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Biomedical subjects

E H Burger

Publications and source records attributed to E H Burger.

At least 73 records · Page 4Linked to original sources

Gene expression and immunolocalisation of amelogenins in developing embryonic and neonatal hamster teeth.

Amelogenins are a group of related matrix proteins, synthesised and secreted by ameloblasts during the formation of dental enamel. We have examined expression patterns and the tissue distribution of amelogenins by in situ hybridisation and by immunohistochemistry of developing teeth of embryonic (E12-E15) and neonatal (1- to 4-day-old) golden hamsters. Amelogenin expression and (intracellular) immunostaining for amelogenins were first observed in late embryonic stages in E14 incisors and E15 first molars in partially polarised pre-ameloblasts located along a thin layer of predentine before any overt deposition of enamel. Expression of mRNA and protein staining for amelogenins increased with age and early pre-dentine became immunopositive. The highest mRNA levels and substantial immunostaining for amelogenins were noted in neonatal-stage secretory ameloblasts fully engaged in enamel matrix deposition. After completion of the secretory phase, amelogenin gene expression continued at a lower level in post-secretory stages and was seen in transition-phase and maturation-phase ameloblasts. No amelogenin transcripts were observed in odontoblasts at any stage of their development. However, young odontoblasts stained weakly with anti-amelogenin antibodies before they formed the first layer of dentine, although this staining disappeared in odontoblasts at later stages of development. We conclude that amelogenin gene transcription occurs as early as the polarisation stage of pre-ameloblasts and is closely followed by translation of mRNA into amelogenin proteins. Odontoblasts do not transcribe the amelogenin gene and probably endocytose and digest amelogenins from the pre-dentine. Amelogenins are also transcribed but at a low level in post-secretory stages of amelogenesis.

Aging↗

Spatiotemporal expression of the homeobox gene S8 during mouse tooth development.

The murine S8 gene encodes a nuclear homeodomain containing transcription factor that is expressed at sites of epithelial-mesenchymal interactions, including those in cranofacial tissues. The spatiotemporal expression of S8 mRNA was examined in tooth primordia by in situ hybridization. S8 transcripts were found in all stages of tooth development in 13- to 16.5-day-old mouse embryos (E13-E16.5), covering the early bud stage up to the late bell stage. S8 mRNA was found exclusively in the ectomesenchyme and its derivatives that originate from the neural crest: future pulp cells, odontoblast precursors and dental follicle cells. Expression was highest at the late cap and early bud stages and declined at the mid-bell stage, in both first molar and incisor primordia. In E13 jaw explants grown in organ culture for 48 h, S8 mRNA was still present in first and second molar primordia after culture. At E15.5, S8 mRNA was also transiently present in the surrounding osteogenic tissue. It is concluded that the distribution pattern of S8 mRNA during tooth development indicates a role for the gene in defining the identity of dental papilla and follicle cells. It is speculated that the time-restricted expression of S8 in tooth primordia involves establishing the definitive form of the tooth organ.

Animals↗

Orientation of the trabecular pattern of the distal radius around the menopause.

The trabecular pattern depicted on radiographs of the distal radius of women around the menopause is analysed by a new method to measure orientation. It is shown that the trabecular pattern on radiographs of the distal radius is aligned preferentially along the axial direction and to a lesser extent along the transverse direction. Two minima of orientation are found at 60 degrees off the axial direction. A significant correlation is found between the average value of the orientation in the radius and the average lumbar bone mineral density. It is known that the bone mineral content decreases around the menopause. Previous reports on the present images describe several features that were found to be related to bone mineral content and to time. The orientation of the trabecular pattern, however, is found to be independent of time, at least during the two years of the investigation. Therefore, it is concluded that the orientation of the radiographic trabecular pattern tends to remain stable even if some features of the pattern evolve. Previous studies on radiographs of the radius of children and elderly women and men provide evidence for the tendency of orientation to remain stable not only in women around the menopause, but also in men and women during the entire adult life.

Analysis of Variance↗

Pulsating fluid flow stimulates prostaglandin release and inducible prostaglandin G/H synthase mRNA expression in primary mouse bone cells.

Bone tissue responds to mechanical stress with adaptive changes in mass and structure. Mechanical stress produces flow of fluid in the osteocyte lacunar-canalicular network, which is likely the physiological signal for bone cell adaptive responses. We examined the effects of 1 h pulsating fluid flow (PFF; 0.7 +/- 0.02 Pa, 5 Hz) on prostaglandin (PG) E2, PGI2, and PGF2alpha production and on the expression of the constitutive and inducible prostaglandin G/H synthases, PGHS-1, and PGHS-2, the major enzymes in the conversion of arachidonic acid to prostaglandins, using mouse calvarial bone cell cultures. PFF treatment stimulated the release of all three prostaglandins under 2% serum conditions, but with a different time course and to a different extent. PGF2alpha was rapidly increased 5-10 minutes after the onset of PFF. PGE2 release increased somewhat more slowly (significant after 10 minutes), but continued throughout 60 minutes of treatment. The response of PGI2 was the slowest, and only significant after 30 and 60 minutes of treatment. In addition, PFF induced the expression of PGHS-2 but not PGHS-1. One hour of PFF treatment increased PGHS-2 mRNA expression about 2-fold relative to the induction by 2% fresh serum given at the start of PFF. When the addition of fresh serum was reduced to 0.1%, the induction of PGHS-2 was 8- to 9-fold in PFF-treated cells relative to controls. This up-regulation continued for at least 1 h after PFF removal. PFF also markedly increased PGHS activity, measured as the conversion of arachidonic acid into PGE2. One hour after PFF removal, the production of all three prostaglandins was still enhanced. These results suggest that prostaglandins are important early mediators of the response of bone cells to mechanical stress. Prostaglandin up-regulation is associated with an induction of PGHS-2 enzyme mRNA, which may subsequently provide a means for amplifying the cellular response to mechanical stress.

Analysis of Variance↗

OP-1 (BMP-7) affects mRNA expression of type I, II, X collagen, and matrix Gla protein in ossifying long bones in vitro.

In long bone development, a regulating role of OP-1 is suggested by the local correlated expression of both OP-1 ligand and OP-1 binding receptors in developing mouse hind limbs. OP-1 is expressed in the interdigital mesenchyme, whereas OP-1 binding receptors are found in the bordering perichondrium, and both OP-1 ligand and receptors are present in the zone of (pre)hypertrophic chondrocytes. We investigated the role of OP-1 in long bone development experimentally by treating organ cultures of embryonic mouse metatarsals with rhOP-1. The mRNA expression patterns of type I, II, X collagen, and matrix Gla protein (MGP) were studied using in situ hybridization and cell proliferation using [3H]thymidine and BrdU labeling. In the epiphyseal perichondrium, treatment with 40 ng/ml OP-1 enhanced cell proliferation after day 2, while 6-day treatment caused a shift in expression from type I collagen to type II collagen mRNA. This supports previous histochemical findings that OP-1 induced the transition of perichondrium into cartilage. In the center of the rudiment, OP-1 inhibited the expression of type X collagen mRNA, indicating inhibition of chondrocyte hypertrophy. An arrest of differentiation at the (pre)hypertrophic chondrocyte stage was also indicated by the large area of cells expressing MGP mRNA in the OP-1-treated rudiments. We conclude that OP-1 affected the expression of marker genes of chondrocyte differentiation by acting on two steps in endochondral ossification. First, cell proliferation was enhanced, particularly so in the perichondrium where cells started to express the chondrocyte phenotype. Second, the terminal differentiation of mature chondrocytes into hypertrophic chondrocytes was inhibited. These results, combined with the known pattern of OP-1 ligand and BMP receptor expression in the embryo, suggest that OP-1 plays a local role in the cascade of events during endochondral ossification.

Animals↗

Pulsating fluid flow increases prostaglandin production by cultured chicken osteocytes--a cytoskeleton-dependent process.

It has been postulated that the transduction of mechanical stress signals to bone cells occurs via loading-dependent flow of interstitial fluid through the lacuno-canalicular network of bone. We have shown earlier that chicken osteocytes release enhanced amounts of prostaglandin E2 after 1 h treatment with pulsating fluid flow (PFF, 0.5 +/- 0.02 Pa, 5 Hz). Here we study the acute response to PFF on three cell populations derived from fetal chick calvariae, namely periosteal fibroblasts (PF), an osteoblast and osteocyte containing population (OBmix), and osteocytes (OCY), and the involvement of the actin-cytoskeleton in this process. All three cell populations rapidly (OCY: within 5 min, OBmix, PF: within 10 min) increased their release of prostaglandins E2 and I2 in response to PFF, but the response by OCY was 2-4 times higher than that by OBmix or PF. Disruption of the actin-cytoskeleton by cytochalasin B completely abolished the response. We conclude that osteocytes are more sensitive to fluid shear stress than immature bone cells, and that the actin-cytoskeleton is involved in the response to fluid flow.

6-Ketoprostaglandin F1 alpha↗

Effect of (poly)-L-lactic acid on the proliferation and differentiation of primary bone cells in vitro.

A previous study has shown bone formation around poly-L-lactic acid (PLLA) wire in vivo. However, it is still unknown how bone cells are stimulated to form bone around PLLA wire. The effect of PLLA on primary bone cells in vitro is the subject of this study. Osteoprogenitor and osteoblastic cells derived from neonatal mouse calvaria were cultured after addition of PLLA wire or L-lactide to the culture medium. Alkaline phosphatase (AP) activity, as a parameter of bone cell differentiation, and DNA content, to assess cell growth were measured. In osteoblast-enriched cell cultures PLLA wire did not affect DNA content, but AP activity was increased by 28%. In osteoprogenitor-enriched cell cultures PLLA wire decreased DNA content by 10%, but AP activity of the cells was not affected. L-Lactide enhanced the DNA content of osteoblastic cell cultures at 0.1 mM by 19%, but not at higher concentrations, and did not affect cell differentiation. In osteoprogenitor cell cultures, L-lactide had no effect at all. These results indicate that the proliferation and differentiation of bone cells in vitro can be modulated by PLLA, dependent on their stage of differentiation. L-Lactide at concentrations up to 10 mM did not adversely affect cell growth or differentiation, while PLLA wire upregulated the expression of the osteoblastic phenotype. These data support the observation of newly formed bone around poly-lactic acid wire in vivo in other the studies.

Alkaline Phosphatase↗

Prostaglandin mediated modulation of transforming growth factor-beta metabolism in primary mouse osteoblastic cells in vitro.

Prostaglandins and transforming growth factor-beta (TGF-beta) are both important local regulators of bone metabolism, but their actions on bone are complex. Prostaglandins mediate bone loss due to immobilization, but prostaglandin E2 (PGE2) treatment stimulates bone formation in vivo. TGF-beta may have both anabolic and catabolic effects on bone in vitro. In this study, we tested the effects of PGE2 on TGF-beta release and on TGF-beta messenger RNA (mRNA) levels in neonatal mouse calvarial cell cultures. We also examined the relationship between endogenous prostaglandin production as a result of mechanical stress and the release of TGF-beta. Addition of PGE2 (10(-8)-10(-6)M) to the culture medium stimulated the release of TGF-beta peptide (active plus latent) after 24 and 48 h in a dose-related manner. This upregulation was paralleled by an increased expression of TGF-beta mRNA levels. Mechanical stimulation by 1 h treatment with pulsating fluid flow (producing a shear stress of 0.5 +/- 0.02 Pa at 5 Hz) resulted 1 h posttreatment in increased production of PGE2, prostaglandin l2 (PGI2), and prostaglandin F2a. In addition, the release of TGF-beta activity but not TGF-beta peptide was decreased 24 h after PFF treatment. Addition of indomethacin, which blocks endogenous prostaglandin production, neutralized the effect of PFF treatment on TGF-beta activity, indicating that the effect of stress was mediated by endogenous prostaglandins. These results suggest that PGE2 and other prostaglandins (probably PGI2 and/or PGF2a) have opposite effects on TGF-beta metabolism in bone cells, as PGE2 upregulates TGF-beta expression and synthesis while other prostaglandins downregulate TGF-beta activation.

Animals↗

1,25-dihydroxyvitamin D3-mediated transforming growth factor-beta release is impaired in cultured osteoblasts from patients with multiple pituitary hormone deficiencies.

To evaluate the osteoblastic function in patients with multiple pituitary hormone deficiencies (M-PHD) and with isolated growth hormone deficiency (I-GHD), bone cells were cultured and the effects of 10(-8) M 1,25-dihydroxyvitamin D3 (1,25[OH]2D3) on parameters of cell proliferation, osteoblastic differentiation, and local paracrine regulation were measured. Three days of 1,25(OH)2D3 treatment increased alkaline phosphatase activity and osteocalcin release but inhibited [3H]thymidine incorporation in all cell cultures from patients as well as from controls. In addition, 1,25(OH)2D3 increased the release of both total and active transforming growth factor-beta (TGF-beta) in bone cells from controls by, respectively, 4.9- and 3.2-fold and in bone cells from I-GHD by 5.1- and 1.5-fold, respectively. However, in bone cells from M-PHD, the stimulation of total TGF-beta release was significantly lower (1.3-fold) than in control and I-GHD cells, and active TGF-beta release was not stimulated at all. One year of supplementation with human growth hormone did not improve this deficient TGF-beta release in bone cells from M-PHD. We conclude that cultured bone cells from I-GHD and M-PHD show a normal response to 1,25(OH)2D3 regarding cell proliferation and osteoblastic differentiation, which implicates a normal 1,25(OH)2D3-receptor function. In cells from controls and I-GHD, 1,25(OH)2D3 enhanced both total and active TGF-beta release. However, bone cells from M-PHD showed a deficient TGF-beta response to 1,25(OH)2D3. These results suggest that the regulation of TGF-beta production is a major paracrine factor involved in hypopituitarism.

Adult↗

DNA fragmentation during bone formation in neonatal rodents assessed by transferase-mediated end labeling.

To study the fate of bone cells, we used the transferase-mediated, biotin-dUTP nick end-labeling (TUNEL) assay to detect DNA fragmentation during the formation of intramembranous and endochondral bone in newly born hamsters, mice, and rats. In alveolar bone forming around the developing tooth crowns, DNA fragmentation was found in three cell types: TRAP-negative mononuclear cells at the bone surface, osteocytes, and some but not all nuclei of TRAP-positive osteoclasts. Osteoblasts did not undergo DNA fragmentation. A strong positive correlation was found between contacts of TUNEL-positive osteocytes and osteoclasts. Extracellular bone matrix also stained occasionally for the presence of DNA fragments. During endochondral bone formation, TUNEL staining was detected in late hypertrophic chondrocytes of the epiphyseal growth plate. During rapid longitudinal growth of long bones, TUNEL-positive hypertrophic chondrocytes were found coincident with or slightly after invasion of blood vessels from the diaphysis. However, during slow longitudinal growth and in secondary ossification centers, DNA fragmentation was seen in hypertrophic chondrocytes still located within their lacunae. We conclude that some of the osteocytes in deeper layers of bone die within their lacuna and disperse nuclear fragments over the extracellular matrix, that a majority of the osteocytes are phagocytosed and degraded by osteoclasts at sites of intense bone resorption, and that during endochondral ossification, substantial numbers of late hypertrophic chondrocyte cells undergo cell death.

Acid Phosphatase↗

The influence of transforming growth factor beta 1 on the development of embryonic mouse long bones.

Transforming growth factor beta (TGF-beta) is an important regulator of bone metabolism, and is found in large quantities in embryonic and adult bone tissue. The influence of TGF-beta 1 on chondro-osteogenesis was studied. In organ cultures of developing long bone rudiments of embryonic mice, growth and development of the various cartilaginous and osseous compartments were investigated by morphometric analysis and autoradiography after [3H]-thymidine labelling. TGF-beta 1 (1 ng/ml) inhibited both chondrogenesis and osteogenesis, and also inhibited matrix calcification. The effect was greatest in cell populations with the highest proliferation rate. It was noticed that the bone collar formation was inhibited. This may be due to an inhibition of osteoblast proliferation or differentiation, but it seems more likely to be an inhibition of the manufacture of matrix substance. These data suggest that TGF-beta 1 may be an important regulator of embryonic bone development.

Animals↗

Pulsating fluid flow increases nitric oxide (NO) synthesis by osteocytes but not periosteal fibroblasts--correlation with prostaglandin upregulation.

Osteocytes are extremely sensitive to fluid shear stress, a phenomenon that may be related to mechanical adaptation of bone (FASEB J 9:441,1995). Here we examined the effect of pulsating fluid flow (PFF, 0.5 +/- 0.02 Pa, 5 Hz, 0.4 Pa/sec) on the release of NO, in relation with upregulation of prostaglandin E2 (PGE2). Chicken calvarial osteocytes, but not periosteal fibroblasts, as well as mouse calvarial cells responded to PFF with a rapid and transient 2 to 3-fold stimulation of NO release. The effect was maximal after 5 min and leveled off thereafter. PFF also stimulated PGE2 release. This effect was significant after 10 min and continued throughout 60 min PFF treatment. Inhibition of NO release by NG-monomethyl-L-arginine prevented the effect of PFF on NO as well as PGE2 release. These results suggest that NO is a mediator of mechanical effects in bone, leading to enhanced PGE2 release. They further strengthen the hypothesis that fluid flow through the osteocyte canalicular network provides the physical stimulus for mechanosensation in bone.

Animals↗

Decreased mineralization and increased calcium release in isolated fetal mouse long bones under near weightlessness.

Mechanical loading plays an important role in the development and maintenance of skeletal tissues. Subnormal mechanical stress as a result of bed rest, immobilization, but also in spaceflight, results in a decreased bone mass and disuse osteoporosis, whereas supranormal loads upon extremities result in an increased bone mass. In this first in vitro experiment with complete fetal mouse cartilaginous long bones, cultured under microgravity conditions, we studied growth, glucose utilization, collagen synthesis, and mineral metabolism, during a 4-day culture period in space. There was no change in percent length increase and collagen synthesis under microgravity compared with in-flight 1x gravity. Glucose utilization and mineralization were decreased under microgravity. In addition, mineral resorption, as measured by 45Ca release, was increased. These data suggest that weightlessness has modulating effects on skeletal tissue cells. Loss of bone during spaceflight could be the result of both impaired mineralization as well as increased resorption.

Animals↗

Mechanical loading stimulates the release of transforming growth factor-beta activity by cultured mouse calvariae and periosteal cells.

We have shown earlier that mechanical stimulation by intermittent hydrostatic compression (IHC) inhibits bone resorption and stimulates bone formation in cultured fetal mouse calvariae (Klein-Nulend et al., 1986, Arthritis Rheum., 29: 1002-1009). The production of soluble bone factors by such calvariae is also modified (Klein-Nulend et al., 1993, Cell Tissue Res., 271:513-517). Transforming growth factor-beta (TGF-beta) is an important local regulator of bone metabolism and is produced by osteoblasts. In this study, the release of TGF-beta activity as a result of mechanical stress was examined in organ cultures of neonatal mouse calvariae, in primary cultures of calvariae-derived osteoprogenitor (OPR) cells, and in more differentiated osteoblastic (OB) cells. Whole calvariae and calvariae-derived cells were cultured in the presence or absence of IHC for 1-7 days and medium concentrations of active as well as total TGF-beta were measured using a bioassay. IHC (maximum 13 kPa, maximal pressure rate 32.5 kPa/sec) was generated by intermittently (0.3 Hz) compressing the gas phase above the cultures. We found that mechanical loading by IHC stimulated the release of TGF-beta activity from cultured calvariae by twofold after 1 day. IHC also stimulated the release of TGF-beta activity from calvariae-derived cells after 1 and 3 days. The absolute amounts of TGF-beta activity released were lower in OPR cells than in OB cells, but the stimulatory effect of IHC was greater in OPR cells. Total TGF-beta (active and bound) released into the medium was not affected by IHC. IHC did not change the dry weight of the organ cultures, nor the DNA or protein content of the cell cultures. These data show that mechanical perturbation of bone cells, particularly OPR cells, enhances the activation of released TGF-beta. We conclude that modulation of TGF-beta metabolism may be part of the response of bone tissue to mechanical stress.

Animals↗

Mechanical stimulation by intermittent hydrostatic compression promotes bone-specific gene expression in vitro.

In a previous study of the cellular mechanism underlaying Wolff's law we showed that mechanical stimulation by intermittent hydrostatic compression (IHC) increases bone formation in cultured fetal mouse calvariae compared to non-stimulated cultures. To test whether mechanical stimuli may modulate bone-specific gene expression, we studied the effect of IHC on alkaline phosphatase (AP) expression and enzyme activity as well as collagen and actin mRNA levels in neonatal mouse calvariae and calvarial bone cells. Two cell populations, one resembling osteoprogenitor (OPR) cells and another resembling osteoblasts (OB) were obtained from calvariae by sequential digestion. IHC was applied by intermittently (0.3 Hz) compressing the gas- phase of a closed culture chamber (peak stress 13kPa, peak stress rate 32.5 kPas-1). In control cultures of calvariae as well as OB and OPR cells, AP activity and AP-, collagen-, and actin-mRNA levels all decreased after one or more days, with the exception of OPR cell collagen expression which increased during culture. IHC treatment upregulated AP, collagen and actin expression and AP activity in calvariae and OB cells, but decreased collagen expression in OPR cells. These results suggest that treatment with IHC promotes the osteoblastic phenotype in bone organ cultures and in osteoblasts. Osteoprogenitor cells seem to react somewhat differently to mechanical stress than osteoblasts. The loss of bone-specific gene expression under control culture conditions, in the absence of mechanical stimuli, suggests that the mechanical environment is important in maintaining the differentiated phenotype of bone cells, and that IHC treatment partially restores this environment in bone cell- and organ cultures.

Actins↗

Intermittent compression stimulates cartilage mineralization.

The effects of intermittent hydrostatic compressive force (ICF; 13 kPa applied at 0.3 Hz frequency), as a substitute for moderate loading in vivo, on ossifying bone organ cultures, were evaluated by means of (histo)-morphometry. In earlier studies, biochemical tests have shown an increased 45Ca intake and an increased alkaline phosphatase activity in bone organ cultures that received ICF, suggesting that ICF promoted matrix mineralization. The purpose of this study was to examine whether an effect of ICF on mineralization can be described by means of histomorphometrical analysis. Fetal mouse metatarsal bone rudiments were cultured for 5 days in serum-free medium, with (experimental) or without (control) ICF. Linear measurements taken during culture demonstrated that the dark zone in the center of the rudiment, representing mineralized hypertrophic cartilage, became significantly longer in the group that received ICF when compared with the control group. This finding was in conformation with the former studies. Histological sections of the rudiments, stained with Goldner's trichrome method were used to study changes at the cellular level and to describe the position and relative amount of mineralizing cartilage matrix (defined as Goldner-positive matrix [GPM]). Histomorphometry demonstrated that ICF treatment significantly increased the length of the hypertrophic cartilaginous zone and enhanced the amount of GPM between the mineralizing hypertrophic chondrocytes. However, the total length of the zone containing GPM was not increased, nor was the future bone collar, consisting of a thin osteoid seam, lengthened by ICF. These data indicate that the cellular processes involved in chrondrocyte hypertrophy were accelerated by ICF, as well as the extracellular processes leading to matrix mineralization. The study supports the earlier conclusion that embryonic bone rudiments are sensitive to mechanical stimulation and that moderate loading promotes their ossification in vitro.

Analysis of Variance↗

Longitudinal analysis of radiographic trabecular pattern by image processing.

To describe structural and textural changes in bone structure, the radiographic trabecular pattern of children, aged 4-14 years, was examined using a digital image processing system. This investigation is based on data from the Nijmegen Growth Study, a mixed-longitudinal growth study comprised of three birth cohorts, which were observed for five years. Of 3075 left hand-forearm radiographs of 426 children, a standard area of 10 x 10 mm of the radius was digitized and the trabecular pattern was described by ten image features. It is demonstrated that all image features show significant changes during the observation period (multivariate analysis of variance of p < 0.01). The age of the children correlates significantly with the image features (Spearman's Rho = 0.4; p < 0.05). Because of cohort effects, the three cohorts were studied separately. Highest correlations between age and trabecular pattern were shown in the cohort of the youngest children. The changes of the image features during the observation period seems to indicate that the fine trabecular pattern of young children changes into a coarser adult pattern. It is concluded that digital quantification of the radiographic trabecular pattern is appropriate to describe changes in trabecular bone.

Adolescent↗

Function of osteocytes in bone--their role in mechanotransduction.

Although osteocytes are by far the most abundant cell type of bone, they are least understood in terms of function and regulation. Previous studies have concentrated on their possible role as mobilizers of bone calcium, via the process of osteocytic osteolysis. Currently, however, their possible involvement in mechanical adaptation, the process whereby bone tissue maintains maximal functional strength with minimal bone mass, is discussed. We have recently obtained experimental evidence that osteocytes are the mechanosensory cells of bone, involved in the transduction of mechanical loads into biochemical signals. Our results support the hypothesis that flow of fluid through the lacunar-canalicular system as a result of loading provides the physical signal that activates the cells.

Adaptation, Physiological↗