Effect of chronic undernutrition on body mass and mechanical bone quality under normoxic and altitude hypoxic conditions.

Both undernutrition and hypoxia exert a negative influence on both growth pattern and bone mechanical properties in developing rats. The present study explored the effects of chronic food restriction on both variables in growing rats exposed to simulated high-altitude hypoxia. Male rats (n 80) aged 28 d were divided into normoxic (Nx) and hypoxic (Hx) groups. Hx rats were exposed to hypobaric air (380 mmHg) in decompression chambers. At T0, Nx and Hx rats were subdivided into four equal subgroups: normoxic control and hypoxic controls, and normoxic growth-restricted and hypoxic growth-restricted received 80 % of the amount of food consumed freely by their respective controls for a 4-week period. Half of these animals were studied at the end of this period (T4). The remaining rats in each group continued under the same environmental conditions, but food was offered ad libitum to explore the type of catch-up growth during 8 weeks. Structural bone properties (strength and stiffness) were evaluated in the right femur midshaft by the mechanical three-point bending test; geometric properties (length, cross-sectional area, cortical mass, bending cross-sectional moment of inertia) and intrinsic properties of the bone tissue (elastic modulus) were measured or derived from appropriate equations. Bone mineralisation was assessed by ash measurement of the left femur. These data indicate that the growth-retarded effects of diminished food intake, induced either by food restriction or hypoxia-related inhibition of appetite, generated the formation of corresponding smaller bones in which subnormal structural and geometric properties were observed. However, they seemed to be appropriate to the body mass of the animals and suggest, therefore, that the bones were not osteopenic. When food restriction was imposed in Hx rats, the combined effects of both variables were additive, inducing a further reduction of bone mass and bone load-carrying capacity. In all cases, the mechanical properties of the mineralised tissue were unaffected. This and the capacity of the treated bones to undergone complete catch-up growth with full restoration of the biomechanical properties suggest that undernutrition, under either Nx or Hx conditions, does not affect bone behaviour because it remains appropriate to its mechanical functions.

Effect of physical consistency of food on the biomechanical behaviour of the mandible in the growing rat.

This study investigated the effect of a soft diet, given to growing rats, on the biomechanical behaviour of the mandible. Female rats, 30 d of age, received an ordinary diet in the form of pellets (i.e. hard-diet group), and another group of female rats received the same diet, but ground and mixed with water, forming a paste (i.e. soft-diet group). The experiment lasted 8 wk. Body-weight and body-length gains were not affected by the consistency of the diet. No significant differences were found between groups concerning the length, height, and area of the right hemimandible. Mechanical properties of the right hemimandibles were determined using a three-point bending test, in which bones were stressed on a perpendicular line immediately posterior to the posterior face of the third molar. Structural properties (load at yielding, load at fracture, structural stiffness, and elastic energy absorption) and geometric properties of the fracture section (cross-sectional area, cortical area, and moment of inertia) were significantly lower in hemimandibles of rats of the soft-diet group than in those of rats of the hard-diet group. Material properties of the mandibular bone tissue (elastic modulus and maximal elastic stress), which were estimated through appropriate equations, did not differ between groups. It was concluded that the reduced physical consistency of the diet, possibly associated with a reduced masticatory load, diminished the skeletal load-bearing capacity of the mandible in growing rats. This observed reduction in the bone structural behaviour was attributed to changes occurring at the level of bone mass and its geometrical properties because intrinsic properties of the bone material tissue were unaffected.

Growth inhibition in rats fed inadequate and incomplete proteins: repercussion on mandibular biomechanics.

This study describes the effects of feeding growing rats with a diet containing inadequate and incomplete proteins on both the morphological and the biomechanical properties of the mandible. Female rats aged 30 d were fed freely with one of two diets, control (CD, 301 Cal/100g) and experimental (ED, 359 Cal/100g). CD was a standard laboratory diet, while ED was a synthetic diet containing cornflower supplemented with vitamins and minerals. Both diets had the same physical characteristics. Control (C) and experimental (E) animals were divided into 4 groups of 10 animals each. C40 and E40 rats were fed CD and ED, respectively, for 40 d; C105 were fed the CD for 105 d; and E105 were fed the ED for 40 d and then the CD for the remainder of the experimental period (65 d). Mandibular growth was estimated directly on excised and cleaned bones by taking measurements between anatomical points. Mechanical properties of the right hemimandible were estimated by using a 3-point bending test to estimate the structural properties of the bone. Geometric properties of both the entire bone and the cross-section were determined. Bone material properties were calculated from structural and geometric properties. The left hemimandibles were ashed and the ash weight obtained. Rats fed the ED failed to achieve normal body weight gain. Complete catch-up was observed at the end of nutritional rehabilitation. Mandibular weight and length were negatively affected by the ED, as were the cross-sectional area, the mineralized cortical area, and the cross-sectional moment of inertia. All of these parameters showed incomplete catch-up. The structural bone mechanical properties indicative of strength and stiffness were negatively affected. Intrinsic material properties, as assessed by the modulus of elasticity and maximal elastic stress, were within normal values. In summary, the experimental bone was weaker than the control and structurally incompetent. The bone considered was smaller than the control bone, showing a significant reduction in the cross-sectional area and the moment of inertia. However, material properties as well as the ash fraction and degree of mineralization were similar in E and C bones. Therefore, the E bone was weaker than the C bone because of its smaller bone mass, which appears to have been negatively influenced by the ED in relation to its effects on overall body mass.

Bone mineral density and bone strength from the mandible of chronically protein restricted rats.

UNLABELLED: The present investigation was performed to assess the biomechanical repercussion of protein malnutrition imposed on rats between the 26th and 135th days of postnatal life on the mandible, which is not a weight-bearing bone but supports the loads related to the masticatory activity. Female Wistar rats aged 26 d (n = 14) were placed on either a 4%-protein diet (ICN 960254, P4 group) or a 20%-protein diet (ICN 960260, P20 group) and killed 111 d later. Both body weight and length were recorded regularly. The mandibles were dissected and cleaned of adhering soft tissue. Mandibular growth was estimated directly by taking measurements between anatomical points. Areal Bone Mineral Density (BMD) was estimated using a bone densitometer (LUNAR DPX-L). Mechanical properties of the right hemimandible were determined using a three-point bending mechanical test to obtain the load/deformation curve and estimate the structural properties of the bone. Results were summarized as means +/- SD. Comparisons between parameters were performed by Student's t test. A 75% reduction in body weight and a 32% reduction in body length were observed in P4 rats. Like body size, mandibular weight, length, height and area (index of mandibular size) were negatively affected by P4 diet, as was the posterior part of the bone (posterior to molar III). The anterior part (alveolar and incisor alveolar process) was not affected by age or diet. The "load capacity" extrinsic properties of the mandible (load fracture, stiffness, yielding point) were between 43% and 64% of control value in protein restricted rats. BMD was similar in both groups of animals. CONCLUSION: 1) Chronic protein malnutrition imposed on rats from infancy to early adulthood reduces the growth of the posterior part of the mandible without inducing changes in the anterior part, which produces some deformation of the bone in relation to age-matched rats; and 2) the significant reduction of strength and stiffness of the mandible seem to be the result of an induced loss of gain in bone structural properties as a consequence of a correlative loss of gain in both growth and mass, yet not in bone material properties.

Increased intrinsic stiffness and mineralization in femurs of adult rats after high intensity treadmill running training.

BACKGROUND: Physical activity plays a tremendous role in determining bone mechanical behavior, which is superimposed to gravidity. OBJECTIVE: Compare the geometric and material responses of the rat femur to a high intensity treadmill running training of a relatively short duration, as assessed by 3-point mechanical test. METHODS: Mature male rats (180.0 ± 30 g) were assigned (7 rats/group) to no exercise (NE) or treadmill exercise (EX). After a preconditioning period, the running speed was set at 45 cm.seg-1 during 2 wks, frequency 5 d/wk, 2-hour sessions/day. Body weight and weight of the crural quadriceps were registered at euthanasia. The right femur was mechanically tested through 3-point bending. The left femur was ashed to estimate bone mineral content. Geometric and material bone properties were estimated directly or calculated by appropriate equations. RESULTS: 1) Final body weight was 14% reduced in EX rats, while the crural quadriceps was 47% increased. Yield and fracture loads, and structural stiffness were significantly higher in the EX rats, as were the apparent elastic modulus, the bone mineral content and the degree of mineralization. Geometric properties were not affected. CONCLUSIONS: High intensity treadmill running training increases bone strength and stiffness by increasing material stiffness and mineralization, without affecting geometric bone parameters.

Synthetic erythropoietic proteins: tuning biological performance by site-specific polymer attachment.

Chemical synthesis in combination with precision polymer modification allows the systematic exploration of the effect of protein properties, such as charge and hydrodynamic radius, on potency using defined, homogeneous conjugates. A series of polymer-modified synthetic erythropoiesis proteins were constructed that had a polypeptide chain similar to the amino acid sequence of human erythropoietin but differed significantly in the number and type of attached polymers. The analogs differed in charge from +5 to -26 at neutral pH and varied in molecular weight from 30 to 54 kDa. All were active in an in vitro cell proliferation assay. However, in vivo potency was found to be strongly dependent on overall charge and size. The trends observed in this study may serve as starting points for the construction of more potent synthetic EPO analogs in the future.

Mandibular bone stiffening and increased bone calcium mass in rats permanently stunted by hypophysectomy.

OBJECTIVE: This investigation was designed to obtain information on the changes induced by hypophysectomy on biometric parameters, bone calcium mass, and material and architectural properties during ontogenesis of the rat mandible. DESIGN: Female Sprague-Dawley rats were hypophysectomised (HX) at 30 days of age. A "basal control group" (BC) was sacrificed on the same day surgery was performed. An "age-matched intact control group" (CON) was also included. HX and CON rats were sacrificed when aged 6 months. Body weight was monitored weekly. Mandibular growth was estimated directly on the right hemimandible by taking measurements between stable anatomical points. Its mechanical properties were determined using a three-point bending mechanical test. Load was applied transversely to the bone axis at a point immediately posterior to the posterior surface of the third molar. The left hemimandibles were ashed in a muffle furnace at 600 degrees C for 18h and the ash weight obtained. Calcium content in the ashes was determined by atomic absorption spectrophotometry. It was taken as the mandibular calcium mass. Histomorphometric studies were performed on decalcified hemimandibles: total interradicular bone, bone volume, and height of the periodontal ligament were measured. CONCLUSIONS: Morphometric studies indicated that hypophysectomy in juvenile rats induced mandibular growth cessation, which was limited to the posterior part of the bone. Thus, the mandible maintained its juvenile proportions and showed an important deformation relative to age. In spite of the reduced bone size, both the mandibular weight and the calcium bone mass increased more than two times in ontogenia. Histomorphometric studies revealed that the interradicular bone volume was markedly increased. These findings strongly suggest that the bone that forms the mandible of the hypophysectomised rat under the conditions of the present study showed a higher than normal density. As evidenced from biomechanical studies, these bone properties, plus the significant stiffening of bone material tissue, were presumably responsible for the unnecessary and marked increment in the "load capacity" suffered by the mandible of the hypophysectomised rat during ontogenesis.

Structural and Material Mechanical Quality of Femoral Shafts in Rats Exposed to Simulated High Altitude from Infancy to Adulthood.

The growth of the body and bone mass and the mechanical properties of appendicular bone are impaired in immature rats exposed to different simulated high altitudes (SHA) (1850-5450 m) between the 32nd and the 74th days of postnatal life. Now, we report the effects of exposure to 4100 m on the above cited variables in female rats from infancy (age: 1 month) to adulthood (age: 8 months) to define the occurrence of catch up and to establish whether the effects of altitude are transient or permanent. The ex vivo right femur was mechanically tested in three-point bending. Body weight and length, and structural (loads at yielding and fracture, and stiffness) and architectural (diaphyseal cross-sectional area, cortical area, and cross-sectional moment of inertia) properties were measured at 2, 4, 6, and 8 months of exposure to SHA. The negative influence of hypoxia on all variables was similar at different ages or, in other words, the difference among ages was maintained at any extent of hypoxia. Hypoxia did not affect the elastic modulus, thus suggesting that the mechanical properties of the bone tissue were maintained. Catch up did not occur. The resulting osteopenic bone remained appropriate to its mechanical function during the entire exposure to SHA.

Enhanced erythropoietin production during hypobaric hypoxia in mice under treatments to keep the erythrocyte mass from rising: implications for the adaptive role of polycythemia.

Stress erythropoiesis is usually considered as a compensatory effort to counteract tissue hypoxia. Its homeostatic importance in anemic hypoxia has not been questioned, but researchers, clinicians, and mountain climbers have had second thoughts on polycythemia as to its appropriateness for hypoxic or altitude hypoxia (HA). Therefore, polycythemia, one of the responses to HA seen in nongenetically adapted mammals, could or could not be considered beneficial. The present study was thus performed to obtain further information on the importance of HA polycythemia on acclimation of mice to HA. To this end, the development of polycythemia was prevented by experimental manipulations (administration of 20 mg/kg/d of the hemolytic drug phenylhydrazine or removal of 0.225 mL/d of blood), and the degree of tissue hypoxia was evaluated from plasma erythropoietin (pEPO) concentration, as determined by immunoassay, in adult female mice exposed to air maintained at 506 mbar (380 mmHg) in a simulated HA (SHA) chamber during at least 23.5 h/d for 9 d. Plasma EPO concentration in those treated hypoxic mice whose hematocrit values remained almost unchanged was between 5.55 and 7.89 times higher (depending on the experimental designs) than in control hypoxic mice allowed to develop HA polycythemia. These results, plus the finding of an inverse relationship between the hematocrit value and pEPO concentration in both the polycythemic and normocythemic SHA-exposed mice indicate that HA polycythemia is highly effective in ameliorating tissue hypoxia under SHA conditions, thus giving support to the concept of the important role of the increased hemoglobin mass in nongenetically adapted animals, whereas a left-shifted oxyhemoglobin dissociation curve confers a good degree of adaptation to HA in genetically adapted animals.

Evidence from catch-up growth and hoarding behavior of rats that exposure to hypobaric air lowers the body-mass set point.

The depression of body growth rate and the reduction of body mass for chronological age and gender in growing experimental animals exposed to hypobaric air (simulated high altitude = SHA) have been associated with hypophagia because of reduced appetite. Catch-up growth during protein recovery after a short period of protein restriction only occurs if food intake becomes super-normal, which should not be possible under hypoxic conditions if the set-point for appetite is adjusted by the level of SHA. The present investigation was designed to test the hypothesis that growth retardation during exposure to SHA is due to an alteration of the neural mechanism for setting body mass size rather than a primary alteration of the central set-point for appetite. One group of female rats aged 35 d were exposed to SHA (5460m) in a SHA chamber for 27 d (HX rats). Other group was maintained under local barometric pressure conditions (NX rats). One half of both NX and HX rats were fed a protein-free diet for the initial 9 d of the experimental period. From this time on, they were fed a diet containing 20% protein, as were the remaining rats of both groups during the entire experimental period. The growth rates of both mass and length of the body were significantly depressed in well-nourished rats exposed to SHA during the entire observation period when compared to normoxic ones. At its end, body mass and body length were 24% and 21% less in HX than in NX rats. Growth rates were negatively affected by protein restriction in both NX and HX rats. During protein recovery, they reached supernormal values in response to supernormal levels of energy intake that allowed a complete catch-up of both body mass and length. The finding that energy intake during the period of protein rehabilitation in HX rats previously stunted by protein restriction was markedly higher than in HX control ones at equal levels of hypoxia demonstrates that the degree of hypoxia does not determine directly the degree of appetite and energy intake. Furthermore, the finding that catch-up growth in the stunted HX rats returns the animal only to the stunted size appropriate for the hypoxic animal supports the hypothesis that hypoxia lowers the set-point for body mass size, which is reached by inhibition of appetite. Confirmation of the hypothesis was done by assessment of the set-point of body mass by the behavioral method of the weight threshold to hoard food. It was lowered by 17.0% in HX rats.

Effect of dexamethasone on mandibular bone biomechanics in rats during the growth phase as assessed by bending test and peripheral quantitative computerized tomography.

Long-term glucocorticoid administration to growing rats induces osteopenia and alterations in the biomechanical behavior of the bone. This study was performed to estimate the effects of dexamethasone (DTX), a synthetic steroid with predominant glucocorticoid activity, on the biomechanical properties of the mandible of rats during the growth phase, as assessed by bending test and peripheral quantitative computed tomographic (pQCT) analysis. The data obtained by the two methods will provide more precise information when analyzed together than separately. Female rats aged 23 d (n=7) received 500µg.kg-1 per day of DXT for 4 weeks. At the end of the treatment period, their body weight and body length were 51.3% and 20.6% lower, respectively, than controls. Hemimandible weight and area (an index of mandibular size) were 27.3% and 9.7% lower, respectively. The right hemimandible of each animal was subjected to a mechanical 3-point bending test. Significant weakening of the bone, as shown by a correlative impairment of strength and stiffness, was observed in experimental rats. Bone density and cross-sectional area were measured by pQCT. Cross-sectional, cortical and trabecular areas were reduced by 20% to 30% in the DTX group, as were other cortical parameters, including the bone density, mineral content and cross-sectional moment of inertia. The "bone strength index" (BSI, the product of the pQCT-assessed xCSMI and vCtBMD) was 56% lower in treated rats, which compares well with the 54% and 52% reduction observed in mandibular strength and stiffness determined through the bending test. Data suggest that the corticosteroid exerts a combined, negative action on bone geometry (mass and architecture) and volumetric bone mineral density of cortical bone, which would express independent effects on both cellular (material quality) and tissue (cross-sectional design) levels of biological organization of the skeleton in the species.

Effect of protein-energy malnutrition in early life on the dimensions and bone quality of the adult rat mandible.

Morphological and biomechanical features of the mandible are negatively affected by protein-energy malnutrition, whose effects are apparently dependent on the time of life of application. The aim here was to investigate, in neonatal rats nursed by dams put on a protein-free diet to depress milk production and thus create a state of protein-energy malnutrition in the offspring, subsequent growth and long-term effects by analyzing mandibular dimensions and bone quality in adulthood. Pregnant Wistar rats were fed a 20% protein diet (control) or a protein-free diet (malnourished) to obtain normal or subnormal milk production, respectively. After weaning, the offspring (males) were fed a 20% protein diet for 70 days. The dimensions of their excised mandibles were measured directly between anatomical points; the geometry and material quality of mandibular bone were assessed by peripheral quantitative computed tomography. Pups suckling from malnourished dams weighed 49.4% of those suckling from control dams at weaning; the actual difference between control and malnourished pups was 25.1g, which persisted until day 91 of age, indicating the absence of catch-up growth. As with body size, the mandibular base length, height and area (an index of mandibular size) were significantly smaller in malnourished than control rats at the end of the study. The mandibular cortical area, volumetric cortical bone mineral content and volumetric cortical bone mineral density assessed by peripheral quantitative computed tomography were similar in both groups of rats at the end of the observation period, but there was a significant reduction in the cortical axial moment of inertia in malnourished rats at this time of postnatal life. These findings suggest that catch-up growth was incomplete in rats malnourished during the suckling period and that the adaptation of mandibular bone architecture to body growth was apparently insufficient to attain normal values, thus not allowing complete compensation in mechanical competence at the end of the study because of an inadequate spatial distribution of resistive material through its cross-section rather than qualitative or quantitative impairment of cortical bone.

Enhanced hypoxia-stimulated erythropoietin production in mice with depression of erythropoiesis induced by hyperoxia.

Current evidence suggests that a modulatory action on O(2)-dependent EPO secretion is exerted by the erythroid/precursor cell population in the erythropoietic organs through a negative feedback system. The hypothesis is based on studies of stimulated-EPO secretion performed in mice in whom the erythropoietic rates were either enhanced or depressed in the presence of normal plasma EPO half-lives. Since erythropoietic depression was elicited by cyclophosphamide administration, which could have altered EPO production directly, the aim of the present investigation was to estimate hypoxia-stimulated EPO secretion in a mouse model of functional depressed erythropoiesis induced by exposure to normobaric hyperoxia. Females CF#1 mice aged 70 d were divided into control (C) and experimental (E) groups. The former was maintained in plastic cages in a normal environment, while the latter was placed in an environment of 60% O(2)/40% N(2) in an 85-dm(3) atmospheric chamber with air flow of 1 L/min. Erythropoiesis was evaluated by either 24-h RBC-(59)Fe uptake or iron kinetics performed 3 h after IV injection of a tracer dose of (59)Fe. Both indexes of the red cell production rate were significantly depressed in E mice. Plasma disappearance of exogenous EPO in C mice, as well as in E mice exposed to hyperoxia for 4 d, was estimated by injecting (125)I-rHuEPO intravenously. Linear regression analysis indicated that neither the differences between the slopes of both curves nor the Y-intercepts were significant. Hypobaric hypoxemia was used as stimulus for EPO production. Plasma immuno-EPO titer after a 4-h exposure to hypobaric air was 73% higher in mice with hyperoxia-induced hypoerythropoiesis than in control mice with normal erythropoiesis. Data support the concept that the rate of erythropoiesis, perhaps through the number of the erythroid progenitor/precursor cell population, modulates O(2)-dependent EPO secretion.

Failure of polycythemia-induced increase in arterial oxygen content to suppress the anorexic effect of simulated high altitude in the adult rat.

The anorexic effect of exposure to high altitude may be related to the reduction in the arterial oxygen content (Ca(O2)) induced by hypoxemia and possibly the associated decreased convective oxygen transport (COT). This study was then performed to evaluate the effects of either transfusion-induced polycythemia or previous acclimation to hypobaria with endogenously induced polycythemia on the anorexic effect of simulated high altitude (SHA) in adult female rats. Food consumption, expressed in g/d/100 g body weight, was reduced by 40% in rats exposed to 506 mbar for 4 d, as compared to control rats maintained in room air. Transfusion polycythemia, which significantly increased hematocrit, hemoglobin concentration, Ca(O2), and COT, did not change the anorexic response to the exposure to hypobaric air. Depression of food intake during exposure to SHA also occurred in rats fasted during 31 h before exposure and allowed to eat ad libitum for 2 h during exposure. Body mass loss was similar in 48-h fasted rats that were either hypoxic or normoxic. Body mass loss was similar in normoxic and hypoxic rats, the former eating the amount of food freely eaten by the latter. Hypoxia-acclimated rats with endogenously induced polycythemia taken to SHA again had diminished food intake and lost body mass at rates that were very close to those found in nonacclimated ones. Exposure to SHA also led to a decrease in food consumption, body weight, and plasma leptin in adult female mice. Analysis of data suggest that body mass loss that accompanies SHA-induced hypoxia is due to hypophagia and that experimental manipulation of the blood oxygen transport capacity cannot ameliorate it. Leptin does not appear to be an inducer of the anorexic response to hypoxia, at least in mice and rats.

The development of bone mass and bone strength in the mandible of the female rat.

The present study provides baseline data for a number of mandibular growth dimensions, specially on bone mass and bone strength, that were collected between the 21st and the 180th days of postnatal life, which are intended as a reference for researchers designing experimental studies, specially on mandibular catch-up growth, and as an aid for clinicians who must evaluate results from published animal studies for validity and potential extrapolation to the human clinical situation. Fifty weanling female Wistar (Hsd:Wi) rats were fed ad libitum a diet previously shown to allow normal, undeformed mandibular growth. Five of them were randomly selected at different times between 21 and 180 d of life. Mandibular growth was estimated directly on the right hemimandible by taking measurements between anatomical points; mandibular bone mass (calcium mass) was estimated from the mg of calcium, determined by atomic absorption spectrophotometry, present in the ashes of the left hemimandible; and mechanical properties of the right hemimandible were determined using three-point bending mechanical test. Dimensions, bone calcium mass and bone strength of the female rat mandible increased linearly from day 21 to approximately day 90. Bone growth, as expected, was more than twice when assessed from bone weight than when derived from mandibular area, length or height when the parameters were expressed as the relative increase from the mean infant condition. The growth rate of the posterior part of the mandible (behind the third molar) was almost five times greater than that of the anterior part. The rates of growth of the studied parameters showed a marked decline after day 90. ANOVA indicated that no statistical differences were found between day 90 and day 120 values. It could be concluded that the female rat mandible attains its adult size, peak bone calcium mass and bone structural mechanical properties at some point between 90 and 120 d of postnatal life. Because of the extremely high positive correlation between mandibular bone calcium mass and both mandibular area and mandibular weight, it was possible to calculate the mandibular peak bone mass from the relations 7.69 mgCa/cm2 and 0.19 mgCa/mg bone.

Gestational and early postnatal exposure to simulated high altitude does not modify postnatal body mass growth trajectory in the rat.

Postnatal hypoxia blunts body mass growth. It is also known that the quality of the fetal environment can influence the subsequent adult phenotype. The main purpose of the study was to determine whether gestational hypoxia and early postnatal hypoxia are able to blunt growth when the offspring is raised under normoxia. Hypobaric hypoxia was induced in simulated high altitude (SHA) chambers in which air was maintained at 380 mmHg (5450 m). Mature Sprague-Dawley rats of both sexes were divided in normoxic (NX) and hypoxic (HX) groups and, in the case of the HX group, maintained for 1 month at 5450 m. Mating was then allowed under NX or HX conditions. Offspring were NX-NX, NX-HX, HX-HX, or HX-NX: the first term indicates NX or HX during both gestation and the first 30 days of life; the second term indicates NX or HX during postnatal life between days 30 and 133. Body mass (g) was measured periodically and body mass growth rate (BMGR, g/d) was estimated between days 33 and 65 of postnatal life. Results can be summarized as follows: 1) BM was significantly higher in NX than in HX rats at weaning; 2) BMGR was not significantly different between NX-NX and HX-NX rats, and between HX-HX and NX-HX animals; and 3) BMGR was significantly higher in rats living under NX conditions than in those living under HX conditions during postnatal life. Data suggest that that hypobaric hypoxia during gestational and early postnatal development of rats does not alter the regulation of body mass growth in rats when compared to that seen under sea-level conditions.

Biomechanical properties of the mandible, as assessed by bending test, in rats fed a low-quality protein.

OBJECTIVE: The present study describes the effects of feeding growing rats with diets containing increasing concentrations of wheat gluten (a low quality protein, G) on both the morphometrical and the biomechanical properties of the mandible. DESIGN: Female rats were fed one of six diets containing different concentrations (5-30%) of G between the 30th and 90th days of life. Control rats were fed a diet containing 20% casein (C), which allows a normal growth and development of the bone. Mandibular growth was estimated directly on excised and cleaned bones by taking measurements between anatomical points. Mechanical properties of the right hemimandibles were determined by using a three-point bending mechanical test to obtain a load/deformation curve and estimate the structural properties of the bone. Bone material properties were calculated from structural and geometric properties. The left hemimandibles were ashed and the ash weight obtained. Calcium content was determined by atomic energy absorption. Results were summarised as means±SEM. Comparisons between parameters were performed by ANOVA and post-test. RESULTS: None of the G-fed groups could achieve a normal growth performance as compared to the C-fed control group. Like body size, age-related increments in mandibular weight, length, height and area (index of mandibular size) were negatively affected by the G diets, as was the posterior part of the bone (posterior to molar III). The cross-sectional geometry of the mandible (cross-sectional area and rectangular moment of inertia) as well as its structural properties (yielding load, fracture load, and stiffness) were also severely affected by the G diets. However, material properties (Young's modulus and maximum elastic stress) and calcium concentration in ashes and the degree of mineralisation were unaffected. CONCLUSIONS: The differences in strength and stiffness between treated and control rats seemed to be the result of an induced loss of gain in bone growth and mass, in the absence of changes in the quality of the bone mineralised material.

Mechanical testing at the whole-bone level of the femur in immature rats stunted by cornstarch consumption.

Both body weight and somatic muscle forces are the main "mechanical factors" in the determination of bone strength in the "weight-bearing bones". However, other "non-mechanical factors", such as dietary proteins, also exist, which modulate bone physiology. This study was designed to explore the mechanical behavior of the femur in post-weaning female rats stunted by feeding on cornstarch. Forty female rats aged 30 days were fed freely with one of the two diets: control (CD) and experimental (ED). CD was the standard rat laboratory diet, whereas ED was cornstarch supplemented with vitamins and minerals. Control (C) and experimental (E) animals were divided into 4 groups: C40 and E40 rats were given CD and ED, respectively, for 40 days; C105 were fed the CD for 105 days; and E40-105 were given the ED for 40 days and then the CD for the remaining experimental period (65 days). Growth of rats was assessed following Parks' model. The biomechanical structural properties of the right femur middiaphysis were estimated using a 3-point bending test. The geometric properties of both the entire bone and the cross-section were determined. The left femur was ashed and both the Ca mass and the Ca concentration were determined. Rats fed the ED failed to achieve normal weight gain. Complete catch-up was observed at the end of a 65 day period of nutritional rehabilitation. The femoral weight and length were negatively affected by the ED, as were the mid-diaphyseal cross-sectional area, the mineralized cortical area, and the cross-sectional moment of inertia. All of these parameters showed incomplete catch-up. The structural bone mechanical properties indicative of strength and stiffness were seriously negatively affected. Intrinsic material bone properties, as assessed by the modulus of elasticity and the maximal elastic stress, were within normal values. In summary, the experimental bone was weaker than the control and structurally incompetent. The considered bone was smaller than the control one, showing a significant reduction in the cross-sectional area and the moment of inertia. However, material properties as well as the ash fraction and Ca concentration were similar in E and C bones. Therefore, E bone is weaker than the C one because of its smaller bone mass, which appears to have been negatively influenced by the ED in relation to its effects on overall body mass.

Static biomechanics in bone from growing rats exposed chronically to simulated high altitudes.

Biomechanical behavior of bone is related to the amount (bone mass) and its architectural distribution, as well as the mechanical quality of bone material. This investigation reports the effects of exposure to different simulated high altitudes (SHA) (1850, 2900, 4100, and 5450 m) on femur biomechanical properties in female growing rats exposed to SHA (22-23 h/d) between the 32° and the 74° days of life. The ex vivo right femur was mechanically tested in three-point bending. The left femur was ashed at 600°C and the ash weight obtained. Final body weight and structural (loads at yielding and fracture, stiffness, and elastic energy absorption) and architectural (diaphyseal cross-sectional area, cortical area, and cross-sectional moment of inertia) were negatively affected in the animals exposed to the two highest SHA. Material properties of the mineralized tissue (Young's modulus and limit elastic stress) and the degree of mineralization were unaffected. In conclusion, hypoxic bone is weaker than normoxic one because of its smaller bone mass, which appear to have been negatively influenced by SHA in relation to its effects on overall body mass.

Biomechanical properties of the mid-shaft femur in middle-aged hypophysectomized rats as assessed by bending test.

Both stiffness and strength of bones are thought to be controlled by the "bone mechanostat". Its natural stimuli would be the strains of bone tissue (sensed by osteocytes) that are induced by both gravitational forces (body weight) and contraction of regional muscles. Body weight and muscle mass increase with age. Biomechanical performance of load-bearing bones must adapt to these growth-induced changes. Hypophysectomy in the rat slows the rate of body growth. With time, a great difference in body size is established between a hypophysectomized rat and its age-matched control, which makes it difficult to establish the real effect of pituitary ablation on bone biomechanics. The purpose of the present investigation was to compare mid-shaft femoral mechanical properties between hypophysectomized and weight-matched normal rats, which will show similar sizes and thus will be exposed to similar habitual loads. Two groups of 10 female rats each (H and C) were established. H rats were 12-month-old that had been hypophysectomized 11 months before. C rats were 2.5-month-old normals. Right femur mechanical properties were tested in 3-point bending. Structural (load-bearing capacity and stiffness), geometric (cross-sectional area, cortical sectional area, and moment of inertia), and material (modulus of elasticity and maximum elastic stress) properties were evaluated. The left femur was ashed for calcium content. Comparisons between parameters were performed by the Student's t test. Average body weight, body length, femur weight, femur length, and gastrocnemius weight were not significantly different between H and C rats. Calcium content in ashes was significantly higher in H than in C rats. Cross-sectional area, medullary area, and cross-sectional moment of inertia were higher in C rats, whereas cortical area did not differ between groups. Structural properties (diaphyseal stiffness, elastic limit, and load at fracture) were about four times higher in hypophysectomized rats, as were the bone material stiffness or Young's modulus and the maximal elastic stress (about 7×). The femur obtained from a middle-aged H rat was stronger and stiffer than the femur obtained from a young-adult C rat, both specimens showing similar size and bone mass and almost equal geometric properties. The higher than normal structural properties shown by the hypophysectomized femur were entirely due to changes in the intrinsic properties of the bone; it was thus stronger at the tissue level. The change of the femoral bone tissue was associated with a high mineral content and an unusual high modulus of elasticity and was probably due to a diminished bone and collagen turnover.