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A L Boskey

Publications and source records attributed to A L Boskey.

At least 73 records · Page 4Linked to original sources
PubMed1996

Persistence of complexed acidic phospholipids in rapidly mineralizing tissues is due to affinity for mineral and resistance to hydrolytic attack: in vitro data.

Acidic phospholipids, complexed with calcium and inorganic phosphate, are components of extracellular matrix vesicles. Both the complexed acidic phospholipids and matrix vesicles have previously been shown to serve as hydroxyapatite (HA) nucleators in solution and when implanted in a muscle pouch. The present study supplies evidence that complexed acidic phospholipids can persist in mineralizing tissues both because of their affinity for HA and because of their resistance to hydrolysis by phospholipase A2. Calcium-phosphatidylserine-phosphate complex (CPLX-PS) synthesized with 14C-labeled phosphatidylserine (PS) was used to measure CPLX-PS affinity for HA using a Langmuir adsorption isotherm model. The affinity was shown to be higher and more specific than that of PS itself (K = 8.66 ml/mumol; N, the number of binding sites = 20.4 mumol/m2 as compared with previously reported values for PS of K = 3.33 ml/mumol, and N = 4.87 mumol/m2). Incorporated into synthetic liposomes and incubated in a calcium phosphate solution in which mineralization is induced by an ionophore, CPLX-PS showed behavior distinct from free PS. As previously reported, PS in these liposomes totally blocked HA formation. On the other hand, CPLX-PS in similar concentrations had a varied response, having no effect, slightly inhibiting, or actually promoting HA formation. CPLX-PS was also shown to be a poorer substrate for phospholipase A2 than PS, with Km = 4.63 mM for CPLX-PS and Km = 0.27 mM for PS; and Vmax = 0.029 ml/minute for CPLX-PS and Vmax = 0.066 ml/minute for PS. These data explain how complexed acidic phospholipids may persist in the growth plate and facilitate initial mineral deposition.

Calcification, Physiologic↗
PubMed1996

Viable cells are a requirement for in vitro cartilage calcification.

It is a common belief that chondrocyte death must precede calcification in the growth plate. To challenge this dogma, cell devitalization was induced in an in vitro model that mimics in situ cartilage calcification. Chick limb-bud mesenchymal cells, plated in micromass culture, differentiate to form a cartilaginous matrix which mineralizes in the presence of inorganic or organic phosphate. The mineral formed resembles physiologic mineral in crystal size, composition, and distribution. Killing cells by water lysis, ethanol fixation, freeze-thawing, trypsinization, or impairing their function by oligomycin treatment prior to the time at which mineralization commenced, prevented mineral deposition. In contrast, devitalizing cells by any of these techniques after mineralization commenced resulted in dystrophic calcification (excessive, randomly distributed mineral of larger than physiologic crystal size). Based on analyses of 45Ca uptake, FT-IR microscopy, X-ray diffraction, and transmission electron microscopy, it is concluded that the presence of viable cells is obligatory for physiologic cartilage calcification in the differentiating chick limb-bud mesenchymal cell culture system.

Animals↗
PubMed1996

Pyridoxine deficiency affects biomechanical properties of chick tibial bone.

The mechanical integrity of bone is dependent on the bone matrix, which is believed to account for the plastic deformation of the tissue, and the mineral, which is believed to account for the elastic deformation. The validity of this model is shown in this study based on analysis of the bones of vitamin B6-deficient and vitamin B6-replete chick bones. In this model, when B6-deficient and control animals are compared, vitamin B6 deficiency has no effect on the mineral content or composition of cortical bone as measured by ash weight (63 +/- 6 vs. 58 +/- 3); mineral to matrix ratio of the FTIR spectra (4.2 +/- 0.6 vs. 4.5 +/- 0.2), line-broadening analyses of the X-ray diffraction 002 peak (beta 002 = 0.50 +/- 0.1 vs. 0.49 +/- 0.01), or other features of the infrared spectra. In contrast, collagen was significantly more extractable from vitamin B6-deficient chick bones (20 +/- 2% of total hydroxyproline extracted vs. 10 +/- 3% p < or = 0.001). The B6-deficient bones also contained an increased amount of the reducible cross-links DHLNL, dehydro-dihydroxylysinonorleucine, (1.03 +/- 0.07 vs. 0.84 +/- 0.13 p < or = 0.001); and a nonsignificant increase in HLNL, dehydro-hydroxylysinonorleucine, (0.51 +/- 0.03 vs. 0.43 +/- 0.03, p < or = 0.10). There were no significant changes in bone length, bone diameter, or area moment of inertia. In four-point bending, no significant changes in elastic modulus, stiffness, offset yield deflection, or fracture deflection were detected. However, fracture load in the B6-deficient animals was decreased from 203 +/- 35 MPa to 151 +/- 23 MPa, p < or = 0.01, and offset yield load was decreased from 165 +/- 9 MPa to 125 +/- 14 MPa, p < or = 0.05. Since earlier histomorphometric studies had demonstrated that the B6-deficient bones were osteopenic, these data suggest that although proper cortical bone mineralization occurred, the alterations of the collagen resulted in changes to bone mechanical performance.

Animals↗
PubMed1996

Fourier transform infrared microspectroscopic analysis identifies alterations in mineral properties in bones from mice transgenic for type X collagen.

Type X collagen has been implicated in the morphogenetic events of endochondral ossification (EO), including the calcification of hypertrophic cartilage and trabeculae prior to their replacement by bone and marrow. Recently, transgenic mice, which expressed a truncated collagen X protein, were reported to exhibit morphologic alterations in all tissues arising through EO. Specifically, the growth plates were compressed within the zone of cartilage hypertrophy, and the number and size of calcified trabeculae were reduced. The condition in the mouse is comparable to Schmid metaphyseal chondrodysplasia in humans for which, to date, 20 defined type X collagen mutations have been reported. The transgenic mouse showed no alterations in mineralization by conventional histology, however, it did show a decrease in newly formed bony trabeculae, and a thinning of periosteal bones. Fourier transform infrared (FTIR) spectroscopy has previously been shown to provide quantitative and qualitative information about the relative amount of mineral and carbonate present, mineral composition, and crystal perfection. To determine whether the expression of abnormal collagen X molecules had an effect on mineral properties, the "quality" of mineral crystals was analyzed in thin sections of tibia from day 17 and day 25 genotypically negative (normal) and positive (mutant) mice from several independent transgenic mouse lines showing varying degrees of the mutant phenotype, by means of Fourier transform infrared microscopic analysis (FTIRM). The results indicate definite differences between normal and transgenic mice calcified cartilage mineral, both in the amount present and the "quality" of the crystals. Calcified cartilage mineral from transgenic mice exhibited less crystallinity and higher acidic phosphate content than the corresponding mineral from normal specimens.

Animals↗
PubMed1996

Polarized FT-IR microscopy of calcified turkey leg tendon.

Polarized Fourier transform infrared microscopy (FT-IRM) was used to assess the orientation of mineral and matrix components of the normally calcified turkey leg tendon. Two groups of tendon, < 16 weeks of age (young) and > 60 weeks of age (old), were analyzed. Linear sequences from calcified, non-calcified, and transitional regions of the tendons were examined. Spectra collected in the "parallel polarization" mode were acquired with the electric vector of the infrared radiation parallel to the collagen fiber axis whereas spectra collected in the "perpendicular polarization" mode were acquired with the electric vector of the infrared radiation perpendicular to this axis. The v2 carbonate (850-890 cm-1) and v1, v3 phosphate (900-1180 cm-1) contours of the tendon mineral as well as the collagen amide I, II, and III bands of the extracellular matrix all displayed marked dichroism. The CO3(2-) ions substituted for PO4(3-) (878 cm-1, type B substitution) in the tendon mineral displayed parallel dichroism while the CO3(2-) ions substituted for OH (871 cm-1, type A substitution) in the tendon mineral displayed perpendicular dichroism. These orientational effects for both sites of carbonate substitution were greater in the older animals. The polarization properties of the v1, v3 phosphate contour were analyzed by use of an empirical anisotropy parameter (A), the value of which monitors the degree of orientation. This index significantly increased in the older animals indicating that aging produces a more highly oriented mineral. The amide I, II, and III contours of the collagen extracellular matrix also exhibited marked dichroism. The amide I component exhibits perpendicular dichroism while the amide II and III components exhibit parallel dichroism. The current study demonstrates the ability of polarized FT-IRM to assess the orientation of the mineral and matrix components of calcified tissue at the microscopic level.

Amides↗
PubMed1996

Mineral changes in a mouse model of osteogenesis imperfecta detected by Fourier transform infrared microscopy.

Osteogenesis imperfecta (OI) is a heritable disease characterized by skeletal deformities and brittle bones. In the current study, the nature of the mineral in long bones of a mouse model of OI (oim/oim, a mutant which produces an alpha 1(I) collagen homotrimer) was examined by Fourier transform infrared microscopy. The mineral:matrix ratio of oim/oim cortical bone was greater than that of the heterozygous oim/+ and of the normal +/+ bones, probably as a result of reduced collagen content. The molecular environments of the apatitic phosphates differed among the oim/oim and the oim/+ and the +/+ bones. This was attributable to several factors, including dissimilar mineral-matrix interactions and differences in the chemical composition of the mineral. It was concluded from these data that the defective collagen matrix leads to abnormal mineral formation at the molecular level and thus results in tissues with reduced mechanical properties.

Animals↗
PubMed1996

Matrix proteins and mineralization: an overview.

There is a wealth of information on the mineral and matrix components in bones and teeth, in the exoskeletons of invertebrates, and in dystrophic calcific deposits. This information includes detailed characterization of their physical and chemical composition and details on the gene localization and regulation of gene expression for the major and minor protein constituents. The reason that mineral deposition occurs in some tissues and not in others remains unclear. In this review, studies in solution, cell culture studies, and investigations in mutant animals will be surveyed to indicate which matrix proteins may affect mineralization. Most of the molecules that appear to be involved in initiation and regulation of biological mineral formation are anionic; they have structural features that facilitate interaction with mineral, cells, and other matrix molecules, and they can have more than one function. Despite extensive data it is not yet clear which of these molecules is absolutely essential for physiologic calcification in each of the calcified tissues.

Animals↗
PubMed1996

The role of type X collagen in endochondral ossification as deduced by Fourier transform infrared microscopy analysis.

Type X collagen has been implicated in the morphogenetic events of endochondral ossification (EO), including the calcification of hypertrophic cartilage and trabeculae prior to their replacement by bone and marrow. Recently, transgenic mice which expressed a truncated collagen X protein were reported to exhibit morphologic alterations in all tissues arising through EO. Fourier Transform InfraRed (FTIR) spectroscopy has previously been shown to provide quantitative and qualitative information about the relative amount of mineral and carbonate present, mineral composition, and crystal perfection. To determine the role of collagen X in mineralization, the "quality" of mineral crystals was analyzed in thin sections of calcified cartilage from tibia obtained from several independent transgenic mouse lines showing varying degrees of the mutant phenotype and mice without type X collagen expression, by means of Fourier Transform InfraRed microscopy (FTIRM). In the present paper, the term "mineral quality" is employed to describe crystallinity/crystal maturation, and acid phosphate content. The results indicate significant differences between normal and transgenic mice bone mineral, both in the amount present and the "quality" of the crystals. In contrast, the analysis of the mineral in mice without type X collagen expression was not different from their age/sex-matched controls.

Animals↗
PubMed1995

Osteopontin and related phosphorylated sialoproteins: effects on mineralization.

Osteopontin is one of a family of phosphorylated sialoproteins found in the extracellular matrices of mineralized connective tissues. Solution studies from a variety of laboratories have shown that while some of these proteins (e.g., osteopontin, dentin sialoprotein) inhibit calcium phosphate production, others, such as bone sialoprotein, can act as a nucleator of calcium phosphate formation. The differences in the behaviors of these proteins can be related to their interactions with mineral crystal nuclei and crystal growth sites. The specificity of these interactions, in turn, can be related to differences in the primary structures of the sialoproteins and to the extent to which they are phosphorylated. In vitro systems for the study of osteopontins and osteopontin-related protein effects on mineral deposition are described, along with mechanisms explaining the contrasting abilities of the polyglutamate-containing bone sialoprotein to act as a nucleator, while the polyaspartate-containing osteopontin and dentin sialoproteins inhibit calcium phosphate formation and growth.

Amino Acid Sequence↗
PubMed1995

Diminished material properties and altered bone structure in rat femora during pregnancy.

Pregnancy and lactation are known to cause structural and mechanical changes in bone, but the effects of pregnancy alone have not been evaluated thoroughly. This study used radiographic measurements, torsion testing, mineral analyses, and histological evaluation to determine whether there are changes in bone material and geometric properties during pregnancy in the growing rat, as implied by earlier biochemical and histological studies. The bones of pregnant 9 to 12-week-old rats and controls that were not pregnant and were matched by age (but not weight) were evaluated at times corresponding to 5, 10, 15, and 20 days of the 23-day gestation period to address the following questions: (a) How is the growth of whole bone affected by pregnancy in the growing rat (as determined by radiographic analyses)? (b) How are the mechanical properties (structural and material) of whole bone affected by pregnancy (as assessed by torsion testing)? (c) Are there changes in the characteristics of bone mineral during pregnancy (as determined by measurement of mineral content and x-ray diffraction analyses)? and (d) Are there detectable morphological or ultrastructural differences between the bones of pregnant and control rats (as assessed by analyses based on histology and back-scattered electron imaging)? The presence of statistically significant differences in this study was determined initially on the basis of a two-factor analysis of variance. In general, significant differences were noted only at late gestation (day 20), when the bones were longer and had a greater outer radius and cortical thickness; this indicates that more growth occurred during pregnancy.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1995

Effect of abnormal mineralization on the mechanical behavior of X-linked hypophosphatemic mice femora.

The Hyp mouse is an established animal model of X-linked hypophosphatemia, one of the most common genetic forms of metabolic bone disease in humans. This study describes the first determination of whole bone mechanical behavior in the heterozygous male and female Hyp mouse. Femora from 12-week-old mice were tested in torsion. The contribution of structural and material properties to mechanical behavior was determined by geometrical evaluation prior to testing and by analysis of the diaphyseal mineral after testing. The male and female Hyp femora were found to undergo significantly more angular deformation at failure than the same sex normal femora (82.49 +/- 24.37 vs. 22.63 +/- 8.02 rad/m [corrected] for the females and 128.90 +/- 37.05 vs. 22.79 +/- 7.24 rad/m [corrected] for the males) and to have a significantly lower structural stiffness (0.373 +/- 0.130 x 10(-3) vs. 1.33 +/- 0.380 x 10(-3) [corrected] [N-m/(rad/m)] for the females and 0.167 +/- 0.104 x 10(-3) vs. 1.60 +/- 0.502 x 10(-3) [corrected] [N-m/(rad/m)] for the males). The male Hyp femora had a significantly lower failure torque than male normal femora (1.58 +/- 0.62 x 10(-2) vs. 3.44 +/- 1.57 x 10(-2) N-m). Because the polar movement of inertia, a geometrical property that affects torsional behavior, was not significantly different between the Hyp femora and the same sex normals, differences in mechanical behavior were attributed to material properties.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1995

Subcutaneous microvascular (capillary) calcification. Another basis for livedo-like skin changes?

A morbidly obese woman with acute renal failure, high serum phosphorus and slightly depressed serum calcium levels, developed areas of induration in the subcutis with associated livedo reticularis. Later, the subcutis became necrotic and the skin ulcerated. The arterioles, and notably widened occluded capillaries, were found to contain calcium and phosphorus as determined by energy dispersive spectrometry. X-ray diffraction of the subcutis in an early stage of this lesion showed that the mineral was most likely a poorly crystalline hydroxyapatite.

Adult↗
PubMed1994

Adenosine 5'-triphosphate promotes mineralization in differentiating chick limb-bud mesenchymal cell cultures.

When chick limb-bud mesenchymal cells are plated in micromass culture, they differentiate to form a mineralizable cartilage matrix. Previous studies have demonstrated that, when the total inorganic phosphate concentration of the medium is adjusted to 3-4 mM by adding inorganic phosphate to the basal medium, the mineralized matrix formed resembles that of chick calcified cartilage in ovo. When the high-energy phosphates adenosine 5'-triphosphate (ATP) or creatine phosphate are used as supplements in place of inorganic phosphate, the mineralized matrix as analyzed by electron microscopy and Fourier transform infrared microscopy is also similar to that in ovo. This is in marked contrast to the mineralized matrix formed in the presence of 2.5-5 mM beta-glycerophosphate, where mineral deposition is random and mineral crystal sizes in general are larger. This is also in contrast to the known ability of ATP to inhibit mineral deposition in solution in the absence of cells. In the differentiating mesenchymal cell culture system, ATP does not alter the rate of cell proliferation (DNA content), the rate of matrix synthesis (3H-leucine uptake), the mean crystallite length, or the rate of mineral deposition (45Ca uptake) when contrasted with cultures supplemented with inorganic phosphate. However, ATP does increase the mineral to matrix ratio, especially around the edge of the culture, where a type I collagen matrix is presented. It is suggested that ATP promotes mineral deposition by providing a high-energy phosphate source, which may be used to phosphorylate extracellular matrix proteins and to regulate calcium flux through cell membranes.

Adenosine Triphosphate↗
PubMed1994

Vitamin B6 deficiency experimentally-induced bone and joint disorder: microscopic, radiographic and biochemical evidence.

In the present study the effect of pyridoxine deficiency on the ultrastructure and morphology of bone and its metabolism was examined in the rapidly growing chick. Pyridoxine-deficient animals had tibias of reduced dry weight and cortical thickness. Histomorphometry demonstrated a disproportionately high eroded surface, lower amount of osteoid tissue and reduced mineralized trabecular width. Anterior-posterior radiographs of the tibiotarsometatarsal joint showed reduced secondary ossification centres and coarse trabeculation. Decalcified metaphyseal cartilage showed irregular trabeculas and a markedly reduced amount of Fast-green counterstain matrix suggesting that there is less collagen present and in turn less availability for matrix to be laid down for later calcification. Plasma activity of the bone alkaline phosphatase isoenzyme (EC 3.1.3.1) was decreased. Plasma Ca and PO4 levels did not vary. The present bone study referring to a pseudo-lathyritic state in which collagen maturation is not completely achieved supports the hypothesis that pyridoxine is an essential nutrient for the connective tissue matrix.

Alkaline Phosphatase↗
PubMed1994

Intramedullary deposits of hydroxyapatite crystals in a patient with rheumatoid arthritis.

A 63-year-old women with a 12-year history of rheumatoid arthritis (RA) had calcific deposits in the periarticular soft tissues with subsequent involvement of the medullary canals of adjacent bones. Except for the RA, there was no evidence of other abnormalities of the bones or mineral homeostasis. Chemical analyses of the deposits identified the crystalline material as apatite. Although the deposition of apatite in the soft tissues of RA is known to occur rarely, this is the first known case of intraosseous apatite deposition.

Arthritis, Rheumatoid↗
PubMed1993

Retinoic acid induces rapid mineralization and expression of mineralization-related genes in chondrocytes.

Numerous studies of experimental hypo- and hypervitaminosis A have long suggested that retinoic acid (RA) is involved in chondrocyte maturation during endochondral ossification and skeletogenesis. However, the specific and direct roles of RA in these complex processes remain unclear. Based on recent studies from our laboratories, we tested the hypothesis that RA induces the expression of genes associated with the terminal mineralization phase of chondrocyte maturation and promotes apatite deposition in the extracellular matrix. Cell populations containing chondrocytes at advanced stages of maturation were isolated from the upper portion of Day 18 chick embryo sterna and grown for 2 weeks in monolayer until confluent. The cells were then treated with low doses (10-100 nM) of RA for up to 6 days in the presence of a phosphate donor (beta-glycerophosphate) but in the absence of ascorbic acid. Within 4 days of treatment, RA dramatically induced expression of the alkaline phosphatase (APase), osteonectin, and osteopontin genes, caused a several-fold increase in APase activity, and provoked massive mineral formation while it left type X collagen gene expression largely unchanged. The mineral had a mean Ca/Pi molar ratio of 1.5; Fourier transform infrared spectra confirmed that it represented hydroxyapatite. Mineralization was completely abolished by treatment with parathyroid hormone; this profound effect confirmed that RA induced cell-mediated mineralization and not nonspecific precipitation. When cultures were treated with both RA and ascorbic acid, there was a slight further increase in APase activity and increased calcium accumulation. The effects of RA were also studied in cultures of immature chondrocytes isolated from the caudal portion of sternum; however, RA only had minimal effects on mineralization and gene expression in these cells. Thus, RA appears to be a rapid, potent, maturation-dependent, ascorbate-independent promoter of terminal maturation and matrix calcification in chondrocytes.

Alkaline Phosphatase↗
PubMed1993

Gallium nitrate inhibits alkaline phosphatase activity in a differentiating mesenchymal cell culture.

The effect of gallium nitrate on alkaline phosphatase activity in a differentiating chick limb-bud mesenchymal cell culture was monitored in order to gain insight into the observation that rachitic rats treated with gallium nitrate failed to show the expected increase in serum alkaline phosphatase activity. Cultures maintained in media containing 15 microM gallium nitrate showed drastically decreased alkaline phosphatase activities in the absence of significant alterations in total protein synthesis and DNA content. However, addition of 15 microM gallium nitrate to cultures 18 h before assay for alkaline phosphatase activity had little effect. At the light microscopic and electron microscopic level, gallium-treated cultures differed morphologically from gallium-free cultures: with gallium present, there were fewer hypertrophic chondrocytes and cartilage nodules were flatter and further apart. Because of altered morphology, staining with an antibody against chick cartilage alkaline phosphatase appeared less extensive; however, all nodules stained equivalently relative to gallium-free controls. Histochemical staining for alkaline phosphatase activity was negative in gallium-treated cultures, demonstrating that the alkaline phosphatase protein present was not active. The defective alkaline phosphatase activity in cultures maintained in the presence of gallium was also evidenced when cultures were supplemented with the alkaline phosphatase substrate, beta-glycerophosphate (beta GP). The data presented suggest that gallium inhibits alkaline phosphatase activity in this culture system and that gallium causes alterations in the differentiation of mesenchymal cells into hypertrophic chondrocytes.

Alkaline Phosphatase↗