Serum OC Research Articles

Phenolic constituents of Trifolium resupinatum var. minus: Protection against rosiglitazone induced osteoporosis in type 2 diabetic male rats

Thiazolidinediones, an antidiabetic drug, promote bone loss and provoke fractures, suggesting a protective role of phytoestrogens rich herbal supplement along with rosiglitazone. The aqueous methanol extract (AME) of Trifolium resupinatum L. var. minus Boiss. (Fabaceae) and defatted AME (DAME) protective effects were assessed against rosiglitazone (Ros) inducing osteoporosis in type 2 diabetic male rats. The bioactive extract was investigated for its phenolics. Eighty male Albino rats (10/group) were used; G1: control, G2: diabetic rats + Ros (10 mg/kgbw/d for 30 days; Ros-rats), G3-G5 and G6-G8: Ros-rats + AME or DAME (24, 12, 6 mg/kgbw/d, each). OPG, RANKL and β2- microglobulin serum levels, femur BMD (Dexa) and OC, COL and ACP5 osteogenic genes mRNA (qRT-PCR) were evaluated. Phenolics were isolated and characterized by chromatographic, chemical and spectral techniques. Their HPLC profiling was also analyzed. Ros-rats + DAME (24 mg/kgbw) revealed the most significant effects on bone biomarkers. It modulated OPG (ng mL-1), RANKL (pg mL-1) and β2- microglobulin (μg mL-1) serum levels with 2.46 ± 0.44, 96.640 ± 06.12, 0.22 ± 0.04, compared to those of Ros--rats (1.62 ± 0.26, 162.42 ± 11.34, 0.46 ± 0.05), respectively. Also, it improved BMD (mg cm-2) of proximal, distal and total femur bones (102.8 ± 8.6, 104.6 ± 10.2, 106.0 ± 9.2), compared to Ros-rats (82.3± 6.4, 81.2 ± 7.2, 82.1 ± 4.8), respectively and up-regulated OC & COL and down-regulated ACP5 expressions. From DAME, 12 phenolics were identified and HPLC profiling showed major phenolics; chlorogenic acid > formononetin> quercetin> pseudobaptigenin. DAME in combination therapy with rosiglitazone provides protective effect on bone, possibly due to its phenolics.

Vaspin and selected indices of bone status in girls with anorexia nervosa.

In vitro studies indicate that vaspin may act as a regulator of bone metabolism. The aim of the study was to evaluate the relationship between vaspin and bone metabolism in girls with anorexia nervosa (AN), as well as the potential involvement of OPG and RANKL in this relationship. Serum vaspin, OC, CTx, OPG, and sRANKL were determined by ELISA in 50 girls with AN and in 30 healthy controls aged 13 to 17 years. Girls with AN exhibited significant reduction in body weight, BMI, and Cole index as well as a significant increase in serum level of vaspin compared to healthy participants. These changes were associated with a significant decrease in serum OC and CTx levels and a significant increase in OPG and sRANKL, while the OPG/sRANKL ratio was significantly decreased. BMI and Cole index correlated negatively and significantly with CTx levels in the control group (C), girls with AN, and all study participants (C+AN). Girls with AN showed a significant negative correlation between BMI, the Cole index, and OPG levels. The combination group (C+AN) showed a significant positive correlation between BMI, Cole index, and the OPG/sRANKL ratio. In this group of girls vaspin levels correlated positively and significantly with sRANKL and negatively with body weight, BMI, Cole index, and OPG/sRANKL ratio. Girls with AN showed a significant negative correlation between vaspin levels and the OPG/sRANKL ratio. Undernourishment and associated deficit of adipose tissue may result in inadequate vaspin production and bone metabolism disorders in girls with AN. Vaspin acts as a coordinator of the dynamic balance between bone formation and resorption processes; its action is affected by the cytokines of the RANKL/RANK/OPG system. Changes in the relationships between vaspin, bone markers, OPG, and RANKL might contribute to the development of osteoporosis in girls with AN. (Endokrynol Pol 2016; 67 (6): 599-606).

Soy isoflavone supplementation improves longitudinal bone growth and bone quality in growing female rats

ObjectivesThe aim of this study was to investigate the effects of soy isoflavone on tibia length, bone mineral density (BMD), and structural parameters in growing female rats. MethodsThree-week-old female Sprague-Dawley rats were randomly assigned to four experimental groups: control (CON: distilled water gavage); low-dose isoflavone (low-IF: 10 mg/kg body weight [BW]/d gavage); high-dose isoflavone (high-IF: 50 mg/kg BW/d gavage); and 17 β-estradiol (E2: subcutaneous injection of 10 μg). All animals received a soy-free diet and vaginal opening was monitored daily. After an 8-wk treatment period, bone-related parameters (alkaline phosphatase [ALP], osteocalcin [OC], N-terminal telopeptide [NTx], bone length, failure load, stiffness, BMD, and structural parameters) were analyzed. ResultsSerum ALP levels of the high-IF group were higher than those of the CON group (P < 0.05); however, serum OC levels of the high-IF group were lower than those of the CON, low-IF, and E2 groups (P < 0.05). The tibias and femurs of the low-IF group were longer than those of the CON and high-IF groups (P < 0.05). Bone volume, trabecular number, and BMD of trabecular bone of the high-IF and E2 groups were higher than those of the CON and low-IF groups (P < 0.05). The trabecular thickness of the high-IF group was higher than that of the CON and low-IF groups (P < 0.05). The failure load of the high-IF group was higher than those of the CON group (P < 0.05). Age and body weight at vaginal opening of the E2 group were significantly lower than those of the CON, low-IF, and high-IF groups (P < 0.05). ConclusionsThis study suggests that 8 wk of low-dose supplementation with soy isoflavone stimulates longitudinal bone growth. Additionally, high-dose supplementation with soy isoflavone may improve bone quality (BMD and structural parameters) in growing female rats.

TGF-β1, bone metabolism, osteoprotegerin, and soluble receptor activator of nuclear factor-kB ligand in girls with anorexia nervosa.

Numerous investigations, and especially in vitro studies, indicate that TGF-β1 may act as an important regulator of bone remodelling. Thus, it could be expected that disturbances of this cytokine production observed by several researchers might play a role in the mechanism leading to the development of osteoporosis in girls with anorexia nervosa (AN). The aim of the study was to determine whether 1) girls with AN exhibited a relationship between TGF-β1 and bone metabolism (as assessed based on serum OC and CTx concentrations) and 2) whether OPG and sRANKL might modify the possible relationship between TGF-β1 and bone metabolism. Serum concentrations of TGF-β, OC, CTx, OPG, and its soluble ligand sRANKL were determined by ELISA in 60 girls with AN and in 20 healthy controls (C). All study participants were aged 13 to 17 years. Body weight, BMI, BMI-SDS and the Cole index, serum TGF-β1, OC, CTx, and the OPG/sRANKL ratio were significantly reduced, while OPG and sRANKL levels were significantly increased, in girls with AN compared to healthy participants. BMI and the Cole index correlated negatively and significantly with serum CTx and OPG (AN group) or CTx only (groups C and C + AN). Girls with AN showed a positive and significant correlation between the Cole index and serum TGF-β1. The combination group (C + AN) showed a positive and significant correlation between BMI, the Cole index, and the OPG/sRANKL ratio and TGF-β1 concentration, while TGF-β1 correlated positively and significantly with OC concentrations and the OPG/sRANKL ratio. The Cole index and BMI were identified to be significant and independent predictors of CTx (C, AN, and C+AN groups) and OPG (AN group); the Cole index, BMI, and TGF-β1 independently predicted the OPG/sRANKL ratio (C, AN, and C + AN groups); TGF-β1 was found to be an independent predictor of OC (C + AN group). Changes in bone markers, OPG, and/or OPG/sRANKL ratio observed in girls with AN are associated with changes in serum TGF-β1 concentrations. TGF-β1 suppression in girls with AN might lead to disturbances in the relationship between bone metabolism and the OPG/sRANKL system, which, in turn, might compromise the mechanism compensating for bone remodelling disturbances. (Endokrynol Pol 2016; 67 (5): 493-500).

Association between omentin-1, bone metabolism markers, and cytokines of the RANKL/RANK/OPG system in girls with anorexia nervosa.

Omentin-1, secreted by visceral adipose tissue, has been indicated in the regulation of bone metabolism in girls with anorexia nervosa (AN). The aim of the study was to evaluate the relationship between omentin-1 and bone metabolism in girls with AN as well as the potential involvement of OPG and RANKL in this relationship. Serum omentin-1, OC, CTx, OPG, and sRANKL were determined by ELISA in 49 girls with AN and in 30 healthy controls, aged 13 to 17 years. Girls with AN exhibited significant reduction in body weight, BMI, and Cole index as well as a significant increase in serum omentin-1 levels, compared to healthy participants. These changes were associated with a significant decrease in serum OC and CTx levels and a significant increase in OPG and sRANKL while the OC/CTx and OPG/sRANKL ratios were significantly decreased. BMI and the Cole index correlated negatively and significantly with omentin-1 levels, positively with CTx levels and the OC/CTx ratio in the control group (C), girls with AN, and all study participants (C + AN). Girls with AN showed a significant negative correlation between BMI, the Cole index, and OPG levels. The combined group (C + AN) showed a significant positive correlation between BMI, the Cole index, and the OPG/sRANKL ratio. Omentin-1 levels correlated negatively and significantly with OC and CTx levels as well as with the OC/CTx and OPG/sRANKL ratios in the C, AN, and C + AN groups. The relationship between omentin-1, bone markers, and the OC/CTx and OPG/sRANKL ratios observed in girls with AN indicates the involvement of this adipokine in the regulation of dynamic balance between bone formation and resorption processes. Omentin-1 might exert a negative effect on bone remodelling in girls with AN by inhibiting both bone formation and resorption. The OPG/sRANKL system plays an important role in the latter.

Serum Osteocalcin and Serum Osteopontin Levels in Osteoporotic Postmenopausal Women with and without Vertebral Fractures

Background: Osteoporosis is a progressive systemic skeletal disorder characterized by low bone mass and micro-architectural deterioration of bone tissue spatially in postmenopausal women and its major complication fractures.Objective: The aim of this study was to find out the significance of serum OC and serum OPN levels with the incidence of osteoporosis and its major complication (fractures).Patients and Methods: Eighty-five postmenopausal women (PMW) whose ages were fifty years and over categorized into three groups: osteoporosis PMW without VFs (n=30), osteoporosis PMW with VFs (n=28), and healthy PMW (n=27). Sera samples were analyzed for alkaline phosphatase, calcium and phosphorous by using spectrophotometric kit. Serum OC and serum OPN levels were measured by ELISA kits.Results: Bone Mineral Density (BMD) and T-score were significantly lower in osteoporotic PMW with and without VFs as compared with healthy PMW (p=0.0001, p=0.0001, respectively). Serum OC levels and serum OPN levels were elevated significantly in osteoporotic PMW with and without VFs as compared with healthy PMW (p=0.0001, p=0.0001, respectively), the levels of serum OC and OPN showed a significant positive correlation with age in osteoporotic PMW. There is a significant positive correlation between serum OC levels and serum OPN levels in osteoporotic PMW and non-significant correlation was found in healthy PMW.Conclusion: The levels of serum OC and OPN can be used as a biochemical indicator in the diagnosis of postmenopausal osteoporotic women

Calorie restriction aggravated cortical and trabecular bone architecture in ovariectomy-induced estrogen-deficient rats

We hypothesized that calorie restriction (CR) and estrogen deficiency (ovariectomy [OVX]) would aggravate bone biomarkers and structural parameters in rats. Seven-week-old female Sprague-Dawley rats were randomized to sham-operated groups and fed either an ad libitum diet (SHAM-AL) or a CR diet (SHAM-CR); ovariectomy-operated groups were fed an ad libitum diet (OVX-AL) or a CR diet (OVX-CR). For 8 weeks, the OVX-AL and SHAM-AL groups were fed the same diet, whereas CR groups were fed a diet containing 50% fewer calories. Bone-related biomarkers and structural parameters (OC; deoxypyridinoline [DPD]; N-terminal telopeptide, NTx; architecture and mineralization; and microcomputed tomography images) were analyzed at the end of the experiment. The serum OC levels of calorie-restricted groups (SHAM-CR and OVX-CR) were significantly lower than those of the AL groups (SHAM-AL and OVX-AL) (P < .05). Urinary DPD levels of calorie-restricted and ovariectomized groups were higher than those of their counterparts (P < .05), whereas urinary NTx levels of calorie-restricted groups were higher than those of AL groups (P < .05). In regard to trabecular bone, the calorie-restricted and ovariectomized groups had lower values of bone volume to total volume, trabecular number, and bone mineral density, but higher values of trabecular separation than those of their counterparts (P < .05). Regarding cortical bone, the calorie-restricted groups had reduced values of bone volume, mean polar moment of inertia, and cortical thickness compared to the AL groups (P < .05). In conclusion, severe CR with or without OVX during the growth period in rats is equally detrimental to bone; CR has detrimental effects on trabecular and cortical bone; and estrogen deficiency only had an effect on trabecular bone.

Gastric bypass in obese rats causes bone loss, vitamin D deficiency, metabolic acidosis, and elevated peptide YY

BackgroundMetabolic bone disease and bariatric surgery have long been interconnected. The objective of this study is to better understand the mechanisms of bone mass loss after Roux-en-Y gastric bypass (RYGB) surgery. We evaluated mineral homeostasis and bone mass in diet-induced obese (DIO) rats after RYGB or sham surgery. MethodsTwelve DIO male Sprague Dawley rats underwent RYGB (n = 8) or sham (n = 4) surgery at 21 weeks of age. Postoperatively, animals ate an ad libitum 40% fat, normal calcium diet and were euthanized 22 weeks later. Serum and urine chemistries, insulin, leptin, bone turnover markers (BTM), and calciotropic and gut hormones were measured before and 22 weeks after surgery. Femurs were analyzed using microcomputed tomography (µCT). ResultsCompared to sham, RYGB animals had lower serum bicarbonate, calcium, 25-hydroxyvitamin D, insulin, and leptin levels with higher serum parathyroid hormone, peptide YY, and urinary calcium at 43 weeks of age. Sham control rats gained weight and had coupled decreases in formation (P1NP and OC) and unchanged resorption (CTX) BTMs. Comparatively, RYGB animals had higher serum CTX and OC but even lower P1NP levels than controls. µCT revealed lower trabecular bone volume, number, and thickness and lower cortical bone volume, thickness, and moment of inertia relative to controls. ConclusionIn rats with DIO, long-term RYGB-associated bone resorption appears to be driven in part by vitamin D malabsorption and secondary hyperparathyroidism. Other mechanisms, such as chronic acidosis, changes in fat-secreted hormones, and persistently elevated gut-derived hormone peptide YY, may also contribute to observed bone mass differences. Further investigation of these potential contributors to bone loss may lead to new targets for skeletal maintenance after RYGB.

Preventive Effects of Eleutherococcus senticosus Bark Extract in OVX-Induced Osteoporosis in Rats

Eleutherococcus senticosus (Siberian ginseng), has been used as a powerful tonic herb with an impressive range of health benefits. This medicinal herb has been commonly used to treat bone metabolism diseases due to its traditional Korean medicine use to strengthen muscle and bone. This study was conducted to investigate prevention of bone loss by a standardized extract of dried E. senticosus stem bark in an ovariectomized (OVX) rat model of osteoporosis. The OVX groups were divided into five groups treated with distilled water, 17β-estradiol (E2 10 μg/kg, once daily, i.p) and dried stem bark of E. senticosus extracts (DES 10, 30, and 100 mg/kg, once daily, p.o) for eight weeks, respectively. After eight weeks of treatments, the femur bone mineral density of the 100 mg/kg DES-treated group was significantly higher than that of the OVX-control group (16.7%, p < 0.01) without affecting the body, organs, and uterus weights, and serum estradiol levels. Additionally, bone markers such as serum ALP, CTx, and OC levels were significantly decreased in the DES 100 mg/kg treated group. These results show that DES is able to prevent OVX-induced in bone loss without the influence of hormones such as estrogen.

Identification of a Vibration Regime Favorable for Bone Healing and Muscle in Estrogen-Deficient Rats

Numerous whole-body vibration (WBV) devices of various forces are available on the market, although their influence on the musculoskeletal system is not yet understood. The effect of different WBVs on bone healing and muscle function was evaluated in rats ovariectomized at 3 months of age. 2 months after ovariectomy, bilateral metaphyseal tibia osteotomy and T-plate osteosynthesis were performed. Rats were divided into groups: intact, OVX, and OVX exposed to vertical WBVs of 35, 50, 70, or 90 Hz (experiment 1) or horizontal WBVs of 30, 50, 70, or 90 Hz (experiment 2) 5 days after osteotomy (0.5 mm, 15 min/day for 30 days). The tibia and gastrocnemius and soleus muscles were collected. Vertical vibrations (>35 Hz) improved cortical and callus densities, enlarged callus area and width, suppressed the tartrate-resistant acid phosphatase gene, enhanced citrate synthase activity, accelerated osteotomy bridging (35 and 50 Hz), upregulated the osteocalcin (Oc) gene (70 Hz), and increased relative muscle weight (50 Hz). Horizontal vibrations reduced cortical width (<90 Hz) and callus density (30 Hz), enhanced alkaline phosphatase (Alp) gene expression (50 Hz), decreased the size of oxidative fibers (35 and 70 Hz), and increased capillary density (70, 90 Hz). Biomechanical data; serum Oc, Alp, and creatine kinase activities; body weight; and food intake did not change after WBVs. Vertical WBVs of 35 and 50 Hz produced more favorable results than the higher frequencies. Horizontal WBV showed no positive or negative effects. Further studies are needed to elucidate the effects of WBV on different physiological systems, and precautions must be taken when implementing WBV in the treatment of patients.

Serum under-carboxylated osteocalcin levels in women with polycystic ovary syndrome: weight-dependent relationships with endocrine and metabolic traits

BackgroundUnder-carboxylated osteocalcin (ucOC), the precursor substrate of bone biomarker OC is a potent regulator of energy metabolism by promoting insulin production and adiponectin synthesis and decreasing fat stores. The aim of the present study was to point out the potential role of ucOC in the physiopathology of polycystic ovary syndrome (PCOS), a common disorder defined by the constellation of anovulation, insulinresistance, hyperinsulinemia, obesity and androgen excess.MethodsIn this prospective case–control investigation, 78 young premenopausal women, i.e. 52 PCOS patients and 26 age- and body mass index (BMI)-matched healthy controls, were successively enrolled. Recruitment of PCOS patients was performed according to Androgen Excess-Polycystic Ovary Syndrome (AE-PCOS) Society 2006 criteria. All study participants were subjected to clinical examination, whole-body composition assessment and measurements of serum ucOC, OC (1-49), glucose and lipids, insulin, total testosterone (TT), estradiol, sex-hormone binding globulin (SHBG), high-sensitivity C-reactive protein (Hs-CRP) and β-CrossLaps.ResultsBMI-stratified multivariate analysis revealed significantly higher ucOC levels in PCOS vs. controls in lean (p = 0.001) but not overweight and obese study participants (p = 0.456). Notably, a positive correlation between ucOC and TT (p = 0.018), calculated free testosterone (cFT, p = 0.028) and serum insulin (p = 0.036), respectively, was found to be confined to the lean analysis subgroup. Furthermore, in stepwise multiple regression models, β-CrossLaps and cFT were able to predict 46.71% of serum ucOC variability. (1-43/49)OC failed to be significantly associated to any PCOS trait.ConclusionsCirculating ucOC concentration is related to key endocrine PCOS characteristics in a weight-dependent manner. Within the bone-pancreas loop, high ucOC may favor insulin release in lean hyperandrogenic women to compensate for impaired insulin sensitivity.

Dehydroepiandrosterone sulfate, osteoprotegerin and its soluble ligand sRANKL and bone metabolism in girls with anorexia nervosa

Only scarce data exist concerning the relationship between dehydroepiandrosterone (DHEA) and/or its sulfate form DHEAS and bone status in adolescents with anorexia nervosa (AN). We investigated whether a relationship existed between DHEAS and bone metabolism (as assessed based on serum osteocalcin [OC], and collagen type I cross-linked carboxy-terminal telopeptide [CTx]). We also aimed to establish whether the above mentioned relationship might be affected by osteoprotegerin (OPG) and its soluble ligand sRANKL. Fifty-six female patients with AN and 21 healthy female subjects aged 13 to 16 years participated in the study. Serum DHEAS, OC, CTx, OPG and sRANKL were measured by ELISA. Our female patients with AN demonstrated significant suppression of DHEAS and bone markers, an increase in OPG and sRANKL levels, and a reduction of the OPG/sRANKL ratio. DHEAS, CTx and the OPG/sRANKL ratio correlated positively with BMI. A significant positive correlation was also observed between DHEAS and the OPG/sRANKL ratio, OC and the OPG/sRANKL ratio, and CTx and sRANKL. The correlation was negative in the case of DHEAS and CTx, DHEAS and sRANKL, CTx and the OPG/sRANKL ratio, and sRANKL and the OPG/sRANKL ratio. DHEAS suppression in girls with anorexia nervosa was associated with a decrease in the levels of bone markers, an increase in OPG and sRANKL concentrations and a significant decrease in the OPG/sRANKL ratio. DHEAS suppression in girls with anorexia nervosa might have a harmful effect on their bone tissue, probably via a shift in the OPG/RANKL ratio toward a functional excess of sRANKL.

Opuntia humifusa Supplementation Increased Bone Density by Regulating Parathyroid Hormone and Osteocalcin in Male Growing Rats

We investigated the effect of Opuntia humifusa (O. humifusa) supplementation on bone density and related hormone secretion in growing male rats. Sixteen six-week-old male Sprague-Dawley rats were randomly divided into two groups; control diet group (CG, n = 8), and experimental diet group (EG, n = 8). The rats in the CG were given a control diet and those in the EG were given 5% O. humifusa added to the control diet for eight weeks. The serum OC level of the EG was significantly higher than that of the CG, and the serum parathyroid hormone (PTH) level of EG was significantly lower than that of the CG. In addition, the femoral and tibial BMD of the EG were significantly higher values than those of the CG, and the tibial BMC of the EG was significantly higher than that of the CG. These results suggest that O. humifusa supplementation has a positive effect on bone density by suppressing PTH and increasing the OC level in growing male rats.

RANKL/RANK/OPG system and bone status in females with anorexia nervosa

Minimal data exist concerning the relationship between osteokines of the RANKL/RANK/OPG system, especially RANKL, and bone status in females with anorexia nervosa (AN). For this reason we investigated the relationship between bone metabolism (as assessed based on serum levels of OC and CTx), and OPG and sRANKL concentrations in females with AN. Ninety-one female patients with AN and 29 healthy female subjects aged 13 to 18 years of age participated in the study. Serum OC, CTx, OPG and sRANKL were measured by ELISA. The female patients with AN demonstrated an essential suppression of OC and CTx, increased OPG and sRANKL levels, and a reduced OPG/sRANKL ratio. OC, CTx and the OPG/sRANKL ratio correlated positively with body mass and BMI in these patients, whereas in the case of OPG and sRANKL the relationship was negative. A significant positive correlation was observed between OPG and sRANKL and also between bone markers and the OPG/sRANKL ratio, and negative between CTx and sRANKL. In female patients with AN, the OPG/RANKL ratio was a significant and independent predictor of osteocalcin, a bone formation marker - OC (R(2)=0.065, p=0.012) whereas the OPG/sRANKL ratio and BMI were significant and independent predictors of a bone resorption marker - CTx (R(2)=0.095; p=0.012). In conclusion, the body mass, BMI values, and bone markers suppression observed in female patients with AN might be associated with an increase in OPG and sRANKL levels and a significant decrease of the OPG/sRANKL ratio. Although higher OPG levels may compensate for excessive bone resorption in female patients with AN, the lower OPG/sRANKL ratio seems to indicate that some inadequacies exist regarding this compensation effect, which might contribute to low bone density in these patients. The OPG/sRANKL ratio might prove a more relevant marker to predict bone metabolism in female patients with AN than sRANKL and/or OPG alone.

Genetic analysis of serum osteocalcin and bone mineral in multigenerational Afro-Caribbean families

Osteocalcin is a major component of bone matrix. Concentrations of total, carboxylated, and uncarboxylated osteocalcin, are highly heritable and genetically correlated with bone mineral content (BMC) within African ancestry families. Osteocalcin (OC) is a protein constituent of bone matrix and a marker of bone formation. We characterized the heritability of serum OC measures and identified genomic regions potentially involved in the regulation of OC via high-density genome-wide linkage analysis in African ancestry individuals. African ancestry individuals (n = 459) were recruited, without regard to health status, from seven probands (mean family size = 66; 4,373 relative pairs). Residual heritability of serum OC measures was estimated and multipoint quantitative trait linkage analysis was performed using pedigree-based maximum likelihood methods. Residual heritabilities of total OC, uncarboxylated OC, carboxylated OC and percent uncarboxylated OC were 0.74 ± 0.10, 0.89 ± 0.08, 0.46 ± 0.10 and 0.41 ± 0.09, respectively. All OC measures were genetically correlated with whole body BMC. We obtained strong evidence of bivariate linkage for percent uncarboxylated OC and whole body BMC on chromosome 17 (logarithm of the odds [LOD] = 3.15, 99 cM). All forms of OC were highly heritable and genetically correlated with total body BMC in these African ancestry families. The identified linkage region contains several candidate genes for bone and energy metabolism including COL1A1 and TNFRSF11A. Further studies of this genomic region may reveal novel insight into the genetic regulation of OC and bone mineralization.

Integrin αvβ3 as a PET Imaging Biomarker for Osteoclast Number in Mouse Models of Negative and Positive Osteoclast Regulation

The goal of this study was to determine the specificity of ⁶⁴Cu-CB-TE2A-c(RGDyK) (⁶⁴Cu-RGD) for osteoclast-related diseases, such as Paget's disease or rheumatoid arthritis. C57BL/6 mice were treated systemically with osteoprotegerin (OPG) for 15 days or RANKL for 11 days to suppress and stimulate osteoclastogenesis, respectively. The mice were then imaged by positron emission tomography/computed tomography using ⁶⁴Cu-RGD, followed by determination of serum TRAP5b and bone histology. Standard uptake values were determined to quantify ⁶⁴Cu-RGD in bones and other tissues. Mice treated with OPG showed decreased bone uptake of ⁶⁴Cu-RGD at 1, 2, and 24 h post-injection of the tracer (p < 0.01 for all time points) compared to vehicle controls, which correlated with a post-treatment decrease in serum TRAP5b. In contrast, mice treated with RANKL showed significantly increased bone uptake at 2 h post-injection of (⁶⁴Cu-RGD (p < 0.05) compared to the vehicle control group, corresponding to increased serum TRAP5b and OC numbers as determined by bone histology. These data demonstrate that ⁶⁴Cu-RGD localizes to areas in bone with increased osteoclast numbers and support the use of ⁶⁴Cu-RGD as an imaging biomarker for osteoclast number that could be used to monitor osteoclast-related pathologies and their treatments.

Change of BMD after Weaning or Resumption of Menstruation in Chinese Women with Different FokI VDR-genotypes: A Randomized, Placebo-controlled, Calcium Supplementation Trial

Objective To investigate the effect of calcium supplementation on bone mineral density (BMD) in Chinese women with different FokI vitamin D receptor (VDR) genotypes (FF, Ff, and ff) after weaning or resumption of menstruation during lactation. Methods A total of 40 subjects with the same FokI VDR genotype were randomly divided into two groups: one received calcium tablet (600 mg once daily as CaCO 3) and the other placebo tablet once daily for 1 year. At baseline, BMD was measured by dual-energy X-ray absorptiometry at lumbar spine (L2-L4) and at left hip whereas serum PICP, serum OC, and urinary CTX, serum 25(OH)VitD 3, and serum estradiol were measured at weaning and 1 year thereafter. Results After the intervention, BMD at lumbar spine and at left hip increased significantly in all these women with a trend among different FokI VDR genotypes such as FF > Ff > ff ( P<0.05, <0.01, and <0.001, respectively). BMD at lumbar spine in women with FF VDR genotype increased much more rapidly than in those with ff VDR genotype ( P<0.05). Compared with the control group women with the FF genotype regained more BMD after calcium supplementation ( P<0.05). Conclusion Daily calcium 600 mg supplementation has beneficial effect on the bone health of women with FF VDR genotype.

The preventive effects of pulsed electromagnetic fields on diabetic bone loss in streptozotocin-treated rats

The present study was the first report demonstrating that pulsed electromagnetic field (PEMF) could partially prevent bone strength and architecture deterioration and improve the impaired bone formation in streptozotocin-induced diabetic rats. The findings indicated that PEMF might become a potential additive method for inhibiting diabetic osteopenia or osteoporosis. Diabetes mellitus (DM) can cause various musculoskeletal abnormalities. Optimal therapeutic methods for diabetic bone complication are still lacking. It is essential to develop more effective and safe therapeutic methods for diabetic bone disorders. Pulsed electromagnetic field (PEMF) as an alternative noninvasive method has proven to be effective for treating fracture healing and osteoporosis in non-diabetic conditions. However, the issue about the therapeutic effects of PEMF on diabetic bone complication has not been previously investigated. We herein systematically evaluated the preventive effects of PEMF on diabetic bone loss in streptozotocin-treated rats. Two similar experiments were conducted. In each experiment, 16 diabetic and eight non-diabetic rats were equally assigned to the control, DM, and DM + PEMF group. DM + PEMF group was subjected to daily 8-h PEMF exposure for 8 weeks. In experiment 1, three-point bending test suggested that PEMF improved the biomechanical quality of diabetic bone tissues, evidenced by increased maximum load, stiffness, and energy absorption. Microcomputed tomography analysis demonstrated that DM-induced bone architecture deterioration was partially reversed by PEMF, evidenced by increased Tb.N, Tb.Th, BV/TV, and Conn.D and reduced Tb.Sp and SMI. Serum OC analysis indicated that PEMF partially prevented DM-induced decrease in bone formation. In experiment 2, no significant difference in the bone resorption marker TRACP5b was observed. These biochemical findings were further supported by the dynamic bone histomorphometric parameters BFR/BS and Oc.N/BS. The results demonstrated that PEMF could partially prevent DM-induced bone strength and architecture deterioration and improve the impaired bone formation. PEMF might become a potential additive method for inhibiting diabetic osteoporosis.

Musculoskeletal Response to Whole-Body Vibration During Fracture Healing in Intact and Ovariectomized Rats

This study investigated the effect of vibration on bone healing and muscle in intact and ovariectomized rats. Thirty ovariectomized (at 3 months of age) and 30 intact 5-month old female Sprague-Dawley rats underwent bilateral metaphyseal osteotomy of tibia. Five days later, half of the ovariectomized and of the intact rats were exposed to whole-body vertical vibration (90 Hz, 0.5 mm, 4 × g acceleration) for 15 min twice a day during 30 days. The other animals did not undergo vibration. After decapitation of rats, one tibia was used for computed tomographic, biomechanical, and histological analyses; the other was used for gene expression analyses of alkaline phosphatase (Alp), osteocalcin (Oc), tartrate-resistant acid phosphatase 1, and insulinlike growth factor 1. Serum Alp and Oc were measured. Mitochondrial activity, fiber area and distribution, and capillary densities were analyzed in M. gastrocnemius and M. longissimus. We found that vibration had no effect on body weight and food intake, but it improved cortical and callus densities (97 vs. 99%, 72 vs. 81%), trabecular structure (9 vs. 14 trabecular nodes), blood supply (1.7 vs. 2.1 capillaries/fiber), and oxidative metabolism (17 vs. 23 pmol O2/s/mg) in ovariectomized rats. Vibration generally increased muscle fiber size. Tibia biomechanical properties were diminished after vibration. Oc gene expression was higher in vibrated rats. Serum Alp was increased in ovariectomized rats. In ovariectomized rats, vibration resulted in an earlier bridging; in intact rats, callus bridging occurred later after vibration. The chosen vibration regimen (90 Hz, 0.5 mm, 4 × g acceleration, 15 min twice a day) was effective in improving musculoskeletal tissues in ovariectomized rats but was not optimal for fracture healing.

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.