Bone Growth Rate Research Articles

Accelerated hypertrophic chondrocyte kinetics in GDF‐7 deficient murine tibial growth plates

The Growth/Differentiation Factors (GDFs) are a subgroup of the Bone Morphogenetic Proteins (BMPs) well known for their role in joint formation and chondrogenesis. Mice deficient in one of these signaling molecules, GDF-5, have recently been shown to exhibit a decreased rate of endochondral bone growth in the proximal tibia due to a significantly longer hypertrophic phase duration. GDF-7 is a related family member, which exhibits a high degree of sequence identity with GDF-5. The purpose of the present study was to determine whether GDF-7 deficiency also alters the endochondral bone growth rate in mice and, if so, how this is achieved. Stereologic and cell kinetic parameters in proximal tibial growth plates from 5-week-old female GDF-7 -/- mice and wild type control littermates were examined. GDF-7 deficiency resulted in a statistically significant increase in growth rate (+26%; p = 0.0084) and rate of cell loss at the chondrosseous junction (+25%; p = 0.0217). Cells from GDF-7 deficient mice also exhibited a significantly shorter hypertrophic phase duration compared to wild type controls (-27%; p = 0.0326). These data demonstrate that, in the absence of GDF-7, the rate of endochondral bone growth is affected through the modulation of hypertrophic phase duration in growth plate chondrocytes. These findings further support a growing body of evidence implicating the GDFs in the formation, maturation, and maintenance of healthy cartilage.

Disturbed Synthesis of Type II Collagen Interferes with Rate of Bone Formation and Growth and Increases Bone Resorption in Transgenic Mice

Transgenic mice carrying an internally deleted human type II collagen gene (COL2A1) were used to study bone growth and development. This mutation has previously been shown to disturb the development of collagen fibrils in articular cartilage, causing chondrodysplasia and osteoarthritis. Type II collagen expression in bones was investigated with immunohistochemistry. The development and mineralization of the skeleton and anthropometric measurements on bones were evaluated using X-rays and dynamic histomorphometry. Type II collagen was expressed in the cartilage of developing bones. The bones of transgenic mice were smaller compared with the controls. The bone mass remained almost unchanged in transgenic mice after 1 month of age, leading to differences of 47% in trabecular bone volume (P = 0.012) and 40% in trabecular thickness (P < 0.01) at the age of 3 months compared with controls. At the age of 3 months the eroded surface per bone volume was 31% greater in transgenic mice compared with controls (P < 0.05). Trabecular thickness correlated positively with body weight (R = 0.71, P < 0.001). Interestingly, body weight correlated with bone volume in control mice (R = 0.27, P < 0.01), but no correlation was observed in transgenic mice. The disturbed synthesis of cartilage-specific type II collagen in growing transgenic mice retarded bone development, increased bone resorption, and altered tissue properties. This led to a negative net bone balance and small bone size. The results support the idea that an altered synthesis of cartilage-specific molecule(s) can disturb postnatal bone development and growth.

Catch-Up Growth after Hypothyroidism Is Caused by Delayed Growth Plate Senescence

Catch-up growth is defined as a linear growth rate greater than expected for age after a period of growth inhibition. We hypothesized that catch-up growth occurs because growth-inhibiting conditions conserve the limited proliferative capacity of growth plate chondrocytes, thus slowing the normal process of growth plate senescence. When the growth-inhibiting condition resolves, the growth plates are less senescent and therefore grow more rapidly than normal for age. To test this hypothesis, we administered propylthiouracil to newborn rats for 8 wk to induce hypothyroidism and then stopped the propylthiouracil to allow catch-up growth. In untreated controls, the growth plates underwent progressive, senescent changes in multiple functional and structural characteristics. We also identified genes that showed large changes in mRNA expression in growth plate and used these changes as molecular markers of senescence. In treated animals, after stopping propylthiouracil, these functional, structural, and molecular senescent changes were delayed, compared with controls. This delayed senescence included a delayed decline in longitudinal growth rate, resulting in catch-up growth. The findings demonstrate that growth inhibition due to hypothyroidism slows the developmental program of growth plate senescence, including the normal decline in the rate of longitudinal bone growth, thus accounting for catch-up growth.

Measurement of rest and activity in newborn lambs using actigraphy: studies in term and preterm lambs

The present study used actigraphy to monitor rest-activity cycles in lambs. We employed an Actiwatch Activity Monitor, which was secured on the lamb's neck in 13 term lambs and six preterm lambs. Activity measurements began on the day of delivery and lasted for 7.3+/-0.7 days. All lambs exhibited bouts of activity, lasting from approximately 2 to 60 min, separated by periods of inactivity of about equal duration. There was a progressive increase in the frequency and intensity of activity bouts with age, and a decrease in duration. In relation to postnatal age, preterm lambs had a significantly lower frequency and intensity of activity bouts compared with term lambs and significantly longer mean active bout duration. However, in relation to post-conceptual age, preterm animals were less active at birth, but thereafter the trajectory for activity development was steeper compared with the term lambs. These differences between term and preterm lambs may be due to several factors including differences in: (1) the lengths of time the two groups spent in utero and as neonates as a proportion of the perinatal period, which could influence the rate of muscle and bone growth; (2) prenatal and postnatal hormonal profiles; and (3) maternal care. We also found differences in postnatal motility in male and female lambs, with the trajectory of activity increasing in males at Days 4-5, which could be due, in part at least, to sex differences in both prenatal and postnatal hormonal profiles.

Bone modeling response to voluntary exercise in the hindlimb of mice

The functional adaptation of juvenile mammalian limb bone to mechanical loading is necessary to maintain bone strength. Diaphyseal size and shape are modified during growth through the process of bone modeling. Although bone modeling is a well-documented response to increased mechanical stress on growing diaphyseal bone, the effect of proximodistal location on bone modeling remains unclear. Distal limb elements in cursorial mammals are longer and thinner, most likely to conserve energy during locomotion because they require less energy to move. Therefore, distal elements are hypothesized to experience greater mechanical loading during locomotion and may be expected to exhibit a greater modeling response to exercise. In this study, histomorphometric comparisons are made between femora and tibiae of mice treated with voluntary exercise and a control group (N = 20). We find that femora of exercised mice exhibit both greater bone growth rates and growth areas than do controls (P < 0.05). The femora of exercised mice also have significantly greater cortical area, bending rigidity, and torsional rigidity (P < 0.05), although bending and torsional rigidity are comparable when standardized by bone length. Histomorphometric and cross-section geometric properties of the tibial midshaft of exercised and control mice did not differ significantly, although tibial length was significantly greater in exercised mice (P < 0.05). Femora of exercised mice were able to adapt to increased mechanical loading through increases in compressive, bending, and torsional rigidity. No such adaptations were found in the tibia. It is unclear if this is a biomechanical adaptation to greater stress in proximal elements or if distal elements are ontogenetically constrained in a tradeoff of bone strength of distal elements for bioenergetic efficiency during locomotion.

Cell fate specification during calvarial bone and suture development

In this study we have addressed the fundamental question of what cellular mechanisms control the growth of the calvarial bones and conversely, what is the fate of the sutural mesenchymal cells when calvarial bones approximate to form a suture. There is evidence that the size of the osteoprogenitor cell population determines the rate of calvarial bone growth. In calvarial cultures we reduced osteoprogenitor cell proliferation; however, we did not observe a reduction in the growth of parietal bone to the same degree. This discrepancy prompted us to study whether suture mesenchymal cells participate in the growth of the parietal bones. We found that mesenchymal cells adjacent to the osteogenic fronts of the parietal bones could differentiate towards the osteoblastic lineage and could become incorporated into the growing bone. Conversely, mid-suture mesenchymal cells did not become incorporated into the bone and remained undifferentiated. Thus mesenchymal cells have different fate depending on their position within the suture. In this study we show that continued proliferation of osteoprogenitors in the osteogenic fronts is the main mechanism for calvarial bone growth, but importantly, we show that suture mesenchyme cells can contribute to calvarial bone growth. These findings help us understand the mechanisms of intramembranous ossification in general, which occurs not only during cranial and facial bone development but also in the surface periosteum of most bones during modeling and remodeling.

Relationships between bone growth rate, body mass and resting metabolic rate in growing amniotes: a phylogenetic approach

We explored the factors that explain the variation in resting metabolic rates (RMR) in growing amniotes by using the phylogenetic comparative method. For this, we measured raw RMR (mL O2 h−1), body mass, body mass growth rate, and periosteal bone growth rate in a sample of 44 growing individuals belonging to 13 species of amniotes. We performed variation partitioning analyses, which showed that phylogeny explains a significant fraction of the variation of mass-specific RMR (mL O2 h−1 g−1), and that the cost of growth is much higher than the cost of maintenance. Moreover, we tested the hypothesis of the independence of energy allocation, and found that maintenance metabolism and growth rates are not significantly related. Finally, we calculated the statistical parameters of the relationship between geometry-corrected RMR (mL O2 h−1 g−0.67) and bone growth rate. This relationship could potentially be used in palaeobiology to infer RMR from bone tissue samples of fossil species by assuming Amprino’s rule (according to which bone tissue types reflect bone growth rates). These estimates would be especially interesting for Mesozoic non-avian theropod dinosaurs and Permian and Triassic therapsids to investigate, respectively, the origin of avian and mammalian endothermy. © 2007 The Linnean Society of London, Biological Journal of the Linnean Society, 2007, 92, 63–76.

Administration of ghrelin to young uraemic rats increases food intake transiently, stimulates growth hormone secretion and does not improve longitudinal growth

Ghrelin administration stimulates appetite and growth hormone (GH) secretion. Whether these effects are preserved in young individuals with chronic renal failure (CRF) and their potential benefit on growth is questioned. Three experiments were performed in subtotally nephrectomized young rats (Nx). (i) Food intake was monitored in CRF rats receiving saline intraperitoneally or a ghrelin dose (30 nmol) shown to increase food intake over 2 and 24 h in rats with normal renal function. (ii) Plasma levels of GH were measured after a dose of intravenous ghrelin (3 nmol) was given to three groups of five rats each: Nx, sham-operated fed ad libitum (SAL) and sham-operated pair-fed with Nx (SPF). (iii) Growth of Nx rats treated with intraperitoneal ghrelin (3 nmol) for 7 days (Nx-Ghr) was compared with that of SAL and Nx groups receiving saline (n=8-10 per group). In CRF rats, the dose of 30 nmol of ghrelin increased food consumption for 2 h (1.3+/-0.2 g vs 0.5+/-0.2 g, P<0.05) but not 24-h food intake (12.5+/-0.6 g vs 12.2+/-0.5 g). Ghrelin (3 nmol) increased plasma levels of GH, which were maximal 10 min after injection, no differences being observed among groups (SAL: 666.2+/-104.6 ng/ml; Nx: 691.6+/-90.7 ng/ml; SPF: 577.8+/-125.4 ng/ml). Return to basal GH levels was delayed in Nx. Ghrelin did not improve body length and weight gains, longitudinal bone growth rate or food intake in the Nx-Ghr group. In young uraemic rats, ghrelin increases appetite but not 24-h food intake, stimulates GH secretion and does not improve growth.

Effect of nutrition and physical activity on bone health of children and young adults

The development and maturation of the skeletal system starts in utero. The bone-growth process starts with the addition of cells; these cells grow and incorporate materials that increase bone strength. During puberty, this process is accelerated to the degree that approximately 25% of the total bone mass is accumulated. Failure to meet the increased demand for bone-building materials during an accelerated growth period may increase fracture tendency. The greater is the bone mass at the start of puberty; the better is the final outcome at full maturity. This is in accordance with the tracking theory (keeping bone growth along a steady percentile). Nutrition and physical activity will facilitate bone accumulation from an early age. Therefore, a program for bone health that includes physical activity and nutrition should be introduced early in the prepubertal age. There are differences in the timing and rate of bone-mass accumulation of different parts of the skeleton so that measurement of one bone may not represent the whole skeleton. In addition, bone measurements using different devices cannot be compared. Therefore, until a universal standard is reached, repeated measurements of one individual should be done using the same apparatus. The choice of the apparatus should be made so that the relevant part of the skeleton is measured. Repeated measurements of bone may detect bone mass loss much earlier than the osteoporosis line is reached. This suggests that a dynamic measure of bone growth rate similar to height velocity should be adopted, since both parameters in an optimal setting follow a steady percentile and falling of the percentile in both parameters needs an etiological diagnosis.

Variation in estradiol level affects cortical bone growth in response to mechanical loading in sheep

Although mechanical loading can stimulate cortical bone growth, little is known about how individual physiology affects this response. This study demonstrates that in vivo variation in estradiol (E2) level alters osteoblast sensitivity to exercise-induced strains, affecting cortical bone responses to mechanical loading. Subadult sheep were divided into treatment groups that varied in terms of circulating E2 levels and loading (exercised and sedentary). After 45 days, periosteal cortical bone growth rates and cross-sectional properties were measured at the midshafts of hindlimb bones and compared with strain data. The results indicate significant interactions between E2 and strain. Cortical bone growth in exercised animals with elevated E2 levels was 27% greater in the femur, 6% greater in the tibia, and 14% greater in the metatarsal than in exercised animals with lower E2 levels, or sedentary animals regardless of E2 dose (P<0.05). There was also a trend toward greater resistance to deformation in the tibia, but not the metatarsal, in the exercised, high-E2 group compared to the other treatment groups. These results demonstrate that E2 plays a role in mediating skeletal responses to strain, such that physiological variation in E2 levels among individuals may lead to differential growth responses to similar mechanical loading regimes. Efforts to model the relationship between environmental strain and bone morphology should include the effects of physiological variation in hormone levels.

Update on rosacea and anti-inflammatory-dose doxycycline

Approximately 13 million individuals in the United Sates suffer from rosacea, a recurrent disease that may require long-term therapy. Topical and oral antibiotics have been used to treat rosacea; however, high-dose antibiotics or long-term, low-dose antibiotics commonly used for the treatment of rosacea flares or for rosacea maintenance therapy, respectively, can lead to the development of antibiotic-resistant organisms. The first oral medication approved by the U.S. Food and Drug Administration for the treatment of rosacea in the United States is Oracea (CollaGenex Pharmaceuticals Inc., Newtown, PA, USA). Oracea is a 40 mg capsule of doxycycline monohydrate, containing 30 mg immediate-release and 10 mg delayed-release doxycycline beads ("anti-inflammatory-dose doxycycline"). Anti-inflammatory-dose doxycycline is not an antibiotic and does not lead to the development of antibiotic-resistant organisms. Each capsule of anti-inflammatory-dose doxycycline contains a total of 40 mg of anhydrous doxycycline as 30 mg of immediate-release and 10 mg of delayed-release beads. In contrast to other oral therapies, anti-inflammatory-dose doxycycline is taken once daily, which may increase treatment compliance. The results of two phase III trials have been encouraging, leading to the recent release (summer 2006) of Oracea for the treatment of rosacea in the United States. Anti-inflammatory-dose doxycycline should not be used by individuals with known hypersensitivity to tetracyclines or increased photosensitivity, or by pregnant or nursing women (anti-inflammatory-dose doxycycline is a pregnancy category-D medication). The risk of permanent teeth discoloration and decreased bone growth rate make anti-inflammatory-dose doxycycline contraindicated in infants and children. However, when used appropriately in patients with rosacea, anti-inflammatory-dose doxycycline may help prolong the effectiveness and life span of our most precious antibiotics.

The structural architecture of adult mammalian articular cartilage evolves by a synchronized process of tissue resorption and neoformation during postnatal development

During postnatal development, mammalian articular cartilage acts as a surface growth plate for the underlying epiphyseal bone. Concomitantly, it undergoes a fundamental process of structural reorganization from an immature isotropic to a mature (adult) anisotropic architecture. However, the mechanism underlying this structural transformation is unknown. It could involve either an internal remodelling process, or complete resorption followed by tissue neoformation. The aim of this study was to establish which of these two alternative tissue reorganization mechanisms is physiologically operative. We also wished to pinpoint the articular cartilage source of the stem cells for clonal expansion and the zonal location of the chondrocyte pool with high proliferative activity. The New Zealand white rabbit served as our animal model. The analysis was confined to the high-weight-bearing (central) areas of the medial and lateral femoral condyles. After birth, the articular cartilage layer was evaluated morphologically at monthly intervals from the first to the eighth postnatal month, when this species attains skeletal maturity. The overall height of the articular cartilage layer at each juncture was measured. The growth performance of the articular cartilage layer was assessed by calcein labelling, which permitted an estimation of the daily growth rate of the epiphyseal bone and its monthly length-gain. The slowly proliferating stem-cell pool was identified immunohistochemically (after labelling with bromodeoxyuridine), and the rapidly proliferating chondrocyte population by autoradiography (after labelling with (3)H-thymidine). The growth activity of the articular cartilage layer was highest 1 month after birth. It declined precipitously between the first and third months, and ceased between the third and fourth months, when the animal enters puberty. The structural maturation of the articular cartilage layer followed a corresponding temporal trend. During the first 3 months, when the articular cartilage layer is undergoing structural reorganization, the net length-gain in the epiphyseal bone exceeded the height of the articular cartilage layer. This finding indicates that the postnatal reorganization of articular cartilage from an immature isotropic to a mature anisotropic structure is not achieved by a process of internal remodelling, but by the resorption and neoformation of all zones except the most superficial (stem-cell) one. The superficial zone was found to consist of slowly dividing stem cells with bidirectional mitotic activity. In the horizontal direction, this zone furnishes new stem cells that replenish the pool and effect a lateral expansion of the articular cartilage layer. In the vertical direction, the superficial zone supplies the rapidly dividing, transit-amplifying daughter-cell pool that feeds the transitional and upper radial zones during the postnatal growth phase of the articular cartilage layer. During postnatal development, mammalian articular cartilage fulfils a dual function, viz., it acts not only as an articulating layer but also as a surface growth plate. In the lapine model, this growth activity ceases at puberty (3-4 months of age), whereas that of the true (metaphyseal) growth plate continues until the time of skeletal maturity (8 months). Hence, the two structures are regulated independently. The structural maturation of the articular cartilage layer coincides temporally with the cessation of its growth activity--for the radial expansion and remodelling of the epiphyseal bone--and with sexual maturation. That articular cartilage is physiologically reorganized by a process of tissue resorption and neoformation, rather than by one of internal remodelling, has important implications for the functional engineering and repair of articular cartilage tissue.

Focus on Diagnosis: The Alkaline Phosphatase Level: Nuances of a Familiar Test

1. Sarah D. Corathers, MD* 1. *Chief Resident for Internal Medicine, University of Cincinnati, Cincinnati, Ohio. Dr Corathers wrote this article while a resident in internal medicine and pediatrics at the Cincinnati Children’s Medical Center Alkaline phosphatase is a commonly encountered laboratory value that is included in the panel of liver function tests. Although elevated concentrations generally are attributed to either liver or bone sources, the enzyme has been identified as a biomarker for a diverse range of diseases and physiologic processes. Elevated alkaline phosphatase may have implications ranging from indicating periodontal disease to predicting preterm labor. Functionally, alkaline phosphatase removes a phosphate group from nucleotides and proteins. As the name suggests, the enzyme works optimally at basic pH levels. It is found throughout the body in a variety of isoenzymes unique to the tissue of origin. Highest concentrations are in liver and bone, but the enzyme is present in lesser amounts in placenta, kidney, intestines, and leukocytes. Increased concentrations of alkaline phosphatase are derived from tissues that are either functionally disturbed (obstructed liver) or greatly stimulated (growing bone). Abnormally low concentrations are far less common and more likely are related to a genetic condition or nutritional deficiency. Normal levels can vary with age, sex, hormonal status, and blood type. In children, serum alkaline phosphatase concentrations are considerably higher than in adults and correlate with the rate of bone growth. In adults, values are slightly higher in men than in women, but after age 60 years, the enzyme value is equal or higher in women. Concentrations are increased during puberty and pregnancy and after menopause. Measurement of alkaline phosphatase activity in the laboratory involves the release of 1 mmol/min of phosphate from a standard compound (p-nitrophenylphosphate). The total amount of enzyme from all …

Estimating peak strains associated with fast exercise in Thoroughbred racehorses

The third metacarpal bones (MC3) of racing Thoroughbreds are a common site for bone damage. The midshaft dorsal cortex (DC) of MC3 thickens in response to fast exercise. It is not clear if it changes to a shape and size that limits the peak bone strains to a range associated with normal loading in other species. To relate the proportionate size of the DC cortex in racehorses exercising at racing speed to surface strains, and test whether the DC reached a plateau that was sustained through subsequent exposures to racing speed exercise. Standardised lateral MC3 radiographs were taken weekly for 2-5 years in 40 racehorses in race training (16-19 m/sec). DC, medulla (M), and palmar cortex (PC) thickness were measured. at 2.5 cm distal to the nutrient foramen. An index (RI) of the relative thickness of the DC was calculated for each radiograph (RI = [DC+PC]/ M multiplied by [DC/ PC]) and used to calculate strains at 12 m/sec from an equation published previously. Mean time to plateau in DC thickness was 501 days, mean increase in DC was 3.9 mm on the left and 3.7 mm on the right. Rate of change was 8.8 microm/day on the left, and 8.4 microm/day on the right during this time. In most horses the medulla width did not change between the first and last measurements so these bone growth rates reflect periosteal bone growth on the DC surface. No further significant change in DC or RI was found, once they had reached a plateau. Mean DC thickness at the plateau was 14.7 mm on the left and 14.9 mm on the right. Mean RI at the plateau was 3.7. This equated to peak microstrains at 12 m/sec of -2492 and suggests that strains much greater than 3000 microstrains occur at racing speed in most horses. Experienced racehorses are likely to be exposed to peak strains beyond the normal limit for adult mammalian bone to resist without damage and strains associated with very fast exercise may not be sufficient stimulus to induce further bone modelling in these horses. Strains occur in the bones of experienced racehorses at a higher level than normal for other mammalian bones. Hence there may be other mechanisms operating in this particular bone in racehorses that protects the bone from failure when exposed to the high strains associated with fast exercise speeds. Investigation of such mechanisms may provide an insight on how to reduce the likelihood of damage to this bone during very fast exercise.

Polymorphisms in vitamin D receptor, osteopontin, insulin‐like growth factor 1 and insulin, and their associations with bone, egg and growth traits in a layer – broiler cross in chickens

Bone strength traits in chickens are gaining importance due to economic losses and welfare concerns associated with bone fractures and other abnormalities. A chicken F2 resource population was generated from layer and broiler genetic lines, and traits relating to bone strength, egg production, egg quality and growth rate were measured in approximately 500 F2 hens. Four biological candidate genes (vitamin D receptor, VDR; insulin, INS; insulin-like growth factor 1, IGF1; and osteopontin, SPP1) were selected for investigation. Single nucleotide polymorphisms (SNPs) were identified for each candidate gene by comparing sequences between grandparent lines. Polymerase chain reaction restriction-fragment length polymorphism or SNaPshot assays were developed to genotype the F2 population and to evaluate associations between each SNP genotype and multiple phenotypes. Significant associations (P < 0.0125) were found between VDR and bone mineral content of the humerus at 35 weeks of age; between IGF1 and SPP1 and 5-week body weight; and between INS and 55-week body weight.

Can the growth factors PTHrP, Ihh and VEGF, together regulate the development of a long bone?

Endochondral ossification is the process of differentiation of cartilaginous into osseous tissue. Parathyroid hormone related protein (PTHrP), Indian hedgehog (Ihh) and vascular endothelial growth factor (VEGF), which are synthesized in different zones of the growth plate, were found to have crucial roles in regulating endochondral ossification. The aim of this study was to evaluate whether the three growth factors PTHrP, Ihh and VEGF, together, could regulate longitudinal growth in a normal human, fetal femur. For this purpose, a one-dimensional finite element (FE) model, incorporating growth factor signaling, was developed of the human, distal, femoral growth plate. It included growth factor synthesis in the relevant zones, their transport and degradation and their effects. Simulations ran from initial hypertrophy in the center of the bone until secondary ossification starts at approximately 3.5 months postnatal. For clarity, we emphasize that no mechanical stresses were considered. The FE model showed a stable growth plate in which the bone growth rate was constant and the number of cells per zone oscillated around an equilibrium. Simulations incorporating increased and decreased PTHrP and Ihh synthesis rates resulted, respectively, in more and less cells per zone and in increased and decreased bone growth rates. The FE model correctly reflected the development of a growth plate and the rate of bone growth in the femur. Simulations incorporating increased and decreased PTHrP and Ihh synthesis rates reflected growth plate pathologies and growth plates in PTHrP-/- and Ihh-/- mice. The three growth factors, PTHrP, Ihh and VEGF, could potentially together regulate tissue differentiation.

Local Simvastatin Effects on Mandibular Bone Growth and Inflammation

Simvastatin has been shown to increase bone growth when applied topically to murine bone; however, it causes considerable soft tissue inflammation at high doses (2.2 mg), making future clinical use problematic. This study evaluated the effect of lower simvastatin doses and cyclooxygenase (COX) synthase inhibitors on tissue inflammation and bone growth in rats and gene expression in mice. Adult female rats were untreated or treated with a single dose of 0.1, 0.5, 1.0, 1.5, or 2.2 mg simvastatin in methylcellulose gel in a polylactic acid membrane (SIM) on the lateral aspect of the mandible. The contralateral mandible side was implanted with methylcellulose gel/polylactic acid membrane alone (GEL), and five rats in each dose pairing were evaluated histomorphometrically after 3, 7, and 24 days. Subsequent rats were similarly treated with 0.5 mg simvastatin (optimal dose) and daily intraperitoneal injections of COX-2 inhibitor (NS-398; 1 mg/kg x 7 days; N = 16), general COX inhibitor (indomethacin; 1 mg/kg x 7 days; N = 16), or no inhibitor (N = 10) and evaluated histomorphometrically after 7 or 24 days by analysis of variance (ANOVA). Gene arrays were also used to evaluate osteogenic gene expression from 0.5 mg simvastatin in murine calvaria (N = 12). There was a 45% increase in bone area with 0.5 mg simvastatin versus gel control (P <0.001; similar to the 2.2-mg dose), and clinical swelling was reduced compared to the high simvastatin dose (P <0.05). The 0.1-mg simvastatin dose failed to stimulate significant bone growth. NS-398 and indomethacin reduced inflammation and bone growth. Simvastatin significantly upregulated procollagen, fibronectin, and matrix metalloproteinase-13 genes. Reducing the simvastatin dose from 2.2 to 0.5 mg reduced inflammation to a more clinically acceptable level without sacrificing bone-growth potential, but COX-associated inflammation appears to be necessary for in vivo bone growth.

Fabrication of a Highly Porous Bioactive Glass–Ceramic Scaffold with a High Surface Area and Strength

A bioactive glass–ceramic scaffold was fabricated, in which two interlaced three‐dimensional (3‐D) glass–ceramic walls could serve as active surfaces for the formation of a bond to bone. For its fabrication, a 3‐D graphite network acting as a template was prepared using the rapid prototyping method, and then uniformly dip coated with a melt‐derived glass slurry. The resultant sample was heat treated at 900° or 1000°C for 5 h to remove the graphite network and densify the glass–ceramic walls. The fabricated sample exhibited an ultra‐high porosity (∼95%), high compressive strength (0.2±0.03 MPa), as well as a tightly controlled pore structure. In addition, this sample had a high surface area owing to its unique structure, which was expected to enhance the rate of bone growth.

Relationship between bone growth rate and the thickness of calcified cartilage in the long bones of the Galloanserae (Aves).

The histological features of mineralized tissues can be preserved for hundreds of millions of years, and are therefore important potential sources of information for reconstructing the life history traits of extinct species. Bone growth rates and the duration of the growth period have recently been estimated in fossil archosaurs from periosteal ossification (a mechanism responsible for bone diametral growth). Similarly, data on endochondral ossification (the mechanism responsible for bone longitudinal growth) may also yield information on growth duration and rate among extinct vertebrates, as long as potentially informative structures are preserved. However, in order to carry out palaeobiological estimations of growth rate and/or the duration of growth, it is first necessary to quantify in extant species the relationship between these life history traits and the histological features of endochondral ossification that are potentially preserved in the fossil record. Here we analyse the ontogenetic variation of both bone longitudinal growth rate and the thickness of the calcified cartilage in the femora of two Galloanserae (Aves) and find a significant positive relationship between these variables in both species. We discuss possible factors underlying interspecific differences in this relationship, and conclude that it could be applied with caution to draw palaeobiological inferences.

Biochemical Markers of Bone Formation in Thai Children and Adolescents

The measurement of biochemical markers of bone turnover is essential in the study of skeletal metabolism in health and diseases. Due to variations in the rate of bone growth in different age groups and possible ethnic differences, age-specific reference ranges for biochemical markers should be established in a particular pediatric population. In this study, biochemical markers of bone formation, bone-specific alkaline phosphatase (BAP), and osteocalcin (OC) in healthy Thai children and adolescents aged 9 to 18 years were evaluated in relation to their ages and pubertal development. Serum BAP levels in boys increased with age and peaked at about 12 to 13 years. In contrast, there was a progressive decline of serum BAP levels with advancing age in girls older than 9 years. Serum OC also increased with age and reached a peak at ages 12 and 13 years in girls and boys, respectively. In addition, both serum BAP and OC levels also varied with pubertal stages. The BAP levels in boys increased sharply at pubertal stage 3 and decreased at pubertal stage 5. In girls, the BAP levels showed a fairly constant high level up to stage 3, followed by a remarkable decrease thereafter. The OC levels in boys increased sharply at pubertal stage 4 and decreased thereafter. In girls, OC started to increase at pubertal stage 3 with no subsequent changes. The levels of serum BAP and OC were higher in boys than in girls at pubertal stages 3 to 5 and at stages 2, 4, and 5, respectively. Moreover, only serum BAP level showed significant positive correlation with height velocity in both genders. In multiple regression analyses, gender, age, and pubertal stage were consistently correlated with both serum BAP and OC levels. In summary, male and female adolescents have different patterns of changes in biochemical markers of bone formation.