Bone loss induced by dietary magnesium reduction to 10% of the nutrient requirement in rats is associated with increased release of substance P and tumor necrosis factor-alpha.

Dietary Mg intake has been linked to osteoporosis. Previous studies have demonstrated that severe Mg deficiency [0.04% of nutrient requirement (NR)] results in osteoporosis in rodent models. We assessed the effects of more moderate dietary Mg restriction (10% of NR) on bone and mineral metabolism over a 6-mo experimental period in rats. At 2, 4 and 6 mo, serum Mg, Ca, parathyroid hormone (PTH), 1,25-dihydroxy-vitamin D, alkaline phosphatase, osteocalcin and urine pyridinoline were measured. Femurs and tibiae were collected for measurement of mineral content, microcomputerized tomography, histomorphometry, and immunocytochemical localization. By 2 mo, profound Mg deficiency had developed as assessed by marked hypomagnesemia and up to a 51% reduction in bone Mg content. These features continued through 6 mo of study. Serum Ca was slightly but significantly higher in Mg-deficient rats than in controls at all time points. At 2 mo, serum PTH was elevated in Mg-deficient rats but was significantly decreased at 6 mo in contrast to control rats in which PTH rose. Serum 1,25-dihydroxy-vitamin D was significantly lower than in controls at 4 and 6 mo. A significant fall in both serum alkaline phosphatase and osteocalcin suggested decreased osteoblast activity. Histomorphometry demonstrated decreased bone volume and trabecular thickness. This was confirmed by microcomputerized tomography analysis, which also showed that trabecular volume, thickness and number were significantly lower in Mg-deficient rats. Increased bone resorption was suggested by an increase in osteoclast number over time compared with controls as well as surface of bone covered by osteoclasts and eroded surface, but there was no difference in osteoblast numbers. The increased bone resorption may be due to an increase in TNF-alpha because immunocytochemical localization of TNF-alpha in osteoclasts was 199% greater than in controls at 2 mo, 75% at 4 mo and 194% at 6 mo. The difference in TNF-alpha may be due to substance P, which was 250% greater than in controls in mononuclear cells at 2 mo and 266% at 4 mo. These data demonstrated that a Mg intake of 10% of NR in rats causes bone loss that may be secondary to the increased release of substance P and TNF-alpha.

Semi-quantitative analysis of cytokines in MM THR tissues and their relationship to metal particles.

Cytokines associated with osteolysis have been demonstrated in tissues surrounding failed metal-metal (MM) total hip replacements (THRs). The objective of the present study was to semi-quantify the amounts of inflammatory cytokines in tissues from 28 failed MM THRs, and determine their relationship with the quantity of metal particles. Paraffin sections were immunohistochemically stained with monoclonal antibodies: anti-IL-1-beta, anti-IL-6 and anti-TNF-alpha. Cytokines and metal particles were rated in 10 fields per tissue using standard light microscopy. Because of the use of light microscopy, only relatively large particles or agglomerations of particles were visible. Therefore, a polarized light and a semi-quantitative scheme based on the discoloration of cell cytoplasms induced by the presence of particles were used to evaluate the quantity of metal particles. Results showed an overall higher amount of IL-6 than IL-1beta while TNF-alpha remained at very low levels. For each patient, the average IL-1beta and IL-6 ratings decreased when the average particle rating increased, following a linear regression, with relatively high correlation factors (r=-0.69 for IL-1beta and r=-0.57 for IL-6). IL-1beta decreased about twice as fast as IL-6. TNF-alpha, remaining at very low levels, did not demonstrate any correlation with particle rating. When multiple tissues were available for the same patient, the correlation factors between the average cytokine and particle ratings were highly variable between samples, demonstrating the heterogeneity between the tissues from the same patient. At the cellular level, there was an even higher correlation between the quantity of metal particles and the production of IL-1beta and IL-6 (r=-0.99), while TNF-alpha did not demonstrate any correlation, remaining at very low levels. In conclusion, this study showed that tissues surrounding failed MM THRs with low to moderate quantities of metal particles can induce the production of potentially osteolytic cytokines. However, the overall number of cells producing these cytokines tended to be lower than that typically seen in tissues surrounding metal-polyethylene THRs.

Magnesium deficiency: effect on bone mineral density in the mouse appendicular skeleton.

BACKGROUND: Dietary magnesium (Mg) deficiency in the mouse perturbs bone and mineral homeostasis. The objective of the present study was to evaluate bone mineral density of the femur in control and Mg-deficient mice. METHODS: BALB/c mice aged 28 days at study initiation were maintained on a normal or Mg deficient (0.0002% Mg) diet, and at time points 0, 2, 4 or 6 weeks bones were harvested for bone mineral density analysis. Peripheral quantitative computed tomography (pQCT) was used to assess the trabecular metaphyseal compartment and the cortical midshaft. RESULTS: Although mean total bone density of the femoral midshaft in Mg deficient mice did not differ significantly from controls throughout the study, the trabecular bone compartment showed significantly decreased mineral content after 4 (p < 0.001) and 6 weeks (p < 0.001) of Mg depletion. CONCLUSIONS: This study demonstrates the profound effect of Mg depletion on the trabecular compartment of bone, which, with its greater surface area and turnover, was more responsive to Mg depletion than cortical bone in the appendicular skeleton of the mouse.