Rat Model Of Postmenopausal Osteoporosis Research Articles

A Novel Vitamin D Receptor Agonist, VS-105, Improves Bone Mineral Density without Affecting Serum Calcium in a Postmenopausal Osteoporosis Rat Model.

Background and objectives:VS-105, a novel vitamin D receptor agonist with significantly less hypercalcemic side effects than calcitriol, is a useful tool to investigate whether or not a vitamin D receptor agonist at non-hypercalcemic doses could improve bone mineral density (BMD).Methods:VS-105 and calcitriol were evaluated in an ovariectomized (OVX) osteoporosis rat model and in calvariae bone organ culture.Results:Treatment of OVX rats by VS-105 (0.1, 0.2 or 0.5 μg/kg, intraperitoneal, 3×/week, for 90 days) significantly improved BMD in the L3 lumbar vertebra in a dose-dependent manner (sham vs. OVX/vehicle: 324 ± 14 vs. 279 ± 10 mg/cm2; VS-105 at 0.1, 0.2 and 0.5 μg/kg: 306 ± 9, 329 ± 12, and 327 ± 10 mg/cm2, respectively) without affecting serum calcium (Ca). Calcitriol at 0.1 μg/kg significantly increased BMD but it also increased serum Ca. VS-105 and calcitriol at the test doses significantly suppressed serum parathyroid hormone and promoted tibia bone growth. With respect to biomarkers of bone remodeling, calcitriol and VS-105 both significantly elevated serum osteocalcin. In the calvariae bone organ culture, net Ca release was significantly less in VS-105-treated groups (vs. calcitriol).Conclusions:VS-105 is efficacious in improving BMD in a dose range that does not affect serum Ca in OVX rats; the improvement in BMD by VS-105 is attributable to increased osteoblastic activity and reduced osteoclastic bone resorption.

Morphometric Effects of Whole-Body Vibration on the Bone in a Rat Model of Postmenopausal Osteoporosis

ObjectiveThe purpose of this study was to analyze the morphometric effects of mechanical vibration with a duration of 4 or 8 weeks on the femur of oophorectomized Wistar rats. MethodsSixty-four female rats were submitted to oophorectomy or a sham operation, and each of those 2 groups were randomized into 4 groups: untreated and euthanized at week 12, untreated and euthanized at week 16, treated for 4 weeks and euthanized at week 12, and treated for 8 weeks and euthanized at week 16. The vibration treatment was performed for 10 min/d, with a frequency of 60 Hz, 3 d/wk. The rats were then euthanized and the right femur dissected. Subsequently, histomorphometric analysis was performed on the proximal epiphysis and diaphysis of the spongy and cortical bone, respectively. ResultsAs expected, the oophorectomy groups presented reduction of spongy and cortical bone tissue. Further, the vibration therapy of 4 and 8 weeks’ duration in the oophorectomized groups led to increased bone mass, observed as an increased percentage of spongy tissue, and increased thickness and percentage of cortical tissue. However, the variables of femoral neck diameter, mean area of the shaft, and number of osteocytes were not altered by oophorectomy and vibration. ConclusionThe mechanical vibration was effective in increasing the bone mass of the femur of oophorectomized Wistar rats, observed by increasing the percentage of spongy bone and increasing the percentage and thickness of cortical bone.

Estrogen Deficiency Influences Hematological Parameters Following Bone Fracture in OVX Rats

Abstract Objectives Estrogen deficiency in postmenopausal women contributes to the development of osteoporosis. Furthermore, estrogen deficiency is associated with significant changes in hematological parameters including inflammatory markers. Since inflammation contributes to the pathophysiology of osteoporosis, the relationship between bone fracture, estrogen deficiency, and hematological parameters warrant investigation. Using an ovariectomized (OVX) rat model of postmenopausal osteoporosis, the objective of this research was to investigate the extent to which bone fracture in conditions of low estrogen affects hematological parameters compared to bone fracture in estrogen sufficient conditions. Methods 12-month old Sprague Dawley rats were randomized into one of four groups (1. OVX + Fracture, 2. OVX + no fracture, 3. Sham + fracture, 4. Sham + no fracture). Rats randomized into the OVX + fracture and sham + fracture groups underwent fracture of the right fibula. After sacrificing the animals, blood was collected for analysis of hematological parameters. Results All data are presented as mean ± S.E. As expected, OVX induced significant changes in hematological parameters compared to Sham. OVX compared to sham increased WBC (5.78 ± 0.93 vs. 1.73 ± 0.93 P < 0.01) and EOS (2.75 ± 0.39 vs. 1.50 ± 0.39 P < 0.05) and decreased PLT (357 ± 61 vs. 627 ± 61 P < 0.01). No changes were observed in hematological parameters of sham + fracture compared to sham. However, OVX + fracture increased MPV (6.33 ± 0.07 vs. 6.03 ± 0.09 P < 0.5) and tended to increase WBC (4.80 ± 0.66 vs. 2.73 ± 0.86 P = 0.06) and EOS (1.73 ± 0.28 vs. 0.81 ± 0.36 P = 0.55). Conclusions These results are in accordance with previous reports indicating that OVX causes changes in hematological parameters. We also demonstrated that under conditions of bone fracture, OVX causes further changes in hematological parameters compared to sham. However, the implications in these changes for bone healing in conditions of low estrogen such as postmenopausal osteoporosis still need further investigation. Funding Sources USDA.

Resveratrol promotes osteoblastic differentiation in a rat model of postmenopausal osteoporosis by regulating autophagy

ObjectiveResveratrol is a natural polyphenolic compound that ameliorates postmenopausal osteoporosis by activating the estrogen receptor. Research has shown that resveratrol exhibits some type of estrogen receptor agonist activity, reducing the risk of breast cancer. However, its mechanism of action remains largely unknown. This study aims to investigate the effect of resveratrol on osteoblastic and osteoclastic differentiation and its potential role in the regulation of autophagy.MethodsSprague Dawley (SD) rats underwent ovariectomies (OVX) and were administered resveratrol (at 10, 20 or 40 mg/kg/d) for 8 weeks. The calcium content and the bone mineral density (BMD) were measured in the lumbar vertebrae (L3) and the right distal femur-tibia bone region. The osteoblasts and osteoclasts were isolated from rat lumbar vertebrae by enzyme digestion and bone marrow induction, respectively. The cells were then cultured with resveratrol in combination with bafilomycin or leupeptin to inhibit or activate autophagy, respectively. Western blotting was used to assess the differentiation markers and autophagy-related genes in the osteoblasts and osteoclasts.ResultsCompared to the sham group, the bone calcium content and BMD were significantly decreased in the OVX group (p < 0.05), while resveratrol attenuated these in a dose-dependent manner. In the osteoblasts, vascular endothelial growth factor (VEGF), and alpha-1 type I collagen (COL1A1) were markedly decreased, and in osteoclasts, the receptor activator of nuclear factor-κB ligand (RANKL) was increased in the OVX group, while resveratrol reversed this pattern in a dose-dependent manner. The inhibition of autophagy in osteoblasts and its activation in osteoclasts was observed in the OVX group. However, with resveratrol, this was reversed in a dose-dependent manner.ConclusionOverall, resveratrol promotes osteoblastic differentiation and suppresses osteoclastic differentiation in a rat model with postmenopausal osteoporosis by regulating autophagy.

Anti-Osteoporotic Activity of an Edible Traditional Chinese Medicine Cistanche deserticola on Bone Metabolism of Ovariectomized Rats Through RANKL/RANK/TRAF6-Mediated Signaling Pathways.

Given the limitations of existing therapeutic agents for treatment of postmenopausal osteoporosis, there still remains a need for more options with both efficacy and less adverse effects. Cistanche deserticola Y. C. Ma is known as a popular tonic herb traditionally used to treatment deficiency of kidney energy including muscle weakness in minority area of Asian counties. Based on the theory of “kidney dominate bone,” an ovariectomized (OVX) rat model of postmenopausal osteoporosis was used to evaluate the therapeutic effect of C. deserticola extract (CDE) on bone loss. Forty eight female Sprague-Dawley rats, aged about 12 weeks, were randomly assigned into six groups including sham group orally administrated with 0.5% carboxymethyl cellulose sodium (CMC-Na) (sham), positive group treated with 1 mg/kg of estradiol valerate (EV), low, moderate, and high dosage groups orally administrated with 200, 400, and 800 mg/kg/day of CDE, respectively. After 3 months of continuous intervention, CDE exhibited significant anti-osteoporotic activity evidenced by the enhanced total bone mineral density, ameliorated bone microarchitecture; increased alkaline phosphatase activity; decreased deoxypyridinoline, cathepsin K, tartrate-resistant acid phosphatase, and malondialdehyde levels; whereas the body, uterus, and vagina weights in OVX rats were not influenced by CDE intervention. In addition, a seemed contradictory phenomenon on levels of calcium and phosphorus between OVX and sham rats were observed and elucidated. Mechanistically, CDE significantly down-regulated the levels of TRAF6, RANKL, RANK, NF-κB, IKKβ, NFAT2, and up-regulated the phosphatidylinositol 3-kinase (PI3K), AKT, osteoprotegerin, and c-Fos expressions, which implied CDE could suppress RANKL/RANK-induced activation of downstream NF-κB and PI3K/AKT pathways, and ultimately, preventing activity of the key osteoclastogenic proteins NFAT2 and c-Fos. All of the data suggested CDE possessed potential anti-osteoporotic activity and this effect was, at least in part, involved in modulation of RANKL/RANK/TRAF6-mediated NF-κB and PI3K/AKT signaling as well as c-Fos and NFAT2 levels. Therefore, CDE may represent a useful promising remedy candidate for treatment of postmenopausal osteoporosis.

Red wine polyphenols modulate bone loss in the ovariectomized rat model of postmenopausal osteoporosis

The aim of this study was to examine the effect of Red Wine Polyphenols (RWPs) extract on bone mineral density (BMD) in the ovariectomized (OVX) rat model. Thirty-five 10-month-old Wistar rats were separated into controls (CTRL), OVX and OVX plus RWPs in their drinking water (dose, 50 mg/kg body weight per day), starting immediately after OVX for 6 months. Βody and uterine weight, BMD of the tibia at baseline, 3 and 6 months post-OVX, and 3-pointing bending of the femur, were examined. Statistical comparison of the total tibia BMD within groups during the study period showed a significant reduction in the OVX and OVX+RWPs groups both from baseline to 3 and 6 months and from 3 to 6 months, whereas in the CTRL group, there was no significant change. For the proximal tibial metaphysis, comparison of BMD percentage changes from baseline to 3 months and 6 months and from 3 to 6 months revealed highly statistical differences between OVX and OVX+RWPs groups (P = 0.000). OVX induced a significant reduction of biomechanical parameters as expected; the 3-point bending test showed that the maximum force before fracture, energy absorption and fracture stress significantly increased in the OVX group treated with RWPs compared with the nontreated OVX rats (P = 0.048, P = 0.002 and P = 0.003, respectively). Dietary intake of RWPs for 6 months significantly prevented trabecular bone loss and improved bone strength in estrogen-deficient ovariectomized rats.

Effect of Swimming on Serum Irisin Levels in Experimentally-Induced Rat Model of Postmenopausal Osteoporosis

Effect of Swimming on Serum Irisin Levels in Experimentally-Induced Rat Model of Postmenopausal Osteoporosis

Development and characterization of supramolecular calcitonin assembly and assessment of its interactions with the bone remodelling process.

Osteoporosis is the most common metabolic bone disease, which poses an immense socio-economic burden on the society. Human calcitonin, though safe, is not considered as a therapeutic option because of its high tendency to self-associate to form amyloid fibrils thereby affecting its potency. To circumvent this issue we harnessed the inherent capacity of aggregation and developed an assemblage of human calcitonin monomers, [Supramolecular Calcitonin Assembly (SCAI)], which releases biologically active calcitonin monomers in a sustained manner for a period of at least three weeks. AFM and FT-IR analysis showed that SCA-I is amorphous aggregates of calcitonin monomers. Both SCA-I and monomer released from it demonstrated superior anti-osteoclast activity and proteolytic stability in-vitro. SCA-I upon single injection significantly improved bone formation markers and reduced bone resorption markers in ovariectomized (OVX) rat model of postmenopausal osteoporosis. Micro-CT analysis revealed that calcitonin released from SCA-I exhibits its beneficial effect on cortical bone more profoundly compared to trabecular bone. This study demonstrates that SCA-I is more effective compared to the human calcitonin monomers on osteoclasts and has site-specific effect on bone in a model of post-menopausal osteoporosis. This approach opens up an innovative way to use and study the function of human calcitonin.

Raman spectroscopy as a predictive tool for monitoring osteoporosis therapy in a rat model of postmenopausal osteoporosis.

Pharmacological therapy of osteoporosis reduces bone loss and risk of fracture in patients. Modulation of bone mineral density cannot explain all effects. Other aspects of bone quality affecting fragility and ways to monitor them need to be better understood. Keratinous tissue acts as surrogate marker for bone protein deterioration caused by oestrogen deficiency in rats. Ovariectomised rats were treated with alendronate (ALN), parathyroid hormone (PTH) or estrogen (E2). MicroCT assessed macro structural changes. Raman spectroscopy assessed biochemical changes. Micro CT confirmed that all treatments prevented ovariectomy-induced macro structural bone loss in rats. PTH induced macro structural changes unrelated to ovariectomy. Raman analysis revealed ALN and PTH partially protect against molecular level changes to bone collagen (80% protection) and mineral (50% protection) phases. E2 failed to prevent biochemical change. The treatments induced alterations unassociated with the ovariectomy; increased beta sheet with E2, globular alpha helices with PTH and fibrous alpha helices with both ALN and PTH. ALN is closest to maintaining physiological status of the animals, while PTH (comparable protective effect) induces side effects. E2 is unable to prevent molecular level changes associated with ovariectomy. Raman spectroscopy can act as predictive tool for monitoring pharmacological therapy of osteoporosis in rodents. Keratinous tissue is a useful surrogate marker for the protein related impact of these therapies.The results demonstrate utility of surrogates where a clear systemic causation connects the surrogate to the target tissue. It demonstrates the need to assess broader biomolecular impact of interventions to examine side effects.

Dietary Natural N-Acetyl-d-Glucosamine Prevents Bone Loss in Ovariectomized Rat Model of Postmenopausal Osteoporosis.

Postmenopausal osteoporosis has seriously affected the life quality of elderly women. A natural polymer, chitin, obtained from shells of crab and shrimp, has been widely used in the biomedical field owing to its nontoxicity, biocompatibility, and biodegradability. In this study, natural N-acetyl-d-glucosamine (NAG) was prepared from liquefied chitin. The protective activities of NAG in postmenopausal osteoporosis were evaluated on Sprague Dawley rats and osteoblast-based models. Results showed that oral administration of NAG boosted trabecular bone volume and trabecular numbers. Additionally, the calcium content in the femur and tibia increased, and femoral biomechanical properties improved. Furthermore, NAG supplementation significantly lowered alkaline phosphatase levels and increased calcium content in the serum of ovariectomized rats. In vitro studies showed that NAG markedly promoted cell proliferation and stimulated osteoblast differentiation of mouse calvaria origin MC3T3-E1 cells with increased alkaline phosphatase activity in a concentration-dependent manner. Moreover, NAG effectively protected osteoblasts from oxidative damage induced by hydrogen peroxide. In conclusion, our data provide an additional foundation for dietary supplementation of NAG, which could protect and reverse osteopenia in postmenopausal women.

Correction to: Long-Term Oral Toxicity and Anti-osteoporotic Effect of Sintered Dicalcium Pyrophosphate in Rat Model of Postmenopausal Osteoporosis

Correction to: Long-Term Oral Toxicity and Anti-osteoporotic Effect of Sintered Dicalcium Pyrophosphate in Rat Model of Postmenopausal Osteoporosis

Evaluation of ostarine as a selective androgen receptor modulator in a rat model of postmenopausal osteoporosis

Selective androgen receptor modulators (SARMs) have shown beneficial effects on muscle wasting, general physical function and bone properties in male mammals. However, data on the effects of SARMs in postmenopausal osteoporotic bone are scarce. We evaluated the effects of the SARM drug ostarine on postmenopausal osteoporotic bone in a rat osteoporosis model. Ovariectomy was performed on 46 of 56 3-month-old female Sprague-Dawley rats. Eight weeks after ovariectomy, ostarine was orally administered daily for 5weeks in dosages of 0.04 (low, OVX + Ost. 0.04), 0.4 (intermediate, OVX + Ost. 0.4), and 4mg/kg (high, OVX + Ost. 4) body weight. Another ovariectomized group received no ostarine. Lumbar vertebrae and femora were removed for biomechanical, gene expression, ashing, and computer tomography analyses. Low dose showed no effects. The effects of intermediate and high doses were comparable overall. Improvements were mainly seen in structural properties such as bone mineral density and bone volume density. However, the effects in femora were superior to effects in vertebrae. Ostarine treatment for 5weeks did not improve significantly biomechanical properties. mRNA expression of the receptor activator of NF-κB ligand decreased after treatment, and uterine weight increased. Serum levels of phosphorus increased following ostarine treatment in intermediate and high-dose groups. Short-term treatment of osteoporotic bone with ostarine leads to improvement of several microstructural bone indices. While we did not observe changes in biomechanics, it is conceivable that longer treatment may also improve biomechanical properties. Further studies are needed to characterize longer time effects and side effects of ostarine in osteoporosis.

The effects of estrogen deficiency on cortical bone microporosity and mineralization.

Recent studies have demonstrated matrix-mineral alterations in bone tissue surrounding osteocytes in estrogen-deficient animals. While cortical bone porosity has been shown to be a contributor to the mechanical properties of bone tissue, little analysis has been done to investigate the effects of estrogen deficiency on bone's microporosities, including the vascular and osteocyte lacunar porosities. In this study we examined alterations in cortical bone microporosity, mineralization, and cancellous bone architecture due to estrogen deficiency in the ovariectomized rat model of postmenopausal osteoporosis. Twenty-week-old female Sprague-Dawley rats were subjected to either ovariectomy or sham surgery. Six weeks post-surgery tibiae were analyzed using high-resolution micro-CT, backscattered electron imaging, nanoindentation, and dynamic histomorphometry. Estrogen deficiency caused an increase in cortical bone vascular porosity, with enlarged vascular pores and little change in tissue mineral density in the proximal tibial metaphysis. Measurements of cancellous architecture corresponded to previous studies reporting a decrease in bone volume fraction, an increase in trabecular separation, and a decrease in trabecular number in the proximal tibia due to estrogen deficiency. Nanoindentation results showed no differences in matrix stiffness in osteocyte-rich areas of the proximal tibia of estrogen-deficient rats, and bone labeling and backscattered electron imaging showed no significant changes in mineralization around the vascular pores. The findings demonstrate local surface alterations of vascular pores due to estrogen deficiency. An increase in cortical vascular porosity may diminish bone strength as well as alter bone mechanotransduction via interstitial fluid flow, both of which could contribute to bone fragility during postmenopausal osteoporosis.

Effects of Labisia pumila on bone turnover markers and OPG/RANKL system in a rat model of post-menopausal osteoporosis

Summary The loss of bone mass due to the increasing age in women is caused by the decline in ovarian function, known as osteoporosis. Numerous studies have evaluated the benefits of Labisia pumila, but not investigated the OPG/RANKL system. The purpose of the present study was to analyze the effects of L. pumila extract on bone turnover markers and OPG/RANKL system in ovariectomized rats. Totally, twenty-five female Wistar rats were divided into five tgroups (n = 5), including the control group, ovariectomized group, ovariectomized group treated with L. pumila extract of various doses (10 mg/kg; 20 mg/kg and 40 mg/kg). L. pumila extract was administered daily for 8 weeks. Serum levels of osteocalcin, urine levels of deoxypiridinoline, serum levels of OPG and RANKL were analyzed by means of ELISA technique. Serum levels of deoxypiridinoline and urine levels of osteocalcin were significantly higher in the ovariectomized group than those of the control group (p 0.05). In conclusion, the ethanol extract of L. pumila normalized osteoblastic bone formation, but could not prevent collagen degradation in bone in a rat model of post-menopausal osteoporosis.

In vitro cell behaviors of bone mesenchymal stem cells derived from normal and postmenopausal osteoporotic rats.

Postmenopausal osteoporosis (PMO) increases bone fragility and the risk of fractures, and impairs the healing procedure of bone defects in aged women. The stromal cell-derived factor-1α (SDF-1α)/CXC chemokine receptor type 4 (CXCR4) axis helps to maintain the biological and physiological functions of bone marrow mesenchymal stem cells (BMSCs) and increase the homing efficiency of BMSCs. The present study aimed to provide insights into the possible association between migration and osteogenic ability and the SDF-1α/CXCR4 axis in BMSCs derived from a rat model of PMO. In order to do this, the general and SDF-1α/CXCR4-associated biological characteristics as well as associated molecular mechanisms in BMSCs isolated from a PMO rat model (OVX-BMSCs) and normal rats (Sham-BMSCs) were investigated and compared. In comparison with Sham-BMSCs, OVX-BMSCs exhibited an impaired osteogenic ability, but a stronger adipogenic activity as well as a higher proliferative ability. In addition, OVX-BMSCs presented a lower chemotactic activity towards SDF-1α, lower expression levels of CXCR4 and reduced levels of phosphorylated AKT (p-AKT). Therefore, the lower expression levels of CXCR4 and p-AKT may be responsible for the impaired osteogenic ability and lower chemotactic activity towards SDF-1α of OVX-BMSCs.

Microstructural changes associated with osteoporosis negatively affect loading-induced fluid flow around osteocytes in cortical bone

Loading-induced interstitial fluid flow in the microporosities of bone is critical for osteocyte mechanotransduction and for the maintenance of tissue health, enhancing convective transport in the lacunar-canalicular system. In recent studies, our group has reported alterations of bone’s vascular porosity and lacunar-canalicular system microarchitecture in a rat model of postmenopausal osteoporosis. In this work, poroelastic finite element analysis was used to investigate whether these microstructural changes can affect interstitial fluid flow around osteocytes. Animal-specific finite element models were developed combining micro-CT reconstructions of bone microstructure and measures of the poroelastic material properties. These models were used to quantify and compare loading-induced fluid flow in the lacunar-canalicular system of ovariectomized and sham-operated rats. A parametric analysis was also used to quantify the influence of the lacunar-canalicular permeability and vascular porosity on the fluid velocity magnitude. Results show that mechanically-induced interstitial fluid velocity can be significantly reduced in the lacunar-canalicular system of ovariectomized rats. Interestingly, the vascular porosity is shown to have a major influence on interstitial fluid flow, while the lacunar-canalicular permeability influence is limited when larger than 10-20m2. Altogether our results suggest that microstructural changes associated with the osteoporotic condition can negatively affect interstitial fluid flow around osteocytes in the lacunar-canalicular system of cortical bone. This fluid flow reduction could impair mechanosensation of the osteocytic network, possibly playing a role in the initiation and progression of age-related bone loss and postmenopausal osteoporosis.

Sclerostin Antibody Reverses Bone Loss by Increasing Bone Formation and Decreasing Bone Resorption in a Rat Model of Male Osteoporosis.

Sclerostin antibody (Scl-Ab) restored bone mass and strength in the ovariectomized rat model of postmenopausal osteoporosis. Increased bone mineral density (BMD) and decreased skeletal fragility fracture risk have been reported in postmenopausal osteoporotic women receiving Scl-Ab. In males, loss of androgen leads to rapid decreases in BMD and an increased risk of fragility fractures. We hypothesized that Scl-Ab could reverse the loss of bone mass and strength caused by androgen ablation in the orchiectomized (ORX) rat model of male osteoporosis. We treated 9-month-old ORX Sprague Dawley rats (3 months after ORX) subcutaneously twice weekly with vehicle or Scl-Ab (5 or 25 mg/kg) for 6 weeks (n = 10 per group). Both doses of Scl-Ab fully reversed the BMD deficit in the lumbar spine and femur and tibia in ORX rats. Microcomputed tomography showed that the bone mass in the fifth lumbar vertebral body, femur diaphysis, and femoral neck were dose-dependently restored by Scl-Ab. The bone strength at these sites increased significantly with Scl-Ab to levels matching those of sham-operated controls and correlated positively with improvements in bone mineral content, demonstrating bone quality maintenance. Dynamic histomorphometry of the tibial diaphysis and second lumbar vertebral body demonstrated that Scl-Ab significantly increased bone formation on periosteal, endocortical, and trabecular surfaces and significantly decreased bone resorption on endocortical and trabecular surfaces. The effects of Scl-Ab on increasing bone formation and decreasing bone resorption led to restoration of bone mass and strength in androgen-deficient rats. These findings support the ongoing evaluation of Scl-Ab as a potential therapeutic agent for osteoporosis in men.

Trabecular Bone Microarchitectural Outcomes of the Mandible and Tibia or Lumbar Vertebrae are Correlated in SHAM but not the OVX Rat Model of Postmenopausal Osteoporosis

In postmenopausal (PM) women, tooth loss and/or low bone mineral density (BMD) of the jaw is correlated with low BMD (i.e. osteoporosis) at other skeletal sites. Moreover, tooth loss, in general, makes it more difficult to consume healthy foods that often require more forceful mastication. Thus, there is a cyclic relationship among oral health, consumption of a nutritious diet, and the risk of osteoporosis. The ovariectomized (OVX) rat model is the FDA‐approved model of PM osteoporosis, and is commonly used to investigate how diet can attenuate the development of osteoporosis. OVX causes a skeletal‐site specific response, such that loss of bone quantity and quality in the long bones is greater than in the lumbar vertebrae (LV) and mandible. However, few studies have evaluated the correlation of trabecular microarchitectural outcomes, as measured by micro‐computed tomography (mCT), between mandible and tibia or LV to determine the fitness of the mandible as a predictor of overall bone health. The objective of this study was to determine if trabecular microarchitectural outcomes of the mandible and the tibia or LV are correlated in this widely used pre‐clinical model of PM osteoporosis, to understand how closely this model mimics changes observed in PM women. Three‐month old Sprague‐Dawley rats were randomized to SHAM (n=47) or OVX (n=49) and were maintained on an ad libitum diet until 6 months of age. Ex vivo mCT imaging of the mandible (alveolar bone) and LV, and in vivo mCT imaging of proximal tibia was conducted. Trabecular microarchitectural measurements, including percent bone volume (BV/TV), trabecular thickness (Tb.Th.), trabecular number (Tb.N.), trabecular separation (Tb.Sp.), and degree of anisotropy were quantified at each bone site. Correlation between these outcomes (mandible vs. tibia; mandible vs. LV) within SHAM or OVX rats was conducted using a Pearson's correlation test. Within rats randomized to SHAM, positive correlations exist between all bone sites for the outcomes BV/TV (p&lt;0.001), Tb.N. (p&lt;0.001), and Tb.Sp. (p&lt;0.05). A negative correlation was observed between the mandible and LV for the outcome Tb.Th. (p&lt;0.05). Within rats randomized to OVX, the sole positive correlation was between the mandible and tibia for Tb.Sp. (p&lt;0.05). As expected, when the percent change between SHAM and OVX groups was assessed at 6 months of age, the most dramatic effect was seen at the tibia (−71% BV/TV), followed by the LV (−36% BV/TV) and mandible (−8% BV/TV). These results demonstrate that OVX has skeletal‐site specific effects on trabecular microarchitecture, and these relative responses may at least partially explain the lack of correlations among mandible and tibia or LV after OVX. In conclusion, positive correlations between BMD and trabecular bone microarchitecture of mandible and tibia or LV are observed in SHAM rats but not after OVX. Therefore, the alveolar bone of the mandible of OVX rats does not closely mimic the bone loss that occurs in PM women and does not reliably predict changes at other skeletal sites in the time frame studied.Support or Funding InformationThe authors acknowledge funding provided by the Natural Sciences and Engineering Research Council of Canada and the Canada Research Chairs program as well as infrastructure support from the Canada Foundation for Innovation.

The effects of Labisia pumila extracts on bone microarchitecture of ovariectomized-induced osteoporosis rats: A micro-CT analysis.

Labisia pumila (LP) is a popular herb used by women over the past few decades. This herb has shown potentials as an alternative agent for treatment and prevention of postmenopausal osteoporosis. It was observed in previous studies that supplementation to ovariectomized rats were associated with increased bone antioxidative enzymes and reduced lipid peroxidation activity. It had also improved bone formation markers in ovariectomized rats. This study aimed to evaluate the effects of giving different forms of LP extracts on the trabecular bone microarchitecture of ovariectomised rats. Forty-eight female Sprague-Dawley rats were randomly divided into sham-operated (Sham), ovariectomized control (OVX), ovariectomized and given estrogen at 64.5 μg/kg (ERT), ovariectomized and given LP aqueous extract (LPaq), LP methanol extract (LPmet) and LP ethanol extract (LPet) at 100 mg/kg, respectively. Treatments were given daily via oral gavages for nine weeks. Following sacrifice, femora were dissected out for bone microarchitectural analysis using an in vitro micro-CT, which provided three dimensional informations on bone microarchitecture. LPaq was the most effective extract found to improve the bone microarchitectural paramaters which comprised ofBone volume fraction (BV/TV), Trabecular separation (Tb.Sp), Trabecular number (Tb.N), Connective density (Conn.dens), Structure model index (SMI) and Degree of anisotropy (DA). LPaq was effective in protecting the bone of postmenopausal osteoporosis rat model against microarchitectural deterioration.

Long-Term Oral Toxicity and Anti-osteoporotic Effect of Sintered Dicalcium Pyrophosphate in Rat Model of Postmenopausal Osteoporosis

Sintered dicalcium pyrophosphate (SDCP), a synthetic pyrophosphate analog, has shown potential for the management of osteoporosis. The long-term oral toxicity and anti-osteoporotic effect of SDCP in a postmenopausal osteoporosis rat model were evaluated in this study. SDCP was orally administered to bilateral ovariectomized (OVX) Wistar rats at a dose of 0.75 mg/kg daily for 24 weeks following by 2 weeks of observation. There were no abnormal findings in clinical signs of toxicity, food consumption, body weight, blood examination, necropsy, and histological inspection attributable to the ingestion of SDCP. The serum level of type I collagen fragments, a bone resorption marker, decreased in SDCP-treated rats, and the bone formation markers alkaline phosphatase, osteocalcin, and osteopontin significantly decreased. These findings indicate that the bone turnover rate decreased in SDCP-treated animals. Relative to OVX rats, the increase in serum tartrate-resistant acid phosphatase 5b level represents an increase in bony tissues in the SDCP-treated rats. Histological examinations of distal femoral metaphyses further revealed that the ingestion of SDCP improved the trabecular bone architecture and decreased bone porosity. Analysis of limb bone ashes showed a significant increase in bone mineral content. Our results show that SDCP inhibits bone resorption to restore bone mass in OVX rats without deleterious effects, and therefore that SDCP has potential in the management of osteoporosis.