Predictive value of desferrioxamine infusion test for bone aluminium deposits in hemodialyzed patients

A transiliac bone biopsy was performed in 16 hemodialyzed patients, and a staining procedure with Aluminon was used to reveal bone aluminium deposits. The baseline serum aluminium levels did not allow to predict the patients with aluminium deposits. In contrast, the peak of serum aluminium, 48 h after a desferrioxamine infusion, was much more reliable for the detection of patients with bone aluminium overload.     

Genetic markers, bone mineral density, and serum osteocalcin levels.

We evaluated five genetic markers for products that contribute to skeletal mineralization including the Sp1 polymorphism for type I collagen Ai (COLIA1), the vitamin D receptor (VDR) translation initiation site polymorphism, the promoter of the osteocalcin gene containing a C/T polymorphism, the estrogen receptor (ER) gene containing a TA repeat, and the polymorphic (AGC)n site in the androgen receptor. These markers were evaluated for their potential relationship with bone mineral density (BMD), measured by dual-energy X-ray densitometry, or its 3-year change. Additionally, potential associations of these genotypes and with baseline osteocalcin concentration or its 3-year change (assessed using radioimmunoassay) were evaluated. The study was conducted in 261 pre- and perimenopausal women of the Michigan Bone Health Study, a population-based longitudinal study of musculoskeletal characteristics and diseases. The polymorphic (AGC)n site in the androgen receptor showed a strong association with BMD of the femoral neck (FN) and lumbar spine and remained highly significant after adjusting for body mass index (BMI), oophorectomy/hysterectomy, oral contraceptive (OC) use and hormone replacement use (p < 0.001). The TA repeat at the 5' end of the ER gene was associated with total body calcium (p < 0.05) after adjusting for BMI, oophorectomy and hysterectomy, and OC use. The frequency of oophorectomy and hysterectomy within selected genotypes explained much of the statistically significant association of the ER genotypes with BMD of the FN and spine. There was no association of measures of BMD or bone turnover with the Sp1 polymorphism for COLIA1, the VDR translation initiation site polymorphism, or the C/T promoter polymorphism of the osteocalcin gene. These findings suggest that sex hormone genes may be important contributors to the variation in BMD among pre- and perimenopausal women.     

Can serum osteocalcin levels predict bone turnover in patients with osteoporosis?

The assessment of bone metabolism by biochemical markers remains difficult problem. Serum osteocalcin, synthesized in bone cells, is now becoming a sensitive indicator of bone turnover in patients with metabolic bone diseases. We measured serum osteocalcin levels by radioimmunoassay in 18 patients with osteoporosis and examined whether they reflect bone formation, resorption or both. We found that serum osteocalcin levels in biopsy-identified osteoporotics were widely ranged (1.8 – 18.8 ng/ml). Tetracycline double labeling method exibited two types of labeling pattern in the iliac bone, that is double labeled or no double labeled (impaired labeled) pattern. Serum concentrations of osteocalcin in patients with double labeling were significantly higher than those with impaired labeling (11.2±4.0 vs 4.1 ±2.1 ng/ml, P<0.01). Furthermore, serum osteocalcin levels showed a significantly positive linear correlation with the parameters reflecting bone formation, but not with bone resorption. These data indicate that serum osteocalcin levels reflect bone formation in osteoporotics and thereby could be a useful indicator of osteoblastic function in osteoporosis.     

卵巢切除大鼠血清三种骨钙素变化与骨丢失

本实验以卵巢切除（ＯＶＸ）大鼠为动物模型，利用血骨钙素（Ｏｓｔｅｏｃａｌｃｉｎ，ＯＣ）能够与羟磷灰石结合的特性，研究了ＯＶＸ大鼠血清中总ＯＣ，与羟磷灰石具有高亲合力ＯＣ（结合型ＯＣ）和低亲合力ＯＣ（游离型ＯＣ）水平随时间变化的特点及与骨丢失的关系。实验结果显示，ＯＶＸ１５天大鼠血中总ＯＣ，结合型ＯＣ和游离型ＯＣ均显著增加，第３腰椎（Ｌ３）矿盐密度显著降低。ＯＶＸ３０天大鼠血清总ＯＣ水平无明显改变，但游离型ＯＣ水平显著增加，结合型ＯＣ水平显著降低，Ｌ３矿盐密度显著降低。ＯＶＸ６０天大鼠血清总ＯＣ和游离型ＯＣ显著增加，Ｌ３矿盐密度显著降低，但结合型ＯＣ无明显改变。实验结果提示，ＯＶＸ大鼠骨盐丢失与骨内ＯＣ羧化作用降低或羧化作用与ＯＣ合成增加间脱偶联有关。血总ＯＣ可用于评价骨转换型的指标，血结合型ＯＣ可用于骨形成的指标，而游离型ＯＣ可用于判断骨丢失的指标。     

Effect of aluminium on osteocalcin production in the rat.

In renal failure, aluminium is an important factor in the development of osteomalacia. The mechanism by which aluminium produces osteomalacia is not clear; it may be toxic to the osteoblast, and as a result of its effect on osteoblasts, impair mineralisation. Another possibility is that aluminium may directly impair mineralisation independent of osteoblast function. Osteocalcin is considered to be a specific marker of osteoblast activity and its production is stimulated by calcitriol. In the present study, calcitriol-stimulated production of osteocalcin was studied as a marker of osteoblast activity in aluminium toxicity. Four groups of rats were evaluated: (1) normals; (2) normal renal function plus aluminium; (3) renal failure; and (4) renal failure plus aluminium. Osteocalcin production was determined by measuring serum osteocalcin at baseline and after stimulation with calcitriol. In rats receiving aluminium, the baseline serum osteocalcin levels were not different from their respective controls. After stimulation with calcitriol, the increase in serum osteocalcin was less in the renal failure group receiving aluminium. However, when corrected for the number of osteoblasts, the increase in serum osteocalcin was not decreased. In summary, although aluminium administration decreased the number of osteoblasts in rats with renal failure, osteocalcin production by the remaining osteoblasts was not decreased.     

Studies of serum bone Al-P isoenzyme and serum osteocalcin in patients on maintenance hemodialysis

This report describes an investigation of 128 patients on maintenance hemodialysis. Their serum bone Al-P isoenzyme (Al-P III) values were measured by the Rosalki method, and simultaneous estimations were made of their serum osteocalcin levels as a diagnostic indicator of renal osteodystrophy (ROD). ROD was evaluated clinically from Jensen's criteria, based on a four-stage classification of the radiographic evidence for subperiosteal resorption: no change (stage 0), minor change (stage I), and definite change (stages IIa and IIb). The serum Al-P III levels in the stage IIa and IIb patients showed significantly elevated values (p less than 0.01), i.e., 4.98 +/- 6.23 and 20.51 +/- 13.46 KAU, respectively. In contrast, their serum osteocalcin levels were not appreciably different. Patients with elevated N-PTH values of 0.30 ng/ml or more were found to have serum Al-P III levels of 20 KAU or more, and all of these patients were categorized under the stage IIb classification. It is concluded therefore that in chronic renal failure patients on hemodialysis, measurements of the serum Al-P III are highly useful for diagnosing ROD, and may also be considered as a critical parameter for assessing the indication for parathyroidectomy (PTX).     

Measurement of serum osteocalcin with a human-specific two-site immunoradiometric assay.

We developed a sensitive and specific two-site radioimmunoassay (IRMA) for human osteocalcin using human osteocalcin as a standard and two monoclonal antibodies raised against human osteocalcin purified from human cortical bone, a solid-phase anti-25-37 region and a tracer anti-5-13 sequence of the molecule. A wide range of osteocalcin levels (up to 300 ng/ml) can be measured with a sensitivity of 0.4 ng/ml. The intra- and interassay coefficients of variation are less than 4 and 6%, respectively. The recovery of human osteocalcin from serum samples ranges from 96 to 103%. IRMA was linear for serial sample dilutions in a wide range of serum osteocalcin levels, even in patients with chronic renal failure on hemodialysis. Depletion of serum in intact osteocalcin demonstrated that IRMA detects, in addition to the intact peptide, a large N-terminal midregion fragment that represents about 50% of total osteocalcin levels in normals and patients with Paget's disease and up to 75% in patients with chronic renal failure. This large fragment, previously unrecognized because it cannot be distinguished from intact osteocalcin with gel filtration chromatography, is not generated in vitro by incubation of the serum up to 26 h. We measured osteocalcin in the serum of 309 healthy adults (180 men and 129 women, age range 20-95 years), 36 patients with Paget's disease, 12 patients with primary hyperparathyroidism, 70 patients with chronic renal failure on hemodialysis, and 10 patients on corticosteroid therapy, simultaneously with human IRMA and with a conventional radioimmunoassay (RIA) based on bovine reagents. A tight correlation (r = 0.889) was observed between the two assays in the normal population, but the values obtained with IRMA were about threefold higher (mean 23.3 +/- 10.5 versus 7.5 +/- 3.4 ng/ml) than those obtained with RIA. Reported as Z scores, that is, number of standard deviations from the predicted normal mean adjusted for sex and age, these two assays (IRMA and RIA) gave concordant results in patients with Paget's disease (4.05 +/- 6.21 versus 2.41 +/- 2.53), primary hyperparathyroidism (4.14 +/- 7.17 versus 2.13 +/- 2.28), chronic renal failure (25.32 +/- 24.49 versus 6.93 +/- 5.48), and glucocorticoid treatment (-1.48 +/- 0.78 versus -1.11 +/- 0.57). However, IRMA was more discriminant from controls for all these metabolic bone diseases because the absolute values of mean Z scores with IRMA were significantly higher than those obtained with the RIA (p < 0.05-0.0001).(ABSTRACT TRUNCATED AT 400 WORDS)     

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

Summary  

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.     

Clinical evaluation of fracture healing by serum osteocalcin and alkaline phosphatase.

The serum osteocalcin (BGP) concentration and alkaline phosphatase (AP) activity were measured prospectively during the healing phases of crural fractures in 15 patients. They were divided into two groups, the time of union of the fracture being under (group 1) or over 16 weeks (group 2). The mean values of BGP and AP were somewhat higher from the outset in the group 1 than in the group 2, but the difference was not significant. A significant increase in BGP and AP (P less than 0.05) was evident in both groups at 6 weeks. In cases with undisturbed healing of fractures (group 1) the values of serum BGP and AP then declined towards the values at the time of accident. Contrary to this, in group 2 both the values of the serum BGP and AP were still at a significantly higher level than those at the day of the fracture. However, no significant difference in the serum BGP or AP was seen between the two groups at 6 or 12 weeks. The results support some earlier ones: the changes in serum BGP and AP may provide a prognostic indicator for consolidation of a fracture.     

Use of the low-dose desferrioxamine test to diagnose and differentiate between patients with aluminium-related bone disease, increased risk for aluminium toxicity, or aluminium overload

BACKGROUND.: Aiming at a safe method in the diagnosis of aluminium-relatedbone disease (ARBD)/aluminium overload the low-dose desferrioxamine(DFO) test was developed. In a multicentre study histologicaland histochemical data and aluminium bulk analysis of bone biopsiesof 77 dialysis patients were correlated with the results ofboth the 5 mg/kg and 10 mg/kg DFO tests. METHODS.: ARBD was considered to be present when > 15% of the bonesurface was positively stained for aluminium and the bone formationrate was below 220 µm²/mm²/day. Patients in which theAluminon® staining was positive (>0%) were consideredat an increased risk for aluminium toxicity independent of thetype of renal osteodystrophy. Patients were considered aluminiumoverloaded when the bone aluminium content was >15 µg/gwet weight and/or the Aluminon® staining was positive (>0%). RESULTS.: Using the proposed criteria 15 patients were found to have ARBD;13 of them presenting with a serum iPTH below 150 ng/l. In conjunctionwith an iPTH measurement the DFO test had a more than acceptablesensitivity and specificity in the diagnosis of ARBD. The testwas considered positive when a post-DFO serum aluminium increment(sAl) above 50 µg/l (5 mg/kg) or 70 µg/l (10 mg/kg)together with a serum iPTH below 150 ng/l was found. Using thesecut-off levels the 5 and 10 mg/kg tests in the diagnosis ofARBD had a sensitivity of 87% and a specificity of 95% and 92%respectively whereas the predictive value for a positive testfor the population under study was 80% (5 mg/kg). Not a singlepatient with a serum iPTH >650 ng/l had a positive staining(>0%) even when the bone aluminium level was elevated (>15 µg/g wet weight). In the detection of patients at riskfor aluminium toxicity sAl thresholds of 50 µg/l (5 mg/kg)and 70 µg/l (10 mg/kg) in combination with a serum iPTH<650 ng/l had a sensitivity of 92% and specificity of 86%and 84% respectively. In the clinical setting of aluminium overload,threshold sAl levels of 50 µg/l (5 mg/kg) and 70 µg/l(10 mg/kg) had a sensitivity of 91% and a specificity of 95%and 90% respectively. CONCLUSIONS.: The low-dose DFO test is a reliable test for the detection ofaluminium overload; however, it is not specific enough to differentiatebetween ARBD, increased risk of aluminium toxicity, and aluminiumoverload unless it is used in combination with a serum iPTHmeasurement. In conjunction with a serum iPTH measurement itis an important tool in the differential diagnosis and may avoidthe necessity of a bone biopsy in the majority of patients.Data obtained in the present study have allowed us to updatethe strategies for monitoring, diagnosis and patient follow-upproposed at the Consensus Conference on Diagnosis and Treatmentof Aluminium Overload in End-Stage Renal Failure; Paris, 1992.     

Evaluation of serum osteocalcin in patients with urolithiasis

Serum osteocalcin, also called bone gla protein, is one of sensitive and specific markers for metabolic bone diseases. Current evidence suggests that the protein may be involved in the regulation of calcium homeostasis in bone. We measured serum osteocalsin levels by radioimmunoassay in 100 patients with urolothiasis, especially in calcium containing stone formers and evaluated the influence of bone metabolism on the formation of calcium-containing stones. Although serum osteocalcin levels in most patients were normal, those of two male and four female patients were high. We considered that they were patients with renal hypercalciuria and secondary hyperparathyroidism and in them the formation of calcium containing stones were influenced by disorder of bone metabolism. In addition, we suggest that serum osteocalcin levels may be available index for the effect of treatment in stone formers with renal hypercalciuria and bone disease.     

Elevation of serum albumin levels in nagase analbuminemic rats by allogeneic bone marrow cell transplantation

We investigated the feasibility of correcting the congenital absence of albumin in Nagase analbuminemic rats (NARs) by allogeneic bone marrow cell transplantation (BMT). Seven-week-old male NARs were used as recipients, and 6- to 8-week-old male Sprague-Dawley (SD) rats were used as allograft donors. NARs were divided into three groups: a BMT group (n=10) in which bone marrow cells were infused into the liver; a hepatocyte transplantation (HCT) group (n=8) in which hepatocytes were transplanted into the liver, and a control group (n=8) in which PBS was injected into the portal vein. Serum albumin levels were measured as an indicator of the function of the grafted cells, and the phenotypic characteristics of the engrafted cells in the recipient's liver were assessed with immunohistochemical and immunofluorescence techniques. At 8 weeks after cell transplantation, the serum albumin levels of the BMT group and HCT group were significantly higher than in the control group. The hepatocyte-like cells derived from bone marrow cells expressed albumin in liver of the NARs. According to this result, bone marrow cells can differentiate into hepatocyte-like cells in vivo. The results show that BMT is an effective treatment for congenital analbuminemia in a rat model and suggest that allogeneic BMT can be used as an efficient therapy for hereditary metabolic diseases.     

骨吸收抑制剂对卵巢切除骨折大鼠脂类代谢及骨钙素的影响

目的 研究骨吸收抑制剂对卵巢切除大鼠脂类代谢及骨钙素的影响.方法 雌性SD大鼠共140只,随机分为5组,每组28只.3月龄时选取其中4组大鼠行双侧卵巢切除建立绝经后骨质疏松症模型(OVX组、OVX+ EE2组、OVX+ Rlx组、OVX+ Aln组),另一组行假手术(Sham组).OVX组及Sham组皮下注射生理盐水,余下3组分别注射阿仑膦酸钠(Aln)、雷洛昔芬(Rlx)、雌激素(EE2),每周注射5次.分别在卵巢切除术后4周、10周及20周测定大鼠血脂、骨钙素及相关生化指标.结果 OVX组与Sham组相比体重增加,总胆固醇升高,甘油三脂降低,差异有统计学意义.应用雌激素及雷洛昔芬可有效调节卵巢切除导致的脂代谢紊乱,表现为体重下降及总胆固醇水平降低,差异有统计学意义(P＜0.05).不同时期OVX组骨钙素水平高于Sham组,差异有统计学意义(P＜0.05),其中阿仑膦酸钠组血清骨钙素水平最低.随着时间推移,Sham组及OVX组骨钙素呈现先上升再下降趋势,而OVX+EE2组、OVX+ Rlx组及OVX+ Aln组骨钙素表现为持续降低.结论 骨吸收抑制剂能够降低骨钙素水平,从而降低卵巢切除导致的高骨转换率,防止骨量丢失.此外,雌激素及雷洛昔芬在预防骨丢失的基础上还能够有效调节卵巢切除引起的脂类代谢紊乱.     

糖尿病肾病患者血清骨钙素的变化的研究

目的：虽然糖尿病的骨量减少的病因学和病理生理学还不清楚，糖尿病并发骨质量丢失已经引起注意。通过检测糖尿病患者反映骨形成的生化标志血清骨钙素以研究糖尿病肾病的骨代谢改变。方法：用放射免疫法测定３１７例糖尿病患者和６０例正常人的血清骨钙素水平，糖尿病患者按Ｍｏｇｅｎｓｅｎ方法将肾脏病变不同阶段分为５组。结果：糖尿病患者血清骨钙素水平显著低于正常对照组（４．０４±１．７４ｖｓ５．４８±１．５１ｎｇ／ｍｌ，ｐ＜０．００１）。按糖尿病肾病程度分组后Ⅰ～Ⅴ组分别为４．１９，４．３５，３．４７，３．３０，３．７４ｎｇ／ｍｌ，随ＤＮ病程的进展逐步降低，但Ⅴ组（氮质血症期）的骨钙素并不低于Ⅲ组（早期糖尿病肾病期）和Ⅳ组（糖尿病肾病期）。结论：糖尿病患者血清骨钙素水平降低提示成骨细胞活性减低，骨形成减少。在糖尿病合并微血管病变，尿白蛋白排出量增加时血清骨钙素水平下降更显著。继发性甲状旁腺机能亢进和骨转换率的增加，可能是造成氮质血症组的骨钙素不低于Ⅲ组和Ⅳ组的原因     

Low osteocalcin/collagen type I bone gene expression ratio is associated with hip fragility fractures

IntroductionOsteocalcin (OC) is the most abundant non-collagenous bone protein and is determinant for bone mineralization.We aimed to compare OC bone expression and serum factors related to its carboxylation in hip fragility fracture and osteoarthritis patients. We also aimed to identify which of these factors were associated with worse mechanical behavior and with the hip fracture event.MethodsIn this case-control study, fragility fracture patients submitted to hip replacement surgery were evaluated and compared to a group of osteoarthritis patients submitted to the same procedure. Fasting blood samples were collected to assess apolipoproteinE (apoE) levels, total OC and undercarboxylated osteocalcin (ucOC), vitamin K, LDL cholesterol, triglycerides and bone turnover markers. The frequency of the apoε4 isoform was determined.Femoral epiphyses were collected and trabecular bone cylinders drilled in order to perform compression mechanical tests. Gene expression of bone matrix components was assessed by quantitative RT-PCR analysis.Results64 patients, 25 submitted to hip replacement surgery due to fragility fracture and 39 due to osteoarthritis, were evaluated. Bone OC/collagen expression (OC/COL1A1) ratio was significantly lower in hip fracture compared to osteoarthritis patients (p < 0.017) adjusted for age, gender and body mass index. Moreover, OC/COL1A1 expression ratio was associated with the hip fracture event (OR ~ 0; p = 0.003) independently of the group assigned, or the clinical characteristics. Apoε4 isoform was more frequent in the hip fracture group (p = 0.029). ucOC levels were higher in the fracture group although not significantly (p = 0.058). No differences were found regarding total OC (p = 0.602), apoE (p = 0.467) and Vitamin K (p = 0.371).In hip fracture patients, multivariate analysis, adjusted for clinical characteristics, serum factors related to OC metabolism and gene expression of bone matrix proteins showed that low OC/COL1A1 expression ratio was significantly associated with worse trabecular strength (β = 0.607; p = 0.013) and stiffness (β = 0.693; p = 0.003). No association was found between ucOC and bone mechanics. Moreover, in osteoarthritis patients, the multivariate analysis revealed that serum total OC was negatively associated with strength (β = − 0.411; p = 0.030) and stiffness (β = − 0.487; p = 0.009).ConclusionWe demonstrated that low bone OC/COL1A1 expression ratio was an independent predictor of worse trabecular mechanical behavior and of the hip fracture event. These findings suggest that in hip fracture patients the imbalance of bone OC/COL1A1 expression ratio reflects disturbances in osteoblast activity leading to bone fragility.     

Low osteocalcin/collagen type I bone gene expression ratio is associated with hip fragility fractures

IntroductionOsteocalcin (OC) is the most abundant non-collagenous bone protein and is determinant for bone mineralization.We aimed to compare OC bone expression and serum factors related to its carboxylation in hip fragility fracture and osteoarthritis patients. We also aimed to identify which of these factors were associated with worse mechanical behavior and with the hip fracture event.MethodsIn this case-control study, fragility fracture patients submitted to hip replacement surgery were evaluated and compared to a group of osteoarthritis patients submitted to the same procedure. Fasting blood samples were collected to assess apolipoproteinE (apoE) levels, total OC and undercarboxylated osteocalcin (ucOC), vitamin K, LDL cholesterol, triglycerides and bone turnover markers. The frequency of the apoε4 isoform was determined.Femoral epiphyses were collected and trabecular bone cylinders drilled in order to perform compression mechanical tests. Gene expression of bone matrix components was assessed by quantitative RT-PCR analysis.Results64 patients, 25 submitted to hip replacement surgery due to fragility fracture and 39 due to osteoarthritis, were evaluated. Bone OC/collagen expression (OC/COL1A1) ratio was significantly lower in hip fracture compared to osteoarthritis patients (p < 0.017) adjusted for age, gender and body mass index. Moreover, OC/COL1A1 expression ratio was associated with the hip fracture event (OR ~ 0; p = 0.003) independently of the group assigned, or the clinical characteristics. Apoε4 isoform was more frequent in the hip fracture group (p = 0.029). ucOC levels were higher in the fracture group although not significantly (p = 0.058). No differences were found regarding total OC (p = 0.602), apoE (p = 0.467) and Vitamin K (p = 0.371).In hip fracture patients, multivariate analysis, adjusted for clinical characteristics, serum factors related to OC metabolism and gene expression of bone matrix proteins showed that low OC/COL1A1 expression ratio was significantly associated with worse trabecular strength (β = 0.607; p = 0.013) and stiffness (β = 0.693; p = 0.003). No association was found between ucOC and bone mechanics. Moreover, in osteoarthritis patients, the multivariate analysis revealed that serum total OC was negatively associated with strength (β = ? 0.411; p = 0.030) and stiffness (β = ? 0.487; p = 0.009).ConclusionWe demonstrated that low bone OC/COL1A1 expression ratio was an independent predictor of worse trabecular mechanical behavior and of the hip fracture event. These findings suggest that in hip fracture patients the imbalance of bone OC/COL1A1 expression ratio reflects disturbances in osteoblast activity leading to bone fragility.     

Influence of bone osteocalcin levels on bone loss induced by ovariectomy in rats

To investigate the role of osteocalcin (OC) in bones, bone parameters in warfarin (WF)-treated rats after ovariectomy (OVX) were compared with those in intact rats. Rats were divided into an intact group and WF-treated group. Warfarin was orally given to rats for 16 weeks, and then OVX was performed and rats in the WF-treated groups continued receiving WF. Twelve weeks after OVX, bone properties were observed. The diaphysial bone OC level in the WF group was 10%–14% of the normal level at the preoperative point and 12 weeks after surgery. On comparison of the intact and WF groups before surgery, no significant differences were noted in bone mass parameters or mechanical properties, but 12 weeks after surgery, the diaphysial bone mineral content (BMC), bone area, and cortical thickness (Cth) were significantly higher in the WF-sham group than in the intact-sham group. Ovariectomy significantly decreased the diaphysial BMC, bone mineral density (BMD), Cth, and maximum load, and increased the endosteal perimeter in the WF group. In the intact group, no such OVX-induced changes were noted, and the metaphysial bone area and the endosteal and periosteal perimeters were increased by OVX. The CO₃/PO₄ ratio in the femur measured by Fourier-transform infrared imaging using reflection preparations was higher in the WF-sham group than the intact-sham group, and higher in the intact-OVX group than the intact-sham group, but no significant difference was noted between the WF-sham and WF-OVX groups. It has been reported that CO₃ ⁻ is contained in new bone and decreases with mineral maturation. These data suggest that long-term reduction in bone OC levels may induce the formation of immature bone, which is easily resorbed with changes in bone metabolism such as OVX, and that OC may be one of the factors affecting bone turnover.