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PubMed1994

Expression of bone morphogenetic proteins in human osteosarcoma. Immunohistochemical detection with monoclonal antibody.

BACKGROUND: Bone morphogenetic proteins (BMP) induce ectopic bone formation in vivo and may play a role in normal bone development. In addition, bone morphogenetic activity, as measured in a bone-forming assay in immunodeficient, athymic nu/nu mice, is present in a proportion of osteosarcomas; this activity, which may be mediated by BMP, is correlated with a poor prognosis. METHODS: The development of a monoclonal antibody against recombinant human BMP-2, AbH3b2/17, has allowed immunohistochemical localization of BMP in tumor tissues. Cryostat sections of osteosarcomas (21 tumor samples), chondrosarcomas (5 samples), and Ewing's sarcomas of bone (5 samples) were examined with AbH3b2/17 using the avidin-biotin-immunoperoxidase method. RESULTS: The authors found AbH3b2/17 immunoreactivity in 12 of the 21 osteosarcoma samples (57% sensitivity) obtained from 20 patients. For one patient, samples of the primary lesion and a subsequent metastasis were tested, and only the latter showed AbH3b2/17 immunoreactivity. The chondrosarcomas and Ewing's sarcomas examined showed no immunoreactivity. In antigen-positive osteosarcomas, AbH3b2/17 immunostaining was localized predominantly in the cytoplasm of tumor cells. Moreover, the proportion of AbH3b2/17-reactive cells varied among osteosarcomas with disparate histologic features. CONCLUSIONS: The authors identified a rapid and widely applicable method for detecting BMP expression in intact tissues, which may complement and enhance the bone-forming assay in nu/nu mice as a prognostic procedure in osteosarcomas.

Adolescent↗

PubMed1990

Carbon fibre composite bone plates. Development, evaluation and early clinical experience.

We compared the mechanical properties of carbon fibre composite bone plates with those of stainless steel and titanium. The composite plates have less stiffness with good fatigue properties. Tissue culture and small animal implantation confirmed the biocompatibility of the material. We also present a preliminary report on the use of the carbon fibre composite plates in 40 forearm fractures. All fractures united, 67% of them showing radiological remodelling within six months. There were no refractures or mechanical failures, but five fractures showed an unexpected reaction; this is discussed.

Adult↗

PubMed2005

Bone mass in childhood is related to maternal diet in pregnancy.

Evidence that birth weight is related to bone mass in later life suggests that the intrauterine environment programs the trajectory of subsequent bone development. To explore this hypothesis, we examined whether maternal diet in pregnancy, as assessed by the maternal food frequency questionnaire (FFQ) completed at 32 weeks gestation, is related to bone mass of the child, as measured by total body DXA carried out at age 9 years in the Avon Longitudinal Study of Parents and Children (ALSPAC). Diet records were linked to DXA scan results for the total body and spine sub-region and pooled between pre- and early pubertal boys and girls (n=4,451). Regression analysis was carried out between DXA values and dietary factors following adjustment for social and other confounding factors. Maternal magnesium intake was related to total body BMC (beta=4.9, 7.4-23.1; g) and BMD (beta=4.9, 2.5-7.3; g/cm2 x10(3)) (standardized regression coefficient with 95% confidence limits; P<0.001). An equivalent relationship was no longer observed after adjusting for the height of the child, to which magnesium intake was also related (beta=0.48, 0.20-0.77; cm; P=0.001). Maternal intake of potassium was related to spinal BMC (beta=1.8, 0.8-2.9; g) and BMD (beta=10.5, 4.9-16.0; g/cm2 x10(3)) (P=0.001), which was no longer observed after adjusting for the weight of the child, to which potassium intake was also related (beta=0.52, 0.16-0.88, P=0.005; kg). A significant association was also observed between maternal dietary folate intake and spinal BMC adjusted for bone area using a linear regression model (beta=0.55, 0.16-0.94; g; P=0.006), which persisted after adjusting for height and weight. Our observation that constituents of maternal diet are related to DXA measures at age 9 is consistent with the hypothesis that the trajectory of bone development in childhood is programmed by early life factors.

Absorptiometry, Photon↗

PubMed1992

Cholinesterase activity in the chondrocranium of normal and exencephalic mouse fetus with trisomy 12.

Cholinesterase (ChE) activity in the chondrocranium of normal and exencephalic trisomy 12 mouse fetuses was studied. Non-specific cholinesterase activity was found strongly in the developing bone cells at the central zone and weakly in the more maturely developed bone cells at the peripheral zone of the chondrocranium of both normal and exencephalic trisomy 12 mouse fetuses. In exencephalic mouse fetuses, the ChE-activity was lesser than in the normal ones which corresponded to hypoplastic chondrocranium. The centrifugal direction of the maturity of individual bone cells could be demonstrated by the activity of cholinesterase. The young bone cells showed strong ChE-activity while the more matured cells showed weak ChE-activity. The enzyme activity disappeared when the definite tissue structure was well developed. From this study, it may be concluded that ChE plays a role in chondrocranium development which is different from its known function in the adult tissue.

Anencephaly↗

PubMed1998

[Development of an in vitro model of culture using mouse fetal long bone].

An in vitro model of culture using mouse fetal long bone was firstly established using self-made turning-equipment. The bone formation, mineralization and resorption were observed simultaneously in cultured bone. It provided a useful model for investigating the relationship between bone, development and the effect of nutrients, toxicants and other factors.

Animals↗

PubMed1993

Developmental expression of osteopontin (OPN) mRNA in rat tissues: evidence for a role for OPN in bone formation and resorption.

Osteopontin (OPN) is a 34-kDa, highly-phosphorylated glycoprotein with cell attachment properties that is a prominent constituent of the bone matrix. To aid in elucidating the function of this protein we have studied the cellular expression of OPN mRNA during the formation, growth and maturation of rat calvarial (membranous) and tibial (endochondral) bone. From Northern hybridization analysis OPN expression was demonstrated in the kidney and gravid uterus as well as in bone tissues. Compared to collagen, the expression of OPN was low in early bone formation but increased subsequently and reached peak levels in 14-day-old bone. However, both the collagen and OPN mRNAs decreased markedly thereafter and remained low in young adult bone. From in situ hybridization studies using a [35S]-labelled rat OPN cRNA probe, OPN mRNA was localized to osteoblastic cells in newly-forming calvariae, jaw bones, and in the metaphyseal and periosteal bone of the tibia. In contrast to bone sialoprotein (BSP), which is expressed almost exclusively by osteoblasts at sites of de novo bone formation, OPN transcripts were present in cells lining both endosteal and periosteal bone surfaces, and in osteocytes. Moreover, expression of OPN persisted during the subsequent growth and remodelling of both membranous and endochondral bone and was expressed at particularly high levels by bone cells and hypertrophic chondrocytes at sites of osteoclastic resorption. In the more mature bone of young adult rats OPN expression was significantly reduced but remained detectable in bone cells lining periosteal and endosteal surfaces and in the primary and secondary spongiosa of the tibia. These studies on the developmental expression of OPN support the concept of a multifunctional role for OPN in bone formation and remodelling. Thus, the expression of OPN by osteoblasts early in bone development is consistent with a role for this protein in the formation of bone matrix, whereas the peak expression of OPN later in bone development, together with high expression at sites of rapid remodelling, indicate that OPN deposited on the surface of mineralized connective tissues may provide a template for osteoclastic resorption.

Animals↗

PubMed1988

Genetic control of mast cell development in bone marrow cultures. Strain-dependent variation in cultures from inbred mice.

A comparison was made of the capacity of bone marrow cells (BM) from genetically distinct strains of mice to develop into mast cells under defined conditions of in vitro culture. In the presence of conditioned media derived from ConA treated spleen cells from normal or Trichinella spiralis-infected mice, mast cell development occurred readily. After 21 days of culture mast cells comprised more than 90% of the total cell population. BM taken from certain strains of mice (SWR and NIH) produced large numbers of mast cells, total cell numbers increasing between 2 and 5 fold; other strains (C57BL/10 [B10] B10 congenics) produced relatively few mast cells, total cell numbers not increasing above the starting concentration or declining during culture. The genetic factors determining the strain-response phenotype (no. of mast cells in culture) were predominantly associated with the background genome. No significant differences in response were noted between the B10 congenic strains B10 [H-2b], B10.G [H-2q] or B10.BR [H-2k], which differ only at the MHC, whereas major differences were seen between B10.G and the other H-2q strains [SWR and NIH]. Response phenotype was not inherited as a simple dominant trait; F1 progeny of high x low responder strains were intermediate between the parental values. The expression of genetic influences upon mast cell response phenotype appears to be at both the level of mast cell precursor cells, as determined from limiting dilution assays of BM from high, low and F1 (high x low) strains, and at the level of mast cell proliferation, as determined by repeated sub-culture of mast cells from these strains.

Animals↗

PubMed2001

Treatment of traumatic bone defects by bone transport.

This study retrospectively evaluates the results of the management using the callus distraction method, of 19 patients who had developed bone defects following acute trauma. Sixteen patients were males, with a mean age of 19.4 years (6-41 years). Thirteen patients developed bone defects as a result of high-velocity gunshot injuries, and six had defects caused by traffic accidents. The location of 12 fractures was the tibia, and seven were in the femur. The mean follow-up period was 23.6 (12-40) months. The mean length of the bone defect was 6.8 cm (4-16 cm), the distraction index was 13.3 days/cm (7.5-18 days/cm) and the external fixation index was 36.6 days/cm (32.5-46 days/cm). Fifteen (9%) pintract infections were observed among those who underwent circular external fixation and four (10%) in cases who underwent unilateral external fixation. There were no deep infections. Refracture was noted in one patient (5%), and two (11%) had delayed union at the target site. One patient developed nonunion at the target site. For the treatment of large bone defects, the callus distraction method may be an alternative option to conventional treatment methods.

Adolescent↗

PubMed2006

Is significant cystic fibrosis-related liver disease a risk factor in the development of bone mineralization abnormalities?

In order to assess the effects of significant cystic fibrosis-related liver disease (CFLD) on bone health, we compared the bone mineral status of older children and adolescents with CFLD to those with cystic fibrosis (CF) alone. Thirteen children (age range, 10-19 years) from our clinical CF services were identified with significant CFLD (9 of these 13 patients had clinical and radiological evidence of portal hypertension). This cohort was then matched by age, gender, and anthropometric measurements with equal numbers of patients with CF alone. All patients had a dual-energy X-ray absorptiometry (DEXA) scan to determine bone mineral content (BMC), bone area (BA), bone mineral density (BMD), and bone mineral apparent density (BMAD) in the region of the lumbar spine. Blood was drawn to determine serum vitamin A, D, E, and K status and liver function tests. The best forced expired volume in 1 sec (FEV1) for each patient in the 12 months around the time of the scan was also documented. Patients with CFLD had slightly worse FEV1 (82 +/- 20% vs. 91 +/- 16%, P = 0.05) and significantly higher alanine aminotransferase (65.5 +/- 35 IU/l vs. 30 +/- 20 IU/l, P = 0.01) than those with CF alone. The mean lumbar spine BA, BMC, BMD, and BMAD were not different between children with CFLD and CF. In conclusion, the presence of significant liver disease in children with CF does not appear to be an additional risk factor for the development of abnormal bone mineralization.

Absorptiometry, Photon↗

PubMed1977

Transmembrane bone morphogenesis across multiple-walled diffusion chambers. New evidence for a diffusible bone morphogenetic property.

Bone generation and regeneration are associated with a bone morphogen that recruits mesenchymal cells for differentiation into bone. Experiments with particulate bone matrix gelatin implanted in multiple-walled diffusion chambers suggest that bone morphogen is a rapidly diffusible molecule, and consists of a noncollagenous bone morphogenetic protein (BMP). When particulate bone matrix gelatin is implanted inside of diffusion chambers constructed of two to five membranes, ranging from 300 to 750 cu micronm in total thickness, large deposits of bone develop on the outside. The volumes of the deposits of new bone are inversely proportional to the thickness (or distance) of transmission of the BMP. Transmission for long distances through interstitial fluid can be accounted for by a low molecular mass hydrophobic BMP, disseminated according to the laws of diffusion.

Animals↗

PubMed2002

L-shell x-ray fluorescence measurements of lead in bone: system development.

This paper reports on the development of an L-shell x-ray fluorescence (XRF) bone lead measurement system. A secondary target gave greater lead x-ray peak signal-to-background ratios than partially plane polarized XRF. Filtration did not improve the lead x-ray peak signal-to-background ratio: the gains in spectrum quality were outweighed by the losses caused by attenuation. There was a substantial matrix effect: the signal from a calcium-rich matrix was far lower than that from a calcium-free matrix. The effect of attenuation was, as expected, profound for the lead L x-rays: detection limits ranged from 18 to 217 microg Pb/g plaster with attenuation equivalent to 0-2.1 mm of skin or 0-3.7 mm of adipose tissue for the Pb Lalpha x-ray group (10.5 keV), and from 16 to 184 microg Pb/g plaster with attenuation equivalent to 0-1.3 mm of skin or 0-2.3 mm of adipose tissue for the Pb Lbeta x-ray group (12.6 keV).

Bone and Bones↗

PubMed1982

Hydroxyurea increases in vitro, antigen-independent B cell development in bone marrow.

The fluorescence-activated cell sorter and Coulter counter were used to study the effect of hydroxyurea in vitro in B cell development in 24 hr cultures of adult mouse bone marrow. This agent, which kills cells in the S phase of the cell cycle, caused a 40-70% increase in the absolute number of B cells in bone marrow cultures as compared with untreated cultures. There was also an increased response to the B cell mitogen, bacterial lipopolysaccharide, as measured by the stimulation index for 3H-thymidine incorporation. Hydroxyurea in vitro increased B cells numbers in fetal liver which contains pre-B cells, but not in lymph nodes or spleen which lack these cells.

Animals↗

PubMed2007

Development of the mouse mandibles and clavicles in the absence of skeletal myogenesis.

In this report we employed double-knock-out mouse embryos and fetuses (designated as Myf5-/-: MyoD-/- that completely lacked striated musculature to study bone development in the absence of mechanical stimuli from the musculature and to distinguish between the effects that static loading and weight-bearing exhibit on embryonic development of skeletal system. We concentrated on development of the mandibles (= dentary) and clavicles because their formation is characterized by intramembranous and endochondral ossification via formation of secondary cartilage that is dependent on mechanical stimuli from the adjacent musculature. We employed morphometry and morphology at different embryonic stages and compared bone development in double-mutant and control embryos and fetuses. Our findings can be summarized as follows: a) the examined mutant bones had significantly altered shape and size that we described morphometrically, b) the effects of muscle absence varied depending on the bone (clavicles being more dependent than mandibles) and even within the same bone (e.g., the mandible), and c) we further supported the notion that, from the evolutionary point of view, mammalian clavicles arise under different influences from those that initiate the furcula (wishbone) in birds. Together, our data show that the development of secondary cartilage, and in turn the development of the final shape and size of the bones, is strongly influenced by mechanical cues from the skeletal musculature.

Animals↗

PubMed1979

Maturation-linked expression of the Fc (IgG) receptor on developing human bone marrow and peripheral blood granulocytes.

Expression of Fc(IgG) receptors, as demonstrated by IgG-coated ox cells and heat-aggregated IgG, was assessed on normal human peripheral blood and bone marrow neutrophils and their precursors. The results indicate that Fc receptor development is confined to post-mitotic granulocytes, being primarily associated with neutrophil segmentation. Fc receptors were not detected on myeloblasts or promyelocytes. A mean value of 79% and 99% Fc positive segmented neutrophils were found in bone marrow and peripheral blood respectively. This highly significant difference (P less than 0.001) appears to represent a morphologically indistinguishable terminal maturation stage. Inhibition studies with IgG show that the neutrophil Fc receptor is inhibited by aggregated but not, in contrast to the corresponding monocyte receptor, by monomeric (7S) IgG. Applications of these findings to the in vivo function of the neutrophil in health and pathology are discussed.

Aged↗

PubMed1992

Determinants of bone density in young women. I. Relationships among pubertal development, total body bone mass, and total body bone density in premenarchal females.

Bone mass accretion during puberty appears to be critical in the development of peak bone mass, which, in turn, is believed to be a major determinant of osteoporosis risk. Although genetics may be the primary determinant of peak bone mass, modifiable secondary factors, such as nutrition and hormone exposure, may significantly affect bone mass accretion during the second decade of life. As part of a longitudinal study of major determinants of bone development during puberty, we obtained cross-sectional measurements from 112 premenarchal caucasian females (mean +/- SD age, 11.9 +/- 0.49 yr at study entry). Total body bone mineral density (TBBMD) and total body bone mineral content (TBBMC) were measured by dual energy x-ray absorptiometry and compared to anthropometric, pubertal development, urinary steroid and gonadotropin levels, and nutrient intake. An integrated estrogen exposure index was developed and used to evaluate the cumulative effect of circulating estrogen levels on both development. Compared to normative reference data for adults, our subjects possessed 90% of adult height, 68% of adult weight, 83% of adult TBBMD, and 53% of TBBMC. The strongest combined predictors of prepubertal TBBMD and TBBMC were body weight, followed by height and pubertal development. Urinary estradiol levels were positively correlated with dietary intake of iron and vitamin B6.

Adolescent↗

PubMed2006

Expression of the calcium receptor in human breast cancer--a potential new marker predicting the risk of bone metastases.

AIMS: This study investigates whether the calcium-sensing receptor (CaR) is commonly expressed in primary breast cancers. The CaR controls secretion of PTHrP in several breast cancer cell lines and PTHrP is known to stimulate osteolysis during metastatic bone resorption. Whether this could explain the propensity of breast cancers to develop bone metastases has not been explored. METHODS: With Ethical Committee approval, immunohistochemistry was performed using a commercially available antiCaR antibody (AffinityBioReagents, Cambridge, UK) on archived histological sections of primary tumours from patients who died with advanced breast cancer. Intensity of CaR expression was assessed by two independent observers on a 6-point scale. RESULTS: One hundred and eight patients with breast cancer were found to have positive bone scans, 42 patients had died. Of the patients with negative bone scans, 23 had liver or lung metastases. Most patients with strongly expressed CaR (score 4-5 on immunohistochemistry) had bone metastases (13/15 patients) compared with 2/23 patients with normal bone scans (p < 0.001, chi(2) test). Other clinical/pathological markers (ER, PR, c-erb B-2, LN status) were not significantly different between patients with CaR-positive or CaR-negative tumours. CONCLUSIONS: CaR expression is common in a selected group of patients with advanced primary breast cancers. A prospective study should investigate if patients with CaR-positive tumours are more likely to develop bone metastases.

Adult↗

PubMed1989

Skeletal deformities in smallmouth bass, Micropterus dolomieui, from southern Appalachian reservoirs.

Smallmouth bass (Micropterus dolomieui) populations in two of five reservoirs sampled in the southern Appalachian Mountains contained high percentages of individuals with lordosis, kyphosis, or scoliosis. Deformities of the vertebral column occurred in several year classes and varied with fish size; they were absent in small fish, present in 25-30% of the fish 241-300 mm long, and then decreased in occurrence with increased length. Because environmental contamination is often responsible for high occurrences of deformed fish, whole-body concentrations of contaminants, bone development characteristics, and blood plasma concentrations of calcium and phosphorus in normal and deformed fish were measured and compared the results with those for fish from reservoirs where no deformities were found. Vertebrae were significantly weaker and more elastic in deformed than in normal fish, but biochemical properties of vertebrae were similar among the groups tested. Concentrations of pesticides and metals were not elevated in deformed fish, and concentrations of calcium and phosphorus in blood plasma were similar in normal and deformed groups. Most environmental contaminants that have been shown to cause fish deformities could be discounted as causative agents on the basis of these results; however, the exact cause was not determined. Further attempts to diagnose the cause of the deformities were limited by the lack of background information on relationships among bone development processes, types of stresses that cause deformities, and types of bone tissue in fish.

Animals↗

PubMed1990

Effect of gestational sex steroid exposure on limb development and endochondral ossification in the pregnant C57Bl/6J mouse: I. Medroxyprogesterone acetate.

Although data supporting the teratogenic potential of intrauterine progestin exposure is lacking, concern persists among some individuals within the scientific community that these drugs have the potential for nongenital teratogenesis, especially with regard to limb reduction defects. Our laboratory has been interested in the ontogeny of steroid receptors in the developing embryo and in the role of steroid-receptor interactions in limb development, particularly the process of endochondral ossification. Since limb reduction defects can be produced from abnormal processes that are operative during organogenesis or during midgestation (vascular disruption) we have designed an animal study whereby embryos were exposed to sex steroids throughout organogenesis and fetal development. The present study assesses the effects of medroxyprogesterone acetate (MPA) on intrauterine endochrondral bone development specifically, as well as overall embryo-fetal development. Primagravid C57Bl/6J mice were treated via subdermal pellets which deliver MPA at dosages of 5.0, 50.0, and 500.0 mg/kg/day on gestational days 7 through 19. These doses were 25-, 250-, and 2,500-fold higher on a mg/kg basis than the human dose equivalent (HDE). No increases in nongenital malformations were noted at any evaluated MPA dosage level. At 25 X the HDE, MPA did not influence endochondral bone development as evidenced by a lack of significant effects on assessed bone growth parameters. In the 250- and 2,500-fold HDE dosage groups, MPA was shown to exert an embryotoxic effect inducing 48 and 100% resorptions respectively. Mean embryo weights/litter were significantly reduced by MPA exposure at 250 X the HDE. Intrauterine exposure to 250 X the MPA HDE induced reductions in humeral and femoral diaphyseal length in proportion to a reduction in overall growth. The data demonstrate that MPA, administered at dosages of up to several orders of magnitude in excess of the HDE and which permitted embryo survival, did not induce increases in the frequency of nongenital teratogenesis at any dose or gestational stage. Importantly, limb reduction defects were not noted even in instances where the dosage of MPA induced an inhibition of endochondral bone growth.

Animals↗