Does exercise modify the effects of zoledronic acid on bone mass, microarchitecture, biomechanics, and turnover in ovariectomized rats?

Regular activity has effects on bone size, shape, and density, resulting in an increase in mechanical strength. The mechanism of action that underlies this improvement in bone strength is mainly linked to an increase in bone formation. Zoledronic acid (Z), in contrast, may prevent bone strength changes in ovariectomized (OVX) rodents by its potent antiresorptive effects. Based on these assumptions we hypothesized that combined effects of exercise (E) and Z may produce higher benefits on bone changes resulting from estrogen deficiency than either intervention alone. At 6 months of age, 60 female Wistar rats were OVX or sham operated (SH) and divided into five groups: SH, OVX, OVX-E, OVX-Z, and OVX-ZE. OVX rats were treated with a single IV injection of Z (20 microg/kg) or vehicle and submitted or not to treadmill exercise (15 m/min, 60 min/day, 5 days/week) for 12 weeks. Whole-body BMD and bone turnover markers were analyzed longitudinally. At sacrifice, femurs were removed. BMD by DXA, three-point bending test, and microCT were performed to study biomechanical and trabecular structure parameters, respectively. After 12 weeks, bone volume fraction decreased in OVX rats, whereas bone turnover rate, trabecular spacing, and structure model index increased compared with those in the SH group (P < 0.05). Zoledronic acid prevented the ovariectomy-induced trabecular bone loss and its subsequent trabecular microarchitectural deterioration. Treadmill exercise running was shown to preserve the bone strength and to induce bone turnover changes in favor of bone formation. However, the combined effects of zoledronic acid and running exercise applied simultaneously did not produce any synergetic or additive effects.

Doping dose of salbutamol and exercise training: impact on the skeleton of ovariectomized rats.

Previous studies in healthy rats have demonstrated a deleterious bone impact of beta-agonist treatment. The purpose of this study was to examine the trabecular and cortical effects of beta(2)-agonists at doping dose on treadmill exercising rats with estrogen deficiency. Adult female rats were ovariectomized (OVX; n = 44) or sham operated (n = 12). Then, OVX rats received a subcutaneous injection of salbutamol (SAB) or vehicle with (EXE) or without treadmill exercise for 10 wk. Bone mineral density (BMD) was analyzed by densitometry. Microcomputed tomography and histomorphometric analysis were performed to study trabecular bone structure and bone cell activities. After 10 wk, SAB rats presented a much more marked decrease of BMD and trabecular parameters. Exercise did not change the high level of bone resorption in OVX EXE SAB compared with OVX SAB group (both on COOH-terminal collagen cross-links and osteoclast number). These results confirm the deleterious effect of beta(2)-agonists on bone quantity (femoral BMD gain: OVX EXE, +6.8%, vs. OVX EXE SAB, -1.8%; P < 0.01) and quality (-8.0% of femoral trabecular thickness in OVX EXE SAB vs. OVX EXE), indicating that SAB suppresses the effect of EXE in OVX rats. Furthermore, we notice that the slight beneficial effect of exercise was mainly localized in the tibia. These findings indicate the presence of a bone alteration threshold below which there is no more alteration in structural bone quantity and quality. The negative effects of SAB on bone observed in this study in trained rats may indicate potential complications in doping female athletes with exercise-induced amenorrhea.

Various effects of antidepressant drugs on bone microarchitectecture, mechanical properties and bone remodeling.

The aim of this study was to evaluate the effects of various drugs which present antidepressant properties: selective serotonin-reuptake inhibitors (SSRIs, fluoxetine), serotonin and noradrenaline-reuptake inhibitors (Desipramine) and phosphodiesterase inhibitors (PDE, rolipram and tofisopam) on bone microarchitecture and biomechanical properties. Twelve female mice were studied per group starting at an age of 10 weeks. During 4 weeks, they received subcutaneously either placebo or 20 mg kg(-1) day(-1) of desipramine, fluoxetine or 10 mg kg(-1) day(-1) of rolipram or tofisopam. Serum Osteocalcin and CTx were evaluated by ELISA. Bone microarchitecture of the distal femur was characterized by X-ray microCT (Skyscan1072). Mechanical properties were assessed by three-point bending test (Instron 4501) and antidepressant efficacy by forced swimming and open field tests. Fluoxetine displayed lower TbTh (-6.1%, p<0.01) and tofisopam higher TbTh (+5.0%, p<0.05) versus placebo. Rolipram and tofisopam treatments induced higher BV/TV than placebo (+23.8% and +18.3% respectively). Desipramine group had significantly higher cortical area (+4.8%, p<0.01) and fluoxetine lower cortical area (-6.1%, p<0.01) compared to placebo. The stiffness and Young's modulus were lower in the fluoxetine group (77+/-13 N mm(-1), 6431+/-1182 MPa) than in placebo (101+/-9 N mm(-1), 8441+/-1180 MPa). Bone markers indicated a significantly higher bone formation in tofisopam (+8.6%) and a lower in fluoxetine (-56.1%) compared to placebo. These data suggest deleterious effects for SSRIs, both on trabecular and cortical bone and a positive effect of PDE inhibitors on trabecular bone. Furthermore tofisopam anabolic effect in terms of bone markers, suggests a potential therapeutic effect of the PDE inhibitors on bone.

Doping dose of salbutamol and exercise: deleterious effect on cancellous and cortical bones in adult rats.

Animal studies suggest that bone remodeling is under beta-adrenergic control via the sympathetic nervous system. To our knowledge, the impact of beta-agonist substances, at doping doses, has not been studied in adult rats. The purpose of this study was to examine the effects of salbutamol injections with or without treadmill exercise on trabecular and cortical bone in adult rats. Adult (36 wk of age) female Wistar rats (n = 56) were treated with salbutamol (3 mg.kg(-1).day(-1) sc, 5 days/wk) or vehicle (sham) with or without subsequent treadmill exercise (13 m/min, 60 min/day, 5 days/wk) for 10 wk. Tibial and femoral bone mineral density was analyzed by dual-energy X-ray absorptiometry. Metaphysic trabecular bone structure was analyzed by micro-CT at the time of the animals' death. Bone cell activities were assessed histomorphometrically. After 10 wk, the increase in bone mineral density was less in salbutamol-treated than in sham rats (+3.3% vs. +12.4%, P < 0.05), and trabecular parameters were altered and bone resorption was increased in salbutamol-treated rats compared with controls. The negative effect on bone architecture in salbutamol-treated rats persisted, even with treadmill exercise. These results confirm the deleterious effect of beta(2)-agonists on bone mass during chronic treatment and describe its effects on bone mechanical properties in adult rats. Bone loss occurred independently of a salbutamol-induced anabolic effect on muscle mass and was equally severe in sedentary and exercising rats, despite a beneficial effect of exercise on the extrinsic and intrinsic energy to ultimate strain. These bone effects may have important consequences in athletes who use salbutamol as a doping substance.

Effect of specimen conditioning on the microarchitectural parameters of trabecular bone assessed by micro-computed tomography.

The best way to preserve the mechanical properties of bone specimens is hydration in NaCl, whereas the reference process in microCT analysis is defatting. However, for finite element modelling (FEM) it is necessary to use the same bone specimens for biomechanical testing and 3D imaging. This study aimed to evaluate the effect of sample conditioning on trabecular bone microarchitectural parameters. Trabecular bones were analysed by microCT under three successive conditions: first, the fatted samples were analysed immersed in NaCl (process N); second, they were hydrated for 24 h then imaged without immersion (process H); third, the samples were defatted before analysis (process D). The microarchitectural parameters bone volume/tissue volume (BV/TV), trabecular spacing (Tb.Sp), number (Tb.N) and thickness (Tb.Th) were calculated. Except for BV/TV, there was no significant difference between the processes N and D. In process H, BV/TV, Tb.Th and Tb.N were higher and BS/BV and Tb.Sp were lower than in process D. Results showed that the process D may be replaced by the process N. The process H induced significant differences in microarchitectural parameters when compared to process D. Nevertheless, this sample conditioning should be used to develop FEM when microCT images are to be acquired during compressive testing.

Effects of acute salbutamol intake during a Wingate test.

To investigate the impact of acute salbutamol intake on performance and selected hormonal and metabolic variables during supramaximal exercise, 13 recreational male athletes performed two 30-second Wingate tests after either placebo (PLA, lactose) or salbutamol (SAL, 4 mg) oral administration, according to a double-blind and randomized protocol. Blood samples collected at rest, end of the Wingate test, recovery (5, 10, 15 min) were tested for growth hormone (GH), insulin (INS), blood glucose (GLU), and lactate determination. We found the peak and mean power performed significantly increased after SAL vs. PLA (PPSAL: 896 +/- 46; PPPLA: 819 +/- 57 W; MPSAL: 585 +/- 27; MPPLA: 534 +/- 35 W, p < 0.05), whereas no change was observed in the fatigue index. Blood glucose and INS were significantly increased by SAL at rest, at the end of the Wingate test, and during the 5 first minutes of recovery (p < 0.05). Plasma GH was significantly decreased by SAL (p < 0.05) during the recovery whereas end-exercise and recovery blood lactate tended but were not significantly increased after SAL vs. PLA. From these data, acute salbutamol intake at therapeutical dosage did appear to improve peak power and mean power during a supramaximal exercise, but the mechanisms involved need further investigation.

Effects of short-term salbutamol ingestion during a Wingate test.

The effects of a chronic salbutamol intake (SAL, 12 mg/d during 3 weeks) on changes in body composition, metabolic indices and performance during a 30-second Wingate test were determined in 8 strength-trained male athletes (T) and 7 sedentary male (UT) subjects, according to a double-blind, randomized, cross-over protocol. Blood samples were collected both at rest, at the end of the test, and during passive recovery (5 min, 10 min, 15 min) for leptin (at rest), and blood lactate measurements. No significant changes in lean body mass, fat mass, and leptin were observed with SAL treatment in either group during the trial. Peak power was significantly increased (p < 0.05) after SAL intake in all subjects (T: 11.9 %; UT: 8.3 %) with a decrease in time to peak power with SAL compared to placebo (PLA) (p < 0.05). There was no change in total work performed and in fatigue indices with SAL compared to PLA. Blood lactate was significantly increased after SAL vs. PLA during the recovery (p < 0.05) in all subjects. The data demonstrate that the chronic administration of therapeutic levels of salbutamol increases maximal anaerobic power in man, irrespective of the subjects' training status. This study also rules out any implication of an anabolic effect in this improvement in performance during supramaximal exercise. Further studies are necessary to clarify the mechanisms involved.