A comparison of the effects of two anabolic agents (fluoride and PTH) on ash density and bone strength assessed in an osteopenic rat model.

The aim of this investigation was to compare the effects of sodium fluoride (NaF) and parathyroid hormone (PTH) on ash density and strength in an osteopenic rat model. The study comprised 66 female virgin rats divided into the following 11 groups, each comprising six animals: baseline controls; baseline ovariectomized (ovx); intact controls (5 and 16 weeks), ovx controls (5 and 16 weeks); ovx-treated with PTH (0.02 mg/kg per day, 5 and 16 weeks); ovx treated with NaF (10 mg/kg per day, 5 and 16 weeks); ovx-treated with NaF (1.0 mg/kg per day, 16 weeks). Ovariectomy was performed at 12 weeks of age, 14 weeks prior to start of treatment. Ash density, bone fluoride content, and biomechanical analyses were performed on femoral cortical bone, the right femoral neck, and the sixth lumbar vertebral body. ovx had no effect on cortical bone, whereas the femoral neck displayed a significantly lower bone strength in ovx baseline animals compared with intact baseline rats (p < 0.05). Vertebral ash density was found to be significantly decreased in ovx rats after 5 and 16 weeks (p < 0.05). Treatment with fluoride had little effect on the osteopenic rat skeleton. Cortical ash density was significantly lower than ovx and intact groups in the high-dose-treated rats after 5 (p < 0.01) but not after 16 weeks. High doses of fluoride for 16 weeks induced a significant increase in maximum load and normalized strength in cortical bone when compared with intact animals (p < 0.05), but not at the other bone sites. Cortical bone strength was not different from the ovx animals at either timepoint. In fluoride-treated animals, femoral neck bone strength, vertebral body bone strength, bone quality, and ash density were found to be at about ovx levels and, in the vertebral body, significantly lower than intact animals (p < 0.05, p < 0.01). In contrast, treatment with PTH increased ash density, bone strength, and bone quality to above ovx levels (p < 0.01), and above the level of the intact animals also, although significant values were reached for cortical bone strength only (p < 0.01). Additionally, biomechanical competence and ash density measurements were significantly higher in PTH-treated rats compared with fluoride-treated rats. In conclusion, this study has shown that PTH has a highly anabolic effect and is capable of effectively restoring ovx-induced loss of bone mass and biomechanical competence. In addition, in this osteopenic rat model, PTH proved much more advantageous than treatment with fluoride, which failed to restore the ovx-induced loss of bone strength.

Parathyroid hormone monotherapy and cotherapy with antiresorptive agents restore vertebral bone mass and strength in aged ovariectomized rats.

Previous studies have shown that parathyroid hormone (PTH) monotherapy and cotherapy with estrogen or risedronate augment vertebral bone mass and bone strength in young, ovariectomized (OVX) rats. The current study was designed to determine whether PTH has similar bone anabolic effects in aged OVX rats at a much later stage of estrogen depletion. Female Sprague Dawley rats were subjected to sham surgery or bilateral ovariectomy at three months of age and maintained untreated for one year after surgery to allow for the development of vertebral osteopenia in OVX rats. Groups of baseline control and OVX rats were sacrificed at the end of this pretreatment period. The remaining OVX rats were then treated for ten weeks with vehicle, antiresorptive agents alone (estrogen, risedronate, or calcitonin), or PTH alone. Other groups of OVX rats were treated concurrently with PTH and each of the antiresorptive agents. The first and fourth lumbar vertebral bodies were processed undecalcified for quantitative bone histomorphometry and biomechanical testing, respectively. As expected, bone mass and compressive strength were decreased in the lumbar vertebral body of baseline OVX rats compared to baseline control rats. This bone loss was associated with decreases in trabecular number and width and an increase in trabecular separation. Treatment with estrogen, risedronate, or calcitonin alone failed to reverse the changes in bone mass, structure, and strength induced by ovariectomy. In contrast, treatment of OVX rats with PTH alone restored vertebral cancellous bone volume and ash density to the level of vehicle-treated control rats and increased vertebral maximum load, stress, and normalized load to well above this level.(ABSTRACT TRUNCATED AT 250 WORDS)

[Loss of trabecular bone strength and bone quality after 5 years of fluoride therapy for osteoporosis]. / Tab af trabekulaer knoglestyrke og knoglekvalitet efter fem års fluoridbehandling for osteoporose.

In order to evaluate the effect of sodium fluoride (NaF) on bone biomechanical competence, iliac crest biopsies were taken before and after one year of treatment in 12 osteoporotic patients, and before and after five years of treatment in 14 patients. Bone fluoride content had increased significantly after both one and five years of treatment, indicating that the administered fluoride had been ingested. After one year of treatment, no difference was observed in iliac crest trabecular bone ash content. A general trend for decreased bone strength and bone quality was observed, but this was insignificant. After five years of fluoride treatment, an insignificant decrease in iliac crest trabecular bone ash content was observed. A significant reduction of 45% was found in trabecular bone strength (p < 0.05), and an even more pronounced reduction of 58% was found in trabecular bone quality (p < 0.01). The results of this study indicate that long-term administration of sodium fluoride may be detrimental to bone quality, at least as measured in non-loaded iliac crest trabecular bone.

The anabolic effects of parathyroid hormone on cortical bone mass, dimensions and strength--assessed in a sexually mature, ovariectomized rat model.

The aim of the study was to determine the effect of parathyroid hormone (PTH), the antiresorptive agents estrogen and bisphosphonate (risedronate), and also the combination of PTH with these antiresorptive drugs on femoral cortical bone mass, dimensions and strength in a sexually mature, ovariectomized rat model. A total of 138, 3-month-old Sprague-Dawley rats were randomized into seven groups: 1--sham operated (control); 2--ovariectomized (OVX); 3--OVX plus estrogen; 4--OVX plus bisphosphonate (risedronate [NE]; 5--OVX plus hPTH (1-34); 6--OVX plus hPTH (1-34) and estrogen; 7--OVX plus hPTH (1-34) and risedronate. Treatment regimens were initiated 4 weeks after OVX and were continued for 5 and 15 weeks for each treatment group. Changes in bone mass (ash content), cross-sectional area, cortical thickness and dimensions and bone strength were assessed in middiaphyseal, femoral specimens. The results revealed that ovariectomy had no effect on cortical bone mass and biomechanical competence. OVX rats treated with estrogen and also OVX rats treated with risedronate showed no significant difference from either OVX or control groups. After only 5 weeks of treatment, hPTH monotherapy increased ash content, cross-sectional area, cortical thickness and compressive bone strength (load) significantly. After 15 weeks of treatment, OVX rats treated with PTH monotherapy or PTH in combination with risedronate showed identical load-values. These values were significantly higher than those seen in both control and OVX rats. However, PTH in combination with estrogen failed to augment cortical bone strength over control or OVX levels after therapy.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of fluoride on rat vertebral body biomechanical competence and bone mass.

For more than 30 years, sodium fluoride has been a commonly used therapeutic agent for established osteoporosis because of its repeatedly documented anabolic effect on trabecular bone mass. Recent clinical and experimental studies have, however, indicated a possible detrimental effect of fluoride on bone strength. Thus, the efficacy of fluoride therapy remains a controversial issue. The aim of this study was to investigate the effect of fluoride on both vertebral bone mass and quality in rats. Twenty-nine 3-month-old, female rats were randomized into three groups. One group served as a control group, and the other two groups received fluoridated water at different doses (100 ppm and 150 ppm). The rats were followed for 90 days. Three lumbar vertebrae were obtained from each rat, and changes in bone fluoride content, bone mass and biomechanical competence were assessed. The results revealed a significant increase in bone fluoride content, ash density and trabecular bone volume after fluoride treatment. Directly obtained load values and load corrected for cross-sectional area were constant. Load corrected for ash content, which is a measure of bone quality, decreased significantly after fluoride therapy. It is concluded that the increase in bone mass during fluoride treatment does not translate into an improved bone strength and that the bone quality declines. This investigation thereby supports the hypothesis of a possible negative effect of fluoride on bone quality.

Spine deformity index in osteoporotic women: relations to forearm and vertebral bone mineral measurements and to iliac crest ash density.

Bone densitometric measurements are widely used for monitoring therapeutic regimens for osteoporosis. However, it is a matter of debate which measurement site is most appropriate for prediction of individual fracture risk. The aim of this cross-sectional study was to investigate the relationship between bone mineral measurements at various sites and spine deformity index (SDI) in osteoporotic women. The SDI was determined in 37 osteoporotic women aged 56-87 years (mean 70.9 years). Peripheral (single-photon absorptiometry of the distal forearm, and iliac crest ash content) and axial (dual-photon absorptiometry of the lumbar spine) bone mass measurements were obtained. SDI increased with age (r = 0.34, p < 0.05), whereas forearm BMC (r = -0.52, p < 0.002) and forearm BMD (r = -0.62, p < 0.0001) decreased. No significant age-related changes were observed in lumbar BMC or iliac crest ash content in these osteoporotic women. A highly significant correlation was found between SDI and lumbar BMC (r = -0.60, p < 0.01). A significant, but less pronounced correlation was found between SDI and forearm BMC (r = -0.37, p < 0.05), whereas no relation was revealed between SDI and forearm BMD or iliac crest ash content. In a multiple regression model, the relationship between lumbar BMC and SDI remained significant after adjusting for the influence of age, whereas the relationship to forearm BMC disappeared. Furthermore, a multiple regression analysis was performed in order to evaluate the ability of all four bone mass measurements and age to predict variations in SDI.(ABSTRACT TRUNCATED AT 250 WORDS)

Marked decrease in trabecular bone quality after five years of sodium fluoride therapy--assessed by biomechanical testing of iliac crest bone biopsies in osteoporotic patients.

Sodium fluoride has for more than 2 decades been a commonly used therapeutic agent for established osteoporosis because of a repeatedly documented anabolic effect on trabecular bone mass. Recently, however, three controlled trials have failed to demonstrate any therapeutic advantage of NaF over placebo with respect to vertebral fracture rate. Also, there have been several reports of an increased incidence of nonvertebral fractures during fluoride administration. Thus, the efficacy of fluoride therapy remains a controversial issue. The aim of this longitudinal study was to investigate the effect of sodium fluoride (40-60 mg per day), calcium (45 mmol), and vitamin D2 (18,000 IU) on trabecular bone strength, assessed before and after 1 or 5 years of treatment for osteoporosis. Iliac crest biopsies were taken before and after 1 year of treatment in 12 patients, and before and after 5 years of treatment in 14 patients. Measurements were made of biomechanical competence, ash content, and bone fluoride content, and bone strength parameters were normalized for ash content, thereby obtaining a measure of trabecular bone quality. Bone fluoride content was significantly increased after both 1 and 5 years of treatment, indicating that the administered fluoride had been ingested. After 1 year of treatment, no difference was observed in iliac crest trabecular bone ash content. A general trend for decreased bone strength and bone quality was observed, but this was insignificant. After 5 years of fluoride treatment, an insignificant decrease in iliac crest trabecular bone ash content was observed.(ABSTRACT TRUNCATED AT 250 WORDS)

PTH has a more pronounced effect on vertebral bone mass and biomechanical competence than antiresorptive agents (estrogen and bisphosphonate)--assessed in sexually mature, ovariectomized rats.

The aim of the study was to determine the effect of PTH, the antiresorptive agents estrogen and bisphosphonate (Risedronate), and also the combination of PTH with the antiresorptive drugs on vertebral bone mass and biomechanical competence in a sexually mature, ovariectomized rat model. A total of 138 3-month-old Sprague-Dawley rats were randomized into seven groups: (1) sham operated (control); (2) ovariectomized (OVX); (3) OVX plus estrogen; (4) OVX plus bisphosphonate [Risedronate (NE)]; (5) OVX plus hPTH (1-34); (6) OVX plus hPTH (1-34) and estrogen; and, finally, (7) OVX plus hPTH (1-34) and Risedronate. Treatment regimens were initiated 4 weeks after OVX and were continued for 5 and 15 weeks for each treatment group. Changes in bone mass (ash content) and biomechanical competence were assessed on lumbar vertebral body L4. The results revealed that the Risedronate-treated OVX animals had a higher vertebral bone mass than the OVX group. hPTH (1-34) on its own had a pronounced anabolic effect and increased bone mass 20-25% and bone strength 70-80% over control levels. Neither combination therapy with estrogen nor with Risedronate provided any further advantage. The combination of PTH with Risedronate, though, seemed to allow a continued increase in both bone mass and strength during the whole treatment period.(ABSTRACT TRUNCATED AT 250 WORDS)

The effect of long-term exercise on vertebral and femoral bone mass, dimensions, and strength--assessed in a rat model.

The rat model has previously been used to test the effect of ovariectomy and of PTH administration on vertebral bone mass, size, and biomechanical competence. In this study, we used the same "biomechanical rat model" to assess the effect of long-term exercise on vertebral bone mass and quality and also on femoral bone mass, dimensions, and strength. Sixty female Fischer rats were randomized into four groups. Two groups were exercised for 5 days a week on a treadmill with a running distance of 2 km per day. The exercise program was initiated at the age of 2 months. The two exercise groups were investigated after 4 and 10 months. Two sedentary groups (observed for 4 and 10 months) served as controls. At death, three lumbar vertebral bodies (L4-L6) and the left femur were obtained from each rat, and changes in bone mass (ash density, trabecular bone volume [BV/TV]), bone size, and biomechanical competence were assessed. The results revealed an age-related (4-10 months) increase in vertebral bone mass and strength. The additional effect of exercise on the vertebral bodies was an increase in cross-sectional area and bone biomechanical competence. In the femoral bone specimens, an age-related increase in bone mass, size, and strength was also disclosed, and while exercise by itself had no significant influence on biomechanical parameters it did reduce cortical-endosteal bone resorption. The study has demonstrated an anabolic effect of a light exercise regimen on both femoral cortical bone and vertebral bodies (mainly trabecular bone).(ABSTRACT TRUNCATED AT 250 WORDS)

Long-term exercise of young and adult female rats: effect on femoral neck biomechanical competence and bone structure.

The present study was designed to examine the effect of exercise on femoral neck bone structure and strength in female rats after 4 and 10 months of exercise. Female Fischer rats aged 2 months were exercised for 4 h a day, 5 days a week on a motor-driven treadmill the speed of which was gradually increased until a daily distance of 2 km was reached. The training level was maintained for 4 months (n = 15) and 10 months (n = 15). Sedentary age-matched rats served as controls (n = 30). At death the proximal third of both femora was obtained from each rat. The left femoral neck was tested in a materials-testing machine, and the right was used for either trabecular bone mass measurement (BV/TV) or scanning electron microscopy (SEM). Biomechanical testing revealed a significant training-induced increase in femoral neck bone strength after 4 months of exercise, which although not accelerated was preserved after 10 months of exercise. Histologic investigation revealed a significant training-induced increase in BV/TV, accounted for by a significantly smaller proportion of marrow space. No difference in cortical area was found. Both histology and SEM revealed a tendency to an earlier closure of the growth line in the exercised animals. However, the exercised animals aged 6 months had a significantly increased total femoral length compared with the sedentary rats. No difference was found in total femoral length between sedentary and exercised rats aged 12 months. On the basis of this study, it is concluded that exercise has a positive effect on femoral neck bone strength.(ABSTRACT TRUNCATED AT 250 WORDS)

The positive effect of parathyroid hormone on femoral neck bone strength in ovariectomized rats is more pronounced than that of estrogen or bisphosphonates.

This study elucidates the effect of PTH, estrogen, and the bisphosphonate Risedronate (NE-58095) on femoral neck bone strength in ovariectomized (OVX) rats aged 90 days at the beginning of the investigation. Furthermore, the effects of these monotherapies were compared with those of concurrent treatment with PTH plus estrogen or PTH plus bisphosphonate. Four weeks after surgery the rats were randomized into a sham-operated vehicle-treated group, an OVX vehicle-treated group, and the various treatment groups and followed for 5 and 15 weeks. The proximal one-third of the left femur was then subjected to geometrical measurements and biomechanical testing. Neither ovariectomy nor the different treatment regimens influenced femoral neck geometry. OVX rats exhibited a decrease in femoral neck bone strength compared with control rats. This was most evident after 5 weeks. Treatment of OVX rats with Risedronate or estrogen alone tended to increase bone strength to control level, though these findings were nonsignificant. In contrast, treatment with PTH showed a highly significant increase in femoral neck biomechanical competence. Concurrent treatment with PTH plus estrogen or PTH plus Risedronate also significantly increased the femoral neck bone strength, but neither showed any advantage over treatment with PTH alone. It is concluded that treatment with PTH increases the strength of the femoral neck in estrogen-depleted rats in a highly significant manner, and that this effect is much more pronounced than the effect of the two antiresorptive agents estrogen or Risedronate. Thus, these findings provide further support for the anabolic effect of PTH and add weight to the argument for the promising potential of PTH in the treatment of postmenopausal osteoporosis.

Normal age-related changes in fluoride content of vertebral trabecular bone--relation to bone quality.

In several clinical osteoporosis studies, fluoride treatment has been shown to have a positive effect on bone mass but without a concomitant decrease in vertebral fracture rate. In contrast, some studies have shown that increases in spinal BMD are also paralleled by decreased vertebral fracture incidence. We have previously demonstrated, in a pig model, that 6-month treatment with fluoride increased bone mass but decreased bone quality. The aim of the present study was to elucidate whether normal age-related fluoride accumulation in human bone per se influences bone quality. From 73 normal individuals, aged 20-91 years (36 females, 37 males) two trabecular bone cylinders were obtained from the central part of L3. Biomechanical competence, ash density, and fluoride content were assessed in one cylinder, and trabecular bone volume was determined in the other. The results showed an age-related decrease in bone mass for both men and women. Bone strength normalized for bone mass (bone quality also identical with bone material strength) also showed an age-related decrease in men and women. Bone fluoride concentration increased significantly in both sexes (range 463-4000 ppm). Multiple regression analyses disclosed that fluoride by itself had no influence on bone quality, in this study with a limited number of cases, when the influence of sex and age were taken into account. It is concluded that normal age-related accumulation of fluoride in vertebral trabecular bone does not seem to affect the quality of bone. Whether this is also the case during fluoride therapy has to be assessed.

Evaluation of the skeletal effects of combined mild dietary calcium restriction and ovariectomy in Sinclair S-1 minipigs: a pilot study.

A pilot study was conducted to investigate the combined effects of ovariectomy (OVX) with preceding and concomitant mild dietary calcium restriction on the minipig skeleton. Minipigs 4 months old were fed diets containing 0.9, 0.75, or 0.5% calcium (Ca). At 10 months, the 0.75 and 0.5% pigs were OVX and the 0.9% were either sham operated or OVX. All pigs were maintained on their respective diets for an additional 6 months. Excised lumbar vertebrae and long bones were evaluated by densitometry and histomorphometry, and vertebral cancellous bone samples were tested biomechanically. In pigs fed the 0.9% Ca diet, OVX alone effected decreases of 6% in vertebral bone mineral density (BMD), 15% in trabecular bone volume (BV/TV), and 13% in trabecular number (Tb.N), an increase of 15% in trabecular separation (Tb.Sp), and a nonsignificant increase (p < 0.056) in vertebral cancellous final erosion depth (F.E.De) compared with the 0.9% Ca sham-operated group. Decreasing dietary Ca to 0.5% in combination with OVX effected an 8% reduction in vertebral BMD that was not associated with any significant alterations in parameters of vertebral cancellous bone microstructure or remodeling compared with the 0.9% Ca sham-operated pigs. Increases in serum PTH noted in the 0.5% Ca OVX group were generally paralleled by increases in calcitriol. In OVX pigs fed a diet containing 0.75% Ca, a 10% reduction in vertebral BMD was observed. This was associated with significant increases in F.E.De and vertebral marrow star volume (Ma.St.V) compared with the 0.9% Ca sham-operated pigs and the other OVX groups. In addition, Tb.Sp was increased and Tb.N decreased compared with the 0.9% Ca sham-operated pigs. Increases in serum PTH in this group were not accompanied by increases in calcitriol. Midradial and midfemoral BMD values were reduced in the 0.75 and 0.5% Ca OVX groups compared with the 0.9% Ca sham-operated pigs. Histomorphometric analyses of cortical bone suggested the reduction in cortical bone mass in the 0.75% Ca OVX group may have been largely due to net loss on the endocortical surface versus possible failure to accrue bone in the 0.5% Ca OVX group. Ash density and biomechanical parameters for vertebral cancellous bone decreased progressively in the 0.9% sham-operated, 0.9% Ca OVX, and 0.75% Ca OVX groups and then increased in the 0.5% Ca OVX group. After normalization for bone mass (ash), mechanical changes were still apparent, particularly for the 0.75% Ca OVX group compared with other OVX groups, reflecting that structural changes had taken place in the trabecular network.(ABSTRACT TRUNCATED AT 400 WORDS)

Calcium-restricted ovariectomized Sinclair S-1 minipigs: an animal model of osteopenia and trabecular plate perforation.

The ovariectomized rat model has now been generally accepted as a useful model for screening different therapeutic agents, but there is a major requirement to identify reliable large animal models for osteoporosis research. In this study, the calcium restricted, ovariectomized minipig has been thoroughly investigated in order to define a large animal model with trabecular and cortical bone remodeling which would be reliable for further testing of agents that had shown promise of efficacy during the screening procedure. Twenty six female, 4-month old minipigs were randomized into four groups and fed either normal diet (0.90% calcium (Ca.)) or diet with restricted calcium content (0.75%, 0.50%). At the age of ten months, 3 groups were ovariectomized (OVX) while one group on normal diet was shamoperated. The groups were followed for six months after the operation. At death, bone mass was determined by densitometry and by ashing. Additionally, biomechanical competence was assessed in trabecular bone cores from the vertebral bodies. Finally, histomorphometry (static and dynamic parameters) and structural analyses (star volume) were performed on the vertebral bodies. The study revealed an OVX-related decline of 6% in vertebral bone mineral density (BMD) and a decline of 15% in trabecular bone volume (BV/TV). In contrast, a 15% increase in mean trabecular plate separation (Tb.Sp.) and a small increase in marrow space star volume (Ma. Star V.) were detected. The structural changes became more pronounced when OVX was combined with mild Ca. restriction (0.75% Ca.) with an increase in Ma. Star V. to 164%.(ABSTRACT TRUNCATED AT 250 WORDS)

The anabolic effects of human parathyroid hormone (hPTH) on rat vertebral body mass are also reflected in the quality of bone, assessed by biomechanical testing: a comparison study between hPTH-(1-34) and hPTH-(1-84).

PTH has a proven anabolic effect on bone mass, as has been shown in several animal models and treatment regimens. The aim of this study was to ascertain whether the positive effect on bone mass is also reflected in the quality of bone formed. The study was performed in a rat model using human PTH (hPTH). One hundred and twenty male Wistar rats, divided into 10 groups, were given either hPTH-(1-34) or hPTH-(1-84) in daily sc doses. Dose levels ranged from 0.11-3.00 nmol/100 g BW.day for a period of 30 days. At death all six lumbar vertebrae were obtained from each rat. A combination of methods was applied to these vertebral bodies in order to ascertain the effect of hPTH on vertebral bone volume, density, trabecular structure, and biomechanical competence. The results revealed a dose-dependent increase in total volume, dry weight, ash weight, and trabecular bone volume. Also, a dose-dependent increase in load values could be demonstrated. The increase in bone strength remained significant after normalization for both cross-sectional area and bone mass. This indicates that the increase in bone size and bone mass was not achieved at the expense of the quality of the bone present. When the two treatments were administered at the same molar dose level, no difference between hPTH-(1-34) and hPTH-(1-84) was revealed. On the basis of this study, it is concluded that PTH could prove to be a promising treatment in the management of osteopenic states.