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Cellular and molecular events during embryonic bone development.

The cellular events underlying the processes of bone maintenance, remodeling, and repair have their basis in the embryonic production of bone. Although it has been suggested that the cartilaginous model which resides in the center of developing limbs provides the scaffolding for new bone formation, our detailed morphological and histochemical studies indicate otherwise. In particular, there exists a layer of four to six cells, referred to as Stacked Cells, which surrounds a prechondrogenic core of undifferentiated cells in the limb. These Stacked Cells give rise to the lineage of osteogenic cells responsible for the formation of all structural bone. This bone is fabricated outside of the cartilage core, and the core is not replaced by bone, but rather, is replaced by marrow and vascular elements. Several generalizations arise from analysis of the cellular and molecular events of embryonic osteogenesis, and they are as follows: 1) osteogenesis and chondrogenesis are independent events which are programmed early in development, 2) bone formation is a vascular driven phenomenon which is characterized by the directional nature of osteoid secretion, and 3) the fundamental events of bone formation are consistent in chick, rat, and human long bones.

Animals

Long-term effects of tissue expansion on cranial and skeletal bone development in neonatal miniature swine: clinical findings and histomorphometric correlates.

Progressive tissue expansion induces significant gross, histologic, and bony changes in skulls and long bones of neonatal miniature swine. These bony changes consist of erosion underlying tissue expanders, with bony lipping and bone deposition at the periphery of the expander. Cranial suture lines underneath expanders appear effaced and convoluted. Serial CT scans reveal decreased bone thickness and volume (p less than 0.02) but identical bone density (p = 0.60) beneath expanders. Increased bone volume and thickness occur at the periphery of expanders (p less than 0.02). Bone density (CT number) is unaffected by tissue expansion in both cranial and long bones. These findings have histomorphometric correlates: Osteoclastic bone resorption occurs underneath expanders with periosteal reaction at the periphery of expanders. Cranial sutures are similarly affected, but no cranial synostosis results. No changes to the inner table of the skull or stigmata of increased intracranial pressure were observed either in CT scans or in behavioral changes in long-term animals. The pathophysiology of bony changes is a remodeling effect, not one of simple pressure deformation. Increased bone resorption and complete inhibition of bone formation occur until the pressure is removed. Cranial bone is significantly more affected than long bone. After removal of the expanders, reparative bone remodeling begins within 5 days and nearly complete healing of the cranial defects occurs within 2 months (p less than 0.02). No plagiocephaly results despite early coronal suture changes. On the basis of this study, we conclude that tissue expansion causes significant but reversible effects, readily monitored by high-resolution CT scans, on neonatal and infant cranial and long bones.

Animals

Comparison of the effects of cassava (Manihot esculenta Crantz) organic cyanide and inorganic cyanide on muscle and bone development in a Nigerian breed of dog.

Effects of cassava (Manihot esculenta Crantz)-borne organic cyanide and inorganic cyanide in the form of sodium cyanide on bone and muscle development were investigated in eighteen dogs of Nigerian breed. After 16 weeks of stabilization in the laboratory from the time of purchase when the dogs were fed on the same diet, they were randomly assigned to three experimental groups of six dogs each. The control group was fed on rice while the other two groups were fed on either cassava (gari) or rice plus cyanide. The three diets were made isoenergetic and isonitrogenous by varying the quantity of meat incorporated into them. The results obtained after 14 weeks of feeding the respective diets indicated that there was retardation of muscle development in the gari-fed dogs. This may have resulted from gluconeogenesis from muscle protein associated with suppression of production of insulin by the pancreas in this group. The results indicated also that the effects of inorganic dietary cyanides on muscle development were different. Both forms of dietary cyanides, however, had no adverse effect on bone development.

Animals

Vascularization and endochondral bone development: changes in plasminogen activator activity.

Changes in plasminogen activator activity were studied during the sequential developmental stages of matrix-induced cartilage, bone, and bone marrow development. The morphological transitions were correlated with biochemical parameters. Morphologic evidence of vascularization of calcified hypertrophic cartilage was accompanied by a concomitant rise in plasminogen activator activity. Thereafter, a steady decline during mineralization and deposition of new bone was observed. Maximal plasminogen activator activity occurs at approximately the same time as peak activity of alkaline and acid phosphatase. These results imply a role for plasminogen activator during angiogenesis, vascular invasion, and attendant bone differentiation.

Animals

[Effect of vitamin D over-dosage on the tooth and bone development of rabbits].

The paper investigated the changes of rabbit's tooth and jaw tissues by vitamin-D poisoning experiment. The results showed the dentin, periodontium, jaw bone and long bone of rabbit were all changed. X-ray showed subperiosteum absorbation and periosteum reaction. The mineral content of the rabbit's epiphysis and long bone diaphysis in experiment group were lower than that of control group. The histopathological findings were irregular hyperplasia of dentin, arrange disturbance of periodontal fibers, absorbation of the alveolar bone and mal-ossification of long bones. The above changes were more significant during 30-45 days of rabbit poisoning, after 60 days of poisoning the above signs were gradually released.

Animals

Abnormal bone development and lameness associated with secondary copper deficiency in young cattle.

Lameness in a group of 5- to 12-month-old calves was found to be clinically, radiographically, and pathologically associated with abnormal bone development of the distal growth plates of the metacarpus and metatarsus. Copper concentrations in serum and liver were low. Serum calcium, phosphorus, and alkaline phosphatase values were normal. In pasture forage samples, sulfate, zinc, and molybdenum concentrations were high, whereas copper, calcium, and phosphorus concentrations were normal. Red blood cell (RBC) counts and hemoglobin values were within normal limits. Radiographic findings included a widened zone of cartilage and lipping of the medial and lateral areas of the physeal plate. Histologic findings included focal widenings of the growth plate consisting of tongues of uncalcified cartilage.

Animal Feed

Characterization of a cDNA for chicken osteopontin: expression during bone development, osteoblast differentiation, and tissue distribution.

The chicken bone phosphoprotein (approximately 66-kDa BPP) is a major noncollagenous component of bone and is the major phosphoprotein synthesized by cultured chicken embryo osteoblasts [Gotoh, Y., Gerstenfeld, L. C., & Glimcher, M. J. (1990) Eur. J. Biochem. 87, 49-58]. A cDNA clone for this protein was isolated from an expression library made from embryonic chicken bone mRNA. The complete primary protein sequence of 264 amino acids was deduced from the cDNA sequence inclusive of a 16 amino acid signal peptide sequence and terminated by 4 in-frame stop sequences. A sequence alignment indicated an approximate 35% overall similarity in protein sequence between the avian approximately 66-kDa BPP and the mammalian protein osteopontin, while at the nucleotide level 60% similarity was observed. Features of this sequence which showed the greatest similarity to mammalian osteopontin included a region in which seven of nine consecutive residues are aspartic acid, a recognition sequence for integrin-mediated cell binding (-Arg-Gly-Asp), and four possible recognition sequences for phosphorylation by casein kinase II. Hybridization analysis indicated a message of 1.5 kb found predominantly in bone and kidney. The mRNA was inducible in phorbol ester treated primary cultures of chondrocytes which show no expression under normal growth conditions. A temporal induction was seen during osteoblastic differentiation both in vivo and in vitro, thus suggesting that regulation of the approximately 66-kDa BPP is under transcriptional control during osteoblast development. In summary, both the protein's primary structure and its biological features suggest that it is the avian homologue to mammalian protein osteopontin.

Amino Acid Sequence

Effects of dietary amino acid levels on bone development in broiler chickens.

Studies with several species suggest that dietary protein may influence bone calcification. Six pens of six male broilers in two consecutive trials were fed from 1 to 21 days either .5 or 1.0% Ca in diets with 80, 90, 100, 110, or 120% of the amino acid (AA) standards suggested by Thomas et al. in 1986 at a constant .46% nonphytate P. The BW and feed conversion rate (FCR) were determined, and bone measurements were taken. Both AA and Ca levels influenced BW with an interaction of the two factors. The Ca level, but not AA level, influenced FCR. The lower Ca level inhibited feed intake. Increasing AA levels reduced bone ash but did not affect tibia breaking strength. There was an interaction of AA and Ca on tibia ash, tibia weight, and tibia length. The higher AA levels supported normal linear growth of the tibia but decreased rate of calcification, especially in the diets with .5% Ca. In diets with marginal Ca levels, higher AA levels may reduce bone calcification.

Amino Acids

Purification and molecular cloning of a novel group of BMPs and localization of BMP mRNA in developing bone.

Using a novel purification scheme, we have characterized BMP as a 30kD fraction. After reduction, which destroys biological activity, this fraction was shown to be comprised of individual polypeptides with molecular weights of 30kD, 18kD and 16kD. Molecular cloning of these polypeptides resulted in the identification of 4 previously undescribed genes (BMP-1, BMP-2A, BMP-2B, and BMP-3) each of which is capable of directing de novo cartilage formation in vivo. While BMP-1 appears to be unrelated to other known growth factors, the derived amino acid sequence of BMP-2A, 2B, and 3 indicate that they are new members of the TGFb gene family. BMP-1, 2A and 2B are expressed in rat embryos during morphogenesis and can be localized by in situ hybridization to developing limb buds. BMP-3 localizes to neural ectoderm and later on in development to newly forming periosteum. Comparisons to other members of the TGFb family suggest that these newly identified BMPs are involved in pattern formation during early skeletal development.

Amino Acid Sequence

Long bone development in extrinsic fetal akinesia: an experimental study in rat fetuses subjected to oligohydramnios.

The transverse growth of long bones during intrauterine development was studied in rat fetuses subjected to experimental oligohydramnios in order to determine whether the skeletal changes, if any, in extrinsic fetal akinesia were similar to those observed in curarized rat fetuses with the fetal akinesia deformation sequence. Oligohydramnios was induced by daily extraction of amniotic fluid from day 17 of gestation until term. Experimental fetuses were compared with a sham-operated control group. The total area and perimeter, the absolute and relative amount of periosteum and bone trabeculae, the major and minor axes, and the elongation factor were measured in histological cross sections of the femoral metaphysis and diaphysis with an IBAS 1 image analysis system. Rat fetuses in the experimental group showed multiple articular contractures, redundant skin, and lung hypoplasia, a phenotype consistent with the oligohydramnios sequence. No alterations in femoral shape and transverse growth of the metaphysis and diaphysis were noted in these fetuses. These results suggest that the main mechanical factor related to fetal bone modeling is muscular strength, while motion would be mainly involved in fetal joint development.

Animals