Magnesium deficiency results in an increased formation of osteoclasts

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Magnesium deficiency results in an increased formation of osteoclasts Marina M. Belluci, Ton Schoenmaker, Carlos Rossa-Junior, Silvana R. Orrico, Teun J. de Vries, Vincent Everts Journal of Nutritional Biochemistry Volume 24, Issue 8, Pages 1488-1498 (August 2013) DOI: 10.1016/j.jnutbio.2012.12.008 Copyright © 2013 Elsevier Inc. Terms and Conditions

Fig. 1 (A) Morphological aspects of TRACP+-MNCs formed from precursors obtained from long bone marrow after 6 days of culture on plastic in four different Mg2+ concentrations in the presence of M-CSF and RANKL (n=6). The cells show an elongated shape in all Mg2+ concentrations. A higher number of nuclei per cell were observed under Mg2+ deprivation. (B–D) Number of TRACP+-MNCs with 3–5 nuclei (B), 6–10 nuclei (C), >10 nuclei (D) and total number of TRACP+-MNC (E) formed under the four Mg2+ culture regimens (C). Data are expressed as mean±S.D. ⁎P<.05; ⁎⁎P<.01; ⁎⁎⁎P<.0001. Journal of Nutritional Biochemistry 2013 24, 1488-1498DOI: (10.1016/j.jnutbio.2012.12.008) Copyright © 2013 Elsevier Inc. Terms and Conditions

Fig. 2 (A) Morphological aspects of TRACP+-MNCs formed by long bone marrow cells after 6 days of culture on bone in four different Mg2+ concentrations in the presence of M-CSF and RANKL (n=6). TRACP+-MNCs on bone had a round shape in all Mg2+ concentrations. A higher number of nuclei per cell were observed under Mg2+ deprivation (B–D). Number of TRACP+-MNCs with 3–5 nuclei (B), 6–10 nuclei (C), > 10 nuclei (D) and total number of TRACP+-MNC (E) formed under the four Mg2+ culture regimens (C). Data are expressed as mean±S.D. ⁎P<.05; ⁎⁎P<.01. Journal of Nutritional Biochemistry 2013 24, 1488-1498DOI: (10.1016/j.jnutbio.2012.12.008) Copyright © 2013 Elsevier Inc. Terms and Conditions

Fig. 3 Number of TRACP+-MNCs formed by jaw marrow cells after 6 days of culture on plastic (A–B) and on bone (C–D) in four different Mg2+ concentrations in the presence of M-CSF and RANKL (n=6). Number of TRACP+-MNCs with 6–10 nuclei (A) and >10 nuclei (B) cultured on plastic. Number of TRAP+-MNCs with 6–10 nuclei (C) and >10 nuclei (D) cultured on bone. Data are expressed as mean±S.D. ⁎P<.05; ⁎⁎P<.01. Journal of Nutritional Biochemistry 2013 24, 1488-1498DOI: (10.1016/j.jnutbio.2012.12.008) Copyright © 2013 Elsevier Inc. Terms and Conditions

Fig. 4 (A) Mg2+ concentration in the medium of cultures with and without cells (n=6). Cells were cultured on plastic or on bone for 6 days. (B) The Ca2+ concentration in the medium with and without cells cultured on plastic or on bone for 6 days (n=6). Data are expressed as mean±S.D. ⁎P<.05. Journal of Nutritional Biochemistry 2013 24, 1488-1498DOI: (10.1016/j.jnutbio.2012.12.008) Copyright © 2013 Elsevier Inc. Terms and Conditions

Fig. 5 (A) The released amount of AK was not altered under the different Mg concentrations. Triton X-100 was added to the wells as a positive control. (B) The amount of DNA of cells from long bone marrow on day 0, 3 and 6 (n=6). ⁎⁎⁎P<.0001. Data are expressed as mean±S.D. Journal of Nutritional Biochemistry 2013 24, 1488-1498DOI: (10.1016/j.jnutbio.2012.12.008) Copyright © 2013 Elsevier Inc. Terms and Conditions

Fig. 6 Gene expression of c-fms (A), RANK (B), DC-STAMP (C), c-fos (D), TNF-α (E) and IL1-β (F) by precursor cells from long bone marrow after 3 days of culture on plastic (n=6). The gene expression was normalized for the housekeeping gene PBGD. Data are expressed as mean±S.D. ⁎P<.05; ⁎⁎P<.01; ⁎⁎⁎P<.0001. Journal of Nutritional Biochemistry 2013 24, 1488-1498DOI: (10.1016/j.jnutbio.2012.12.008) Copyright © 2013 Elsevier Inc. Terms and Conditions

Fig. 7 Gene expression of c-fms (A), RANK (B), DC-STAMP (C), c-fos (D), TNF-α (E) and IL1-β (F) by precursor cells from long bone marrow after 3 days of culture on bone (n=6). The gene expression was normalized for the housekeeping gene PBGD. Data are expressed as mean±S.D. ⁎P<.05. Journal of Nutritional Biochemistry 2013 24, 1488-1498DOI: (10.1016/j.jnutbio.2012.12.008) Copyright © 2013 Elsevier Inc. Terms and Conditions

Fig. 8 Gene expression of TRPM7 (A, C) and MRS2 (B, D) by precursor cells from long bone after 3 days of culture. Gene expression was assessed from cells cultured on plastic (A–B) and on bone (C–D), respectively (n=6). The gene expression was normalized for the housekeeping gene PBGD. Data are expressed as mean±S.D. ⁎P<.05; ⁎⁎P<.01. Journal of Nutritional Biochemistry 2013 24, 1488-1498DOI: (10.1016/j.jnutbio.2012.12.008) Copyright © 2013 Elsevier Inc. Terms and Conditions

Fig. 9 (A) Bone resorption after 8 days of culture. (B) Bone resorption corrected for the number of osteoclasts. (C) Levels of TRACP enzyme secreted in the medium. (D) Levels of TRACP corrected for the number of osteoclasts. Data are expressed as mean±S.D. ⁎P<.05; ⁎⁎P<.01. Journal of Nutritional Biochemistry 2013 24, 1488-1498DOI: (10.1016/j.jnutbio.2012.12.008) Copyright © 2013 Elsevier Inc. Terms and Conditions