Stereological measures of trabecular bone structure: comparison of 3D micro computed tomography with 2D histological sections in human proximal tibial bone biopsies.

Stereology applied on histological sections is the 'gold standard' for obtaining quantitative information on cancellous bone structure. Recent advances in micro computed tomography (microCT) have made it possible to acquire three-dimensional (3D) data non-destructively. However, before the 3D methods can be used as a substitute for the current 'gold standard' they have to be verified against the existing standard. The aim of this study was to compare bone structural measures obtained from 3D microCT data sets with those obtained by stereology performed on conventional histological sections using human tibial bone biopsies. Furthermore, this study forms the first step in introducing the proximal tibia as a potential bone examination location by peripheral quantitative CT and CT. Twenty-nine trabecular bone biopsies were obtained from autopsy material at the medial side of the proximal tibial metaphysis. The biopsies were embedded in methylmetacrylate before microCT scanning in a Scanco microCT 40 scanner at a resolution of 20 x 20 x 20 microm3, and the 3D data sets were analysed with a computer program. After microCT scanning, 16 sections were cut from the central 2 mm of each biopsy and analysed with a computerized method. Trabecular bone volume (BV/TV) and connectivity density (CD) were estimated in both modalities, whereas trabecular bone pattern factor (TBPf) was estimated on the histological sections only. Trabecular thickness (Tb.Th), number (Tb.N) and separation (Tb.Sp), and structure model index (SMI) were estimated with the microCT method only. Excellent correlations were found between the two techniques for BV/TV (r = 0.95) and CD (r = 0.95). Additionally, an excellent relationship (r = 0.95) was ascertained between TBPf and SMI. The study revealed high correlations between measures of bone structure obtained from conventional 2D sections and 3D microCT data. This indicates that 3D microCT data sets can be used as a substitute for conventional histological sections for bone structural evaluations.

Determinants of the size of incident vertebral deformities in European men and women in the sixth to ninth decades of age: the European Prospective Osteoporosis Study (EPOS).

UNLABELLED: More severe vertebral fractures have more personal impact. In the European Prospective Osteoporosis Study, more severe vertebral collapse was predictable from prior fracture characteristics. Subjects with bi-concave or crush fractures at baseline had a 2-fold increase in incident fracture size and thus increased risk of a disabling future fracture. INTRODUCTION: According to Euler's buckling theory, loss of horizontal trabeculae in vertebrae increases the risk of fracture and suggests that the extent of vertebral collapse will be increased in proportion. We tested the hypothesis that the characteristics of a baseline deformity would influence the size of a subsequent deformity. METHODS: In 207 subjects participating in the European Prospective Osteoporosis Study who suffered an incident spine fracture in a previously normal vertebra, we estimated loss of volume (fracture size) from plane film images of all vertebral bodies that were classified as having a new fracture. The sum of the three vertebral heights (anterior, mid-body, and posterior) obtained at follow-up was subtracted from the sum of the same measures at baseline. Each of the summed height loss for vertebrae with a McCloskey-Kanis deformity on the second film was expressed as a percentage. RESULTS AND CONCLUSIONS: In univariate models, the numbers of baseline deformities and the clinical category of the most severe baseline deformity were each significantly associated with the size of the most severe incident fracture and with the cumulated sum of all vertebral height losses. In multivariate modeling, age and the clinical category of the baseline deformity (crush > bi-concave > uni-concave > wedge) were the strongest determinants of both more severe and cumulative height loss. Baseline biconcave and crush fractures were associated at follow-up with new fractures that were approximately twice as large as those seen with other types of deformity or who previously had undeformed spines. In conclusion, the characteristics of a baseline vertebral deformity determines statistically the magnitude of vertebral body volume lost when a subsequent fracture occurs. Because severity of fracture and number of fractures are determinants of impact, the results should improve prediction of the future personal impact of osteoporosis once a baseline prevalent deformity has been identified.

Detecting microdamage in bone.

Fatigue-induced microdamage in bone contributes to stress and fragility fractures and acts as a stimulus for bone remodelling. Detecting such microdamage is difficult as pre-existing microdamage sustained in vivo must be differentiated from artefactual damage incurred during specimen preparation. This was addressed by bulk staining specimens in alcohol-soluble basic fuchsin dye, but cutting and grinding them in an aqueous medium. Nonetheless, some artefactual cracks are partially stained and careful observation under transmitted light, or epifluorescence microscopy, is required. Fuchsin lodges in cracks, but is not site-specific. Cracks are discontinuities in the calcium-rich bone matrix and chelating agents, which bind calcium, can selectively label them. Oxytetracycline, alizarin complexone, calcein, calcein blue and xylenol orange all selectively bind microcracks and, as they fluoresce at different wavelengths and colours, can be used in sequence to label microcrack growth. New agents that only fluoresce when involved in a chelate are currently being developed--fluorescent photoinduced electron transfer (PET) sensors. Such agents enable microdamage to be quantified and crack growth to be measured and are useful histological tools in providing data for modelling the material behaviour of bone. However, a non-invasive method is needed to measure microdamage in patients. Micro-CT is being studied and initial work with iodine dyes linked to a chelating group has shown some promise. In the long term, it is hoped that repeated measurements can be made at critical sites and microdamage accumulation monitored. Quantification of microdamage, together with bone mass measurements, will help in predicting and preventing bone fracture failure in patients with osteoporosis.

Determinants of incident vertebral fracture in men and women: results from the European Prospective Osteoporosis Study (EPOS).

The aim of this analysis was to determine the influence of lifestyle, anthropometric and reproductive factors on the subsequent risk of incident vertebral fracture in men and women aged 50-79 years. Subjects were recruited from population registers from 28 centers across Europe. At baseline, they completed an interviewer-administered questionnaire and had lateral thoraco-lumbar spine radiographs performed. Repeat spinal radiographs were performed a mean of 3.8 years later. Incident vertebral fractures were defined morphometrically and also qualitatively by an experienced radiologist. Poisson regression was used to determine the influence of the baseline risk factor variables on the occurrence of incident vertebral fracture. A total of 3173 men (mean age 63.1 years) and 3402 women (mean age 62.2 years) contributed data to the analysis. In total there were 193 incident morphometric and 224 qualitative fractures. In women, an age at menarche 16 years or older was associated with an increased risk of vertebral fracture (RR = 1.80; 95%CI 1.24, 2.63), whilst use of hormonal replacement was protective (RR = 0.58; 95%CI 0.34, 0.99). None of the lifestyle factors studied including smoking, alcohol intake, physical activity or milk consumption showed any consistent associations with incident vertebral fracture. In men and women, increasing body weight and body mass index were associated with a reduced risk of vertebral fracture though, apart from body mass index in men, the confidence intervals embraced unity. For most variables the strengths of the associations observed were similar using the qualitative and morphometric approaches to fracture definition. In conclusion our data suggest that modification of other lifestyle risk factors is unlikely to have a major impact on the population occurrence of vertebral fractures. The important biological mechanisms underlying vertebral fracture risk need to be explored using new investigational strategies.

[Age- and gender-related distribution of bone mineral density and mechanical properties of the proximal humerus]. / Alters- und geschlechtsabhängige Knochenmineraldichteverteilung und mechanische Eigenschaften des proximalen Humerus.

PURPOSE: To evaluate age- and gender-related mechanical properties and bone mineral density (BMD) of the proximal humerus at different levels and regions. MATERIALS AND METHODS: Mechanical indentation testing, DXA, QCT, pQCT and the radiogrammetry (Cortical Index, CI) were carried out in 70 freshly harvested humeri from 46 human cadavers (23 females, 23-males; median age 70.5 years). RESULTS: In the female group, a high correlation between age and BMD was found (rho = 0.62 to -0.70, p < 0.01) with statistically significant differences between specimens of patients 69 years or younger, and 70 years or older (p < 0.05). In the group of female specimens of age 70 years or older, BMD values were found to be significantly lower compared to their male counterparts (p < 0.05). Regardless of the specimen's age, the highest BMD and bone strength were found in the proximal aspect and in the medial and dorsal regions of the proximal humerus. CONCLUSION: These findings provide an insight into the fracture mechanism of the proximal humerus and should be the basis for designing structure-oriented implants with improved implant-bone stability in osteoporotic patients.

Incidence of vertebral fracture in europe: results from the European Prospective Osteoporosis Study (EPOS).

Vertebral fracture is one of the major adverse clinical consequences of osteoporosis; however, there are few data concerning the incidence of vertebral fracture in population samples of men and women. The aim of this study was to determine the incidence of vertebral fracture in European men and women. A total of 14,011 men and women aged 50 years and over were recruited from population-based registers in 29 European centers and had an interviewer-administered questionnaire and lateral spinal radiographs performed. The response rate for participation in the study was approximately 50%. Repeat spinal radiographs were performed a mean of 3.8 years following the baseline film. All films were evaluated morphometrically. The definition of a morphometric fracture was a vertebra in which there was evidence of a 20% (+4 mm) or more reduction in anterior, middle, or posterior vertebral height between films--plus the additional requirement that a vertebra satisfy criteria for a prevalent deformity (using the McCloskey-Kanis method) in the follow-up film. There were 3174 men, mean age 63.1 years, and 3,614 women, mean age 62.2 years, with paired duplicate spinal radiographs (48% of those originally recruited to the baseline survey). The age standardized incidence of morphometric fracture was 10.7/1,000 person years (pyr) in women and 5.7/1,000 pyr in men. The age-standardized incidence of vertebral fracture as assessed qualitatively by the radiologist was broadly similar-12.1/1,000 pyr and 6.8/1,000 pyr, respectively. The incidence increased markedly with age in both men and women. There was some evidence of geographic variation in fracture occurrence; rates were higher in Sweden than elsewhere in Europe. This is the first large population-based study to ascertain the incidence of vertebral fracture in men and women over 50 years of age across Europe. The data confirm the frequent occurrence of the disorder in men as well as in women and the rise in incidence with age.

Bone modeling and structural measures of complexity.

We test sensitivity and powerfulness of recently suggested Structure Measures of Complexity (SMC) with simulated test objects, represented by simple structures or modelled on the basis of a real bone image. We check how these SMC reflect the local and global disordering processes, as well as a deterioration of the bone structure. We show that applications of SMC provide additional information about any changes of the bone structure in comparison to bone mineral density (BMD), and that they can be potentially helpful in the diagnosis of osteoporosis.

Bone architecture assessment with measures of complexity.

Architectural changes in trabecular bone by osteoporosis were utilized as a model for the changes which probably occur in human bone while exposed to microgravity conditions. Although there are many concerns about microgravity-induced bone loss, little is known about the impact of microgravity on the three-dimensional architecture of the skeleton. 50 (level L3) and 57 (level L4) vertebral bones harvested from human cadavers were investigated by computed tomography (CT) and quantified in terms of bone mineral density (BMD). Based on the symbol-encoded transformed CT-images, five measures of complexity were developed which quantify the structural composition of the trabecular bone. This quantification determines the bone architecture as a whole. Depending on the specific measure of complexity and its relation to BMD, a 5-10% change of BMD is related to a 5-90% change in structural composition. The method requires a non-invasive CT-procedure of the lumbar spine resulting in a radiation exposure of about 30 microSv effective dose. The technique is useful for the evaluation of the bone status of space-flying, personnel as well as for patients on ground. Grant numbers: BMH1-CT92-0296.

Osteoporosis changes the amount of vertebral trabecular bone at risk of fracture but not the vertebral load distribution.

STUDY DESIGN: A finite-element study to investigate the amount of trabecular bone at risk of fracture and the distribution of load between trabecular core and cortical shell, for healthy, osteopenic, and osteoporotic vertebrae. OBJECTIVES: To determine differences between healthy, osteopenic, and osteoporotic vertebrae with regard to the risk of fracture and the load distribution. SUMMARY OF BACKGROUND DATA: The literature contains no reports on the effects of osteopenia and osteoporosis on load distribution in vertebral bodies, nor any reports on the amount of trabecular bone at risk of fracture. METHODS: Computed tomography data of vertebral bodies were used to construct patient-specific finite-element models. These models were then used in finite-element analyses to determine the physiologic stresses and strains in the vertebrae. RESULTS: For all three classes of vertebrae the contribution of the trabecular core to the total load transfer decreased from about 70% near the endplates to about 50% in the midtransverse region. The amount of trabecular bone that is at risk of fracture was about 1% for healthy vertebrae, about 3% for osteopenic vertebrae, and about 16% for osteoporotic vertebrae. CONCLUSIONS: Our finite-element models indicated that neither osteopenia nor osteoporosis had any effect on the contribution of the trabecular core to the total load placed on the vertebra. The trabecular core carried about half the load. Our finite-element models indicated that osteoporosis had a significant effect on the amount of trabecular bone at risk of fracture, which increased from about 1% in healthy vertebrae to about 16% for osteoporotic vertebrae.

Prevalent vertebral deformity predicts incident hip though not distal forearm fracture: results from the European Prospective Osteoporosis Study.

The presence of a vertebral deformity increases the risk of subsequent spinal deformities. The aim of this analysis was to determine whether the presence of vertebral deformity predicts incident hip and other limb fractures. Six thousand three hundred and forty-four men and 6788 women aged 50 years and over were recruited from population registers in 31 European centers and followed prospectively for a median of 3 years. All subjects had radiographs performed at baseline and the presence of vertebral deformity was assessed using established morphometric methods. Incident limb fractures which occurred during the follow- up period were ascertained by annual postal questionnaire and confirmed by radiographs, review of medical records and personal interview. During a total of 40348 person-years of follow-up, 138 men and 391 women sustained a limb fracture. Amongst the women, after adjustment for age, prevalent vertebral deformity was a strong predictor of incident hip fracture, (rate ratio (RR) = 4.5; 95% CI 2.1-9.4) and a weak predictor of 'other' limb fractures (RR = 1.6; 95% CI 1.1-2.4), though not distal forearm fracture (RR = 1.0; 95% CI 0.6-1.6). The predictive risk increased with increasing number of prevalent deformities, particularly for subsequent hip fracture: for two or more deformities, RR = 7.2 (95% CI 3.0-17.3). Amongst men, vertebral deformity was not associated with an increased risk of incident limb fracture though there was a nonsignificant trend toward an increased risk of hip fracture with increasing number of deformities. In summary, prevalent radiographic vertebral deformities in women are a strong predictor of hip fracture, and to a lesser extent humerus and 'other' limb fractures; however, they do not predict distal forearm fractures.

A statistical method to minimize magnification errors in serial vertebral radiographs.

Incident vertebral deformities are commonly defined by observed changes in height between measurements on two consecutive radiographs. However, conventional radiographs are subject to magnification, and this magnification may differ between films, leading to artifactual changes in height. In order to minimize this effect, it is common practice to record the spine-film and film-focus distances, and from this to calculate a magnification factor for each film. We present a simple statistical method for correcting for differences in magnification between two films if the spine-film and film-focus distances are unknown. This method is shown to reduce the variance of the magnification differences in vertebral heights by 14%, considerably more than is possible using the spine-film distance. Using the statistical method, the number of vertebrae that showed not only a reduction in one or more height of 15%, but were also judged clinically to be free from any incident deformity by an expert radiologist, was reduced from 100 to 46. The number showing a reduction of 20% that were judged fracture-free was reduced from 15 to 9. In the subset of subjects for whom the spine-film distance was known, the reduction in false positives was similar, whichever method was used to correct for magnification. There was no difference in the number of confirmed incident fractures detected when magnification correction by either method was employed. It is concluded that correcting for magnification differences using the statistical method outlined here reduces the number of false positive deformities very substantially and by a similar extent as correcting the magnification using reliable, measured spine-film and film-focus distances. A further advantage of this method is that it can be used retrospectively.

Serum leptin levels, body fat deposition, and weight in females with anorexia or bulimia nervosa.

The objective of this retrospective study was to investigate the relation between serum leptin level and fat deposition in patients with eating disorders. 40 female inpatients with anorexia (n=24) or bulimia nervosa (n=16) were assessed for leptin level, body mass index (BMI), and percentage body fat by dual-energy X-ray absorbometry (DXA). The results show that percentage body fat is a better predictor for leptin level and clinical findings in eating disordered patients than BMI. We discuss the necessity for DXA measurements in anorectic patients for prognostic and research purposes.

[Bone densitometry by dual energy methods]. / Knochendichtemessung mit Zwei-Spektren-Methoden.

Osteoporosis is mainly diagnosed by means of bone densitometry. Dual X-ray absorptiometry examinations represent the basis for a highly reproducible and correct measurement. At present, densitometry is the only method at our disposal capable of assessing material-related fracture risk. The calculation of general fracture risk is dependent on a number of varios factors and is, therefore, not to be deduced from bone density values only. Reference values are necessary in order to estimate bone strength. The most sensible way to achieve this is to compare measured values with a normal, healthy population (T score). Material-related fracture risk increases with the decrease of bone density.

Measurement imprecision in vertebral morphometry of spinal radiographs obtained in the European Prospective Osteoporosis Study: consequences for the identification of prevalent and incident deformities.

Several algorithms are currently in use for evaluating vertebral deformities from plain lateral radiographs of the lumbar and thoracic spine. However, the effects of measurement imprecision as well as uncertainties over image magnification on the correct identification of prevalent and incident vertebral deformities with these algorithms has been little studied. In a pilot study for the European Prospective Osteoporosis Study (EPOS), plain radiographs were submitted to a single central evaluating centre for measurement of vertebral height from T4 to L4. The thoracic and lumbar spines were imaged on separate films, and we have assessed the precision of measurement of vertebral heights and height ratios. The standard deviation of the differences between films of each of three height measurements ranged from 1.1 to 1.2 mm. A two-stage strategy for identifying incident deformities was devised. This required that the vertebra be a prevalent deformity at the time of the second radiograph and also that at least one of the vertebral ratios should have changed significantly since the first radiograph. The second stage removed all but two of the 18 vertebrae flagged positive in the first stage but not considered to be certain incident fractures by clinical reading of the radiographs.

Bone densitometry: applications in sports-medicine.

Physical exercise and sports increase muscular mass and the remodelling process of bones. The increment of bone depends on the type and the quality of sport. Short-term high-performance activities such as sprint, tennis, fencing lead to increased bone mineral density as well as weight lifting or heavy athletics. Swimming, bicycling, walking are associated with good musculature conditioning without an increase of bone mass. The effects on the bone by performing endurance activities are controversially discussed. Excessive sport leads to an increase of fatigue fractures. Low bone mass may result from hormonal disregulation in female athletes.

[Morphometric analysis of roentgen images of the spine for diagnosis of osteoporosis-induced fracture]. / Morphometrische Analyse von Röntgenbildern der Wirbelsäule zur Diagnose einer osteoporotischen Fraktur.

The European Vertebral Osteoporosis Study (EVOS) is one of the largest studies to investigate the prevalence of osteoporosis related vertebral fractures in a population based cross-sectional study. One of the main tasks was to create standards and logistics to obtain uniform and comparable radiographs in all 36 European centers. Furthermore the central reading of the X-rays and the morphometry of the vertebral bodies were 2 important challenges. This paper describes the standardized patient positioning and making of the radiographs as well as their morphometry, reading, and evaluation.

[Prevalence of vertebral spinal deformities in women and men in Germany. EVOS group in Germany]. / Prävalenz der vertebralen Wirbelkörperdeformationen bei Frauen und Männern in Deutschland. EVOS-Gruppe in Deutschland.

BACKGROUND: The prevalence of radiographically defined vertebral deformities, as a marker of vertebral osteoporosis, was calculated in a population based cross-sectional survey in Germany. METHOD: Lateral spine X-rays were taken according to a standardized protocol and evaluated centrally. Three thousand nine hundred and eighty subjects (2064 male and 1916 female) aged 50 to 79 years, have been examined in 8 German centers. RESULTS: Based on McCloskey's method of deformity definition the age-standardized mean prevalence of vertebral deformities in Germany was 10.2% for males and 10.5% for females. Based on the definition by Melton/Eastell a significant higher prevalence was calculated (males 17.8%, females 18.7%). The prevalence increased with age in both sexes with a steeper increase in females. There was no difference in East and West German populations. There were substantial variations between the centers regarding the prevalence of deformities in females and males.

Measures of complexity for cancellous bone.

The problem of quantifying the structure of cancellous bone has been addressed in the past by histomorphometry and more recently by imaging techniques using X-ray attenuation. The current approaches compute and describe parts of the construction of the trabecular net. We developed a new technique which quantifies cancellous bone of human lumbar vertebrae as a whole. The interactions, transactions, and interrelationships of all parts of the structural composition of the trabeculae are accounted for and quantified. The method is based on the concept of structural complexity within the framework of nonlinear dynamics. The methodology was developed by using axial high resolution computed tomography images. The technique was transferred to quantitative computed tomography images and is based on the non-invasive assessment of 50 human L3 specimens. The value of Houndsfield units per pixel representing trabecular bone of the vertebrae was transformed into color-encoded and alphabet-encoded symbols. The procedure of transformation of the X-ray attenuation pixels into symbols was necessary as a basis on which measures of complexity were introduced to assess the composition of symbols within the images. The development of a generalization of symbolic dynamics, a mathematical method, to work with two-dimensional images was a prerequisite. The results of this study demonstrate that the structural composition of cancellous bone declines more rapidly than bone mineral density during the loss of bone. This outcome strongly suggests an exponential relationship between bone mineral density and the architectural composition of cancellous bone. Normal trabecular bone has a complex ordered structure. The structural composition during the osteopenic phase of bone loss is characterized by lower structural complexity and a significantly higher level of architectural disorder. A high grade of osteoporosis leads again to an ordered structure, although its structural complexity is minimal.

[Vertebral morphometry with DXA/MXA equipment--an equipment comparison. Dual x-ray absorptiometry/morphometric x-ray absorptiometry]. / Wirbelkörpermorphometrie mit DXA/MXA-Geräten--Ein Gerätevergleich.

PURPOSE: The accuracy and reproducibility of morphometric measurements (Morphometric X-ray Absorptiometry = MXA) of vertebrae were determined. The significant difference of the change in height of vertebral bodies in follow-up studies was computed for MXA methods and digitised spinal radiographs as well. MATERIAL AND METHODS: The measurements were carried out on two new Dual X-ray Absorptiometry (DXA) devices (device A = Expert, Lunar Corp., device B = QDR 2000 Plus, Hologic Inc.). The data were obtained by using the European Spine Phantom (ESP) and lumbar spine specimens. RESULTS: The accuracy of vertebral morphometry performed on radiographs is 2.0%, on the device A 2.3%, and on the device B 4.9%. Measurements taken with the ESP showed a reproducibility of 1.0 to 3.0%, whereas measurements of fractured vertebrae resulted in 5.1 to 6.0%. CONCLUSION: The results of the morphometric measurements demonstrate that a reliable fracture analysis in phantoms and specimens is possible. Further in-vivo studies are necessary.

[Systematics of osteodensitometric methods and acronyms]. / Systematik osteodensitometrischer Methoden und Akronyme.

The radiological diagnosis of osteoporosis is primarily based on osteodensitometry. The continuous development of new methods requires persistent analysis of its potential and limits. The knowledge of the differences of osteodensitometric methods is essential for understanding patient reports. The acronyms are internationally established, their correct usage is advocated.