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PubMed1991

Early mineral deposition in calcifying tendon characterized by high voltage electron microscopy and three-dimensional graphic imaging.

Extracellular matrix organization and the spatial relationship between collagen fibrils, vesicular structures, and the first deposits of mineral in the calcifying leg tendon from the domestic turkey, Meleagris gallopavo, have been investigated by high voltage electron microscopy and three-dimensional computer graphic imaging of serial thick tissue sections. The work demonstrates that the tendon extracellular matrix is a complex assembly of somewhat flexible, highly aligned collagen fibrils with different diameters and occasionally opposite directionality. Smaller collagen fibrils appear to branch from larger fibrils or to aggregate to form those of greater size. While the matrices are dominated by fibrils, space exists between adjacent packed fibrils. The three-dimensional perspective indicates that approximately 60% of the total tendon volume is extrafibrillar over the regions examined. The first observable mineral in this tissue is extrafibrillar and appears to derive from vesicles. This view of three-dimensional matrix-mineral spatial relations supports earlier two-dimensional results that mineral is initially associated with membrane-invested vesicles and is deposited between collagen fibrils, but it is distinct in showing the mineral at different depths in the matrix rather than at a single depth as deduced from two-dimensional conventional electron microscopy. These results are important in the onset and development of tendon calcification in that they suggest, first, that collagen fibrils appear to be aligned three-dimensionally such that their hole zones are in contiguous arrangement. This situation may create channels or grooves within the collagen volume to accommodate extensive mineral deposition in association with the fibrils. Second, the results indicate that there are widely dispersed sites of vesicle-mediated mineralization in the tendon matrix, that the bulk of mineralization in this tissue is collagen-mediated, and that, while vesicles may possibly exert some local influence temporally on mineralization of neighboring collagen, vesicle- and collagen-mediated mineralization arise at spatially and structurally distinct sites by independent nucleation phenomena. Such concepts are fundamental in considerations of possible mechanisms of mineralization of tendon and potentially of other normally calcifying vertebrate tissues in general.

Animals↗
PubMed2002

Enhancing mineral content in plant food products.

Plant foods can serve as dietary sources of all essential minerals required by humans. Unfortunately, mineral concentrations are low in some plants, especially many staple food crops; thus, efforts are underway to increase the mineral content of these foods as a means to ensure adequate attainment of dietary minerals in all individuals. While these efforts have included classical breeding approaches in the past, it is clear that future progress can be made by utilizing the tools of biotechnology to effect directed changes in plant mineral status. Reviewed are the short- and long-distance mineral transport mechanisms responsible for the root acquisition and whole-plant partitioning of mineral ions in crop plants. This background is used to discuss different transgenic strategies with the potential to enhance mineral content in vegetative and/or reproductive tissues. Due to various constraints imposed by plant transport systems on whole-plant mineral movement, it is argued that modifications designed to increase the supply of minerals to edible organs should have the highest chance for success. Examples of previous efforts to manipulate plant mineral nutrition through the introduction of novel transgenes are presented to demonstrate the utility of these approaches.

Biotechnology↗
PubMed1999

Matrix GLA protein is a developmental regulator of chondrocyte mineralization and, when constitutively expressed, blocks endochondral and intramembranous ossification in the limb.

Matrix GLA protein (MGP), a gamma-carboxyglutamic acid (GLA)-rich, vitamin K-dependent and apatite-binding protein, is a regulator of hypertrophic cartilage mineralization during development. However, MGP is produced by both hypertrophic and immature chondrocytes, suggesting that MGP's role in mineralization is cell stage-dependent, and that MGP may have other roles in immature cells. It is also unclear whether MGP regulates the quantity of mineral or mineral nature and quality as well. To address these issues, we determined the effects of manipulations of MGP synthesis and expression in (a) immature and hypertrophic chondrocyte cultures and (b) the chick limb bud in vivo. The two chondrocyte cultures displayed comparable levels of MGP gene expression. Yet, treatment with warfarin, a gamma-carboxylase inhibitor and vitamin K antagonist, triggered mineralization in hypertrophic but not immature cultures. Warfarin effects on mineralization were highly selective, were accompanied by no appreciable changes in MGP expression, alkaline phosphatase activity, or cell number, and were counteracted by vitamin K cotreatment. Scanning electron microscopy, x-ray microanalysis, and Fourier-transform infrared spectroscopy revealed that mineral forming in control and warfarin-treated hypertrophic cell cultures was similar and represented stoichiometric apatite. Virally driven MGP overexpression in cultured chondrocytes greatly decreased mineralization. Surprisingly, MGP overexpression in the developing limb not only inhibited cartilage mineralization, but also delayed chondrocyte maturation and blocked endochondral ossification and formation of a diaphyseal intramembranous bone collar. The results show that MGP is a powerful but developmentally regulated inhibitor of cartilage mineralization, controls mineral quantity but not type, and appears to have a previously unsuspected role in regulating chondrocyte maturation and ossification processes.

1-Carboxyglutamic Acid↗
PubMed1990

Mineral nutrition of the sow: a review.

The sow varies greatly in her mineral requirements, which largely reflects the nutritional demands during different phases of her reproductive cycle. Minerals are required for conceptus product formation, mammary secretions, and growth and maintenance. Although the sow's mineral requirements are highest during late gestation and lactation, those that are supplied from the diet and tissue reserves both contribute in meeting these nutritional needs. The sow is fairly resilient to borderline mineral insults, but longevity in the herd may be compromised under these conditions. Specific mineral deficiencies vary, but the reproducing sow will deplete the pool of tissue minerals before litter size, congenital abnormalities of developing fetuses or milk mineral composition are affected. Inadequate mineral supplies may prolong the duration of parturition, increase the number of stillbirths and result in a higher occurrence of skeletal problems. Tissue mineral reserves can be increased in various tissues with higher dietary fortification, but subsequent mineral availability from various pools differ. Because feed intake during lactation is low, daily mineral intake during lactation often is below NRC (1988) requirements.

Animal Nutritional Physiological Phenomena↗
PubMed2006

Mineral bioavailability and metabolism determined by using stable isotope tracers.

Definitive data on mineral bioavailability in humans and animals can be obtained by using isotopic tracers. The use of stable isotope tracers to study important issues in mineral nutrition has expanded rapidly in the past two decades, particularly in human nutrition studies. Stable isotopes have a number of advantages over radioisotopes. There is no exposure to radiation with stable isotopes, and some minerals have no radioisotope that can be used satisfactorily as a tracer. Multiple stable isotopes of one mineral and isotopes of multiple minerals can be administered simultaneously or sequentially. The analytical methods of choice for stable isotopes are thermal ionization mass spectrometry and inductively coupled plasma mass spectrometry (ICPMS). Thermal ionization mass spectrometry offers the greatest precision and accuracy, but it is slower, more labor intensive, and more costly than ICPMS. Bioavailability data are critical to establishing reliable dietary mineral requirements and recommendations. Combined with a computer program for compartmental modeling, mineral kinetics can be studied, including mineral turnover, pool sizes, and transfer rates between compartments. Our laboratory conducts studies using stable isotopes of Zn, Cu, Fe, Ca, Mg, and Mo. We have studied the effect of the amount of dietary intake of minerals on bioavailability and use, pregnancy and aging, and interactions among minerals. The research resulted in establishing new dietary recommendations for Cu and Mo and developing compartmental models for these minerals. Although stable isotopes have been used more extensively to date in humans than in animals, the techniques applied to humans can be used to study a number of issues important to optimizing feeding strategies for animal production.

Animal Nutritional Physiological Phenomena↗
PubMed1983

[Clinical study on bone mineral mass in metabolic bone disorders--I-125 photon absorptiometry].

Changes in bone mineral mass were investigated by 125I-photon absorptiometry in 5 groups of patients; postmenopausal osteoporosis (n = 50), after partial or total gastrectomy (n = 54), thyroid disorders (n = 28), parathyroid disorders (n = 6) and healthy subjects (n = 214) as control. Radial bone mineral content (BMC) was measured at the distal 1/3 and 1/6 sites of the radius and the densitometric patterns were analized . The results obtained were as follows: Age variation in bone mineral mass and bone mineral change in pathological status were more sensitively detectable at the distal 1/6 site of the radius (predominantly trabecular) than the distal 1/3 site of the radius (predominantly cortical). In osteoporosis there was a diminution in radial 1/6 BMC and a significant decrease of radial trabecular bone index ( RTBI ) estimated from densitometric patterns, as compared with age-matched control. Sequential changes of bone mineral mass were evaluated during the treatment. There was no significant change in bone mineral mass during the first 3 months of 1 alpha-OH-D3 administration. Bone mineral mass then decreased about 20% in radial 1/6 BMC in the course of the treatment. An increase in bone mineral mass at the distal 1/6 site of the radius was revealed at 1 year treatment with 1 alpha-OH-D3. But at the distal 1/3 site of the radius only a little change in bone mineral mass was recognized during the treatment. Total gastrectomy caused a significant deficit of radial BMC as compared to partial gastrectomy. 1 alpha-OH-D3 was also effective on bone mineral loss after gastrectomy. Loss of radial 1/6 BMC occurred in hyperthyroidism (high turn over osteoporosis) but not in hypothyroidism. Striking bone mineral loss was observed in hyperparathyroidism, which recovered after surgical treatment.

Adolescent↗
PubMed1996

Cell-mediated mineralization in culture at low temperature associated with subtle thermogenic response.

In both the growth plate and in marrow stromal cell cultures cell-mediated mineralization is preceded by characteristics of anaerobic and low efficiency energy metabolism. Reagents that increase mineralization like malonate and dexamethasone (DEX) also increase the mitochondrial membrane potential (MtMP) especially 1 week after DEX stimulation. Contrarily, levamisole, which decreases mineralization, also decreases MtMP. Modulation of MtMP and energy metabolism could be linked to regulation of mineralization by the uncoupling of oxidative phosphorylation. This uncoupling should be associated with thermogenesis in cells that induce mineralization. We examined whether cold temperature affects mineralization, and whether cellular thermogenesis takes place at cold temperature in parallel to changes in MtMP. Osteoprogenitor cells (OPC) induced, in DEX stimulated rat marrow stroma, higher mineralization at 33 degrees C than at 37 degrees C. Increased mineralization by cold temperature required long incubation since incubation in the cold during short intervals, 3-4 days, did not increase mineralization relative to (37 degrees C) controls. Marrow stromal cells in the presence of valinomycin responded to incubation at 33 degrees C by retaining all the vital dye after 4 h, unlike the cells at 37 degrees C; however, after 24 h the level of dye retention at 33 degrees C was the same as at 37 degrees C. The delayed response of the temperature-dependent (> 37 degrees C) K+ ionophor to incubation in the cold indicated that certain cells may respond to low temperature by local intracellular heating, and by heat conduction to the plasma membrane. DEX-stimulated stromal cells, unlike unstimulated cells, showed increased mitochondrial rhodamine 123 retention in the presence of valinomycin after 24 h in the cold, which corresponds to day 4 of OPC induction. This is consistent with the concept that valinomycin-induced cell damage is mediated by (cold-induced) local heating. The mechanism of this cell damage should selectively prefer nonthermogenic (rhodamine retaining) over thermogenic (rhodamine leaking) cells such as OPC. At cold temperature DEX-stimulated stromal cells showed the best anti-OPC selection under exposure to valinomycine between days 3-7, concurrent with the period of rhodamine leakage from the mitochondria. These results indicate that thermogenesis is enhanced during the period of low MtMP in mineralizing cells, and prolonged exposure to cold increases mineralization also due to induction of subtle thermogenesis.

Animals↗
PubMed2000

Calcium and phosphate supplementation promotes bone cell mineralization: implications for hydroxyapatite (HA)-enhanced bone formation.

Organic phosphate, in particular beta-glycerophosphate (beta-GP), has been used to induce mineralization in cell culture systems. It serves as a source of inorganic phosphate when hydrolyzed by alkaline phosphatase. This study examined the effect of supplemental calcium and phosphate as well as the influence of various metabolic inhibitors on mineralization in a rat osteoblast-like cell-culture system. Mineralization was induced by supplementation of 1.8 mM of Ca(+2) and 5 mM of beta-GP or Pi. Mineral deposits associated with in vitro mineralization were revealed under SEM and TEM. Levamisole (10-100 microM) inhibited alkaline phosphatase activity and effectively reduced mineral formation. Actinomycin (500 ng/mL) and cycloheximide (50 microg/mL) also reduced mineral depositions by blocking RNA synthesis and protein synthesis, respectively. Levamisole and beta-GP did not appear to influence DNA synthesis. Spontaneous precipitation of calcium phosphate mineral was not detected in the culture medium with calcium and phosphate supplements in the absence of cell culture. The findings suggest that an elevated concentration of calcium and phosphate is crucial for in vitro mineralization. Furthermore, the mineralization process is associated with biologic events rather than with a spontaneous precipitation of calcium phosphate mineral. In view of the degradation potential of hydroxyapatite (HA)-coated implants, these results may be a viable indication that HA enhances bone formation through a similar mechanism.

Animals↗
PubMed2005

The effect of ligand type and density on osteoblast adhesion, proliferation, and matrix mineralization.

Polystyrene surfaces grafted with a nonfouling interfacial interpenetrating polymer network (IPN) of poly(acrylamide-co-ethylene glycol/acrylic acid) [p(AAm-co-EG/AAc)] were modified with several peptide ligands adapted from bone sialoprotein (BSP). IPNs were modified with both single ligands and ligand blends to study the correlation between a simple metric, ligand-receptor adhesion strength, and the extent of matrix mineralization for osteoblast like cells (rat calvarial osteoblasts). The ligands studied included RGD cell-binding [CGGNGEPRGDTYRAY (l-RGD), CGGEPRGDTYRA (s2-RGD), CGPRGDTYG (lc-RGD), cyclic(CGPRGDTYG) (c-RGD), and CGGPRGDT (s-RGD)], heparin binding (CGGFHRRIKA), and collagen binding (CGGDGEAG) peptides, with the appropriate controls. Adhesion strength scaled with ligand density (1-20 pmol/cm(2)) and was dependent on ligand type with the following trend: l-RGD > s2-RGD approximately c-RGD >> s-RGD approximately lc-RGD >>> FHRRIKA approximately DGEA. Independent of ligand density, % matrix mineralization varied with ligand type resulting in the following trend: lc-RGD > s2-RGD > l-RGD approximately c-RGD >> s-RGD >>> FHRRIKA. The Tyr (Y) residue immediately following the RGD cell-binding domain proved to be critical for stable cell proliferation and mineralization, since removal of this residue resulted in erratic cell attachment and mineralization behavior. The minimum BSP sequence necessary for strong adhesion and extensive mineralization was CGGEPRGDTYRA; the minimal sequence suitable for extensive mineralization but lacking strong adhesion was CGPRGDTYG. The cyclic peptide (c-RGD) had much greater adhesion strength compared to its linear counterpart (lc-RGD). The calculated characteristic adhesion strength (F(70)) obtained using a centrifuge adhesion assay proved to be a poor metric for predicting % mineralized area; however, in general, surfaces possessing a F(70) > 100g promoted extensive matrix mineralization. Percent mineralization and number of mineralized nodules scaled with number of cells seeded suggesting a critical dependence on the initial number of osteoprogenitors in culture. This study demonstrates matrix mineralization dependence on ligand type, ligand density, and adhesion strength. The high-throughput character of these surfaces allowed efficient investigation of multiple ligands at multiple densities providing an excellent tool for studying ligand-receptor interactions under normal cell culture conditions with serum present.

Animals↗
PubMed1992

Factors influencing synthesis and mineralization of bone matrix from fetal bovine bone cells grown in vitro.

This study of the in vitro synthesis and mineralization of bovine bone demonstrates that sheets of mineralized matrix can be produced consistently within 18-24 days of cell isolation. Mineralization surpasses that achieved by other systems with other species: The deposition of mineral extends beyond nodules to form branching trabeculae and then solid wafers of bone. Comparison of the fetal age of the bone source, enzyme digestion methods, seeding density, culture surface, nutritive media, and concentration of fetal calf serum and other additives, including insulin and ascorbic acid, has yielded a set of optimal culture conditions. In the presence of ascorbic acid and beta-glycerol phosphate, insulin has a dose-dependent effect on the morphology of the mineralized bone matrix produced. Quantitative analysis shows that in these cultures calcium accumulates most rapidly between days 6 and 10 after the introduction of mineralization medium but that mineral accretion continues throughout 14-16 days of culture. Alkaline phosphatase levels rise up to 200-fold, concomitant with a rapid increase in the number of cells per culture during the early mineralization phases; both fall as mineralization proceeds. This system has been used to study the induction of mRNA of type I collagen, alkaline phosphatase, and several noncollagenous bone proteins during the course of mineralization. Because of the degree of mineralization achieved with this system, it has many potential applications.

Alkaline Phosphatase↗
PubMed1995

The phosphatidylinositol-glycolipid anchor on alkaline phosphatase facilitates mineralization initiation in vitro.

Alkaline phosphatase (AP) is required for the proper mineralization of cartilage and bone. The enzyme is localized to the outer surface of cells through a phosphatidylinositol-glycolipid anchor, which is covalently attached to the carboxyl terminus of the protein. In calcifying cartilage, AP-rich matrix vesicles (MVs) are released into the matrix from chondrocytes, and apatite formation is initiated within and around these particles. In this paper we examine the role of the AP glycolipid anchor using an in vitro mineralization assay system. AP was purified to homogeneity, and the purified enzyme was used to drive mineral formation in vitro with and without the anchor. Mineral formation was initiated through phosphate release from beta-glycerol phosphate (beta-GP). The amount of PO4(-3) released was similar whether the anchor was present or absent. However, SEM and X-ray microanalysis revealed that the mineral produced by anchored AP was indistinguishable from that produced by MVs and that both of those minerals were more apatite-like than mineral formed by soluble AP or through spontaneous precipitation. Taken together, the data suggest that in addition to providing PO4(-3) to drive mineralization, AP influences the nature of the mineral formed. Further, AP containing its glycolipid anchor produces mineral comparable with that formed by tissue-derived MVs. Thus, in the absence of extracellular matrix, MV mineralization in vitro can be emulated by glycolipid-anchor containing AP.

Alkaline Phosphatase↗
PubMed2002

Mineral-related proteins of sea urchin teeth: Lytechinus variegatus.

Sea urchins have a set of five continuously growing teeth, each of which has a very complex structure. The mineral phase is calcite of varying Mg content, depending on the location within a tooth. The calcium carbonate is present in amorphous, plate-like and rod-like forms. It has been hypothesized that the mineral deposition is a matrix-mediated process, similar to that in vertebrate bone and tooth, wherein certain macromolecules within the organic matrix of the mineralized tissue play an important role in nucleating and controlling the growth habit of the mineral crystals. It has also been hypothesized that the mineral-related macromolecules involved in urchin teeth might bear a direct evolutionary relationship to those of the vertebrate tooth. These hypotheses are explored here by examining the pattern and nature of the mineral distribution, using microCT of intact teeth, and the nature of the mineral-related matrix proteins. The mineral-related proteins were extracted and fractionated by anion exchange chromatography. The relationship of certain fractions to vertebrate matrix proteins was established by immunoblots using antibodies to vertebrate tooth proteins. The antibodies were then used to localize the proteins within the teeth, by immunocytochemistry and histology with specific staining. The microCT data on mineral density has been correlated with the patterns of cellular migration and mineral deposition within the tooth as it grows. It appears that the mineralization within the different tooth compartments might take place under the influence of different matrix proteins. Further studies are in progress to more completely describe the vertebrate-invertebrate immunologically cross-reactive proteins of the urchin teeth.

Animals↗
PubMedDate not supplied

Mineralization of 2,4-dichlorophenoxyacetic acid (2,4-D) in soil inoculated with Pseudomonas cepacia DBO1(pRO101), Alcaligenes eutrophus AEO106(pRO101) and Alcaligenes eutrophus JMP134(pJP4): effects of inoculation level and substrate concentration.

Mineralization of 2,4-dichlorophenoxyacetic acid (2,4-D) by two Alcaligenes eutrophus strains and one Pseudomonas cepacia strain containing the 2,4-D degrading plasmids pJP4 or pRO101 (= pJP4::Tn1721) was tested in 50 g (wet wt) samples of non-sterile soil. Mineralization was measured as 14C-CO2 evolved during degradation of uniformly-ring-labelled 14C-2,4-D. When the strains were inoculated to a level of approximately 10(8) CFU/g soil, between 20 and 45% of the added 2,4-D (0.05 ppm, 10 ppm or 500 ppm) was mineralized within 72 h. Mineralization of 0.05 ppm and 10 ppm 2,4-D by the two A. eutrophus strains was identical and rapid whereas mineralization by P. cepacia DBO1(pRO101) occurred more slowly. In contrast, mineralization of 500 ppm 2,4-D by the two A. eutrophus strains was very slow whereas mineralization by P. cepacia DBO1 was more rapid. Comparison of 2,4-D mineralization at different levels of inoculation with P. cepacia DBO1(pRO101) (6 x 10(4), 6 x 10(6) and 1 x 10(8) CFU/g soil) revealed that the maximum mineralization rate was reached earlier with the high inoculation levels than with the low level. The kinetics of mineralization were evaluated by nonlinear regression analysis using five different models. The linear or the logarithmic form of a three-half-order model were found to be the most appropriate models for describing 2,4-D mineralization in soil. In the cases in which the logarithmic form of the three-half-order model was the most appropriate model we found, in accordance with the assumptions of the model, a significant growth of the inoculated strains.

2,4-Dichlorophenoxyacetic Acid↗
PubMed1979

The mineralization processes in teleost fish scales.

In Teleost fish scales, growth and mineralization are continuous. Different mineralization processes can be distinguished. The external layer of the scale is the first to be mineralized and may be classified as the initial calcifying structure of the scale. The initial calcification loci are matrix vesicles of cellular origin always observed during the formation of this layer. This mineralization process takes place progressively, closely following the elaboration of the organic matrix in the scale periphery. The outer limiting and internal layers of the scale are developed after the external layer has been formed. A mineral substance is deposited without the mediation of matrix vesicles, but in contact with the previously mineralized external layer. This type of mineralization is called subsequential. However, the mineralization of the outer limiting layer closely follows the secretion of a collagen-free organic matrix and is thus different from the mineralization of the internal layer in which the calcification front remains remote from the collagen matrix surface and corresponds to a delayed mineralization process. The isolated calcifications (Mandl's corpuscles) which develop in the unmineralized laminae of the internal layer are mineralized in the absence of matrix vesicles and without making contact with a pre-existing calcified tissue, probably by a heterogeneous nucleation of the collagen fibrils.

Animals↗
PubMed2003

Fourier transform infrared imaging spectroscopy (FT-IRIS) of mineralization in bisphosphonate-treated oim/oim mice.

Fourier transform infrared microscopy (FT-IRM) and imaging spectroscopy (FT-IRIS) are increasingly used to analyze the molecular components of mineralized tissues. A primary advantage of these techniques is the capability to simultaneously image the quantity and quality of multiple components in histological sections at 7 microm spatial resolution. In the current study, FT-IRM and FT-IRIS were used to characterize bone mineralization in a mouse model of osteogenesis imperfecta (OI) after treatment with the bisphosphonate alendronate (ALN). This application is currently relevant since recent studies have demonstrated great promise for the treatment of children with OI with bisphosphonates, but have not identified bisphosphonate-associated bone quality changes. Growing oim/oim mice, a model of moderate-to-severe OI, were treated with ALN (73 microg ALN/kg/day for 4 weeks followed by 26 mg/kg/day for 4 weeks) or saline from 6 to 14 weeks of age, and mineralization was evaluated in femoral cortical and metaphyseal bone. Infrared vibrations of the mineral (a carbonated apatite) and the matrix phases were monitored. The relative amounts of mineral and matrix present (min:matrix), the relative amount of carbonate present in the mineral (carb:min), and the crystallinity of the mineral phase were calculated. In untreated oim/oim bone, the min:matrix was greater and the crystallinity was reduced (indicative of less mature mineral) in the primary versus the secondary spongiosa, most likely due to the presence of calcified cartilage. With ALN treatment, the oim/oim mm:matrix increased in the secondary spongiosa, but the mineral crystallinity was not changed. In the cortical bone, no changes were evident with ALN treatment. These data demonstrate that in this mouse model, ALN treatment results in increased metaphyseal bone mineralization, but does not improve mineral maturity.

Alendronate↗
PubMed1993

Stress relaxation in native and EDTA-treated bone as a function of mineral content.

The relaxation Young's modulus, epsilon(t), of bovine femoral bone was measured as a function of mineral content. Five different specimens with different mineral contents were prepared by the EDTA treatment. The relaxation curves for specimens of different mineral contents were superimposable upon one another by shifting along the log t axis as well as the log epsilon(t) axis. A well-specified master curve of stress relaxation was obtained from a set of relaxation curves for samples with different mineral contents. The mineral content dependence of vertical shift factors, along the log epsilon(t) axis, accorded well with the mineral content dependence of Young's modulus itself, the accordance indicating the plausibility of the vertical shifting procedure. The stress relaxation in bone has been reported to be related to the viscoelastic properties of the collagen matrix. It is considered that the reinforcement of the matrix around the mineral, by the mineral as fillers, increases the average modulus of the matrix and lengthens the characteristic time of the relaxation. This consideration explains the existence of a shift factor along the logt axis and its mineral content dependence. As the structures of bone specimens treated by EDTA are expected to be different from that of normal bone, the conclusion drawn from this experiment cannot be immediately applied to normal bone. It can be, however, concluded that the size of the mineral and also the interface area between mineral and collagen matrix play an important role in the viscoelastic properties of normal bone and the bone specimen treated by EDTA.

Animals↗
PubMed1996

Strontium increases vertebral bone volume in rats at a low dose that does not induce detectable mineralization defect.

Low doses of strontium and fluoride were shown to increase bone formation and trabecular bone density in rodents. To assess whether strontium or fluoride affect the quality of the mineral at doses known to increase bone density, we have determined the effects of low doses of strontium and fluoride on bone formation and bone mineral characteristics in rats. Adult rats were given strontium alone (0.20%), fluoride alone (1 mg/kg per day), or the combined treatment for 8 weeks. Strontium levels in serum and femur were similar in groups treated with strontium alone or in combination, being about 5% of calcium levels. Biochemical and neutron activation analyses in femur showed that calcium and magnesium contents did not differ in the four group of rats, suggesting that strontium was incorporated in the apatite lattice of the bone minerals in the strontium-treated rats. The mineralized bone volume was significantly increased by 17% in the strontium-treated group, by 20% in the fluoride-treated group, and by 19% in rats given with the combined treatment. This was associated with increased osteoid surface, osteoblast surface, and double tetracycline labeled surfaces in the strontium-treated and fluoride-treated groups, showing that the number of bone forming sites was increased. However, the mineral apposition rate, the osteoid thickness, and the mineralization lag time were similar in controls and treated groups, reflecting the lack of deleterious effects of low doses of strontium and fluoride on bone mineralization. The density fractionation analysis measured in the femur also showed that neither strontium, nor fluoride at the low doses used, significantly altered the mineralization profile. The results indicate that treatment with low doses of strontium or fluoride increase the number of bone forming sites and vertebral bone volume in rats, but does not have detectable adverse effects on the mineral profile, bone mineral chemistry or bone matrix mineralization.

Analysis of Variance↗
PubMed2004

Bone mineralization density and femoral neck fragility.

The traditional view of osteoporotic fractures is that they result from a reduction in bone mass combined with alterations in the micro-architecture. Apart from the effects of bone remodeling, the material properties of the remaining bone are thought to be unaffected. To test this, we compared the degree of matrix mineralization in femoral neck biopsies taken from cases of intracapsular hip fracture with age- and sex-matched postmortem controls. Whole femoral neck biopsies from seven female hip fracture cases (72-90 years) and nine controls (68-94 years) were embedded in methylmethacrylate, and sections stained with Solochrome Cyanin R for analysis of osteoid. The blocks were then diamond micro-milled, carbon coated, and analyzed for the degree of matrix mineralization using halogenated dimethacrylate standards for quantitative backscattered electron (qBSE) imaging (20 kV, entire block face, sampling interval 5 microm). The BSE gray scale was adjusted such that 0 corresponds to an electron backscattering coefficient of 0.1159 (approximately 1.70 g/ml) and 255-0.1519 (approximately 2.18 g/ml). Remodeling and mineralization data were analyzed for both the whole biopsy face and on a regional (anterior; inferior, posterior, or superior) basis. Over the whole biopsy, the level of mineralization was lower in the cases than the postmortem controls (-2.8%, P < 0.05). In both cases and controls, cortical mineralization was higher in the inferior (compressive) region compared with superior (tensile) region (P < 0.05). Mineralization was lower in all regions of the cases (inferior: -3.3%; posterior: -3.1%; anterior: -2.7%; superior: -1.6%) compared to the controls. Mineralization density in cancellous bone was not regionally dependent but was lower in the fracture cases (-3.5%; P = 0.001). Although there were weak relationships between osteoid formation (%O.Ar/B.Ar) and the mean level of mineralization in both cortical (P = 0.068) and cancellous (P < 0.01) bone, adjustment for this did not markedly affect the case-control differences. In conclusion, this study has shown that in cases of intracapsular hip fracture, matrix mineralization is reduced in the femoral neck. Unexpectedly, in view of the likely role of mild to moderate vitamin D deficiency osteopathy in hip fracture, this decreased mineralization was independent of osteoid indices and therefore potentially independent of bone age. This raises the possibility that alterations in the bone matrix such as excessive glycation or changes in the composition of the collagen fibrils affect its mineralization in hip fracture cases.

Aged↗