with high resolution spectrometry of long-lived radionuclides (INAA-LLR).

    The study was approved by the Ethical Committee of the Medical Radiological Research Center, Obninsk.

    Materials and Methods

    Thirty-three children, adolescents and adults were included in this study. The subjects were divided into two groups: control and chondrosarcoma. The control group consisted of 27 patients with intact bone (12 females and 15 males, aged from 16 to 49 years, M±SEM 34.1±2.1 years) who had died from various non bone related causes, mainly unexpected from trauma. The intact cortical bone samples of femur, femoral neck, tibia and iliac crest were collected at the Department of Pathology, Obninsk City Hospital. Samples from 16 patients with chondrosarcoma (3 females and 13 males, 8 to 65 years old, M±SEM 34.6±4.6 years) were obtained from open biopsies or after operation from resected specimens. All patients with bone diseases were hospitalized at the Medical Radiological Research Centre. In all cases the diagnosis was confirmed by clinical and histological data.

    A titanium tool was used to cut and to scrape samples33, 34. All bone and tumor tissue samples were freeze dried, until constant mass was obtained, and homogenized. Then samples weighing about 100 mg were wrapped separately in high-purity aluminum foil washed with rectified alcohol beforehand and placed in a nitric acid-washed quartz ampoule.

    To determine contents of the elements by comparison with a known standard, biological synthetic standards (BSS) prepared from phenol–formaldehyde resins and aliquots of commercial, chemically pure compounds were used.Corrected certified values of BSS element contents were reported by us before.35 Ten certified reference material (CRM) IAEA H-5 (Animal Bone) sub-samples and ten standard reference material (SRM) NIST 1486 (Bone Meal) sub-samples weighing about 100 mg were analyzed in the same conditions as bone and tumor samples to estimate the precision and accuracy of the results.

    A vertical channel of the WWR-c research nuclear reactor was applied to determine the mass fraction of Ag, Co, Cr, Fe, Hg, Rb, Sb, Se, and Zn by INAA-LLR. The quartz ampoule with bone samples, tumor samples, standards, CRM, and SRM was soldered, positioned in a transport aluminum container and exposed to a 100-hour neutron irradiation in a vertical channel with a thermal neutron flux about 1013 n×cm-2×s-1. Two months after irradiation the samples were reweighed and repacked. The duration of each measurement was from 1 to 10 hours. To reduce the high intensity of32 P b-particles (T1/2=14.3 d) background, a berillium filter was used. A coaxial 98 cm3 Ge (Li) detector and a spectrometric unit (NUC 8100), including a PC-coupled multichannel analyzer, were used for measurements. The spectrometric unit provided 2.9 keV resolution at the 60Co 1332 keV line. The information of used nuclear reactions, radionuclides, gamma-energies, and other details of the analysis including the quality control of results were reported by us before.22, 24, 25, 26

    A dedicated computer program of INAA mode optimization was used37. Using the Microsoft Office Excel programs, the summary of statistics, arithmetic mean, standard deviation, standard error of mean, minimum and maximum values, median, percentiles with 0.025 and 0.975 levels were calculated for different trace element mass fractions and their selected combinations. The reliability of difference in the results between intact bone and chondrosarcoma tissue was evaluated by Student’s t-test. For the estimation of the Pearson correlation coefficient between different pairs of the trace element mass fractions in intact bone and chondrosarcoma tissue the Microsoft Office Excel program was also used.

    Results

    Figure 1. shows individual data sets for Ag, Co, Cr, Fe, Hg, Rb, Sb, Se, and Zn mass fractions (mg/kg, dry mass basis) in all samples of intact bone (1) and chondrosarcoma (2) Tissue type
    Figure 1.

    Table 1 depicts the basic statistical parameters (arithmetic mean, standard deviation, standard error of mean, minimal and maximal values, median, percentiles with 0.025 and 0.975 levels) for the Ag, Co, Cr, Fe, Hg, Rb, Sb, Se, and Zn mass fraction in intact bone and chondrosarcoma tissue.

    Table 1. Basic statistical parameters for Al, Co, Cr, Fe, Hg, Rb, Sb, Se, and Zn mass fractions (mg/kg, dry mass basis) in tissue of intact bone and chondrosarcoma
    E M SD SEM Min Max Med P0.025 P0.975
    Intact bone, n=27                
    Ag 0.0027 0.0015 0.0005 0.00026 0.0047 0.0028 0.00032 0.0046
    Co 0.0107 0.0070 0.0014 0.00370 0.0345 0.0079 0.00464 0.0288
    Cr 0.274 0.182 0.057 0.110 0.669 0.202 0.117 0.629
    Fe 51.2 46.3 9.3 9.20 173 30.2 9.68 155
    Hg 0.0057 0.0044 0.0014 0.00100 0.0138 0.0052 0.0010 0.0133
    Rb 3.68 1.58 0.48 0.970 6.57 3.30 1.40 6.41
    Sb 0.0151 0.0102 0.0032 0.00600 0.0420 0.0139 0.00600 0.0364
    Se 0.176 0.092 0.029 0.0550 0.358 0.169 0.0633 0.336
    Zn 80.6 15.4 3.0 45.4 115 82.1 51.7 109
    Chondrosarcoma, n=16                
    Ag 0.0030 0.0029 0.00072 0.00043 0.0122 0.0025 0.00051 0.0096
    Co 0.0365 0.0236 0.0061 0.00950 0.0998 0.0282 0.0111 0.0876
    Cr 0.483 0.386 0.097 0.0980 1.51 0.416 0.116 1.41
    Fe 178 126 34 16.5 413 129 29.8 402
    Hg 0.0172 0.0192 0.0048 0.00007 0.0571 0.0080 0.00051 0.0537
    Rb 1.56 1.58 0.40 0.170 6.15 1.33 0.170 5.11
    Sb 0.0171 0.0122 0.0031 0.00510 0.0437 0.0130 0.00518 0.0418
    Se 1.78 1.19 0.31 0.203 4.32 1.70 0.251 3.81
    Zn 118 56 14 41.7 226 108 47.4 225

    E element M arithmetic mean, SD standard deviation, SEM standard error of mean, Min minimum value, Max maximum value, Med median, P0.025 percentile with 0.025 level, P0.975 percentile with 0.975 level

    The ratio of means and the reliability of difference between mean values of Al, Co, Cr, Fe, Hg, Rb, Sb, Se, and Zn mass fractions in tissue of intact bone and chondrosarcoma are presented in Table 2.

    Table 2. Means (M±SEM, mg/kg, dry mass basis), ratio of means and the reliability of difference between mean values of Al, Co, Cr, Fe, Hg, Rb, Sb, Se, and Zn mass fractions in tissue of intact bone and chondrosarcoma
    Element Intact boneM1 Chondrosarcoma M2 M2 / M1Ratio Student’s t-test
    Ag 0.0027±0.0005 0.0030±0.0007 1.08 p ≤ 0.85
    Co 0.0107±0.0014 0.0365±0.0061 3.41 p 0.0022
    Cr 0.274±0.057 0.483±0.097 1.76 p ≤ 0.155
    Fe 51.2±9.3 178±34 3.48 p 0.0055
    Hg 0.0057±0.0014 0.0172±0.0048 3.02 p ≤ 0.128
    Rb 3.68±0.48 1.56±0.40 0.42 p 0.00076
    Sb 0.0151±0.0032 0.0171±0.0031 1.13 p ≤ 0.575
    Se 0.176±0.029 1.78±0.31 10.1 p 0.00069
    Zn 80.6±3.0 118±14 1.46 p ≤ 0.0775

    M arithmetic mean, SEM standard error of mean, bold statistically significant

    Table 3 represents the basic statistical parameters for Co/Zn, Cr/Zn, Fe/Zn, Hg/Zn, Sb/Zn, Co/Rb, Cr/Rb, Fe/Rb, Hg/Rb, Sb/Rb, and Se/Rb mass fractions ratios in the samples of intact bone and chondrosarcoma.

    Table 3. Basic statistical parameters for Co/Zn, Cr/Zn, Fe/Zn, Hg/Zn, Sb/Zn, Co/Rb, Cr/Rb, Fe/Rb, Hg/Rb, Sb/Rb, and Se/Rb mass fractions ratios in tissue of intact bone and chondrosarcoma
    Mass fractions ratio M SD SEM Min Max Med P0.025 P0.975
    Intact bone, n=27                
    (Co/Zn)×103 0.14 0.088 0.017 0.053 0.40 0.10 0.0593 0.364
    (Cr/Zn)×103 2.94 1.78 0.56 1.57 6.37 2.07 1.59 6.27
    Fe/Zn 0.69 0.74 0.15 0.098 3.15 0.33 0.120 2.48
    (Hg/Zn)×104 0.65 0.48 0.15 0.099 1.37 0.59 0.0995 1.36
    (Sb/Zn)×103 0.17 0.11 0.034 0.072 0.45 0.13 0.0753 0.393
    (Co/Rb)×102 0.61 0.99 0.30 0.13 3.56 0.25 0.144 2.87
    (Cr/Rb)×101 0.79 0.41 0.13 0.24 1.47 0.74 0.269 1.45
    Fe/Rb 29 51 15 2.49 178 10.8 2.75 142
    (Hg/Rb)×103 2.3 3.5 1.1 0.30 12.1 1.23 0.308 9.84
    (Sb/Rb)×103 5.2 4.9 1.6 1.48 16.6 3.52 1.52 15.4
    (Se/Rb)×102 7.2 10.6 3.3 2.04 36.9 3.45 2.28 30.4
    Chondrosarcoma, n=16                
    (Co/Zn)×103 0.35 0.30 0.077 0.080 1.04 0.28 0.0960 1.03
    (Cr/Zn)×103 4.61 3.65 0.91 0.85 13.2 3.22 0.927 12.2
    Fe/Zn 1.79 1.66 0.41 0.28 6.42 1.31 0.372 5.76
    (Hg/Zn)×104 1.35 1.53 0.38 0.008 6.08 0.87 0.0689 4.81
    (Sb/Zn)×103 0.18 0.19 0.046 0.043 0.79 0.09 0.0474 0.614
    (Co/Rb)×102 5.3 5.6 1.4 0.46 21.5 3.24 0.553 18.3
    (Cr/Rb)×101 10.0 21.6 5.4 0.61 88.8 3.42 0.645 63.5
    Fe/Rb 266 293 73 22.9 1047 149 25.2 952
    (Hg/Rb)×103 16.2 26.4 6.6 0.22 112 10.5 0.753 79.5
    (Sb/Rb)×103 26.3 29.1 7.3 2.73 115 18.9 2.79 92.2
    (Se/Rb)×102 314 484 121 8.57 1588 148 13.1 1549

    M arithmetic mean, SD standard deviation, SEM standard error of mean, Min minimum value, Max maximum value, Med median, P0.025 percentile with 0.025 level, P0.975 percentile with 0.975 level

    The ratio of means and the reliability of difference between mean values of Co/Zn, Cr/Zn, Fe/Zn, Hg/Zn, Sb/Zn, Co/Rb, Cr/Rb, Fe/Rb, Hg/Rb, Sb/Rb, and Se/Rb mass fractions ratios in tissue of intact bone and chondrosarcoma are presented in Table 4.

    Table 4. Means (M±SEM), ratio of means and the reliability of difference between mean values of Co/Zn, Cr/Zn, Fe/Zn, Hg/Zn, Sb/Zn, Co/Rb, Cr/Rb, Fe/Rb, Hg/Rb, Sb/Rb, and Se/Rb mass fractions ratios in tissue of intact bone and chondrosarcoma
    Mass fractionsratio Intact boneM1 Chondrosarcoma M2 M2 / M1Ratio Student’s t-test
    (Co/Zn)×103 0.140±0.017 0.350±0.077 2.57 p ≤ 0.0224
    (Cr/Zn)×103 2.94±0.56 4.61±0.91 1.57 p ≤ 0.140
    Fe/Zn 0.69±0.15 1.79±0.41 2.59 p ≤ 0.0488
    (Hg/Zn)×104 0.65±0.15 1.35±0.38 2.08 p ≤ 0.174
    (Sb/Zn)×103 0.170±0.034 0.180±0.046 1.08 p ≤ 0.568
    (Co/Rb)×102 0.61±0.30 5.3±1.4 8.69 p ≤ 0.00292
    (Cr/Rb)×101 0.79±0.13 10.0±5.4 12.7 p ≤ 0.0141
    Fe/Rb 29±15 266±73 9.17 p ≤ 0.00253
    (Hg/Rb)×103 2.3±1.1 16.2±6.6 7.04 p ≤ 0.160
    (Sb/Rb)×103 5.2±1.6 26.3±7.3 5.06 p ≤ 0.00615
    (Se/Rb)×102 7.2±3.3 314±121 43.6 p ≤ 0.0497

    M arithmetic mean, SEM standard error of mean, bold statistically significant

    The data of inter-correlation calculations (values of r – coefficient of correlation) including all pairs of the chemical elements identified by us in the intact bone and the chondrosarcoma.tissue are shown in Table 5.

    Table 5. Intercorrelations of pairs of the trace element mass fractions in tissue of intact bone and chondrosarcoma
    Tissue E Co Cr Fe Hg Rb Sb Se Zn
    Intact bone Ag -0.23 0.51 -0.80b -0.02 0.62a 0.31 -0.45 0.38
    n=27 Co - 0.16 0.55b 0.79b -0.10 0.08 0.52 0.17
      Cr 0.16 - -0.48 0.51 0.56a -0.31 -0.08 0.46
      Fe 0.55b -0.48 - 0.09 -0.54 -0.25 0.60a -0.17
      Hg 0.79b 0.51 0.09 - 0.18 -0.13 0.35 -0.14
      Rb -0.10 0.56a -0.54 0.18 - -0.05 -0.06 0.34
      Sb 0.08 -0.31 -0.25 -0.13 -0.05 - 0.04 0.22
      Se 0.52 -0.08 0.60a 0.35 -0.06 0.04 - 0.24
      Zn 0.46 0.46 -0.17 -0.14 0.34 0.22 0.24 -
    Chondrosarcoma Ag 0.14 0.08 0.13 0.12 0.38 -0.24 -0.36 0.20
    n=16 Co - 0.61a 0.38 0.04 0.23 0.34 0.28 -0.08
      Cr 0.61a - 0.11 -0.03 0.06 0.26 0.12 0.11
      Fe 0.38 0.11 - -0.16 -0.01 0.16 0.07 -0.10
      Hg 0.04 -0.03 -0.16 - 0.75b -0.11 0.19 0.64b
      Rb 0.23 0.06 -0.01 0.75b - 0.01 -0.09 0.39
      Sb 0.34 0.26 0.16 -0.11 0.01 - 0.34 0.07
      Se 0.28 0.12 0.07 0.19 -0.09 0.34 - -0.10
      Zn -0.08 0.11 -0.10 0.64b 0.39 0.07 -0.10 -

    E element, statistically significant difference:
    a - p ≤ 0.05,
    b - p ≤ 0.01

    Discussions

    The non-destructive INAA-LLR was used in this research study because this method has many definite advantages over other analytical methods, particularly, in the clinical chemistry. For example, after non-destructive INAA-LLR there is a possibility to check the results for some trace elements and to receive additional information about other trace element contents by destructive analytical methods such as atomic absorption spectrometry, inductively coupled plasma atomic emissionspectrometry, inductively coupled plasma massspectrometry and so on, using the same bone samples. Moreover, if a deep-cooled channel of nuclear reactor is available, the non-destructive INAA-LLR allows determining trace element contents in the fresh bone/tumor samples and combining trace element study with histological investigation. It is also necessary to keep in mind that the non-destructive methods are the current gold-standard solution to control destructive analytical techniques.2 The destructive analytical methods are based on measurements of processed tissue. In such studies tissue samples are ashed and/or acid digested before analysis. There is evidence that certain quantities of chemical elements are lost as a result of such treatment.2, 34, 38 There is no doubt that every method available for the measurement of trace element contents in bone and tumor samples can be used. However, when using destructive analytical methods it is necessary to control for the losses of trace elements, for complete acid digestion of the sample, and for the contaminations by trace elements during sample decomposition, which needs adding some chemicals.

    In our previous study it was shown that the results of mean values for all representative elements of CRM IAEA H-5 (Animal Bone) and SRM NIST1486 (Bone Meal) were in the range of 95% confidence interval (M±2SD) of the certificates’ values.22, 24, 25, 36 Good agreement with the certified data of CRM and SRM indicate an acceptable accuracy for the trace element mass fractions obtained in the study of intact bone and chondrosarcoma tissue presented in Tables 1-5.

    In the control group the mass fractions of Co, Fe and Zn were measured in all samples, but the mass fraction of Rb – in 11 samples and mass fractions of Ag, Cr, Hg, Sb, and Se – in 10 samples Figure 1. In the chondrosarcoma group the mass fraction of all nine trace elements were determined in all samples Figure 1.

    Table 2 shows that in chondrosarcoma tissue the mean mass fraction of Ag, Co, Cr, Fe, Hg, Sb, Se, and Zn is higher while the mean mass fraction of Rb is lower than in the normal bone tissues. However, in chondrosarcoma only the mean mass fractions of Co (p ≤ 0.0022), Fe (p ≤ 0.0055), and Se (p ≤ 0.0055) are significantly increased and the mean mass fraction of Rb (p ≤ 0.00076) is significantly decreased when compared with those in normal bone. Different directions of mass fraction changes suggest potential use of mass fraction ratios of these trace elements as chondrosarcoma markers. A simple ratio of two trace element mass fractions, which change in two direction, can improve the difference between such characteristics of intact bone and chondrosarcoma tissues. These conclusion was the main reason for calculating Co/Zn, Cr/Zn, Fe/Zn, Hg/Zn, Sb/Zn, Co/Rb, Cr/Rb, Fe/Rb, Hg/Rb, Sb/Rb, and Se/Rb mass fractions ratios (Table 3). It was found that higher mean values of all selected mass fraction ratios were typical of chondrosarcoma tissue compared with intact bone (Table 4). However, in chondrosarcoma only the mean mass fractions ratio of Co/Zn (p ≤ 0.0224), Fe/Zn (p ≤ 0.0488), Co/Rb (p ≤ 0.00292), Cr/Rb (p ≤ 0.0141), Fe/Rb (p ≤ 0.00253), Sb/Rb (p ≤ 0.00615), and Se/Rb (p ≤ 0.0497) are significantly increased when compared with those in normal bone.

    In the control group a statistically significant direct correlation was found, for example, between the Fe and Se (r = 0.60, p ≤0.05), Fe and Co (r = 0.55, p ≤0.01), Co and Hg (r = 0.79, p ≤0.01), Rb and Ag (r = 0.62, p ≤0.05), and between Rb and Cr (r = 0.56, p ≤0.05) mass fractions (Table 5). In the same group a pronounced (p ≤ 0.01) inverse correlation was observed between the Fe and Ag (r = - 0.80, p ≤0.05). If some positive correlations between the trace elements were predictable (e.g., Fe–Co), the interpretation of other observed relationships requires further study for a more complete understanding.

    In the chondrosarcoma tissue many significant correlations between trace elements found in the control group are no longer evident, for example, direct correlation between Fe and Se, etc. (Table 5). However, direct correlations between Co and Cr (r = 0.61, p ≤0.05), Hg and Rb (r = 0.75, p ≤0.01) and Hg and Zn (r = 0.64, p ≤0.01) were observed (Table 5). Thus, if we accept the levels and relationships of trace element mass fraction in the intact bone samples of control group as a norm, we have to conclude that with a chondrosarcoma transformation the levels and relationships of trace elements in bone significantly change. No published data referring to correlations between trace elements mass fractions in chondrosarcoma tissue were found.

    The changes in trace element contents of cancerous tissues in comparison with non-cancerous tissues may be attributed to a cause or effect of malignant transformation. Bone is a mineralized connective tissue. It is formed by osteoblast, that deposit collage and release Ca, Mg, and phosphate ions that combine chemically within the collagenous matrix into a crystalline mineral, known as bon hydroxyapatite. On average, bone tissue contains about 10-25% water, 25% protein fibers like collagen, and 50% hydroxyapatit Ca10(PO4)6(OH)2. Many trace elements are bone-seeking elements and they are closely associated with hydroxyapatit.24, 25, 26, 27, 28 Chondrosarcoma is classified as a bone tumor. Our previous findings showed that the means of the Ca and P mass fraction in the chondrosarcoma tissue are lower than in normal bone, but the mean of Ca/P ratio is similar.6 It suggested that chondrosarcoma continues to form bon hydroxyapatit but to a lesser degree than normal bone. Our findings show that the mean of the Fe mass fraction in chondrosarcoma tissue samples was 3.48 times greater than in normal bone tissues (Table 2). It is well known that Fe mass fraction in sample depends mainly from the blood volumes in tissues. Chondrosarcoma at least in grades II and III show increased microvascularity.39 Thus, it is possible to speculate that chondrosarcoma is characterized by an increase of the mean value of the Fe mass fraction because the level of tumor vascularization is higher than that in normal bone. As we found, there is a direct correlation between Fe and Co mass fractions (Table 5). Therefore an increased level of Co in the chondrosarcoma may be closely connected with a high Fe content in tumor tissue (Table 2).

    In the chondrosarcoma tissue the mean Se mass fractions is 10.1 times higher (p ≤ 0.00069) than in normal bone (Table 2). The high Se level was reported in malignant tumors of ovary,40 lung,41 prostate,42 breast43, 44 intestine,45 and in gastric cancer tissue.46 The role played by Se in those tumors remains unknown, but in general it is accepted that certain proteins containing Se can mediate the protective effects against oxidative stress. The literature-based analysis found the association of malignant tissue transformation with local oxidative stress. Studies have shown that oxidative stress conditions play an important role in both the initiation and the progression of cancer by regulating molecules such as DNA, enhancers, transcription factors, and cell cycle regulators.47 However the cause of increased Se in cancerous tissue and particularly in the chondrosarcoma is not completely understood and requires further studies.

    Conclusions

    INAA-LLR is a satisfactory analytical tool to determine non-destructively the elemental content of Ag, Co, Cr, Fe, Hg, Rb, Sb, Se, and Zn in human bone samples and samples of intraosseous lesions weighing about 100 mg. In the chondrosarcoma tissue the mass fractions of Co, Fe, and Se are significantly higher while the mass fraction of Rb is lower than in normal bone tissues. Moreover, significantly higher Co/Zn, Fe/Zn, Co/Rb, Cr/Rb, Fe/Rb, Sb/Rb, and Se/Rb mass fraction ratios are typical of the chondrosarcoma tissue compared to intact bone. In the chondrosarcoma tissue many correlations between trace elements found in the control group are no longer evident. Thus, if we accept the levels and relationships of trace element mass fraction in the intact bone as a norm, we have to conclude that in chondrosarcoma transformed bone tissues the trace element homeostasis is significantly disturbed.

    Ethical Approval

    The study was approved by the Ethical Committee of the Medical Radiological Research Center, Obninsk.

    Acknowledgements

    The authors are grateful to the late Prof. V.A. Bizer, Medical Radiological Research Center, Obninsk, for supplying the chondrosarcoma samples.

    References

    1.Ulmer D D. (1973) Metals - from privation to pollution. Federation. Proc 32, 1758-1762.
    2.Zaichick V. (2006) Medical elementology as a new scientific discipline. , J Radioanal Nucl Chem 269, 303-309.
    3.Zherbin E A, Zaichick V. (1976) Several aspects of applied neutron activation in medicine: The present state and development of activation analysis in the Institute of Medical Radiology. In:. Proceedings of the 2nd Meeting on New Nuclear-Physical Methods Used in Solving Scientific-Technical and National Economic Problems. Join Institute of Nuclear Research , Dubna, Russia, 104–126 (in Russian) .
    4.Kalashnikov V M, Zaichick V. (1977) Bone analysis of N, F and P by photonuclear activation. (in Russian) , Vopr Med Khim 23(1), 122-127.
    5.Zaichick V. (1993) The in vivo neutron activation analysis of calcium in the skeleton of normal subjects, with hypokinesia and bone diseases. , J Radioanal Nucl Chem, Articles 169, 307-316.
    6.Zaichick V. (1994) Instrumental activation and X-ray fluorescent analysis of human bones in health and disease. , J Radioanal Nucl Chem, Articles 179, 295-303.
    7.Zaichick V, Ovchjarenko N N. (1996) In vivo X-ray fluorescent analysis of Ca, Zn, Sr, and Pb in frontal tooth enamel. , J Trace and Microprobe Techniques 14(1), 143-152.
    8.Zaichick V, Morukov B. (1998) In vivo bone mineral studies on volunteers during a 370-day antiorthostatic hypokinesia test. Appl Radiat Isot. 49(5), 691-694.
    9.Zaichick V. (1998) In vivo and in vitro application of energy-dispersive XRF in clinical investigations: experience and the future. , J Trace Elements in Experimental Medicine 11(4), 509-510.
    10.Zaichick V, Ovchjarenko N, Zaichick S. (1999) In vivo energy dispersive X-ray fluorescence for measuring the content of essential and toxic trace elements in teeth. App l Radiat Isot. 50(2), 283-293.
    11.Zaichick V, Snetkov A. (2000) Bone composition in children with rickets-like diseases before and during treatment. In Anke M. et al. (eds): Mengen- und Spurenelemente. 20 Arbeitstagung. Friedrich-Schiller-Universitat , Jena 1109-1117.
    12.Zaichick V, Dyatlov A, Zaihick S. (2000) INAA application in the age dynamics assessment of major, minor, and trace elements in the human rib. , J Radioanal Nucl Chem 244, 189-193.
    13.C S Sastri, Iyengar V, Blondiaux G, Tessier Y, Petri H et al. (2001) Fluorine determination in human and animal bones by particle-induced gamma-ray emission. , Fresenius J Anal Chem 370, 924-929.
    14.Zaichick V, Tzaphlidou M. (2002) Determination of calcium, phosphorus, and the calcium/phosphorus ratio in cortical bone from the human femoral neck by neutron activation analysis. Appl Rad Isotop. 56, 781-786.
    15.Tzaphlidou M, Zaichick V. (2002) Neutron activation analysis of calcium/phosphorus ratio in rib bone of healthy humans. , Appl Rad Isotop 57, 779-783.
    16.Tzaphlidou M, Zaichick V. (2003) Calcium, Phosphorus, Calcium-Phosphorus ratio in rib bone of healthy humans. , Biol Trace Elem. Res 93, 63-74.
    17.Zaichick V, Tzaphlidou M. (2003) Calcium and phosphorus concentrations and calcium/phosphorus ratio in trabecular bone from femoral neck of healthy humans as determined by neutron activation analysis. Appl Rad Isotop. 58, 623-627.
    18.Zaichick V. (2004) INAA application in the age dynamics assessment of Ca. , Cl, K, Mg, Mn, Na, P, and, J Radioanal Nucl. Chem 259, 351-354.
    19.Zaichick V. (2004) Sex and age related Ca/P ratio in trabecular bone of iliac crest of healthy humans. In Anke M. et al. (eds): Macro and Trace Elements. 22 Workshop, Vol.1 , Friedrich-Schiller-Universitat, Jena 248-255.
    20.Tzaphlidou M, Zaichick V. (2004) Sex and age related Ca/P ratio in cortical bone of iliac crest of healthy humans. , J Radioanal Nucl Chem 259, 347-349.
    21.Zaichick V. (2007) INAA application in the assessment of selected elements in cancellous bone of human iliac crest. , J Radioanal Nucl Chem 271, 573-576.
    22.Zaichick V, Zaichick S. (2009) Instrumental neutron activation analysis of trace element contents in the rib bone of healthy men. , J Radioanal Nucl Chem 281, 47-52.
    23.Zaichick V. (2009) Neutron activation analysis of. , J Radioanal Nucl Chem 281, 41-45.
    24.Zaichick S, Zaichick V. (2010) The effect of age and gender on 38 chemical element contents in human iliac crest investigated by instrumental neutron activation analysis. , J. Trace Elem. Med. Biol 24, 1-6.
    25.Zaichick S, Zaichick V. (2010) The effect of age and gender on 38 chemical element contents in human femoral neck investigated by instrumental neutron activation analysis. , Biol. Trace Elem. Res 137, 1-12.
    26.Zaichick S, Zaichick V. (2012) Neutron activation analysis of Ca, Cl, Mg, Na, and P content in human bone affected by osteomyelitis or osteogenic sarcoma. , J Radioanal Nucl Chem 293(1), 241-246.
    27.Zaichick V. (2013) Chemical elements of human bone tissue investigated by nuclear analytical and related methods. , Biol Trace Elem. Res 153, 84-99.
    28.Zaichick V. (2013) Data for the Reference Man: skeleton content of chemical elements. , Radiat Environ Biophys 52, 65-85.
    29.Zaichick V, Zaichick S. (2014) The Ca, Cl, Mg, Na, and P mass fractions in human bone affected by Ewing’s sarcoma. Biol Trace Elem Res. 159(1), 32-38.
    30.Randall R, Gowski. (2005) Grade 1 chondrosarcoma of bone: a diagnostic and treatment dilemma. , J Natl Compr Canc Net 3, 149-156.
    31.H D Dorfman, Czerniak B. (1998) Bone Tumors. Mosby-Yearbook, St Louis.
    32.Berg Slaar Van den, Kroon H, Taminiau A, Hogendoorn. (2008) Results of diagnostic review in pediatric bone tumors and tumorlike lesions. , J Pediatr Orthop 28(5), 561-564.
    33.Zaichick V, Zaichick S. (1996) Instrumental effect on the contamination of biomedical samples in the course of sampling. , J Anal Chem 51(12), 1200-1205.
    34.Zaichick V. (1997) Sampling, sample storage and preparation of biomaterials for INAA in clinical medicine, occupational and environmental health. In: Harmonization of Health-Related Environmental Measurements Using Nuclear and Isotopic Techniques. , IAEA, Vienna 123-133.
    35.Zaichick V. (1995) Application of synthetic reference materials in the Medical Radiological Research Centre. , Fresenius J Anal Chem 352, 219-223.
    36.Zaichick V, Kolotov A V, Dogadkin N. Friedrich Schiller University (2002) Comparative analysis of major and trace elements in bone reference materials. IAEA H-5 (animal bone) and NIST SRM 1486 (bone meal). In Anke M. et al. (eds): Macro and Trace Elements. 21 Workshop , Jena 39-47.
    37.Korelo A, Zaichick V. (1993) Software to optimize the multielement INAA of medical and environmental samples. In: Activation analysis in environment protection. Join Institute of Nuclear Research , Dubna, Russia, 326–32 (in Russian) .
    38.Zaichick V. (2004) Losses of chemical elements in biological samples under the dry ashing process. Trace Elements in Medicine. 5(3), 17-22.
    39.Terek R M. (2002) Angiogenesis in Chondrosarcoma. , Current Opinions in Orthopaedics 13(6), 449-453.
    40.Xue F, Zhang S. (1991) Selenium concentrations in serum, hair and tumor tissue from patients with ovarian tumors. Zhonghua Fu Chan Ke Za Zh. 26(5), 290-292.
    41.Zachara B, Marchaluk-Wiœniewska Maciag, Pepliñski Skokowski, Lambrecht. (1997) Decreased selenium concentration and glutathione peroxidase activity in blood and increase of these parameters in malignant tissue of lung cancer patients. , Lun 175(5), 321-332.
    42.Zachara B, Szewczyk-Golec Tyloch, Wolski Szylberg. (2005) Blood and tissue selenium concentrations and glutathione peroxidase activities in patients with prostate cancer and benign prostate hyperplasia. , Neoplasm 52(3), 248-254.
    43.Raju G JN, Sarita P, Kumar M R, Murty G, Reddy B S. (2006) Trace elemental correlation study in malignant and normal breast tissue by PIXE technique. , Nuclear Instruments & Methods in Physics Research B 247(2), 361-367.
    44.Charalabopoulos Kotsalos, Batistatou Charalabopoulos, Vezyraki. (2006) Selenium in serum and neoplastic tissue in breast cancer: correlation with. 95(6), 674-676.
    45.Kucharzewski Braziewicz, Majewska, Gуzdz. (2003) Selenium, copper, and zinc concentrations in intestinal cancer tissue and in colon and rectum polyps. , Biol Trace Elem. Re 92(1), 1-10.
    46.Charalabopoulos Kotsalos, Batistatou Charalabopoulos, Peschos. (2009) Serum and tissue selenium levels in gastric cancer patients and correlation with CEA. , Anticancer Re 29(8), 3465-3467.
    47.Elera Gupta, Garrett A, Robison R, O’Neill K. (2012) The role of oxidative stress in prostate cancer. , Eur J Cancer Pre 21(2), 155-162.

    Cited by (1)

    1.Zaichick Vladimir, Zaichick Sofia, Davydov German, 2016, Method and portable facility for measurement of trace element concentration in prostate fluid samples using radionuclide-induced energy-dispersive X-ray fluorescent analysis, Nuclear Science and Techniques, 27(6), 10.1007/s41365-016-0133-3