Anabolic and Antiresorptive Drugs Improve Trabecular Microarchitecture and Reduce Fracture Risk following Radiation Therapy

Many patients with symptomatic bone metastases receive radiation therapy, even though radiation is known to have potential adverse effects on bone. We hypothesized that the concurrent use of a bisphosphonate drug (zoledronic acid, ZA) or a combination of ZA plus an anabolic agent (parathyroid hormone, PTH) would lead to improvements in the microarchitecture and mechanical properties of irradiated bone. Human breast cancer cells were injected into the distal femur of 56 female nude mice, which were then divided into four groups: no treatment (0 Gy), radiation administered 4 weeks postinjection (20 Gy), radiation plus ZA (12.5 μg/kg weekly from weeks 4 to 12) (20 Gy + ZA), and radiation followed by ZA (25 μg/kg weekly from weeks 4 to 8) and PTH(1–34) (100 μg μg/kg daily from weeks 8 to 12) (20 Gy + ZA + PTH). Left limbs served as normal control bones. Bone loss over the 12-week study was tracked with serial radiography and bone densitometry. At the end of the study, micro-computed tomography and mechanical testing were used to quantify bone microarchitecture and bone strength. Radiation alone failed to prevent tumor-induced decreases in bone mineral density (BMD), trabecular bone volume, and bone strength. Treatment with 20 Gy + ZA or 20 Gy + ZA + PTH as adjuncts to radiation was effective at preserving trabecular bone architecture and bone strength at normal levels. ZA reduced the risk of mechanical fragility following irradiation of a lytic bone lesion. Supplemental use of PTH did not result in further increases in bone strength but was associated with significant increases in BMD and bone mass, suggesting that it may be beneficial in enhancing bone architecture following radiation therapy.     

Effects of combined and separate intermittent administration of low-dose human parathyroid hormone fragment (1–34) and 17β-estradiol on bone histomorphometry in ovariectomized rats with established osteopenia

Summary To evaluate the potential use of a combination of parathyroid hormone (PTH) and estrogen as therapy for osteoporosis, we examined the effects of combined and separate administration of low-dose PTH and estradiol in ovariectomized rats with established osteopenia. Ovariectomized rats were untreated for 5 weeks after surgery and then injected s.c. with vehicle (Ovx+V), 1–34 hPTH (2.5 g/kg/day) (Ovx+P), 17-estradiol (50 g/kg/day) (Ovx+E), or a combination of these (Ovx+P+E), for a further 4 weeks. We found no differences in serum calcium, tubular reabsorption of phosphate, or 25OHD. 1,25(OH)₂D levels were significantly higher in Ovx+P and lower in Ovx+E, when compared with Ovx+V. Though there was no change in bone mineral density (BMD) in the diaphysis region of femurs, reduction of BMD in the distal region of the femurs in Ovx+V was reversed in Ovx+E and Ovx+P+E. Compared with Ovx+V, Ovx+P and Ovx+P+E had significantly higher cancellous bone volume (Cn-BV/TV) whereas Ovx+E showed a nonsignificant increase. When indices of bone turnover were examined, PTH alone showed a small but not significant improvement in bone formation rate (BFR). Increased osteoclast surface (OCS), as the result of ovariectomy, was inhibited in Ovx+E and Ovx+P+E. Estrogen alone (Ovx+E) severely inhibited BFR, but co-administration of PTH and estrogen (Ovx+P+E) showed an impressive reversal of such inhibition. The changes in BFR were mainly derived from changes in double-labeled surface (dLS), except a small increase in mineral apposition rate was also observed in Ovx+P+E. These results suggest that, after extensive cancellous bone loss in the rat tibia, low doses of PTH function anabolically, especially in situations where the bone formation rate is low. A combination of both estrogen and PTH may provide the best treatment for improving bone mass by decreasing resorption and maintaining a high bone formation rate.     

Intermittent Therapy with 1,25 Vitamin D and Calcitonin Prevents Cyclosporin-Induced Alveolar Bone Loss in Rats

Bone loss associated with cyclosporin A (CsA) therapy can result in serious morbidity to patients. Intermittent administration of 1,25 Vitamin D and calcitonin reduces osteopenia in a murine model of postmenopausal osteoporosis. The purpose of this study was to evaluate the effects of this therapeutic approach on CsA-induced alveolar bone loss in rats. Forty male Wistar rats were allocated to four experimental groups according to the treatment received during 8 weeks: (1) CsA (10 mg/kg/day, s.c.); (2) 1,25 Vitamin D (2 μg/kg, p.o.; in weeks 1, 3, 5, and 7) plus calcitonin (2 μg/kg, i.p.; in weeks 2, 4, 6, and 8); (3) CsA concurrently with intermittent 1,25 Vitamin D and calcitonin administration; and (4) the control treatment group (vehicle). At the end of the 8-week treatment period, serum concentrations of bone-specific alkaline phosphatase, tartrate-resistant acid phosphatase (TRAP-5b), osteocalcin, interleukin (IL)-1β, IL-6, and tumor necrosis factor alpha (TNF-α) were measured and an analysis of bone volume, bone surface, number of osteoblasts, and osteoclasts was performed. CsA administration resulted in significant alveolar bone resorption, as assessed by a lower bone volume and an increased number of osteoclasts, and increased serum bone-specific alkaline phosphatase, TRAP-5b, IL-1β, IL-6, and TNF-α concentrations. The intermittent administration of calcitriol and calcitonin prevented the CsA-induced osteopenic changes and the increased serum concentrations of TRAP-5b and inflammatory cytokines. Intermittent calcitriol/calcitonin therapy prevents CsA-induced alveolar bone loss in rats and normalizes the production of associated inflammatory mediators.     

Effect of alendronate and intermittent parathyroid hormone on implant fixation in ovariectomized rats

Background  Intermittent administration of parathyroid hormone (PTH) leads to bone formation by increasing osteoblast numbers and activity levels. Animal studies have shown that intermittent PTH administration increases implant fixation in normal rats. The purpose of this study was to analyze the osseous incorporation of an implant in osteoporotic rats while treating them with intermittent PTH (1–34) or alendronate. Methods  A total of 36 ovariectomized (OVX) Wistar rats were randomized into three groups. Polymethylmethacrylate cement rods were implanted in one tibia in each rat. The three groups received daily PTH (60 μg/kg body weight [BW]), alendronate (200 μg/kg BW), or saline (0.5 ml/kg BW). A sham-ovariectomized group (n = 12) was treated with saline. After 2 weeks, the area around the implants was analyzed by histomorphometry for bone volume density (BVD) and implant bone contact. Bone mineral density (BMD) was evaluated by dual-energy X-ray absorptiometry. Results  The BVD was higher in the specimens treated with PTH than in the other groups. PTH improved the BVD, BMD, and implant bone contact. Alendronate doubled the implant bone contact compared to the OVX and sham groups but did not improve BVD or BMD. Conclusions  These findings confirm that intermittent PTH enhances implant fixation in osteoporotic bone. The clinical significance of these findings is that application of intermittent PTH may be beneficial for early implant fixation in fractures, nonunions, and prosthetic replacements when bone density is decreased.     

Skeletal effects of parathyroid hormone infusion in ovariectomized rats with or without estrogen repletion.

We employed skeletally matured rats to study changes in biochemical markers of bone turnover, bone mineral density (BMD), and bone biomechanics produced by continuous elevation of parathyroid hormone (PTH) in estrogen-deplete and -replete rodents. Ninety-six 7-month-old virgin female rats were divided randomly into 12 groups (n = 8) and treated as follows. One group was killed on the day of surgery. The remaining groups were either bilaterally ovariectomized (Ovx) or sham-operated and left untreated for 8 weeks, at which point, two groups, one sham and one Ovx, were killed. The remaining nine groups were treated for 2 weeks or 4 weeks. One sham and two Ovx groups received subcutaneous implants of Alzet miniosmotic pumps with vehicle for PTH. Two Ovx groups were given pumps with vehicle as well as a subcutaneous implant of 17beta-estradiol, which delivered 10 microg/kg per day. Two Ovx groups were implanted with rat PTH(1-34) in Alzet miniosmotic pumps, which delivered 30 microg PTH/kg per day. Two Ovx groups were implanted with both estradiol pellets and PTH-loaded pumps. One group of Ovx animals from each treatment was killed after 2 weeks and the other after 4 weeks. Biochemical markers of bone turnover, serum osteocalcin and urinary free pyridinoline, BMD, and mechanical strength of excised bones were measured. As expected, there was a significant increase in N-terminal PTH and serum calcium levels in all PTH infusion groups. Both serum osteocalcin and urinary pyridinoline showed a rapid increase within the first 2 weeks of the PTH infusion and remained elevated at week 4. In estrogen-replete groups, osteocalcin increased by week 2 of PTH infusion but pyridinoline did not increase until week 4. BMD of the distal and proximal femur showed the expected decrease 8 weeks after ovariectomy but did not exhibit any further changes during the 4 weeks of treatment with vehicle. Four weeks of PTH infusion in Ovx animals resulted in BMD loss at the midshaft, distal, and proximal regions of the femur. Estrogen repletion by itself, beginning 8 weeks after ovariectomy, produced no change in BMD at any site when compared with from Ovx vehicle-treated rats. Estrogen repletion in PTH-infused Ovx animals resulted in significant improvements of BMD comparable with sham-operated animals at all three femoral regions. The indentation test at the cancellous bone of the distal femur, three-point bending test at the midshaft femur, and cantilever bending test at the femoral neck showed that the changes in mechanical strength in these sites were consistent to the changes found in BMD. Our results showed that (1) continuously elevated levels of PTH induced additional loss of BMD in estrogen-deficient animals beyond the rapid bone loss phase associated with ovariectomy, (2) estrogen repletion, given by implant, to PTH-infused Ovx animals, reversed these BMD changes increasing BMD to levels comparable with estrogen-sufficient rats, and (3) these changes were reflected in the mechanical strength determined at these sites. These results lend experimental support that hormone replacement therapy may benefit bone health in postmenopausal women with primary hyperparathyroidism (PHPT). In addition, it raises the possibility that a continuous elevation of PTH could exert anabolic effects on skeletal tissue if its catabolic component can be minimized.     

Effects of Clodronate on Cortical and Trabecular Bone in Ovariectomized Rats on a Low Calcium Diet

The aim of this study was to evaluate the contribution of a low calcium diet to the cortical and trabecular osteoporosis seen in ovariectomized rats after 7 weeks on a low calcium diet and to investigate the effects of the bisphosphonate clodronate on this development of osteoporosis. Thirty-six mature, female Wistar rats were randomized into four groups: Ovx−B (bisphosphonate) and Ovx−C (control) were ovariectomized, and Sham−Ca (low calcium) and Sham+Ca (normal calcium) were sham operated. The first three groups were fed a low calcium diet (0.01%) and Sham+Ca normal rat chow (Ca 1.1%). The Ovx−B received 10 mg/kg s.c. clodronate daily for nine weeks, and Ovx−C, Sham−Ca, and Sham+Ca received the same volumes of saline. Bone mineral turnover measured as ⁸⁵Sr-uptake was increased in all low calcium groups compared to Sham+Ca. The Sham+Ca femora had higher dry weight and ash weight than the other groups, and Ovx−C had higher dry weight compared with Ovx−B and Sham−Ca. Calcium content was lower in both Ovx groups compared to both Sham groups. Magnesium was lower in all groups compared to Sham+Ca and higher in Ovx−B compared with Ovx−C. In the femoral shaft, Sham+Ca had significantly higher ultimate bending moment, energy absorption, and deflection compared to the other three groups. Ultimate bending moment was higher in Sham−Ca than in Ovx−C. Stiffness was increased in both Sham+Ca and Ovx−B compared to Ovx−C. The maximum stress in the femoral midshaft was higher in Sham+Ca than in the other groups, and higher in Ovx−B than in Ovx−C. Histomorphometry showed increased medullary area in all low calcium groups compared to Sham+Ca and larger cortical area in Sham+Ca and Ovx−B compared to Ovx−C. Compared to Sham+Ca the trabecular bone volume was decreased to 30% in Sham−Ca and to 9% in Ovx−C, but was unchanged in Ovx−B. The low calcium diet generally increased bone mineral turnover and reduced the tibial bone volume. Femoral changes led to a reduction of cortical fracture strength and maximal stress. Ovariectomy in addition to a low calcium diet reduced femoral strength even more. Daily injections of clodronate to ovariectomized rats on a low calcium diet increased femoral shaft stiffness and maximum stress, and clodronate preserved both trabecular and cortical tibial bone volume completely. Received: 11 June 1996 / Accepted: 5 March 1997     

Vitamin K<Subscript>2</Subscript> Prevents Hyperglycemia and Cancellous Osteopenia in Rats with Streptozotocin-Induced Type 1 Diabetes

The purpose of the present study was to examine the effect of vitamin K₂ on cancellous and cortical bone mass in rats with streptozotocin (STZ)-induced type 1 diabetes. Twenty-seven male Sprague-Dawley rats aged 12 weeks were randomized by the weight-stratified method into the following three groups: age-matched control group, STZ + vehicle group, and STZ + vitamin K₂ group. STZ (40 + 50 mg/kg) was administered intravenously twice during the initial 1-week period. Vitamin K₂ (menatetrenone, 30 mg/kg) was administered orally 5 days a week. After 12 weeks of treatment, the serum glucose concentration and femoral length and weight were measured and histomorphometric analysis was performed on the cancellous and cortical bone of the distal femoral metaphysis and femoral diaphysis, respectively. STZ administration induced hyperglycemia and a decrease in femoral weight. The STZ + vehicle group also showed cancellous osteopenia due to a decrease in the number of osteoblasts/bone surface (N.Ob/BS) and the osteoblast surface (ObS)/BS without any significant changes in bone-resorption parameters, but it did not have a significant decrease in cortical bone mass. Administration of vitamin K₂ to STZ-treated rats prevented the development of hyperglycemia and a decrease in femoral weight. Vitamin K₂ also prevented cancellous osteopenia by inhibiting the decrease in N.Ob/BS and ObS/BS without significantly affecting bone-resorption parameters, but it did not significantly increase cortical bone mass. These results suggest that vitamin K₂ has beneficial effects on glucose concentration and cancellous bone mass in rats with STZ-induced type 1 diabetes.     

Intermittent weekly administration of human parathyroid hormone (1–34) improves bone-hydroxyapatite block bonding in ovariectomized rats

Hydroxyapatite (HA) blocks have been widely used for the reconstruction of bone defects and as a bone substitute. Bone-implant bonding depends on both implant-related factors and patient variables. Intermittent human parathyroid hormone (h-PTH) has a strong anabolic effect on bone formation. The purpose of the present study is to evaluate whether intermittent h-PTH administration enhances bone-HA block bonding in normal versus ovariectomized (OVX) rats. Cancellous bone osteotomy and HA-block implantation were performed on the proximal left tibia in both OVX and sham-operated 7-month-old female Sprague-Dawley rats. Newly formed cancellous bone around the HA block and bone-HA block bonding were evaluated by bone histomorphometry at 8 weeks after the administration of h-PTH (100 μg/kg/week) or its vehicle. The administration of h-PTH significantly increased cancellous bone volume by stimulating bone formation in OVX rats (p < 0.01). Although bone-HA block bonding was significantly decreased in OVX rats compared to that of sham-operated rats (p < 0.01), h-PTH improved the bone-HA block bonding in OVX rats (p < 0.01). These results suggest that intermittent h-PTH treatment may improve bone-HA bonding in osteoporosis by restoring cancellous bone volume and enhancing cancellous bone formation around the HA block.     

A comparative study of the bone-restorative efficacy of anabolic agents in aged ovariectomized rats

Introduction The study was designed to compare the bone anabolic effects of basic fibroblast growth factor (bFGF), a selective agonist for prostaglandin E receptor subtype EP4, and parathyroid hormone (PTH) in aged ovariectomized (OVX) rats with severe cancellous osteopenia. Methods Groups of aged OVX rats were maintained untreated for 1 year postovariectomy (15 months of age) to develop severe tibial cancellous osteopenia. These animals were then treated with bFGF or the EP4 agonist (EP4) for 3 weeks. Other groups of aged OVX rats were treated with EP4 or PTH alone for 11 weeks, or sequentially with bFGF or EP4 for 3 weeks followed by PTH for 8 weeks. Cancellous and cortical bone histomorphometry were performed in the right proximal tibial metaphysis and tibial diaphysis respectively. Results Treatment with bFGF for 3 weeks markedly increased serum osteocalcin, osteoid volume, and osteoblast and osteoid surfaces to a greater extent than EP4. Basic FGF, but not EP4 or PTH, induced formation of osteoid islands within bone marrow. EP4 stimulated cancellous bone turnover, but failed to restore lost cancellous bone in the severely osteopenic proximal tibia after 11 weeks of treatment. In contrast, EP4, much like PTH, increased cortical bone mass in the tibial diaphysis by stimulating both periosteal and endocortical bone formation. Treatment of aged OVX rats with PTH alone tended to partially reverse the severe tibial cancellous osteopenia, whereas sequential treatment with bFGF and PTH increased tibial cancellous bone mass to near the level of vehicle-treated control rats. These findings indicate that bFGF had the strongest stimulatory effect on cancellous bone formation, and was the only anabolic agent to induce formation of osteoid islands within the bone marrow of the severely osteopenic proximal tibia. Therefore, bFGF may be more effective for the reversal of severe cancellous osteopenia. PTH and EP4 increased cortical bone mass to nearly the same extent, but cancellous bone mass was greater by two-fold in PTH-treated OVX rats than in EP4-treated OVX rats. Conclusion These findings in aged OVX rats suggest that PTH is more efficacious than EP4 for augmentation of cancellous bone in the severely osteopenic, estrogen-deplete skeleton.     

Mineral Composition is Altered by Osteoblast Expression of an Engineered G<Subscript>s</Subscript>-Coupled Receptor

Activation of the G_s G protein–coupled receptor Rs1 in osteoblasts increases bone mineral density by 5- to 15-fold in mice and recapitulates histologic aspects of fibrous dysplasia of the bone. However, the effects of constitutive G_s signaling on bone tissue quality are not known. The goal of this study was to determine bone tissue quality in mice resulting from osteoblast-specific constitutive G_s activation, by the complementary techniques of FTIR spectroscopy and synchrotron radiation micro-computed tomography (SRμCT). Col1(2.3)-tTA/TetO-Rs1 double transgenic (DT) mice, which showed osteoblast-specific constitutive G_s signaling activity by the Rs1 receptor, were created. Femora and calvariae of DT and wild-type (WT) mice (6 and 15 weeks old) were analyzed by FTIR spectroscopy. WT and DT femora (3 and 9 weeks old) were imaged by SRμCT. Mineral-to-matrix ratio was 25% lower (P = 0.010), carbonate-to-phosphate ratio was 20% higher (P = 0.025), crystallinity was 4% lower (P = 0.004), and cross-link ratio was 11% lower (P = 0.025) in 6-week DT bone. Differences persisted in 15-week animals. Quantitative SRμCT analysis revealed substantial differences in mean values and heterogeneity of tissue mineral density (TMD). TMD values were 1,156 ± 100 and 711 ± 251 mg/cm³ (mean ± SD) in WT and DT femoral diaphyses, respectively, at 3 weeks. Similar differences were found in 9-week animals. These results demonstrate that continuous G_s activation in murine osteoblasts leads to deposition of immature bone tissue with reduced mineralization. Our findings suggest that bone tissue quality may be an important contributor to increased fracture risk in fibrous dysplasia patients.     

The Effects of Bone Remodeling Inhibition by Alendronate on Three-Dimensional Microarchitecture of Subchondral Bone Tissues in Guinea Pig Primary Osteoarthrosis

We assessed whether increase of subchondral bone density enhances cartilage stress during impact loading, leading to progressive cartilage degeneration and accelerated osteoarthrosis (OA) progression. Sixty-six male guinea pigs were randomly divided into six groups. During a 9-week treatment period, four groups received twice-weekly subcutaneous injections of alendronate (ALN) in two doses: two groups received 10 μg/kg and two groups received 50 μg/kg. The two control groups received vehicle. After 9 weeks, one 10 μg/kg ALN group, one 50 μg/kg ALN group, and one control group were killed. The remaining three groups (17-week groups) were left for an additional 8 weeks, receiving the same treatment regimen before death. The left proximal tibiae were scanned by micro-computed tomography to quantify the microarchitecture of subchondral bone, followed by mechanical testing and determination of collagen and mineral. The control groups had typical OA-related cartilage degeneration at 9 and 17 weeks, whereas the 50 μg/kg ALN group had even worse degeneration in the medial condyle. It is unclear whether there is a direct or a secondary effect of ALN on the cartilage. The 9-week ALN group had significantly greater subchondral plate thickness. The 9- and 17-week groups had similar changes of cancellous bone microarchitecture, with greater volume fraction and connectivity and an extremely plate-like structure. The 9-week ALN group had greater bone mineral concentration, and the 17-week ALN group had reduced collagen concentration and greater mineral concentration. Treatment with ALN did not significantly change the mechanical properties of the cancellous bone.     

Low dose of human PTH(1-34) improved tibial subcortical bone mass without further cortical bone loss in adult intact beagles

Cortical bone loss has not been observed in parathyroid hormone- (PTH-) treated rats, but clinical investigators suggest that it may augment cancellous bone mass at the expense of cortical bone in PTH-treated patients. In this study, the effects of PTH on cancellous and cortical bone mineral density of the tibia were studied in dogs with Haversian cortical bone remodeling by peripheral quantitative computed tomography (pQCT). Sixteen 19- to 20-month-old beagle dogs were randomized into four groups. In group 1, the vehicle control group, saline was injected daily throughout the experimental period. In group 2, the sequential group, 0.375 μg/kg of hPTH(1-34) was injected daily during the first 4 weeks, then stopped for 8 weeks, and this sequence was once repeated (on 4 weeks, off 8 weeks, and on 4 weeks, off 8 weeks). In group 3, the one-time group, the same dose of PTH was injected once per week for 24 weeks. In group 4, the three-time group, the same dose of PTH was given three times per week for 24 weeks. Peripheral QCT measurements were carried out 7 mm distal to the growth plate in the proximal metaphysis of the tibia. A voxel size of 0.295 mm and threshold for cortical and subcortical bone of 0.930 was chosen throughout the experiment. In adult beagles, group 4 had a significantly higher value in subcortical bone mineral density (SubCt.BMD) than control group (852 vs 771 mg/mm³; P < .05). Higher but non-significant increase occurred in the PTH-treated animals (groups 2, 3, 4) in the following parameters: total (Total.BMD), cancellous bone mineral density (Cn.BMD), cortical bone mineral density (Ct.BMD), and cortical thickness (Ct.Th). These findings show that a low dose of PTH improved subcortical bone mineral density without decreasing cortical bone mineral density during this dosage and period. Received: Sept. 8, 1997 / Accepted: Jan. 6, 1998     

Histomorphometric Evaluation of the Recovering Effect of Human Parathyroid Hormone (1–34) on Bone Structure and Turnover in Streptozotocin-Induced Diabetic Rats

The purpose of this study was to determine whether human parathyroid hormone (h-PTH) enhances trabecular bone mass and connectivities that were reduced by streptozotocin (STZ)-induced diabetes in rats. Seven-month-old female Wistar rats were injected with STZ or its vehicle intraperitoneally. All vehicle-injected normal controls were sacrificed 0, 4, 6, 8, 12, 14, and 16 weeks after injection, and one-third of the STZ-injected rats were sacrificed as the baseline controls 4, 6, and 8 weeks after the injection. Eight-week h-PTH (6.0 μg/kg, 6 times a week) or its vehicle treatment by subcutaneous injection for residual diabetic rats was started 4, 6, or 8 weeks after the STZ injection. The rats' proximal right tibiae were processed for undecalcified Villanueva bone staining sections for bone histomorphometry. Furthermore, changes in trabecular connectivities were determined by node-strut analysis. The decreased cancellous bone volume (BV/TV) and turnover in diabetic rats were recovered in all PTH-treated groups. In node-strut analysis, the node-related parameters (N.Nd/TV, NdNd/TV) were significantly increased by PTH when it was administered 4 weeks after STZ injection but were not increased when administration was started after 6 weeks. The results indicated that PTH enhanced bone turnover and bone mass but not trabecular connectivity in the late stage of diabetes in rats. Early treatment of osteoporosis is important in preventing fractures caused by decreased bone strength resulting from decreased trabecular connectivity. Received: 11 May 1999 / Accepted: 20 September 1999     

Bone Mass and Markers of Bone and Calcium Metabolism in Postmenopausal Women Treated with 1,25-Dihydroxyvitamin D (Calcitriol) for Four Years

To evaluate the long-term effect of calcitriol treatment on bone mineral density (BMD) of the femoral neck and lumbar spine and the parameters of calcium and bone metabolism in elderly women, 55 healthy, postmenopausal women, all aged 66 years, were enrolled in the study. Eighteen started a 4-year supplementation with 0.5 μg of calcitriol daily and 37 served as controls. Calcium intake of all the subjects was adjusted to 800 mg daily. In 4 years femoral neck BMD increased by 3.0% in the calcitriol group, but decreased by 1.6% in the control group (P= 0.009). The respective changes in lumbar spine BMD were +2.3% and +0.9% (P= 0.067). Two years' treatment with calcitriol increased the intestinal absorption of strontium by 57% (P < 0.001), doubled the urinary excretion of calcium (P < 0.001), and decreased the mean parathyroid hormone (PTH) level by 32% (P < 0.01). In the calcitriol group the marker of bone formation, serum osteocalcin, decreased by 27% (P < 0.01), and the marker of bone resorption, serum C-telopeptide of type I collagen (CTx), by 33% (P= 0.05) after 2 years. In two subjects the calcitriol dose had to be reduced because of hypercalciuria. We conclude that calcitriol treatment increases bone mass at the femoral neck and lumbar spine, the increases being maintained for up to 4 years. The gain in bone mass results from reduced bone turnover which is partly a consequence of the enhanced intestinal absorption of calcium and suppressed serum PTH levels. Received: 8 January 1999 / Accepted: 29 February 2000     

Parathyroid Hormone Treatment Increases Fixation of Orthopedic Implants with Gap Healing: A Biomechanical and Histomorphometric Canine Study of Porous Coated Titanium Alloy Implants in Cancellous Bone

Parathyroid hormone (PTH) administered intermittently is a bone-building peptide. In joint replacements, implants are unavoidably surrounded by gaps despite meticulous surgical technique and osseointegration is challenging. We examined the effect of human PTH(1–34) on implant fixation in an experimental gap model. We inserted cylindrical (10 × 6 mm) porous coated titanium alloy implants in a concentric 1-mm gap in normal cancellous bone of proximal tibia in 20 canines. Animals were randomized to treatment with PTH(1–34) 5 μg/kg daily. After 4 weeks, fixation was evaluated by histomorphometry and push-out test. Bone volume was increased significantly in the gap. In the outer gap (500 μm), the bone volume fraction median (interquartile range) was 27% (20–37%) for PTH and 10% (6–14%) for control. In the inner gap, the bone volume fraction was 33% (26–36%) for PTH and 13% (11–18%) for control. At the implant interface, the bone fraction improved with 16% (11–20%) for PTH and 10% (7–12%) (P = 0.07) for control. Mechanical implant fixation was improved for implants exposed to PTH. For PTH, median (interquartile range) shear stiffness was significantly higher (PTH 17.4 [12.7–39.7] MPa/mm and control 8.8 [3.3–12.4] MPa/mm) (P < 0.05). Energy absorption was significantly enhanced for PTH (PTH 781 [595–1,198.5] J/m² and control 470 [189–596] J/m²). Increased shear strength was observed but was not significant (PTH 3.0 [2.6–4.9] and control 2.0 [0.9–3.0] MPa) (P = 0.08). Results show that PTH has a positive effect on implant fixation in regions where gaps exist in the surrounding bone. With further studies, PTH may potentially be used clinically to enhance tissue integration in these challenging environments.     

Effect of vitamin K<Subscript>2</Subscript> and growth hormone on the long bones in hypophysectomized young rats: a bone histomorphometry study

The purpose of the present study was to determine whether vitamin K₂ and growth hormone (GH) had an additive effect on the long bones in hypophysectomized young rats. Forty-eight female Sprague–Dawley rats (6 weeks old) were assigned to the following five groups by the stratified weight randomization method: intact controls, hypophysectomy (HX) alone, HX + vitamin K₂ (30 mg/kg, p.o., daily), HX + GH (0.625 mg/kg, s.c., 5 days a week), and HX + vitamin K₂ + GH. The duration of the experiment was 4 weeks. HX resulted in a reduction of the cancellous bone volume/total tissue volume (BV/TV) at the proximal tibial metaphysis, as well as decreasing the total tissue area and cortical area of the tibial diaphysis. These changes resulted from a decrease of the longitudinal growth rate and the bone formation rate (BFR)/TV of cancellous bone, as well as a decrease of the periosteal BFR/bone surface (BS) and an increase of endocortical bone turnover (indicated by the BFR/BS) in cortical bone. Administration of vitamin K₂ to HX rats did not affect the cancellous BV/TV or the cortical area. On the other hand, GH completely prevented the decrease of total tissue area and cortical area in cortical bone, as well as the decrease of marrow area and endocortical circumference, by increasing the periosteal BFR/BS compared with that in intact controls and reversing the increase of endocortical bone turnover (BFR/BS). However, GH only partly improved the reduction of the cancellous BV/TV, despite an increase of the longitudinal growth rate and BFR/TV compared with those of intact controls. When administered with GH, vitamin K₂ counteracted the reduction of endocortical bone turnover (BFR/BS) and circumference caused by GH treatment, resulting in no significant difference of marrow area from that in untreated HX rats. These results suggest that, despite the lack of an obvious effect on bone parameters, vitamin K₂ normalizes the size of the marrow cavity during development of the bone marrow in young HX rats treated with GH.     

Altered parathyroid hormone- or calcitonin-stimulated adenosine 3′,5′-monophosphate release by isolated perfused bone from glucocorticoid-treated rats

Summary The present studies were designed to examinein vivo effects of glucocorticoid on PTH-or calcitonin (CT)-stimulated adenosine 3′,5′-monophosphate (cAMP) release from the isolated perfused bone of rat and to test whether the duration of glucocorticoid administration influenced such effects. We assessed the ability of acute (24 hour) or chronic (2 week) dexamethasone administration to modulate the cAMP response to 5 μg human PTH-(1–34) or 1 μg eel CT. Acute treatment with dexamethasone (1 mg/100 g body wt) increased the cAMP response to PTH, but decreased the response to CT. This enhanced effect on PTH-stimulated cAMP release was not apparent in the presence of phosphodiesterase inhibitor 3-isobutyl-1-methyl-xanthine (IBMX, ImM). In contrast, chronic dexamethasone treatment (0.2 mg daily for 2 weeks) led to a decrease in both PTH- and CT-stimulated cAMP release. Such impaired response of the dexamethasone-treated bones to PTH was also found in rats that underwent parathyroidectomy 24 hours before sacrifice. These data indicate that 1) the duration of glucocorticoid administration may influence the effect of PTH on bone and 2) glucocorticoid may decrease cAMP-mediated CT function, regardless of the duration of treatment.     

Effect of human parathyroid hormone hPTH (1–34) applied at different regimes on fracture healing and muscle in ovariectomized and healthy rats

Three experiments were conducted to investigate the effect of intermittent administration of parathyroid hormone (PTH) (1–34) applied at different regimes on fracture healing and muscle in healthy and ovariectomized (Ovx at 3 months of age) rats. Five-month old rats underwent bilateral transverse metaphyseal osteotomy of tibia and were divided into groups (12 rats each). In Exp 1, Ovx rats were either treated with PTH (7×/w, 1–35d), with oral estradiol-17β-benzoate (0.4 mg/kg BW, 1–35d) or untreated. In Exp. 2, there were 3 groups: healthy untreated or treated with PTH (5×/w, 1–35d or 7–35d). In Exp. 3, there were 7 groups: healthy, Ovx, “healthy PTH 5×/w 7–35d”, “Ovx PTH 5×/w 7–35d, 14–35d or 14–28d”, “Ovx PTH every other day 7–35d”. Single dosage of PTH was 40 μg/kg BW. After 35 days of healing one tibia was analyzed by computed tomographical, biomechanical, histological analyses. The other tibia was used in analyses of Alp, Oc, Trap 1, Igf-1, Rankl, Opg genes (Exp.2, 3). Serum Oc and Alp were measured. Body, uterus weight was recorded. M. gastrocnemius was analyzed for weight (Exp. 2), fiber size and mitochondrial respiratory activity (MRA) (Exp.3). Estrogen enhanced uterus weight, prevented body increase, however, did not improve bone healing in Ovx rats (Exp. 1). PTH administration from days 1 and 7 improved bone parameters in all rats regardless of the application frequency (7, 5×/w or every other day) (Exp. 1, stiffness Ovx: 118 + 13 N/mm, Ovx PTH: 250 ± 20 N/mm) being more effective in healthy rats (Exp. 3, stiffness improvement Healthy: 59 to 174 N/mm, Ovx: 52 to 98 N/mm). Serum Oc level was elevated in PTH treated rats. Application from day 14 proved to be less effective (Exp. 3). PTH had no effect (P > 0.05) on body, uterus and muscle weight, muscle fiber size, MRA and expression of bone markers. PTH promoted bone healing in Ovx and healthy rats, when it is applied during early stage of healing without having any adverse systemic effect. In perspective, PTH may represent a treatment for enhancement of fracture healing. The findings need to be confirmed by follow-up studies on other animals.     

Parathyroid hormone (1–34) increases attachment of PMMA cement to bone

The attachment of an implant material to bone is related to the surface of the implanted material and the ability of the bone to form around the implant. Intermittent parathyroid (PTH) administration increases bone formation by stimulating osteoblastic activity. Little is known about the effect of PTH administration on orthopedic implant incorporation. The present study determined how PTH (1–34) administration influenced bone bonding, i.e., the bone-cement interfacial tensile strength, of vacuum-mixed polymethylmethacrylate (PMMA) bone cement (surface roughness; Ra, 4.8 μm). Bone bonding was evaluated by a detachment test. We used unloaded cement surfaces, which could be detached from the bone. Titanium plates were developed such that a cement fill was contained within a plate that was contained within a titanium holder. Thus, a flat cement surface came into contact with traumatized bone only, and the rest of the plate had no contact with tissue. After implantation of the plate in the left tibia, 20 adult male rats were injected daily with human PTH (1–34) at 60 μg/kg per injection (n = 10) or vehicle (n = 10); the animals were killed after 4 weeks. The plates were detached from the bone by a perpendicular force. PTH treatment increased the median pull-away strength (0.21 MPa), compared with that in the vehicle-treated rats, (0.04 MPa) (P = 0.02). The results suggest that PTH treatment may have the potential to enhance the incorporation of cemented orthopedic implants. Received: September 18, 2000 / Accepted: June 6, 2001