Bone remodeling and bone matrix quality before and after menopause in healthy women.

Acceleration of remodeling activity after menopause leads to bone loss and fragility, however, whether this is associated with modifications of bone matrix quality has been less studied. The impact of variation in bone remodeling rate on bone matrix has been studied mainly in pathologies or anti-osteoporotic treatments. However, in healthy women this has been less studied. We analyzed, at the global level, bone matrix quality in bone biopsies from 3 groups of healthy women (20 per group): 1) before menopause (PreM), 2) 1 year after menopause (PostM, paired biopsies with preM), and 3) 14 (±9) years after menopause (LT-PostM). The mean degree of mineralization (DMB) and heterogeneity index (HI) of mineralization were assessed by X-ray microradiography on whole bone matrix; intrinsic properties (mineral/organic ratio, mineral maturity, mineral crystallinity, collagen maturity) were assessed by Fourier Transform Infrared microspectroscopy, microhardness by microindentation, both at a global level and calculated by mean of several measurements over the whole tissue area. In PostM compared to PreM (bone remodeling rate had doubled), mean DMB measured on the entire bone plane (whole bone matrix) of the sample was not different. HI was increased in trabecular bone indicating a higher heterogeneity of mineralization. However, in PostM, mineral/organic ratio (trabecular) and microhardness (cortical and trabecular) were decreased, whereas mineral/collagen maturation or crystal size/perfection were unchanged. Thus, in PostM, the local mineral content and microhardness were first affected. In LT-PostM (bone remodeling rate was 3 times higher), the mean DMB was still not different. However, the mineral/organic ratio, microhardness, mineral maturity, crystallinity all were lower compared to PreM and PostM, in both cortical and trabecular bone. Bone remodeling rate was negatively correlated with microhardness, DMB, mineral/organic and crystallinity. This suggests that increases in bone remodeling rates after menopause have a direct impact on bone quality by inducing the formation of more extensive "immature" bone areas, but the amount of immature bone does not cause modification of the global DMB.

Bone cortical thickness and porosity assessment using ultrasound guided waves: An ex vivo validation study.

Several studies showed the ability of the cortex of long bones such as the radius and tibia to guide mechanical waves. Such experimental evidence has given rise to the emergence of a category of quantitative ultrasound techniques, referred to as the axial transmission, specifically developed to measure the propagation of ultrasound guided waves in the cortical shell along the axis of long bones. An ultrasound axial transmission technique, with an automated approach to quantify cortical thickness and porosity is described. The guided modes propagating in the cortex are recorded with a 1-MHz custom made linear transducer array. Measurement of the dispersion curves is achieved using a two-dimensional spatio-temporal Fourier transform combined with singular value decomposition. Automatic parameters identification is obtained through the solution of an inverse problem in which the dispersion curves are predicted with a two-dimensional transverse isotropic free plate model. Thirty-one radii and fifteen tibiae harvested from human cadavers underwent axial transmission measurements. Estimates of cortical thickness and porosity were obtained on 40 samples out of 46. The reproducibility, given by the root mean square error of the standard deviation of estimates, was 0.11 mm for thickness and 1.9% for porosity. To assess accuracy, site-matched micro-computed tomography images of the bone specimens imaged at 9 µm voxel size served as the gold standard. Agreement between micro-computed tomography and axial transmission for quantification of thickness and porosity at the radius and tibia ranged from R2=0.63 for porosity (root mean square error RMSE=1.8%) to 0.89 for thickness (RMSE=0.3 mm). Despite an overall good agreement for porosity, the method performs less well for porosities lower than 10%. The heterogeneity and general complexity of cortical bone structure, which are not fully accounted for by our model, are suspected to weaken the model approximation. This study presents the first validation study for assessing cortical thickness and porosity using the axial transmission technique. The automatic signal processing minimizes operator-dependent errors for parameters determination. Recovering the waveguide characteristics, that is to say cortical thickness and porosity, could provide reliable information about skeletal status and future fracture risk.

Genetic algorithms-based inversion of multimode guided waves for cortical bone characterization.

Recent progress in quantitative ultrasound has exploited the multimode waveguide response of long bones. Measurements of the guided modes, along with suitable waveguide modeling, have the potential to infer strength-related factors such as stiffness (mainly determined by cortical porosity) and cortical thickness. However, the development of such model-based approaches is challenging, in particular because of the multiparametric nature of the inverse problem. Current estimation methods in the bone field rely on a number of assumptions for pairing the incomplete experimental data with the theoretical guided modes (e.g. semi-automatic selection and classification of the data). The availability of an alternative inversion scheme that is user-independent is highly desirable. Thus, this paper introduces an efficient inversion method based on genetic algorithms using multimode guided waves, in which the mode-order is kept blind. Prior to its evaluation on bone, our proposal is validated using laboratory-controlled measurements on isotropic plates and bone-mimicking phantoms. The results show that the model parameters (i.e. cortical thickness and porosity) estimated from measurements on a few ex vivo human radii are in good agreement with the reference values derived from x-ray micro-computed tomography. Further, the cortical thickness estimated from in vivo measurements at the third from the distal end of the radius is in good agreement with the values delivered by site-matched high-resolution x-ray peripheral computed tomography.

Measurement of cortical porosity of the proximal femur improves identification of women with nonvertebral fragility fractures.

UNLABELLED: We tested whether cortical porosity of the proximal femur measured using StrAx1.0 software provides additional information to areal bone mineral density (aBMD) or Fracture Risk Assessment Tool (FRAX) in differentiating women with and without fracture. Porosity was associated with fracture independent of aBMD and FRAX and identified additional women with fractures than by osteoporosis or FRAX thresholds. INTRODUCTION: Neither aBMD nor the FRAX captures cortical porosity, a major determinant of bone strength. We therefore tested whether combining porosity with aBMD or FRAX improves identification of women with fractures. METHODS: We quantified femoral neck (FN) aBMD using dual-energy X-ray absorptiometry, FRAX score, and femoral subtrochanteric cortical porosity using StrAx1.0 software in 211 postmenopausal women aged 54-94 years with nonvertebral fractures and 232 controls in Tromsø, Norway. Odds ratios (ORs) were calculated using logistic regression analysis. RESULTS: Women with fractures had lower FN aBMD, higher FRAX score, and higher cortical porosity than controls (all p < 0.001). Each standard deviation higher porosity was associated with fracture independent of FN aBMD (OR 1.39; 95% confidence interval 1.11-1.74) and FRAX score (OR 1.58; 1.27-1.97) in all women combined. Porosity was also associated with fracture independent of FRAX score in subgroups with normal FN aBMD (OR 1.88; 1.21-2.94), osteopenia (OR 1.40; 1.06-1.85), but not significantly in those with osteoporosis (OR 1.48; 0.68-3.23). Of the 211 fracture cases, only 18 women (9%) were identified using FN aBMD T-score < -2.5, 45 women (21%) using FRAX threshold >20%, whereas porosity >80th percentile identified 61 women (29%). Porosity identified 26% additional women with fractures than identified by the osteoporosis threshold and 21% additional women with fractures than by this FRAX threshold. CONCLUSIONS: Cortical porosity is a risk factor for fracture independent of aBMD and FRAX and improves identification of women with fracture.

Bone's Material Constituents and their Contribution to Bone Strength in Health, Disease, and Treatment.

Type 1 collagen matrix volume, its degree of completeness of its mineralization, the extent of collagen crosslinking and water content, and the non-collagenous proteins like osteopontin and osteocalcin comprise the main constituents of bone's material composition. Each influences material strength and change in different ways during advancing age, health, disease, and drug therapy. These traits are not quantifiable using bone densitometry and their plurality is better captured by the term bone 'qualities' than 'quality'. These qualities are the subject of this manuscript.

Bone mineralization: from tissue to crystal in normal and pathological contexts.

Bone is a complex and structured material; its mechanical behavior results from an interaction between the properties of each level of its structural hierarchy. The degree of mineralization of bone (bone density measured at tissue level) and the characteristics of the mineral deposited (apatite crystals) are major determinants of bone strength. Bone remodeling activity acts as a regulator of the degree of mineralization and of the distribution of mineral at the tissue level, directly impacting bone mechanical properties. Recent findings have highlighted the need to understand the underlying process occurring at the nanostructure level that may be independent of bone remodeling itself. A more global comprehension of bone qualities will need further works designed to characterize what are the consequences on whole bone strength of changes at nano- or microstructure levels relative to each other.

Favorable infertility outcomes following anti-tubercular treatment prescribed on the sole basis of a positive polymerase chain reaction test for endometrial tuberculosis.

BACKGROUND: The endometrial tuberculosis (TB) PCR test is now commonly employed for the diagnosis of female genital TB, a common cause of infertility in India. Although treatment in the absence of demonstrable tubal damage may be of doubtful benefit to fertility, the presence of mycobacterial DNA demonstrated by a positive PCR indicates infection by tubercle bacilli causing sub-clinical or latent disease potentially responsible for future clinical manifestations. This study was undertaken to assess the outcome of infertility management following early anti-tubercular treatment (ATT) based only on a positive endometrial TB-PCR test. METHODS: This was an intervention study conducted at an IVF center in northern India in 443 infertile women of whom 169 (38.15%) were found to have positive TB-PCR (Group I), while 274 (61.85%) had negative Mycobacterium tuberculosis (MTB)-PCR (Group II). Infertile women of <40 years of age, without any evidence of tubo-peritoneal or endometrial involvement, who underwent endometrial biopsy for the detection of MTB by PCR, were included. All the TB-PCR positive women were administered standard 6-month anti-tubercular chemotherapy. Additional treatment with assisted reproduction techniques was offered in the case of failure of spontaneous pregnancy after completion of ATT. RESULTS: There were no statistical differences in the two groups in the overall pregnancy rate, 101 (59.8%) versus 167 (60.9%). In Group I, 48 (92.3%) spontaneous conceptions occurred within the first 12 months, i.e. during the period of ATT administration or within 6 months of treatment completion; in Group II, the occurrence of spontaneous conceptions was distributed more evenly in relation to time, i.e. 36 (53.7%) in <12 months as compared with 31 (46.3%) after first year (P< 0.001). CONCLUSION: Infertile women without tubal or endometrial damage given early anti-tuberculosis treatment based on a positive endometrial TB-PCR test had an excellent chance of early spontaneous conception.

Respective roles of organic and mineral components of human cortical bone matrix in micromechanical behavior: an instrumented indentation study.

Bone is a multiscale composite material made of both a type I collagen matrix and a poorly crystalline apatite mineral phase. Due to remodeling activity, cortical bone is made of Bone Structural Units (BSUs) called osteons. Since osteon represents a fundamental level of structural hierarchy, it is important to investigate the relationship between mechanical behavior and tissue composition at this scale for a better understanding of the mechanisms of bone fragility. The aim of this study is to analyze the links between ultrastructural properties and the mechanical behavior of bone tissue at the scale of osteon. Iliac bone biopsies were taken from untreated postmenopausal osteoporotic women, embedded, sectioned and microradiographed to assess the degree of mineralization of bone (DMB). On each section, BSUs of known DMB were indented with relatively high load (~500 mN) to determine local elastic modulus (E), contact hardness (H(c)) and true hardness (H) of several bone lamellae. Crystallinity and collagen maturity were measured by Fourier Transform InfraRed Microspectroscopy (FTIRM) on the same BSUs. Inter-relationships between mechanical properties and ultrastructural components were analyzed using multiple regression analysis. This study showed that elastic deformation was only explained by DMB whereas plastic deformation was more correlated with collagen maturity. Contact hardness, reflecting both elastic and plastic behaviors, was correlated with both DMB and collagen maturity. No relationship was found between crystallinity and mechanical properties at the osteon level.

Female genital tuberculosis: early diagnosis by laparoscopy and endometrial polymerase chain reaction.

OBJECTIVE: To evaluate the efficacy of laparoscopic visual inspection vs. endometrial tuberculosis (TB) polymerase chain reaction (PCR) for an early diagnosis and management of female genital TB (GTB) in India. DESIGN: Observational case study. RESULTS: Both laparoscopy and endometrial (endo) TB-PCR were performed on 162 infertile women: 52 endo TB-PCR-positive patients were diagnosed as definite GTB, of whom 44 (84.6%) also showed laparoscopic findings suspicious of TB (Type I). Subgroup analysis showed a similar PCR positivity of 54.5% (24/44) in the strongly suspicious and 54.1% (20/37) in the mildly suspicious patients. Using the Bayesian approach, the maximum likelihood estimates of the sensitivity and specificity of laparoscopy in diagnosing GTB were 0.96 and 0.93 and those of a positive PCR were respectively 0.59 and 0.92. Of the 52 women who were TB-PCR positive, 16 (30.8%) conceived following treatment. CONCLUSIONS: Endo TB-PCR had high specificity to diagnose GTB, as did laparoscopy. Laparoscopy may therefore be avoided in TB-PCR-positive patients for diagnosis but may still be required to rule out GTB in PCR-negative cases.

Anti-hyperglycemic activity of the leaves of Tetracera scandens Linn. Merr. (Dilleniaceae) in alloxan induced diabetic rats.

AIM OF THE STUDY: The present study was aimed to investigate the anti-diabetic potential of the leaves of Tetracera scandens Linn. Merr. (Dilleniaceae) in vivo with regard to prove its efficacy by local herbalists in the treatment of diabetes frailties. MATERIALS AND METHODS: Crude aqueous (AQ) and methanol (MEOH) extracts of the leaves of T. scandens L. were administered to both normal and alloxan induced diabetic male albino rats (Wistar strain). The blood glucose levels were measured at 0, 2, 4, 6 and 8h after oral administration of AQ and MEOH extracts. RESULTS: Significant reduction in glucose was observed in fasting blood glucose levels in the treated diabetic rats without causing any hypoglycemic effect compared to normal rats. Both polar extracts of the leaves of T. scandens L. exhibited significant anti-hyperglycemic activity at different doses and intervals. The highest anti-hyperglycemic effect (62.5%) was observed by the AQ extract at 0.25 g/kg body weight (b.w.) and MEOH extract (36.5%) at 0.5 g/kg b.w. after 8h. The significant anti-hyperglycemic activity was found to be comparable with a known oral synthetic hypoglycemic drug, glibenclamide 0.25mg/kg b.w. CONCLUSION: This study provides scientific evidence that the leaves of T. scandens L. have anti-diabetic efficacy and justifies its utility by local herbalists. However, more experiments at the clinical levels are required to confirm the utility of this plant by traditional practitioners in the management of diabetes mellitus.

The role of mineralization and organic matrix in the microhardness of bone tissue from controls and osteoporotic patients.

Degree of mineralization of bone (DMB) is a major intrinsic determinant of bone strength at the tissue level but its contribution to the microhardness (Vickers indentation) at the intermediary level of organization of bone tissue, i.e., Bone Structural Units (BSUs), has never been assessed. The purpose of this study was to analyze the relationship between the microhardness, the DMB and the organic matrix, measured in BSUs from human iliac bone biopsies. Iliac bone samples from controls and osteoporotic patients (men and women), embedded in methyl methacrylate, were used. Using a Vickers indenter, microhardness (kg/mm2) was measured, either globally on surfaced blocks or focally on 100 microm-thick sections from bone samples (load of 25 g applied during 10 sec; CV=5%). The Vickers indenter was more suited than the Knoop indenter for a tissue like bone in which components are diversely oriented. Quantitative microradiography performed on 100 microm-thick sections, allowed measurement of parameters reflecting the DMB (g/cm3). Assessed on the whole bone sample, both microhardness and DMB were significantly lower (-10% and -7%, respectively) in osteoporotic patients versus controls (p<0.001). When measured separately at the BSU level, there were significant positive correlations between microhardness and DMB in controls (r2=0.36, p<0.0001) and osteoporotic patients (r2=0.43, p<0.0001). Mineralization is an important determinant of the microhardness, but did not explain all of its variance. To highlight the role of the organic matrix in bone quality, microhardness of both osteoid and adjacent calcified matrix were measured in iliac samples from subjects with osteomalacia. Microhardness of organic matrix is 3-fold lower than the microhardness of calcified tissue. In human calcanei, microhardness was significantly correlated with DMB (r2=0.33, p=0.02) and apparent Young's modulus (r2=0.26, p=0.03). In conclusion, bone microhardness measured by Vickers indentation is an interesting methodology for the evaluation of bone strength and its determinants at the BSU level. Bone microhardness is linked to Young's modulus of bone and is strongly correlated to mineralization, but the organic matrix accounts for about one third of its variance.

An in vitro model to test the contribution of advanced glycation end products to bone biomechanical properties.

We developed an in vitro model which provides the ability to test the effects of advanced glycation end products (AGEs), specifically pentosidine (PEN) and one of its inhibitors, the aminoguanidine (AMG), on cortical bone. This model allows modification of the extent of collagen cross-linking, while controlling other factors known to influence bone strength. In this in vitro model, young bovine cortical bone specimens were incubated in phosphate-buffered saline (PBS)+/-ribose (RIB, an inducer of AGEs formation)+/-AMG for 15 days at 37 degrees C. The mineral and organic matrix as well as biomechanical properties were examined. We found that (i) incubation+/-treatments did not induce collagen denaturation compared to specimens that were not incubated; (ii) neither treatment or incubation time effected the concentration of trivalent enzymatic cross-links pyridinoline and deoxypyridinoline. The non-enzymatic cross-link PEN was undetectable in specimens that were not incubated or that were incubated in PBS or AMG alone. However, PEN concentration increased significantly in specimens incubated with RIB, whereas ribose-induced PEN formation was markedly inhibited by AMG. (iii) Incubation+/-treatments did not change the mineral maturity, crystallinity or microhardness assessed by X-ray diffraction, X-ray microscopy analyses, FTIRM and micro-indentation tests. (iv) PEN concentration was not associated with biomechanical properties assessed by 3-point bending. In conclusion, this in vitro incubation model of young bovine cortical bone induced physiologic concentrations of PEN in the RIB+AMG group and is the first to show that AMG inhibits ribose-induced formation of PEN cross-links in bone while not affecting the organic and mineral phases. AGE concentration did not influence bending mechanical properties; however, the simple 3-point bending test we used was likely inadequate to demonstrate effects of AGEs on mechanical properties.

Short-term and long-term effects of Norplant-2 on plasma lipoproteins and glucose tolerance.

Twenty-five women using the levonorgestrel-containing contraceptive implant, Norplant-2, were studied longitudinally, to see the effects on lipid profile, glucose tolerance and hepatic transaminases with short-term (12 weeks) and long-term (18 to 24 months) use of Norplant-2 [corrected]. Total lipids, cholesterol, phospholipids, triglycerides and their subfractions were evaluated. There was a significant increase in LDL-cholesterol after 12 weeks of use (p less than 0.05), which did not persist at 18-24 months follow-up. The HDL-cholesterol, HDL/LDL-cholesterol ratio, phospholipids, other lipid fractions, hepatic transaminases and glucose tolerance did not alter significantly following short-term or long-term use of Norplant-2.

Ion-exchange equilibrium distribution studies of some metal ions in nitrite media.

Ion-exchange equilibrium distribution of some metal ions has been studied in nitrite media, with cation- as well as anion-exchangers. The distribution coefficients (K(D)) show the utility of a new nitrite medium which is a complexing medium for most transition metal ions giving anionic complexes in aqueous solution. Similar studies have been performed in aqueous ethanol. Increase in the concentration of ethanol led to increased complex formation and changed K(D) values.