Monophasic dose-response curves of betamethasone on geometric and mechanical properties of femur diaphyses in growing rats.

The biomechanical repercussion of the corticoid-induced osteopenia (a severe consequence of long-term glucocorticoid therapy) was studied in cortical bone of small rodents. Growing rats receiving 12.5-3200 micrograms/kg/d of betamethasone (BMS) s.c. for 20 days suffered a log-dose related impairment in body weight gain and in mechanical (fracture load, bending stiffness) and cross-sectional properties (area, moment of inertia) of femur diaphyses. No changes in bone material properties (ability to stand stress, elastic modulus, energy absorption per unit volume) were observed. At variance with the biphasic dose-response curves (positive effects at low-medium doses, negative at high doses) previously obtained with cortisol in a similar model, only negative effects on every variable studied were observed in this experiment. Results suggest that BMS effects on cortical bone biomechanics derived mainly or completely from those induced on bone geometry (biomechanical correlate of corticoid-induced osteopenia) in the assayed conditions. Data are compatible with a BMS-induced change in the setpoint of bone mechanostat. Correlation of bone geometric and biomechanical data with body weight gain showed that the anti-anabolic effects of BMS on bone were proportionally less intense than those exerted on the whole biomass.

Protective effects of disodium etidronate and pamidronate against the biomechanical repercussion of betamethasone-induced osteopenia in growing rat femurs.

To assess the protective effect of bisphosphonates on the biomechanical repercussion of glucocorticoid-induced osteopenia, intraperitoneal doses of 1 or 10 mg/kg/d of disodium etidronate or 1 or 50 mg/kg/day of pamidronate were given to groups of 6 growing rats simultaneously receiving subcutaneous doses of 4.8 mg/kg/day of betamethasone for 20 days. Betamethasone impaired strength and stiffness of femur diaphyses through a reduction of geometric properties, abnormally enhancing bone ability to absorb energy. Both bisphosphonates partially prevented betamethasone effects on diaphyseal stiffness (but not strength) through positive, dose-related effects on material modulus of elasticity and slighter improvements in diaphyseal geometry, avoiding the enhancement of energy-absorbing ability and the subsequent tendency to production of comminute fractures. These results and others obtained treating normal rats with (pamidronate) APD suggest that the sign of bisphosphonate effects on bone biomechanics may depend not only on the type of compound but also on eventual interactions with concomitant treatments.

Biphasic dose-response curves of cortisol effects on rat diaphyseal bone biomechanics.

Doses of 8, 16 (low), 32, 48, 64 (medium), and 150 (high) mg/kg/day of cortisol were administered to groups of 8 growing rats each during 16 days, and their femurs were then submitted to 3-point bending tests at low strain rate. Low doses had no effect. Medium doses, previously shown to improve calcium (Ca) balance and weight gain in the species, augmented diaphyseal elastic and ultimate strength, stiffness, and plastic-to-elastic deformation ratio with respect to untreated controls. This effect was achieved either by enhancing bone mass (volume, sectional moment of inertia, wall/lumen ration) without changes in material quality parameters (32 mg/kg/day) or, conversely, by increasing bone tissue mechanical properties (stress, modulus of elasticity) not affecting bone geometry (48 and 64 mg/kg/day). The highest dose, known to depress Ca balance and weight gain, impaired diaphyseal mechanical performance in controls by substantially reducing bone mass without major variation in bone material properties, that is, developing a true osteopenic state in mechanical terms. The energy elastically absorbed per unit volume (proportional to the risk of comminute fractures) was greater with the highest dose because of enhanced deformability and diminished bone mass. The biphasic dose-response curves obtained, grossly parallel to those previously demonstrated for metabolic actions of cortisol in the same species, showed that biomechanical repercussion of this treatment on bone depends on different, dose-dependent effects which vary independently in temporal course, intensity, and sign.

Characterization and structure of ovarian and testicular LH/hCG receptors.

Ovarian and Leydig cell LH/hCG receptors purified to homogeneity were identified as a single protein of Mr 80,000 and 90,000 respectively. The homogeneity of this protein was confirmed by microsequencing of the first 18 amino acids of the ovarian receptor. The unblocked N-terminal peptide consisted of NH2-R-E-L-S-G-S-R-X-P-E-P-X-D-X-A-P-D-G. These receptors are N-linked sialoglycoproteins which accounts for the size difference between testicular and ovarian receptors and may participate in the interaction with gonadotropin. Crosslinking of pure receptor with hCG with 125I label in either subunit indicated significant interaction of alpha-hCG with the receptor, while beta-hCG seems involved mostly through association and conformational influence on the alpha-subunit. Comparison of Mr derived from SDS with those from FPLC suggested that the native LH receptor are dimers of identical subunits. Autoradiographs of blotted receptors demonstrated that both monomeric and dimeric forms can bind hCG. Receptors from both tissues can be phosphorylated by the catalytic subunit of cAMP-dependent protein kinase and phosphopeptide maps were identical. Receptor occupancy by agonist leads to a conformational change which facilitates its phosphorylation during initial binding and reduces the rate of phosphorylation after more prolonged exposure to gonadotropin. Aggregation or dimerization of the hCG/LH receptors could promote clustering and or crosslinking of receptors in the membrane favouring the initial transduction steps in the action of these hormones.

Biomechanical performance of diaphyseal shafts and bone tissue of femurs from protein-restricted rats.

The aim of this study was to define the biomechanical repercussion of a severe protein restriction on the shaft of long bones and on cortical bone tissue. Femurs from 9 rats fed a protein-free diet from the 30th to the 50th day of age showed a great reduction of bending strength and stiffness with respect to 9 controls. These alterations correlated with severe impairment in the amount and/or the architectural arrangement of bone material (volume, wall/lumen ratio, sectional inertia) and also with reduction of Ca content and modulus of elasticity of bone tissue. No changes were observed, however, in bone elastic stress. The impairment in stiffness derived from reductions in both bone mass and modulus of elasticity led to an increment in energy absorption by bone and bone tissue which in turn induced a high incidence of comminuted fractures. The data provide a biomechanical basis for the interpretation of clinical features of nutritional osteopenia in long bones.