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Zebrafish admp is required to restrict the size of the organizer and to promote posterior and ventral development.

Bone morphogenetic proteins (Bmps) and their roles during early dorsoventral patterning of the vertebrate embryo are well understood. The role and regulation of a more distant member of this family, the anti-dorsalizing morphogenetic protein (Admp), however, are less clear. Here, we report the isolation and characterization of zebrafish admp. Unlike other bmps, admp is exclusively expressed on the dorsal side. Expression starts at blastula stages in the region of the organizer, giving rise to anterior neuroectoderm and axial mesoderm. During the course of gastrulation, both the neuroectodermal and the mesodermal admp transcripts vanish in an anterior-posterior wave. The maintenance of admp expression is positively influenced by Nodal signaling and by Bozozok (Boz), an organizer-promoting homeodomain protein acting as a repressor of early bmp2b expression. Despite the positive effect of boz on admp expression, Boz and Admp have rather opposite effects on zebrafish patterning, as revealed in gain- and loss-of-function experiments. Upon overexpression, admp has Bmp-like activities causing a smaller organizer and enhanced ventral specification, very similar to the phenotype caused by the loss of boz function in mutant embryos. Antisense-based admp knockdown, on the other side, leads to an enlarged organizer and impaired ventral and posterior development, as observed in embryos after boz overexpression. This finding indicates that admp is required for the development of embryonic structures normally suppressed by organizer activities. The seeming discrepancy between the regulative and functional relationship of boz and admp is discussed, and models are proposed according to which Admp might be part of a negative feedback loop to pattern and confine the organizer region.

Animals↗

Impaired development of bone mineral density during chemotherapy: a prospective analysis of 46 children newly diagnosed with cancer.

Osteopenia and osteoporosis are becoming increasingly recognized in children with cancer, though reasons for these changes are poorly understood. The purpose of the present study was to evaluate longitudinal changes in bone mineral density (BMD) and bone turnover in newly diagnosed children with a malignancy. Lumbar spine (L2-L4) and femoral neck bone mineral density (BMDareal, g/cm2) was measured by dual-energy X-ray absorptiometry in 46 children (age 2.9-16.0, median 8.0 years; 15 leukemias, 12 lymphomas, 19 solid tumors) at diagnosis, and after 6 months from the baseline. The apparent volumetric bone mineral density (BMDvol) was calculated to minimize the effect of bone size on BMD. Serum levels of osteocalcin (OC), type I collagen carboxy-terminal propeptide (PICP), and type I collagen carboxy-terminal telopeptide (ICTP) were analyzed at diagnosis, and during a 6-month follow-up. A significant decrease in lumbar BMDvol (-2.1%, p < 0.05), and in femoral BMDareal (-9.9%, p = 0.0001) and BMDvol (-8.5%, p = 0.0001) was observed after 6 months when compared with baseline measurements. The markers of bone formation (PICP, OC) were significantly decreased, and the marker of bone resorption (ICTP) was significantly increased at diagnosis as compared with normal values. By the end the follow-up, the levels of PICP and OC were normalized, whereas the level of ICTP continued to increase indicating that there was a negative balance in bone turnover. A deficient accumulation of bone mass might predispose children with a malignancy to impaired development of peak bone mass. A controlled study determining the benefits of an early intervention on bone turnover should be considered in these patients.

Adolescent↗

The muscle-bone relationship: methods and management - perspectives in glycogen storage disease.

UNLABELLED: Currently bone development is commonly presented as a process leading to the 'accumulation of peak bone mass'. Consequently, the usual approach to a suspected bone disorder in a child is to address the question are this child's bones as heavy as those of a healthy child of the same sex and age? However, from a functional perspective the aim of bone development should not be make bones as heavy as possible but to make them as stable as necessary. A functionally oriented approach should address two different questions: how strong are the bones? are they as strong as they need to be? It is clear that the bone has to be strong enough to withstand the mechanical forces to which it is exposed. CONCLUSION: since the main forces applied to bones are due to muscle action, the strength of a bone should be related to the force of the muscles attached to it.

Bone Density↗

[Compliance with gluten free diet, physical development and bone mineral status in patients with celiac disease].

OBJECTIVES: The aim of the study was the assessment of the influence of a gluten free diet on physical development and bone mineral density (BMD) in patients with celiac disease. MATERIALS & METHODS: 59 patients (40 girls, 19 boys) aged 10-20 years with celiac disease, diagnosed according to ESPGAN criteria were included in the study. Patients were divided in 3 groups: 1--strict gluten free diet, II--not entirely compliant i.e. faults in gluten free diet 1-2 times per week, III--gluten free diet not followed or frequent faults. Daily calcium (Ca) intake and physical activity was assessed. BMD of the lumbar spine L2-L4 was measured by dual-energy-X-ray absorptiometry with LUNAR DPX-IQ. Physical development was assessed by anthropometric measurements: growth and weight. RESULTS: Gluten free diet was strictly followed by 16 (27%) patients (group I), group II consisted of 23 (39%) patients. Diet was not followed by 20 (34%) patients. Thirty five (59%) patients had low Ca intake and they made up the majority of the patients in every group. High physical activity declared 35 (59%) patients. No statistically significant differences in BMD were found between group I and II. BMD was lower in group III in comparison to group I (p = 0.01) and group II (p = 0.003). BMD was higher in patients with high Ca intake (p = 0.002). Physical activity had no significant influence on BMD. There was no statistically significant difference in physical development between groups. CONCLUSIONS: Majority of the patients with celiac disease did not strictly follow gluten free diet. Poor compliance to the diet had no significant influence on physical development. BMD was lower in patients who were not compliant to the gluten free diet, occasional faults in the diet had no influence on the BMD.

Absorptiometry, Photon↗

Maternal hypothyroidism and fetal chondro-osseous development in rats.

The bone development of rat fetuses from female Wistar rats thyroidectomized prior to mating was studied, between 16 and 21 days of gestation. The parameters studied in each rat fetus were fetal weight, number of fetuses per litter, radiological data and frequency of appearance of ossification centers, diaphyseal radiological length of long bones, longitudinal ratio between epiphysis and diaphysis of long bones on cleaned and stained limbs, and volumetric tibial studies on the same limbs. Our results show a delay of fetal chondro-osseous development and a delay of weight increase in fetuses from thyroidectomized rats. After onset of fetal thyroid function, the fetal development begins a catch-up growth, which is not sufficient to completely overcome development delay prior to birth.

Animals↗

Ovine haemopoiesis: the development of bone marrow-derived colony-forming cells in vitro in the presence of factors derived from lymphoid cells and helper T-cells.

Techniques for the development of ovine bone marrow-derived haemopoietic progenitor cells and in situ identification of colony morphology are described. Both mitogen stimulated lymphoid cells and antigen stimulated helper T-cells generated potent colony-stimulating activity in conditioned medium. Monocyte/macrophage, neutrophil, eosinophil, basophil/mast cell, neutrophil/monocyte and mixed phenotype colonies developed in stimulated bone marrow cultures in a conditioned medium dose-dependent manner. Neutrophil, monocyte/macrophage and eosinophil colonies were detected in greater numbers than the other types, with mixed colonies representing only around 1% of the total. Eosinophil colonies were particularly abundant when compared to published reports of the numbers obtained with similar cultures of 'normal' mouse or human bone marrow cells. This culture technique will allow a detailed analysis of both ovine colony-stimulating factors and of the distribution of haemopoietic progenitor cells in vivo.

Animals↗

Regulation of BMP/Dpp signaling during embryonic development.

Bone morphogenetic protein-4 (BMP-4) and its Drosophila ortholog, decapentaplegic (Dpp), are multifunctional developmental regulators. Both gain-of-function and loss-of-function studies demonstrate that the biological activity and signaling range of these morphogens must be strictly regulated to ensure normal embryonic patterning. BMP-4 and Dpp are produced from inactive precursors that are proteolytically cleaved, following which the active ligand is secreted into the extracellular space. Binding of BMP-4 or Dpp to its cognate receptor leads to phosphorylation of intracellular signal-transducing Smad proteins that then form hetero-oligomers, translocate to the nucleus and modulate transcription of target genes. Recent studies have shown that the BMP signal transduction cascade can be modulated at every step of this process.

Animals↗

Cross-sectional reference data for phalangeal quantitative ultrasound from early childhood to young-adulthood according to gender, age, skeletal growth, and pubertal development.

Bone mineral status by phalangeal quantitative ultrasound (QUS, DBM Sonic, IGEA, Carpi, Modena, Italy) was examined in 3044 (1513 males and 1531 females) healthy subjects, aged 2-21 years. The aim of the study was to provide a reference database for phalangeal QUS parameters, amplitude-dependent speed of sound (AD-SoS) and bone transmission time (BTT), both expressed as centiles and Z score, according to gender, age, height, weight, body mass index (BMI), and pubertal stage to be used for estimating bone mineral status in patients with disorders of growth or of bone and mineral metabolism. In both sexes, AD-SoS and BTT increased significantly (P<0.0001) according to all the anthropometric variables. Females showed higher values than males in the age groups 9-14 for AD-SoS (P<0.04-P<0.0001) and in the age groups 11-13 for BTT (P<0.02). Males had higher BTT values than females in the age groups 6-8 and 15-21 (P<0.04-P<0.0001). AD-SoS was higher (P<0.02-P<0.0001) in females than in males at pubertal stages 2, 3, and 4, but it was higher (P=0.001) in males compared with females at pubertal stage 5. BTT was higher in males than females at pubertal stages 1 (P<0.0001), 2 (P<0.01), and 5 (P<0.0001). In both sexes, AD-SoS and BTT were significantly correlated between them (r=0.92, P<0.0001) and with all the anthropometric variables (r=0.53-r=0.85, P<0.0001). Age, weight, BMI, and pubertal stage were independent predictors of AD-SoS in males; age and pubertal stage were independent predictors of AD-SoS in females. In both sexes, height and pubertal stage, and also age only in females, were independent predictors of BTT. In conclusion, our data show that gender, age, height, and timing of sexual maturation are main determinants of bone structure and geometry, and that both these two processes may be captured by phalangeal QUS. It may be a useful tool to assess bone mineral status from early childhood to young-adulthood with a very small confounding effect related to bone sizes and without exposing the subjects to a source of radiation.

Adolescent↗

BMP signaling stimulates cellular differentiation at multiple steps during cartilage development.

Bone morphogenetic proteins (BMPs) play important roles at multiple stages of endochondral bone formation. However, the roles of BMP signaling in chondrocytes in vivo are still contentious. In the present study, we overexpressed a constitutively active BMP receptor 1A (caBmpr1a) in chondrocytes by using two systems: caBmpr1a was directly driven by a rat type II collagen promoter in a conventional transgenic system and indirectly driven in a UAS-Gal4 binary system. CaBmpr1a expression caused shortening of the columnar layer of proliferating chondrocytes and up-regulation of maturation markers, suggesting acceleration of differentiation of proliferating chondrocytes toward hypertrophic chondrocytes. In addition to the acceleration of chondrocyte differentiation, conventional transgenic mice showed widening of cartilage elements and morphological alteration of perichondrial cells, possibly due to stimulation of differentiation of prechondrogenic cells. Moreover, bigenic expression of caBmpr1a rescued the differentiation defect of prechondrogenic cells in Bmpr1b-null phalanges. This finding indicates that BMP signaling is necessary for phalangeal prechondrogenic cells to differentiate into chondrocytes and that signaling of BMP receptor 1B in this context is replaceable by that of a constitutively active BMP receptor 1A. These results suggest that BMP signaling in prechondrogenic cells and in growth plate chondrocytes stimulates their chondrocytic differentiation and maturation toward hypertrophy, respectively.

Animals↗

Epithelial Bmpr1a regulates differentiation and proliferation in postnatal hair follicles and is essential for tooth development.

Bone morphogenetic protein (BMP) signaling is thought to perform multiple functions in the regulation of skin appendage morphogenesis and the postnatal growth of hair follicles. However, definitive genetic evidence for these roles has been lacking. Here, we show that Cre-mediated mutation of the gene encoding BMP receptor 1A in the surface epithelium and its derivatives causes arrest of tooth morphogenesis and lack of external hair. The hair shaft and hair follicle inner root sheath (IRS) fail to differentiate, and expression of the known transcriptional regulators of follicular differentiation Msx1, Msx2, Foxn1 and Gata3 is markedly downregulated or absent in mutant follicles. Lef1 expression is maintained, but nuclear beta-catenin is absent from the epithelium of severely affected mutant follicles, indicating that activation of the WNT pathway lies downstream of BMPR1A signaling in postnatal follicles. Mutant hair follicles fail to undergo programmed regression, and instead continue to proliferate, producing follicular cysts and matricomas. These results provide definitive genetic evidence that epithelial Bmpr1a is required for completion of tooth morphogenesis, and regulates terminal differentiation and proliferation in postnatal hair follicles.

Animals↗

Complement proteins are present in developing endochondral bone and may mediate cartilage cell death and vascularization.

Normal endochondral bone formation follows a temporal sequence: immature or resting chondrocytes move away from the resting zone, proliferate, flatten, become arranged into columns, and finally become hypertrophic, disintegrate, and are replaced by bone. The mechanisms that guide this process are incompletely understood, but they include programmed cell death, a stage important in development and some disease processes. Using immunofluorescence we have studied the distribution of various complement proteins to examine the hypothesis that this sequence of events, particularly cell disintegration and matrix dissolution, are complement mediated. The results of these studies show that complement proteins C3 and Factor B are distributed uniformly in the resting and proliferating zones. Properdin is localized in the resting and hypertrophic zone but not in the proliferating zone. Complement proteins C5 and C9 are localized exclusively in the hypertrophic zones. This anatomically segregated pattern of distribution suggests that complement proteins may be important in cartilage-bone transformation and that the alternate pathway is involved.

Animals↗

Changes in bone density during childhood and adolescence: an approach based on bone's biological organization.

Bone densitometry has great potential to improve our understanding of bone development. However, densitometric data in children rarely are interpreted in light of the biological processes they reflect. To strengthen the link between bone densitometry and the physiology of bone development, we review the literature on physiological mechanisms and structural changes determining bone mineral density (BMD). BMD (defined as mass of mineral per unit volume) is analyzed in three levels: in bone material (BMD(material)), in a bone's trabecular and cortical tissue compartments (BMD(compartment)), and in the entire bone (BMD(total)). BMD(material) of the femoral midshaft cortex decreases after birth to a nadir in the first year of life and thereafter increases. In iliac trabecular bone, BMD(material) also increases from infancy to adulthood, reflecting the decrease in bone turnover. BMD(material) cannot be determined with current noninvasive techniques because of insufficient spatial resolution. BM(compartment) of the femoral midshaft cortex decreases in the first months after birth followed by a rapid increase during the next 2 years and slower changes thereafter, reflecting changes in both relative bone volume and BMD(material). Trabecular BMD(compartment) increases in vertebral bodies but not at the distal radius. Quantitative computed tomography (QCT) allows for the determination of both trabecular and cortical BMD(compartment), whereas projectional techniques such as dual-energy X-ray absorptiometry (DXA) can be used only to assess cortical BMD(compartment) of long bone diaphyses. BMD(total) of long bones decreases by about 30% in the first months after birth, reflecting a redistribution of bone tissue from the endocortical to the periosteal surface. In children of school age and in adolescents, changes in BMD(total) are site-specific. There is a marked rise in BMD(total) at locations where relative cortical area increases (metacarpal bones, phalanges, and forearm), but little change at the femoral neck and midshaft. BMD(total) can be measured by QCT at any site of the skeleton, regardless of bone shape. DXA allows the estimation of BMD(total) at skeletal sites, which have an approximately circular cross-section. The system presented here may help to interpret densitometric results in growing subjects on a physiological basis.

Absorptiometry, Photon↗

Recapitulation of signals regulating embryonic bone formation during postnatal growth and in fracture repair.

A number of proteins have recently been identified which play roles in regulating bone development. One important example is Indian hedgehog (Ihh) which is secreted by the prehyprtrophic chondrocytes. Ihh acts as an activator of a second secreted factor, parathyroid hormone-related protein (PTHrP), which, in turn, negatively regulates the rate of chondrocyte differentiation. Here we examine the expression of these genes and their molecular targets during different stages of bone development. In addition to regulating PTHrP expression in the perichondrium, we find evidence that Ihh may also act on the chondrocytes themselves at particular stages. As bone growth continues postnatally in mammals and the developmental process is reactivated during fracture repair, understanding the molecular basis regulating bone development is of medical relevance. We find that the same molecules that regulate embryonic endochondral ossification are also expressed during postnatal bone growth and fracture healing, suggesting that these processes are controlled by similar mechanisms.

Animals↗

Localization of the expression of types I, III, and IV collagen, TGF-beta 1 and c-fos genes in developing human calvarial bones.

Total RNA extracted from developing calvarial bones of 15- to 18-week human fetuses was studied by Northern hybridization: in addition to high levels of type I collagen mRNAs, the presence of mRNAs for type III and type IV collagen, TGF-beta and c-fos was observed. In situ hybridization of sections containing calvarial bone, overlying connective tissues, and skin was employed to identify the cells containing these mRNAs. Considerable variation was observed in the distribution of pro alpha 1(I) collagen mRNA in osteoblasts: the amount of the mRNA in cells at or near the upper surface of calvarial bone was distinctly greater than that in cells at the lower surface, indicating the direction of bone growth. High levels of type I collagen mRNAs were also detected in fibroblasts of periosteum, dura mater, and skin. Type III collagen mRNA revealed a considerably different distribution: the highest levels were detected in upper dermis, lower levels were seen in fibroblasts of the periosteum and the fibrous mesenchyme between bone spiculas, and none was seen in osteoblasts. Type IV collagen mRNAs were only observed in the endothelial cells of blood capillaries. Immunohistochemical localization of type III and IV collagens agreed well with these observations. The distribution of TGF-beta mRNA resembled that of type I collagen mRNA. In addition, high levels of TGF-beta mRNA were observed in osteoclasts of the calvarial bone. These cells, responsible for bone resorption, were also found to contain high levels of c-fos mRNA. Production of TGF-beta by osteoclasts and its activation by the acidic environment could form a link between bone resorption and new matrix formation.

Blotting, Northern↗

[Analysis of the involvement of the tumour suppressor genes TP53, p16INK4, p21WAF1, RB1 and the drugs metabolizing enzymes in the development of bone tumours in children].

BACKGROUND: Several tumor suppressor genes such as p16INK4, TP53, RB1 y p21WAF1 are involved in cell cycle regulation in response to DNA damage and belong to the complex pathway that regulates cell proliferation and/or differentiation. We have investigated the presence of mutations in those genes and polymorphisms of Drug Metabolizing Enzymes that could be involved in the development of pediatric bone tumors or in their outcome. MATERIALS AND METHODS: By means of PCR-based techniques, we have analyzed the presence of variations in the coding sequence of p16INK4, TP53, RB1 y p21WAF1 and of the Drug Metabolizing Enzymes in a group of 82 osteosarcomas and 47 Ewing's sarcomas as well as in a control group of 115 healthy children. RESULTS: We detected mutations of the TP53 gene in about 25% of the samples analyzed, most frequently in association with tumors of poor prognosis or reduced survival. The p16INK4 gene was homozygously deleted in 18% of the osteosarcomas, also associated with poor prognosis and unfavourable histologic subtypes; RB1 was altered in 21% of the osteosarcomas. We did not detect relevant associations between polymorphisms of the Drug Metabolizing Enzymes or mutation of the p21WAF1 and development of pediatric bone tumors. CONCLUSIONS: Alteration of TP53, p16INK4 and p21WAF1 seems to be involved in the development of pediatric bone tumors and to be an unfavourable prognostic factor in this type of tumors.

English Abstract↗

The effect of high or low dietary calcium on bone and calcium homeostasis in young male rats.

Young male rats (100 g body weight) were fed diets containing varying amounts of calcium. Body weight and bone development were studied together with various endocrine parameters, including blood levels of Ca2+, calcitonin, parathyroid hormone, vitamin D, and gastrin, and the enterochromaffin-like (ECL) cell-related parameters gastric mucosal histidine decarboxylase activity and histamine concentration. A diet containing 0.5% calcium resulted in optimum body weight gain and bone development. A lower calcium intake impaired body weight gain and bone development. The impairment was manifested in reduced bone calcium content whereas the size of the bones was unaffected. The net absorption of calcium seemed to be proportional to the calcium intake. A low calcium diet (0.03%) raised the circulating levels of 1,25(OH)2D and parathyroid hormone and lowered 25(OH)D3 and Ca2+, whereas a high calcium diet (5.46%) raised calcitonin, Ca2+, 25(OH)D3, and 1,25(OH)2D. In addition, the low calcium diet lowered the circulating gastrin concentration and the histidine decarboxylase activity and histamine content of the ECL cells in the gastric mucosa. A high calcium diet raised the circulating gastrin concentration, but the rise was not associated with an increase in the histidine decarboxylase activity and histamine content.

Animals↗

Relative impact of androgen and estrogen receptor activation in the effects of androgens on trabecular and cortical bone in growing male mice: a study in the androgen receptor knockout mouse model.

UNLABELLED: The relative importance of AR and ER activation has been studied in pubertal male AR knockout and WT mice after orchidectomy and androgen replacement therapy, either with or without an aromatase inhibitor. AR activation dominates normal trabecular bone development and cortical bone modeling in male mice. Moreover, optimal periosteal bone expansion is only observed in the presence of both AR and ER activation. INTRODUCTION: Androgen receptor (AR)-mediated androgen action has traditionally been considered a key determinant of male skeletal growth. Increasing evidence, however, suggests that estrogens are also essential for normal male bone growth. Therefore, the relative importance of AR-mediated and estrogen receptor (ER)-mediated androgen action after aromatization remains to be clarified. MATERIALS AND METHODS: Trabecular and cortical bone was studied in intact or orchidectomized pubertal AR knockout (ARKO) and male wildtype (WT) mice, with or without replacement therapy (3-8 weeks of age). Nonaromatizable (dihydrotestosterone [DHT]) and aromatizable (testosterone [T]) androgens and T plus an aromatase inhibitor (anastrazole) were administered to orchidectomized ARKO and WT mice. Trabecular and cortical bone modeling were evaluated by static and dynamic histomorphometry, respectively. RESULTS: AR inactivation or orchidectomy induced a similar degree of trabecular bone loss (-68% and -71%, respectively). Both DHT and T prevented orchidectomy-induced bone loss in WT mice but not in ARKO mice. Administration of an aromatase inhibitor did not affect T action on trabecular bone. AR inactivation and orchidectomy had similar negative effects on cortical thickness (-13% and -8%, respectively) and periosteal bone formation (-50% and -26%, respectively). In orchidectomized WT mice, both DHT and T were found to stimulate periosteal bone formation and, as a result, to increase cortical thickness. In contrast, the periosteum of ARKO mice remained unresponsive to either DHT or T. Interestingly, administration of an aromatase inhibitor partly reduced T action on periosteal bone formation in orchidectomized WT mice (-34% versus orchidectomized WT mice on T), but not in ARKO mice. This effect was associated with a significant decrease in serum IGF-I (-21% versus orchidectomized WT mice on T). CONCLUSIONS: These findings suggest a major role for AR activation in normal development of trabecular bone and periosteal bone growth in male mice. Moreover, optimal stimulation of periosteal growth is only obtained in the presence of both AR and ER activation.

Androgens↗

[Destruction of the alveolar bone during development of periodontal disease].

The papers surveys the mechanisms of alveolar bone destruction in the course of parodontopathy development. The results obtained in the studies carried out so far have shown that human inflamed gingiva contains biologically active substance Interleukin-1 (IL-1), tumor necrosis factor and prostaglandin E2 in concentration which are high above the minimal values that can induce in vitro the bone resorption. It has been generally understood that alveolar bone destruction is a result of the increased osteoclastic activity caused by the synchronous effect of mentioned mediators released from the structural cells and cells of inflamed infiltrate into the bone tissue encircling.

Alveolar Bone Loss↗