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Biomedical subjects

A L Boskey

Publications and source records attributed to A L Boskey.

At least 127 records · Page 7Linked to original sources

The effect of vitamin D deficiency on rat bone lipid composition.

Vitamin D and phosphate deficiency has been shown previously to result in an accumulation of calcium acidic phospholipid phosphate complexes in the epiphysis of immature rats and to increase the cholesterol and decrease the lysophosphatide content throughout bone. To define the effects of vitamin D deficiency and those of combined vitamin D and phosphate deficiency, the lipid composition of the bones of second generation rachitic rats, raised on a diet that was vitamin D deficient was compared to that of normal animals fed a vitamin D-repleted diet and to previously studied vitamin D and phosphate-deficient rats. Animals raised on the vitamin D-deficient diet in utero and during postfetal life were serologically quite different from controls. These animals were significantly hypocalcemic and had no detectable vitamin D metabolites. Similar to the previously studied phosphate-deficient rachitic animals, the vitamin D-deficient animals had significantly elevated total lipid contents (weight percent lipid per demineralized dry weight) throughout their bones as compared to controls (epimetaphysis 9.6 vs 6.7 (P less than or equal to 0.03), diaphysis 6 vs 2 (P less than or equal to 0.01), calvaria 6.1 vs 2.4 (P less than or equal to 0.002). Similar to the vitamin D and phosphate-deficient animals, the second generation vitamin D-deficient rat bones had elevated cholesterol and reduced lysophosphatide contents. Complexed acidic phospholipids were not significantly elevated in the vitamin D-deficient animal bones. The data suggest that the lipid changes seen in these animals are due to vitamin D deficiency and not to phosphate deficiency.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Topographical variations in the morphology and biochemistry of adult canine tibial plateau articular cartilage.

Topographically, there are both morphological and biochemical differences in the articular cartilage of the tibial plateau of normal adult dogs when the cartilage covered by the meniscus is compared with that more centrally placed and not covered by meniscus. Histologically, differences are present in the surface morphology, in intra- and extracellular lipid content, and in the morphology of the mineralization front. Electron microscopy shows, in the covered cartilage, variability in collagen fiber size, with evenly spaced fibers apparently randomly distributed and an orderly relationship between the proteoglycans and collagen, whereas in the uncovered area, the collagen is aggregated into bundles and appears to be dissociated in large part from the proteoglycans. The most striking feature in the biochemistry of the two regions is an increased water content in the uncovered cartilage, as compared with the covered. In addition, there is an increased amount of proteoglycans that can be extracted in the uncovered cartilage. The heterogeneity of the cartilage on the tibial plateau should be taken into account when considering both the histologic and biochemical variations found in osteoarthritic cartilage; and when reflecting on the pathogenesis of osteoarthritis.

Animals↗

Mechanisms of proteoglycan inhibition of hydroxyapatite growth.

Purified bovine nasal cartilage proteoglycans (aggregate and subunit containing fractions) and to a lesser degree, chondroitin 4-sulfate of physiological size, retard seeded hydroxyapatite (HA) growth in vitro. The large hydrodynamic size and high charge density of these macromolecules are believed to be associated with the ability of proteoglycans to inhibit HA formation and growth. We now demonstrate the involvement of the negative charges of proteoglycans in this inhibition by comparing the inhibitory ability of chondroitin 4-sulfate and its desulfated analog, and by comparing the growth of HA seed crystals coated either with proteoglycan aggregates or chondroitin 4-sulfate to that of uncoated crystals. In the desulfation experiments, desulfated chondroitin sulfate was a less efficient HA growth inhibitor than untreated, undesulfated chondroitin sulfate of similar molecular size. Dextran sulfate showed higher inhibitory effectiveness than unchanged neutral dextran. Both experiments suggest that sulfate groups play an important role in the regulation of mineral deposition by proteoglycans. In the coating experiment, precoating of HA seed crystals with proteoglycan aggregates decreased the amount of HA precipitated as a function of time, suggesting proteoglycans may block the active nucleating sites on HA surface and slow down the growth process. Chondroitin 4-sulfate had a similar but weaker coating effect. Neutral dextran, having little affinity for HA, had no effect.

Animals↗

Calcium-acidic phospholipid-phosphate complexes in human atherosclerotic aortas.

Since aortic calcification chemically resembles bone mineralization, we tested the hypothesis that a bone mineral nucleator, the Ca-acidic phospholipid-phosphate complexes, is found in atherosclerotic plaques. Calcium-acidic phospholipid-phosphate complexes were isolated from hydroxyapatite-containing calcified plaques and from adjacent nonmineralized areas of adult human aortas. Neonatal aortas, which served as nonmineralized lesion-free controls, contained negligible amounts of the Ca-acidic phospholipid-phosphate complexes. The concentration of complexed acidic phospholipids in the aortic plaques (5 micrograms/mg demineralized dry wt) was comparable to that found in newly mineralized bone and calcified cartilage. The presence of Ca-acidic phospholipid-phosphate complexes in the nonmineralized regions of the adult aorta, as well as in the mineralized plaques, suggests that this tissue may calcify through mechanisms similar to those involved in bone mineralization.

Adult↗

Mineral and matrix alterations in the bones of incisors-absent (ia/ia) osteopetrotic rats.

The bones of incisors-absent (ia/ia) osteopetrotic rats differ from those of their normal littermates (ia/+) in histologic and radiographic appearance and in mechanical properties. This study examined how the mineral and matrices of osteopetrotic metaphyses and calvaria differed from normal controls. Bones of 11-day-old and 52-day-old osteopetrotic animals had higher ash (mineral) contents that age-matched controls; osteopetrotic metaphyses had elevated hexosamine contents, indicative of the persistence of cartilage. Calcium acidic phospholipid phosphate complexes, involved in initiation of hydroxyapatite formation in vivo and in vitro, were significantly reduced in content in all osteopetrotic bones. These results suggest that in the osteopetrotic rat, where osteoclast activity is defective, new mineral formation is reduced and replaced by accretion of mineral on existing crystals. Fractionation of osteopetrotic bone particles by density centrifugation demonstrated that the osteopetrotic bone was much more mineralized than that of age-matched controls. X-ray diffraction analysis of the mineralized fractions indicated a lack of growth of mineral crystals during maturation of the osteopetrotic animals. The absence of remodeling was apparent from the elevated Ca:P ratios of the highly mineralized osteopetrotic bone fractions, and from the high hexosamine content of these dense fractions. These observations may explain some of the unusual mechanical properties of osteopetrotic bone.

Animals↗

The effect of osteocalcin on in vitro lipid-induced hydroxyapatite formation and seeded hydroxyapatite growth.

Osteocalcin, the bone gamma-carboxy glutamic acid containing protein, is one of the major noncollagenous proteins both synthesized and localized in bony tissue. Previously, investigators have suggested, based on the in vitro and in vivo properties of this protein, that it may be involved in controlling initiation of mineralization and/or hydroxyapatite (HA) growth. In this study, the in vitro effects of osteocalcin on lipid-induced HA formation, and HA seeded growth were compared. Although osteocalcin inhibited the growth of HA, as indicated by the osteocalcin concentration-dependent decreases in the first order rate constant, kCa, osteocalcin had no effect on lipid-induced calcification, kCa remaining constant at .033 h-1. Binding studies revealed that osteocalcin did not associate with the lipid macromolecules tested (phosphatidyl serine, phosphatidyl inositol, and the Ca-acidic phospholipid-phosphate (Ca-PL-PO4) complexes prepared from these phospholipids) although the protein bound to HA with high affinity. These data suggest that a) osteocalcin is quite distinct from the gamma carboxy glutamic acid containing clotting proteins which have a high affinity for both the acidic phospholipids and for HA, and b) that osteocalcin has little effect on the initial Ca-PL-PO4-dependent formation of HA.

Calcium-Binding Proteins↗

Lipid changes in the bones of the healing vitamin D-deficient phosphate-deficient rat.

The bones of vitamin D-deficient, phosphate-deficient rats have a lipid composition that is significantly different from that of normal bones. Specifically, these bones have elevated cholesterol and reduced lysophosphatide and free fatty acid contents. Treatment of these animals with a single dose of vitamin D and phosphate produces healing within 72 h and causes rapid corrections of alterations in growth plate and cancellous bone lipid composition. Healing of the rachitic/osteomalacic state in these animals was demonstrated radiographically and histologically. Histomorphometric measurements showed that the relative osteoid volume of the cancellous bone rapidly approached the 7% value of normal controls, decreasing from 29% in the rachitic animals to 16% by 12 h and 8.5% by 72 h. Significant changes in ash weight, Ca:P ratio, and crystal-lite size and perfection were detectable at 12 h, with these parameters approaching values found in normal animals within 72 h. Calcium-acidic phospholipid-phosphate complexes, which are known to promote hydroxyapatite formation, peaked in concentration at 12 h in epiphysis, cancellous, and cortical bone, returning rapidly to normal values after that time. In untreated animals the complexed acidic phospholipid content of the nonmineralized epiphysis was comparable to that in normal mineralizing epiphysis, whereas the content of the complexes was reduced in the cancellous bones of the untreated animals.

Animals↗

Co-isolation of proteolipids and calcium-phospholipid-phosphate complexes.

This study demonstrates that calcium-phospholipid-phosphate complexes (CPLX) and calcifiable proteolipid are associated in vivo by establishing that they can be co-isolated from calcified bacteria. Both of these membrane constituents, which support apatite formation in vitro, have been isolated independently from Bacterionema matruchotii. However, isolation of proteolipid was preceded by demineralization in 2N formic acid, thereby dissociating bound Ca, whereas isolation of CPLX included sonication of calcified bacteria in 2:1:1.5 chloroform:methanol:Tris buffer, thereby dissociating any protein. Co-isolation is possible by demineralizing the calcified bacteria with 50 mM phthalic acid, pH 5.5, followed by extraction with 2:1 chloroform:methanol, and precipitation of crude phospholipid with acetone. CPLX and proteolipid are present in all Sephadex LH-20 chromatographic fractions of the crude phospholipid and of diethyl ether precipitates of the crude phospholipid. CPLXs contain protein:phospholipid:Ca:Pi but differ in relative composition from each other and from independently isolated CPLX. The Ca:phospholipid:Pi molar ratio of diethyl ether precipitable proteolipid-CPLX is most similar to previously published values for CPLX. The protein content of CPLX accounts for all of the proteolipid apoprotein in each Sephadex LH-20 fraction.

Actinomycetaceae↗

The inhibitory effect of cartilage proteoglycans on hydroxyapatite growth.

The calcification of connective tissues, including cartilage, is under the control of many interacting systems. Proteoglycans are thought to retard the deposition of hydroxyapatite crystals, and modification of the proteoglycans presumably facilitates mineralization in those tissues that are actively calcifying. The mechanism underlying these regulations remains speculative. This study investigates this question by comparing the inhibitory effectiveness of several macromolecules at neutral pH and approximately physiological ionic strengths. Inhibitors tested include bovine nasal proteoglycan monomer A1D1D1 and aggregate-containing A1 fractions, glycosaminoglycan chains (chondroitin 4-sulfate), and neutral dextran (as an uncharged analog). Hydroxyapatite growth was assessed either by measuring the time-dependent decreases in solution calcium and phosphate concentrations, or by determining utilization of hydroxyl ion in a pH-Stat. All species studied inhibit hydroxyapatite growth, and the extent of inhibition for each class is concentration-dependent. The proteoglycan aggregate-containing A1 fraction is more effective than the proteoglycan monomer at the same concentration, and the proteoglycan monomer is more effective than chondroitin 4-sulfate. Neutral dextran inhibits hydroxyapatite growth less effectively than proteoglycans. These results suggest that inhibition of hydroxyapatite growth by proteoglycans critically depends on both status (aggregate, monomer, etc.) and hydrodynamic size of this macromolecule, supporting the hypothesis that modification of proteoglycans in vivo functions to modulate the effectiveness of proteoglycans as a hydroxyapatite growth inhibitor.

Animals↗

Phosphate modulation of calcium transport by a calcium-phospholipid-phosphate complex of calcifying tissues.

The ionophoretic activities of a stable Ca-PS-Pi complex were examined in a three-compartment lipid-aqueous phase transport system. It was found that the complex transported Ca2+ at a slower rate than free PS. Analysis of the transport process, using an equilibrium partitioning model, revealed that Ca2+ binding to the complex was the rate-limiting step. This inhibition was overcome by the addition of exogeneous Pi. It was concluded that the complex acted as an ionophore and that the presence of coordinated Ca and P atoms facilitated Pi modulation of Ca2+ transport.

Animals↗

Calmodulin localization in bone and cartilage.

The localization of calmodulin (CaM), a calcium-dependent regulatory protein, was demonstrated in the following rat skeletal tissues by the indirect immunofluorescence method: a) growth plate cartilage of fetal and juvenile long bones, b) fetal epiphyseal cartilage, c) juvenile hyaline costal cartilage, d) neonatal mandibular condylar cartilage, e) neonatal diaphyseal lamellar bone. CaM was not detected in perichondrial and periosteal cells. Mature and mineralizing chondrocytes demonstrated the highest labelling intensities.

Animals↗

The effects of trifluoperazine on calcifying tissues in the immature rat.

When the antipsychotic drug trifluoperazine is given to immature rats during stages of rapid bone development, severe localized derangement in their growth plate architecture results. In addition, 3 weeks of treatment with doses of 3 mg/kg body wt/day had marked effects on the anatomic distribution of bone mineral, and on the quantitative chemical composition of the bone matrix. Specifically, ash contents and Ca/P ratios were decreased and mineral crystals appeared larger and/or more perfect in the treated animals. The finding of decreased hexosamine content and alterations in bone lipid composition could be interpreted as specific effects of the drug on calmodulin activity.

Animals↗

Cartilage calcification: normal and aberrant.

Hydroxyapatite crystal deposition occurs physiologically in cartilage as a prelude to bone formation via endochondral ossification. Both extracellular molecules and organelles, and the chondrocytes themselves control the initial formation of hydroxyapatite, as well as the growth and orientation of the hydroxyapatite crystals. In this study pathologic calcium containing deposits from 48 patients (27 hydroxyapatite and 21 calcium pyrophosphate dihydrate) were subjected to crystallographic, histologic (light microscopy, scanning electron microscopy, transmission electron microscopy) and chemical analyses and compared to normal controls. It is suggested that both hydroxyapatite and calcium pyrophosphate dihydrate deposition involve elevations in ionic concentrations, exposure of mineral nucleators, and removal of mineral inhibitors. Peculiar to the matrices of pathologic deposits of hydroxyapatite are elevated concentrations of calcium acidic phospholipid phosphate complexes and lower concentrations of hexosamine while collagen (hydroxyprolin) and total lipid contents are not altered. Matrices of deposits of calcium pyrophosphate dihydrate were similar biochemically except that calcium acidic phospholipid phosphate complexes concentrations were not elevated.

Calcification, Physiologic↗

Bone structure, composition, and mineralization.

Bone structure and function are dependent on complex interactions between cells, matrix, cell-derived factors, and systemic factors. The deposition of mineral in bone, which enables the skeleton to function properly, is described as a four-step process of matrix modification, crystal nucleation, crystal growth, and remodeling. Insight into the function of bone components in the mineralization process is provided by in vitro studies and analysis of abnormal calcifications.

Animals↗

Changes in lipids during matrix: induced endochondral bone formation.

The changes in lipids occurring during the process of endochondral ossification have been characterized by studying the discrete phases of matrix-induced endochondral bone formation in the rat. Calcium-acidic phospholipid-phosphate complexes were shown to increase in concentration during cartilage calcification (day 9) and to peak in content during early bone formation (days 11-13), the times during which the rate of mineral deposition, as indicated by the change in ash weight was greatest. These data support the hypothesis that the calcium-acidic phospholipid-phosphate complexes play a role in the in vivo initiation of hydroxyapatite deposition. The overall lipid composition of the induced matrix newly formed cartilage (days 7-9) was comparable to that of normal cartilage, with the phospholipid composition matching that of chondrocyte plasma membranes. Times of vascular invasion and formation of marrow cavities were marked by elevated total lipid and triglyceride contents.

Animals↗

Phospholipids associated with human parotid gland sialoliths.

Ca-acidic phospholipid-phosphate complexes (CPLX) were isolated from 6 whole and 2 pooled human parotid sialoliths containing the mineral phases hydroxyapatite, whitlockite and octacalcium phosphate (ash 78 +/- 16 per cent, n = 7). Total lipids accounted for 25 per cent of the demineralized dry weight of these sialoliths. CPLX was 39 +/- 18 per cent of the lipid phosphorous pool, while diphosphatidyl glycerol, phosphatidyl ethanolamine and phosphatidyl choline were the principal non-complexed phospholipids. The high cholesterol:phospholipid molar ratio (1.0 +/- 0.3) observed in the sialolith lipids suggests a plasma membrane origin for these lipids.

Humans↗

Effect of high levels of corticosteroids on the lipids of the long bones of the mature rabbit.

Corticosteroid administration is associated with the development of nontraumatic osteonecrosis in man. However, the pathogenesis of steroid-induced osteonecrosis is unknown. Osteopenia, occurring as a result of corticosteroid treatment is thought to be one factor contributing to the compromise of vascular space. We have treated rabbits daily with hydrocortisone-acetate (15 mg per 4 kg rabbit for 4 and 9 weeks, respectively). These animals progressively developed osteoporosis. Cholesterol accumulated in both the unwashed cancellous bone and the marrow-free compact bones of the treated rabbits when contrasted with age-matched controls. Elevations in marrow lipid content were also observed in the treated animals. Although none of the treated rabbits developed osteonecrosis during the short time of the study, the findings of elevated cholesterol suggest that alterations in bone cell membranes may lead to cell dysfunction and osteopenia. This osteopenia may eventually cause osteonecrosis by occlusion of subchondral vessels. The elevated marrow lipids may also contribute to the development of osteonecrosis by increasing intramedullary pressure and causing venous stasis.

Animals↗