Reference intervals of bone turnover markers determined by using their curve-fitting valley for adult females in China.

SUMMARY: The reference values for bone turnover markers (BTMs) have a significant role in the diagnosis, monitoring, and treatment of metabolic bone disease. This study proposes that the peak value of bone mineral density and the trough value for the BTM curve can be used to determine the reference range of BTM. INTRODUCTION: The aim of this study is to determine the reference intervals of BTMs for adult females in China with an attempt to reference the peak bone mineral density (BMD) with the corresponding BTM valley. METHODS: This study included 546 premenopausal and 394 postmenopausal women. The levels of several BTMs were determined, and the BMD was measured using a dual-energy X-ray absorptiometry. RESULTS: The BTMs of postmenopausal women were 17-96 % higher than premenopausal women. The change of BTM with age presented an optimal goodness-of-fit according to the cubic regression model (R (2) = 0.074-0.346, all P = 0.000). All kinds of BTM levels were positively correlated with age in premenopausal women aged 27-56 years old (r = 0.167-0.502, P = 0.023-0.000). Except for uCTX, the BTM reference value determined using a curve-fitting valley was significantly lower than the reference values for premenopausal women. The BTM reference values determined in this study were also significantly different from the reference values given by the manufacturers of the reagents used. CONCLUSIONS: This study found that the changes of level with age of BTMs in Chinese women present an optimal goodness-of-fit according to the cubic regression model. The fitting valley corresponds to the BMD fitting peak and may possibly be an effective means of determining the BTM reference intervals.

Pressure dependence of Mn2+ fluorescence in MnS/ZnS core-shell quantum dots.

The high pressure photoluminescence spectra of MnS/ZnS core-shells quantum dots were measured using a diamond anvil cell up to 9.4 GPa. Orange emission at 590 nm from the 4T1 --> 6A1 transition of Mn2+ ions was observed. The Mn2+ emission shifted to red with increasing pressure. The experimental pressure coefficient was -48.3 meV/GPa, which is agreement with the calculated value based on the crystal field theory. The redshift is attributed to the increase of crystal field strength and decrease of Racah parameters during compression.

High pressure luminescence and Raman studies on the phase transition of Gd2O3:Eu3+ nanorods.

The cubic Gd2O3:Eu3+ nanorods were synthesized by a hydrothermal method. The SEM image indicated the nanorods with diameter of 30-35 nm and length of 200-500 nm. The structural stability of Gd2O3:Eu3+ nanorods was investigated by in situ high pressure luminescence and Raman spectra up to 18.9 GPa at room temperature. The results reveals a pressure-induced phase transition from cubic to hexagonal structure at about 11.3 GPa. After releasing pressure, the part of hexagonal structure is retained and the other transfers to monoclinic phase.

Photoluminescence and raman studies of Y2O3:Eu(3+) nanotubes under high pressure.

In this work, the cubic compound Y2O3:Eu(3+) nanotubes with diameter of 70-90 nm and length of 2-3 microm are synthesized by a hydrothermal method. Photoluminescence and Raman spectra of Y2O3:Eu(3+) nanotubes in a diamond anvil cell under high pressure are measured at room temperature. The 5D0 --> 7F(0,1,2) transitions of the Eu(3+) ions exhibit red shifts to higher wavelength with pressure increasing. Above 13.4 GPa, all the Raman active modes disappear. When the pressure is released from 25.6 GPa to ambient pressure, these Raman peaks are not retrieved; this fact indicates that the nanotubes are transformed into amorphous from cubic phase at about 13.4 GPa. It may be related to the collapse of nanotube form under high pressure condition.

Zoledronate reverses mandibular bone loss in osteoprotegerin-deficient mice.

UNLABELLED: To characterize the changes in osteoprotegerin-deficient (OPG-/-) mice mandibles and the possible mandibular bone loss prevention by zoledronate. This preventive effect in the mandible differed from that in the proximal tibia and was independent of the OPG pathway. INTRODUCTION: The study aimed to characterize both the changes in the mandible in osteoprotegerin-deficient (OPG-/-) mice and possible mandibular bone loss prevention by zoledronate. METHODS: Twenty-eight 6-week-old female mice (C57BL/6J), including OPG-/- (n = 21) and wild-type (WT) (n = 7) mice, were assigned to four groups after 2 weeks of acclimatization to local vivarium conditions: wild mice with vehicle (WT group); OPG-/- mice with vehicle (OPG-/- group); and OPG-/- mice that were subcutaneously injected with either 50 or 150 microg/kg zoledronate (Zol-50 and Zol-150 groups, respectively). Mice were sacrificed at 4 weeks after these treatments and after fasting for 12 h. Sera were harvested for biochemical analyses. The right mandible and tibia of each mouse were selected for microCT analysis. Student's t-test was performed for comparisons of bone parameters at different sites in the WT group. Analysis of variance (ANOVA) was used to compare the biomarkers and bone parameters in the different treatment groups. RESULTS: Serum bone-specific alkaline phosphatase (B-ALP) and tartrate-resistant acid phosphatase 5b (TRACP-5b) were significantly decreased in WT mice as compared to the levels in the OPG-/- mice (P < 0.05). Zoledronate treatment decreased the high serum B-ALP activity observed in OPG-/- mice to the levels seen in WT mice, while serum TRACP-5b concentrations were decreased to levels even lower than those in WT mice. There were substantial variations in BMD and microstructure of the mandibular and proximal tibial trabeculae. Mandibular bone loss was less affected by OPG gene deprivation than the proximal tibia was. Both zoledronate groups showed greater BMD, trabecular BV/TV, Tb.Th, Tb.N, and Conn.D and a significant decrease in Tb.Sp and SMI as compared to the findings in OPG-/- mice (P < 0.05). However, higher apparent BMD and more compact plate-like trabeculae were observed in the mandible after treatment with zoledronate as compared to the findings in the proximal tibia. No significant differences were found in any parameter in both zoledronate groups. CONCLUSIONS: The present study showed that zoledronate could reverse the significant bone loss in mice mandibles that was induced by OPG gene deficiency. This preventive effect, which was accompanied with considerable inhibition of bone turnover, differed in the mandible and in the proximal tibia and was independent of the OPG pathway.

Effects of methylprednisolone on bone mineral density and microarchitecture of trabecular bones in rats with administration time and assessed by micro-computed tomography.

BACKGROUND: Little research exists on the dynamic effects of glucocorticoids on bone mineral density (BMD) and microarchitecture of trabecular bones of rats assessed by micro-computed tomography (micro-CT). PURPOSE: To investigate time-related changes in the BMD and microarchitecture of trabeculae in rats exposed to glucocorticoid. MATERIAL AND METHODS: Female Sprague-Dawley rats were recruited into a baseline group, glucocorticoid-treated groups, or control groups. Glucocorticoid-treated rats were given daily subcutaneous injections of methylprednisolone at a dosage of 3.5 mg/kg for 1 or 9 weeks. A high-resolution micro-CT was used to identify the densitometric and microarchitectural properties of trabeculae in both the proximal metaphysis of tibiae and the sixth lumbar vertebrae (L6). RESULTS: Compared with baseline rats, volumetric BMD, tissue BMD, bone volume fraction, trabecular number, and degree of anisotropy of trabeculae from tibiae or L6 increased in control rats and glucocorticoid-treated rats with time; however, changes in the latter group were smaller. Compared with control rats at each time point, a decrease occurred in volumetric BMD, tissue BMD, bone volume fraction, trabecular number, degree of anisotropy, and trabecular connectivity density in trabecular bones from tibiae or L6 in glucocorticoid-treated rats. The decrease was greater in week 9 compared to week 1. Contrarily, an increase was noted in trabecular thickness, trabecular separation, and structure model index in glucocorticoid-treated rats. A time-related analysis within glucocorticoid-treated groups in both skeletal regions showed a decline in bone volume fraction, trabecular connectivity density, trabecular number, and degree of anisotropy with time, but trabecular thickness and trabecular separation were elevated. CONCLUSION: Methylprednisolone can inhibit bone mineralization and bone mass gain with growth in rats. It can also deteriorate microarchitecture of trabeculae in a time-dependent or an accumulative dose-dependent manner. Further, the remaining trabeculae appear to thicken in order to adapt to altered stress.

Effects of the sample size of reference population on determining BMD reference curve and peak BMD and diagnosing osteoporosis.

UNLABELLED: Establishing reference databases generally requires a large sample size to achieve reliable results. Our study revealed that the varying sample size from hundreds to thousands of individuals has no decisive effect on the bone mineral density (BMD) reference curve, peak BMD, and diagnosing osteoporosis. It provides a reference point for determining the sample size while establishing local BMD reference databases. INTRODUCTION: This study attempts to determine a suitable sample size for establishing bone mineral density (BMD) reference databases in a local laboratory. METHODS: The total reference population consisted of 3,662 Chinese females aged 6-85 years. BMDs were measured with a dual-energy X-ray absorptiometry densitometer. The subjects were randomly divided into four different sample groups, that is, total number (Tn) = 3,662, 1/2n = 1,831, 1/4n = 916, and 1/8n = 458. We used the best regression model to determine BMD reference curve and peak BMD. RESULTS: There was no significant difference in the full curves between the four sample groups at each skeletal site, although some discrepancy at the end of the curves was observed at the spine. Peak BMDs were very similar in the four sample groups. According to the Chinese diagnostic criteria (BMD >25% below the peak BMD as osteoporosis), no difference was observed in the osteoporosis detection rate using the reference values determined by the four different sample groups. CONCLUSIONS: Varying the sample size from hundreds to thousands has no decisive effect on establishing BMD reference curve and determining peak BMD. It should be practical for determining the reference population while establishing local BMD databases.

Regionally specific compensation for bone loss in the tibial trabeculae of estrogen-deficient rats.

BACKGROUND: Bone mineral density (BMD) and microstructural variations have been extensively investigated in recent years; however, the compensation for bone loss between different regions is still unclear. PURPOSE: To fully characterize regional variations in bone mineral density (BMD) as well as the microstructure and dynamic changes of rat tibial trabeculae that occur with bone loss associated with estrogen deficiency. MATERIAL AND METHODS: Female Sprague-Dawley rats were ovariectomized (OVX), sham-operated (sham), or left unoperated (baseline control). The left tibiae were harvested at baseline, and at postoperative weeks 3 and 15. High-resolution micro-computed tomography (microCT) was used to identify the densitometric and microstructural properties of trabeculae in the proximal ends of the rat tibia, specifically the epiphysis and metaphysis. RESULTS: Volumetric BMDs at the organ (organ BMD) and tissue (tissue BMD) levels were significantly higher for trabeculae at the epiphysis than metaphysis. Moreover, trabeculae at the epiphysis were thicker, and fewer in number and connectivity than those at the metaphysis, which were more rod like. Trabeculae at the metaphysis were more susceptible to bone loss induced by estrogen deprivation than at the epiphysis, and the regions varied greatly in their adaptation to this loss. At the metaphysis, trabecular tissue BMD and thickness were unexpectedly higher at postoperative week 15 than week 3 or baseline. In contrast, at the epiphysis, tissue BMD did not change with time, but trabecular thickness significantly increased at week 15 compared to baseline and was also greater in OVX compared to sham rats. CONCLUSION: Metaphyseal and epiphyseal trabeculae show regionally specific variations in BMD and microstructure. The former are more susceptible to bone loss induced by estrogen deficiency and would be strengthened by either hypertrophy or hypermineralization, while epiphyseal trabeculae are mainly strengthened by thickening.

Ascorbic acid inhibits osteoclastogenesis of RAW264.7 cells induced by receptor activated nuclear factor kappaB ligand (RANKL) in vitro.

Ascorbic acid (AA) plays a key role in the regulation of differentiation and activation of osteoclast (OCL). It was reported that AA might induce the formation of OCL in cocultures of mouse bone marrow cells and ST2 cells, but it is not clear whether AA has a direct impact on the OCL precursors. The purpose of this study was to examine the effect of AA on the differentiation of OCL precursor RAW264.7 cells, cultured with receptor-activated nuclear factor kappaB ligand (RANKL). The results showed that AA remarkably inhibited the cell proliferation at a higher concentration and RANKL alone is sufficient for osteoclastogenesis. The expression of carbonic anhydrase (CAII) mRNA and protein, the number of tartrate-resistant acid phosphatase (TRAP)-positive multinucleated cells (MNCs), and the percentage area of resorption lacunae induced by RANKL were decreased when AA was added to the cultures. The results demonstrate that AA inhibits RANKL-induced differentiation of OCL precursor cells into mature OCL and reduces the formation of bone resorption pits in vitro.

Membrane-type matrix metalloproteinase-1 (MT1-MMP) is down-regulated in estrogen-deficient rat osteoblast in vivo.

Our previous study showed that estrogen stimulates membrane-type matrix metalloproteinases-1 (MT1-MMP) production in osteoblastic cells culture, but has no effect on MMP-2 and TIMP-2 synthesis. Osteoblast-derived MT1-MMP have been recently implied to play a role in bone metabolism by degrading tumor necrosis factor-alpha (TNF-alpha), resolving extracellular matrix and activating proMMP-2, which requires the process of activation mediated by MT1-MMP/tissue inhibitor of metalloproteinase (TIMP-2) complex on the cell surface. To investigate the mechanism of bone loss following estrogen deficiency, we examined the effects of estrogen on osteoblast synthesis of MT1-MMP, MMP-2 and TIMP-2. In situ hybridization and immunohistochemistry of rat bone samples were used to document the synthesis of MT1-MMP, MMP-2, and TIMP-2 mRNA and protein. Osteoblasts from distal femoral head showed an increase in the pattern of MT1-MMP mRNA and protein production in sham-operated controls and 17beta-estradiol (E2)-treated rats, compared with the ovariectomized group; the synthesis of MMP-2 and TIMP-2 mRNA and protein was unaffected. Our data show a down-regulation of MT1-MMP synthesis by osteoblast in vivo following estrogen withdrawal, and treatment with E2 resulted in induced MT1-MMP expression in vivo. There is evidence suggesting a role for MT1-MMP in the process of bone loss during the pathogenesis of osteoporosis.

A comparison study of the reference curves of bone mineral density at different skeletal sites in native Chinese, Japanese, and American Caucasian women.

To understand the differences among reference curves for bone mineral density (BMD) for Chinese, Japanese, and American Caucasian women, we measured the BMD at the anteroposterior (AP) lumbar spine (L1-L4), lateral lumbar spine (L2-L4), hip (including the femoral neck, trochanter, intertrochanter, Ward's triangle, and total hip), and ultradistal forearm by the dual-energy X-ray absorptiometry (DXA) in a total of 2728 healthy Chinese women, aged 5-96 years. Documented BMD data for Japanese women and device manufacturer's BMD new reference databases (including the NHANES III dataset) for American Caucasian women were also used in this study. The cubic regression model was found to fit best in analyzing the age-associated variations of BMD at various sites in Chinese women, i.e., the equations had the largest coefficient of determination (R2). At the AP/Lat spine, trochanter, intertrochanter, and Ward's triangle, BMD reference curves for Chinese women were lower than those for Japanese or Caucasian women, while at the femoral neck, total hip, and ultradistal forearm, the reference curves for Chinese women were higher than those for Japanese women, with overlaps and crossing of the curves for some age spans in comparing the Chinese and Caucasian women. There were significant differences in the peak BMD (PBMD) at various sites among the Chinese, Japanese, and Caucasian women (P = 0.000). The PBMDs for Chinese women at the lumbar spine and various sites of the hip were 5.7% +/- 2.1% (mean +/- SD, range, 2.7-7.9%) lower than those for Japanese women and 5.1% +/- 2.7% (range, 0.5-7.2%) lower than those for Caucasian women; however, the PBMDs for Chinese women were 26.2% higher than those for Japanese women and 10% higher than those for Caucasian women at the ultradistal forearm. After the PBMD, average T-scores of Chinese women for losses at the AP lumbar spine with increasing age were nearly identical to those for Japanese women, but both were greater than those for Caucasian women. The average T-scores for BMD loss at various sites in Chinese women were higher than those for both Japanese and Caucasian women except at the femoral neck, where the T-scores of Chinese women were exceeded by those of both Japanese and Caucasian women. Estimated from the T-score curve of BMD loss, the age of osteoporosis occurrence at the femoral neck in Chinese women was about 10 years later than that in Japanese or Caucasian women; at the AP spine, Chinese women were similar to Japanese women; at the other sites, the age for occurrence of osteoporosis in Chinese women was about 5-15 years earlier than that in either Japanese or Caucasian women. There are differences in prevalence or odds ratio (OR) of osteoporosis at the same skeletal region for Chinese, Japanese, and Caucasian women aged > or = 50 years or at different skeletal regions in women of the same race. The prevalences of osteoporosis at various regions of the hip in Chinese women are 10.1-19.8% and ORs are 22.0-32.3, of which prevalence at the femoral neck is the lowest (10.1%); the prevalences of osteoporosis in Japanese women are 11.6-16.8% and ORs are 21.1-26.3, of which prevalence at the femoral neck is the lowest (11.6%); and the prevalences of osteoporosis in Caucasian women are 13.0-20.0% and ORs are 19.4-48.9, of which prevalence at the femoral neck is the highest (20%). In conclusion, racial differences in BMD reference curves, prevalences, and risks of osteoporosis at various skeletal sites exist among native Chinese, Japanese, and American Caucasian women.

Age-related variation in quantitative ultrasound at the tibia and prevalence of osteoporosis in native Chinese women.

This study investigated the variations in age-related speed of sound (SOS) at the tibia and prevalence of osteoporosis in native Chinese women, and establishment of a reference database by quantitative ultrasound. SOS at the right midtibia was measured using a quantitative ultrasound device (SoundScan 2000, Myriad Ultrasound Systems, Israel) in 1596 healthy Chinese women ranging from 12 years to 96 years of age. Healthy women were selected on the basis of (1). a detailed questionnaire about their medical history, (2). face to face questioning about their medical history, and (3). a physical examination. Women with a medical condition that required medication that affected bone metabolism or those who had had a pathologic or moderate traumatic fracture were excluded. We followed the diagnostic criteria provided by the instrument's manufacturer and equivalent to the WHO criteria (using the T-score cut-off that diagnoses 30% of the post-menopausal women aged >or =50 years with osteoporosis) as the diagnostic criteria for osteoporosis in this group of women. Data were analyzed in age groups divided by intervals of 5 years. The peak SOS at the tibia of 3991+/-68 m s(-1) (mean+/-SD) occurred in the 35-39 year age group and the T-score precision was 0.99 T-score units. The SOS value increased with age up to 34 years of age and then declined with age after 40 years of age with the rate of decrease at 9.68 m s(-1) per year. The curve representing the SOS change according to age is best fitted by the regression analysis of cubic model, and the cubic equation for SOS=3383+39.9 (age)-0.78 (age)(2)+0.0039 (age)(3) (R(2)=0.505, p=0.000). The T-score cut-off that diagnoses 30% of the post-menopausal women (n=559, mean age 63.2+/-8.97 years) aged >or=50 years with osteoporosis was SOSor=80 years, the prevalences of osteoporosis detected using equivalent to the WHO criteria were 0.39%, 9.27%, 30.3%, 58.4% and 69.0%, respectively. The prevalences detected following the manufacturer's diagnostic criteria (cut-off value: SOS

Effects of projective bone area size of the spine on bone density and the diagnosis of osteoporosis in healthy pre-menopausal women in China.

The aim of this study was to understand the effects of projective bone area (BA) size of the spine on bone density and the diagnosis of osteoporosis. Measurements of BA, bone mineral content (BMC), areal bone density (aBMD) and volumetric bone density (vBMD) at the posteroanterior (PA) lumbar spine (vertebrae L2-L4) followed by a paired PA/lateral spine (L2-L4) were made using a dual-energy X-ray absorptiometry (DXA) fan-beam bone densitometer (Hologic QDR 4500A) in 1436 healthy pre-menopausal women aged from 20 to 56-years-old. At the PA and lateral lumbar spine, there was a significant positive correlation between BA and BMC (r=0.762 and 0.762, p=0.000) and aBMD (r=0.370 and 0.352, p=0.000), but not vBMD (r=0.000 and 0.102, p=0.813 and 0.063). When BA at the PA spine changed by one standard deviation (SD), BMC and aBMD correspondingly changed by 12.6% and 4.3% on the basis of their respective means while vBMD indicated no change. When a variation of 1 SD was observed in BA at the lateral spine, BMC, aBMD and vBMD correspondingly changed by 13.8%, 4.4% and 1.73% on the basis of their respective means. Through an intercomparison among large, intermediate and small BA groups, significant differences were found in the means of subject's height, weight, BMC and aBMD at the PA and lateral spine as well as the detection rate of osteoporosis by aBMD (p=0.000). Detection rates of osteoporosis by aBMD at the PA, lateral spine and vBMD in healthy pre-menopausal women aged from 40 years to 56 years were 4.5%, 16.4% and 9.7%, respectively, in the small BA group; 1.3%, 6.4% and 7.3%, respectively, in the intermediate BA group; and 0, 0 and 5.5%, respectively, in the large BA group. No significant differences were found in the detection rates of osteoporosis by vBMD among the groups. The results of multiple linear regression revealed that the major factors influencing BA of the lumbar spine was height. In healthy pre-menopausal women of the same race and age, the BA size of the lumbar spine would have significant influence upon aBMD and the diagnosis of osteoporosis, i.e. the larger the BA, the greater the aBMD and the lower the osteoporosis detection rate while conversely, the smaller the BA, the smaller the aBMD and the higher the osteoporosis detection rate. Though vBMD does not change with BA sizes of the lumbar spine, it is a sensitive marker for diagnosing osteoporosis.

[Approach to application of laser scanning confocal microscope in bone morphometry].

The laser scanning confocal microscope(LSCM) is the new high distinguishing microscope, which can be used in observation on fluorescence, since 1990. The bone morphometry is an important way to study metabolic bone diseases. Our study on research combination of LSCM with bone morphometry in observing microarchitecture of bone found that LSCM could make the photo of bone trabecula distinctly and its location accurately. Because of its technical advantages, LSCM could link histochemical or immunohistochemical method for the bone morphometry and bone cells at the same time and obserrate the thick-slide. It is a new method and way to study and diagnose metabolic bone diseases.