This study investigated age and sex differences in patterns of porosity distribution in the midshaft of the human femur. Cross-sections were obtained from 168 individuals from a modern Australian population. The sample comprised 73 females and 95 males, aged between 20 and 97 years. Microradiographs were made of 100-microm sections and pore and bone areas were determined using image processing software. Initially the sample was divided by age: young (20-44 years), middle (45-64 years) and old (65+ years), but it was found that analysis on the basis of the ratio of medullary area to total subperiosteal area gave clearer results. The cortex was divided into three rings radially and into octants circumferentially and the porosity of each segment was calculated. Results showed that a pattern with raised porosity in the posterior and anterolateral regions, and with greater porosity in the inner parts of the cortex, becomes more pronounced with age. In males this pattern develops steadily; in females there are much greater differences between the middle and older groups than earlier in life. The patterns observed are consistent with progressive bone loss occurring along a neutral axis of the cortex where bending stress is lowest and the mechanical advantage of the bone is least.

Mineralization density and collagen fibre orientation are two aspects of a bone's microstructural organization that influence its mechanical properties. Previous studies by our group have demonstrated a distinctly non-random, though highly variable, spatial distribution of these two variables in the human femoral cortex. In this study of 37 specimens, these variables are examined relative to one another in order to determine whether regions of bone demonstrating higher or lower mineralization density also demonstrate a prevalence of either transversely or longitudinally oriented collagen fibres. An analysis of rank-transformed collagen fibre orientation (as determined by circularly polarized light) and mineralization density (as determined by backscattered electron microscopy) data sets demonstrated that areas of low mineralization density (predominantly in the anterior-lateral cortex) tended to correspond to regions of higher proportions of longitudinally oriented collagen fibres. Conversely, areas of higher mineralization density (postero-medially) tended to correspond to regions of higher proportions of transversely oriented collagen fibres. High variability in the sample led to generally low correlations between the two data sets, however. A second analysis focused only on the orientation of collagen fibres within poorly mineralized bone (representing bone that was newly formed). This analysis demonstrated a lower proportion of transverse collagen fibres in newly formed bone with age, along with some significant regional differences in the prevalence of collagen fibres of either orientation. Again high variability characterized the sample. These results are discussed relative to the hypothesized forces experienced at the midshaft femur.

Introduction: Femoral neck (FN) fragility has been attributed to age-related bone loss, with increased loss in women. It has been shown that the mechanical properties of a supporting structure will also change with any alteration to the structure’s dimensions. The purpose of this study was to identify the age-related changes that take place in the morphology of the mid cross-section of the FN, and the implications for its mechanical properties in the different regions around the mid FN cross-section. Materials and Methods: Measurements were taken from peripheral quantitative computed tomogram (pQCT) images of 81 cadaveric femurs (36 F, 45 M). The mid FN cross-section was segmented radially into eight regions and the cortical bone thickness (CT) and change of the centroid position (CP) of the FN cross-section were measured. The age-related effects of the corresponding changes in the proportion of cortical bone and the “resistance to bending” (section modulus, (Z)) were also measured. Results: Four femurs were excluded because there were clear signs of OA being present. The maximum difference in regional CT between men and women, was less than 7% (Female: 3.07 ± 0.108mm; Male: 3.28 ± 0.123 mm (mean ± SEM) p =0.21). However, there were regional differences in CT between the young under fifty, (Un50, n=26) and the old, (Abv50), (ANOVAs for young vs old: CT p = 0.001 t 0.01). These effects were attributable to differences in the inferior region, where there was an increase in thickness of the cortical bone of 27% with senior status (Abv50: 3.44 ± 0.09mm; Und50: 2.70 ± 0.12mm. p = 0001) counter balanced by anterior and posterior loss. There was a corresponding change in CP, the distance of the superior, posterior, and superoposterior regions to the FN cross-section’s centroid, 7.6% (Abv50: 20.88 ± 0.28mm; Und50: 19.40 ± 0.47mm; p = 0.005); 6.7% (Abv50: 14.67 ± 0.2mm; Und50: 13.74 ± 0.32mm; p = 0.01); and 8%(Abv50: 17.95 ± 0.24; Und50: 16.61 ± 0.37), respectively. When these two measurements were combined (CP divided by CT) to provide the Local Buckling Ratio (BLR), where the higher the ratio the more unstable the structure, there were significant differences in superoanterior, 30%(Abv50: 15.8 ± 0.52; Und50: 12.1 ± 0.59;p=0.0001); anterior, 20%(Abv50: 10.1 ± 0.32; Und50: 8.3 ± 0.4; p=0.001); inferior, 35%(Abv50: 4.37 ± 0.14; Und50: 5.8 ± 0.34; p=0.0001); inferoposterior 18%(Abv50 8.6 ± 0.27: Und50: 7.36 ± 0.41; p=0.008); posterior, 29%(Abv50: 14.0 ± 0.33; Und50: 10.8 ± 0.5; p=0.0001) and superoposterior, 14%(Abv50: 14.6 ± 0.3; Und50: 12.8 ± 0.4; p=0.001), regions. There was no significant difference in bending resistance nor in the proportion of cortical bone. Conclusions: A more uniform cortical thickness, seen in the young, would optimise fracture resistance to overloading from unusually loaded directions. Ageing was associated with a thickening of the inferior cortex and thinning of the cortex elsewhere. This effects the location of the area that is least susceptible to the loading forces experienced in stance – that is of the FN mid cross-section’s neutral bending axis – as it will be nearer to the inferior region. Such a change in the morphology will produce deterioration in the FN’s capacity to take a load as shown by the detrimental change in the LBR. This change may indicate that the potential for femoral neck fracture increases with age when load is applied in a direction different to normal stance eg through the greater trochanter.

Collagen fiber orientation is one aspect of the microstructure of bone that influences its mechanical properties. While the spatial distribution of preferentially oriented collagen is hypothesized to reflect the effects of loading during the process of aging, its variability in a modern human sample is essentially unknown. In a large sample (n = 67) of autopsied adults, the variability of collagen fiber orientation in the mid-shaft femur was examined in relation to age and sex. Montaged images of entire 100 microm thick cross-sections were obtained using circularly polarized light microscopy (CPLM) under standardized illuminating conditions. An automated image-analyzing routine divided images into 48 segments according to anatomical position. Average gray values (varying with orientation) were quantified for each segment, and one-way ANOVA with Tukey HSD post hoc tests were applied to assess differences between segments. Collagen fiber orientation appeared to be nonrandomly distributed across the mid-shaft femur sample; however, no single "human" pattern was identified. Individual variation, unexplainable by age, sex, or body size, exceeded population-level trends. Differences between age and sex groups suggest there is a strong correspondence between collagen fiber orientation and tissue-type distributions. The minimal consistencies demonstrated here may reflect mechanical forces induced at the femoral mid-shaft. However, the myriad of other factors that may influence collagen fiber orientation patterning, including growth trajectories, metabolic and nutritional status, and disease states, must be explored further. Only then, in conjunction with studies of other structural and material properties of bone, will we be able to elucidate the linkages between microstructure and functional adaptation in the human mid-shaft femur.

The method developed by Yoshino et al. in [Forensic Sci. Int. 109 (2000) 225 and Jpn. J. Sci. Tech. Iden. 5 (2000) 9] and already being applied in Japan utilizes a three-dimensional (3D) physiognomic rangefinder combined with a computer-assisted superimposition system. Facial outlines can be compared between two-dimensional (2D) surveillance images and data extracted from 3D images obtained from the rangefinder. Also, the loci of potentially concordant features can be compared and differences measured. The method is largely objective and gives statistics for false positive/false negative findings. This recently developed method by Yoshino et al. is currently being introduced to the Japanese courts. To enable courts outside Japan to assess the admissibility of this new method, studies of non-Japanese faces have been undertaken and shown to produce similar low error rates. The present authors, therefore, consider the Yoshino method to be applicable in a non-Japanese context. As part of this study a comparison of morphological features between two ethnic groups has been undertaken using 3D measurements for the first time and will serve as the foundation for an anthropological database in the future.

One of several microstructural variables known to affect the mechanical properties of bone is the degree of mineralization of bone matrix. The aim of this study was to examine mineralization density, and its variability with age and sex, from a biomechanical perspective. Histological sections, prepared from mid-shaft femora obtained at autopsy from 40 individuals, were imaged using quantitative backscattered electron microscopy. Each cross-section montage was divided into 48 segments according to anatomical position. Mean grey-level values were quantified for each segment. One-way ANOVA with Tukey HSD post hoc tests were used to test for differences in mineralization between segments, age groups and sexes. Results showed a decrease in overall degree of mineralization density with adult age, but an increase in its coefficient of variation. Degree of mineralization was significantly lower in the periosteal third of the cortex, particularly in the antero-lateral aspect. This pattern was most prevalent amongst the youngest individuals in the sample. Whereas males between ages 45-64 years had a higher average degree of mineralization than females, the opposite was true of the older age group. Mineralization significantly decreased between middle and older age groups in males, but not in females. Despite limited consistencies in the location of high and low average mineralization bone through the cortex, the degree of interindividual variation, even within a single age and sex group, overwhelmed population level trends. The patterns of variability identified in this study are consistent with results of an analysis of collagen fibre orientation using the same sample material.

This study was designed to investigate the relationship between traditional skeletal cephalometric measurement and Fourier analysis of the lateral soft-tissue profile. A random sample of 121 untreated subjects of European descent, with wide ranges of malocclusions and underlying facial patterns, was selected in the Orthodontic Unit at the University of Melbourne. Lateral cephalograms were available for all subjects. Both traditional lateral cephalometric analysis and Fourier soft-tissue profile analysis were carried out. Multivariate statistical analysis among 11 hard-tissue cephalometric measurements and the first 50 Fourier harmonics was then performed. This analysis formed the basis for a subsequently proposed soft-tissue prediction model. From this model, 50 predicted x- and y-harmonics were generated for each subject in the total sample. Calculation of Pearson's correlation coefficients between the actual and predicted harmonics revealed strong relationships for many of the lower-order harmonics. To further test the model, the prediction-coefficients derived from all 121 subjects were then used to make predictions for the first 50 x- and y-harmonics for a subgroup of 10 independent test subjects. Once again, Pearson's correlations between the actual and predicted harmonics of the test model in the lower-order harmonics revealed strong associations. Superimposition of the actual and predicted soft-tissue outlines, however, revealed that much actual detail in the region between the nose and the chin was still lost using the predicted Fourier harmonics. This suggests that soft-tissue prediction based on this Fourier test model, while already useful in Forensic facial reconstruction, may not yet be appropriate for useful diagnosis and planning in clinical disciplines.

The reliability of a morphometrical matching method for identifying disguised faces was examined experimentally using a computer-assisted facial image identification system. The 2D right oblique facial images of three target persons disguised with sunglasses, cap and gauze mask were each compared with each of the 3D facial images of 100 subjects, yielding 900 face-to-face superimpositions. The average perpendicular distance between the facial outlines and the average point-to-point distance of the corresponding landmarks in the 2D image of the disguised face and the 3D facial image, were calculated. As a matching criterion, the sum of the values of the average perpendicular difference of the facial outlines and the average point-to-point difference between the corresponding landmarks was used (abbreviation: average difference). The range of the average difference was 2.3–2.8 mm for the same person (a match) and 4.0–14.6 mm for different (non-matching) people, respectively. The ranges for matching and non-matching faces did not overlap. Even the 3D facial images of the non-matching person showing the closest value of average difference to the average difference for the matching person could be excluded easily. It was concluded that the morphometrical matching method can reliably identify disguised faces and the results produced by this method could be easily understandable by a court of law.

Previous studies of cortical remodeling in the fractured femoral neck indicated that the merging of spatially clustered remodeling osteons could result in the formation of deleteriously large cavities associated with femoral neck fracture. This study aimed to identify whether remodeling osteons in the femoral shaft were also clustered and to assess the influence of age and gender. Microradiographic images of femoral mid-shaft cross-sections from 66 subjects over 21 years of age were analyzed to determine the number, size and location of all Haversian canals. Those most recently remodeled were identified using an edge-detection algorithm highlighting the most marked differential gradients in grey levels. Cluster analysis (JMP software) of these osteons identified the proportion of recently remodeled osteons that were within 0.75 mm clusters. As in the femoral neck, remodeling osteons were significantly more clustered than could occur by chance (real, 59.4%; random, 39.4%; P < 0.0001). The density of these clusters (number/mm(2)) was not significantly associated with subject age or gender but was greatest near the periosteum and decreased toward the marrow cavity (periosteal 0.043 +/- 0.004; mid-cortex 0.028 +/- 0.003; endosteal 0.017 +/- 0.002). Cortical porosity increased with age. The presence of giant canals (diameter >385 microm) was inversely related to the presence of clusters (R(2) = 0.237, P < 0.0001). This data suggest that remodeling osteons tend to be spatially colocalized in the shaft as they are in the neck of the femur and their presence is independent of age or gender. We propose that these remodeling clusters be termed super-osteons. The negative relationship between super-osteons and giant canals raises the intriguing possibility that loss of the control of remodeling depth results in the merging of osteonal systems to form deleteriously large cortical cavities with a marked reduction in bone strength.

Bone is characterised by age‐related morphological and histological changes. We have previously established an automated method of recording bone morphometry and histology from entire transverse sections of cortical bone. Our aim was to determine whether data acquired using this automated system were useful in the prediction of age. Ninety‐six specimens of human femoral middiaphysis were studied from subjects aged 21–92 y. Equations predicting specimen age were constructed using macroscopic data (total subperiosteal area (TSPA), periosteal perimeter (PP), endosteal perimeter (EP), cortical bone area (CA) and moments of area) and microscopic data (the number, size and diversity of pores and intracortical porosity) together with sex, height and weight. Both TSPA and PP were independent predictors of age but the number of pores was not a significant predictor of age in any equation. The age predicted by these equations was inaccurate by more than 8 y in over half the subjects. We conclude that we could not predict age at a clinically acceptable level using data from our automated system. This most likely reflects an insensitivity to regional age‐related changes in bone histology because we recorded data from each entire cortex. Automated bone measurement according to cortical region might be more useful in the prediction of age. The inclusion of TSPA together with PP as independent predictors of age raises the possibility that a future measure of periosteal shape at the femoral diaphysis could also be helpful in the prediction of age. The accuracy reached with the relatively simple methods described here is sufficient to encourage the development of image‐analysis systems for the automatic detection of more complex features.

Modern lifestyle changes may result in site-specific alterations in the skeleton. Our aim was to determine sex and age differences in regional geometry at the mid-femur. Complete cross sections from 113 individuals aged 20-97 years from a modern Australian population were obtained. A further subsample of 24, in whom the precise orientation of specimens was known, was subsequently collected. Microradiographs were made of 100-microm sections and the bone was analyzed using image processing software (Optimas, Media Cybernetics). The periosteal boundary was extracted automatically and the centroid of the periosteal outline was calculated. Fourier shape analysis was used to delineate the endocortical surface. Radial and cortical widths in each quadrant were determined. The posterior was identified by the linea aspera, and the medial and lateral were indistinguishable and therefore grouped together. For analysis, the entire sample was divided into three groups: young (20-40 years), middle (41-60 years), and old (61+ years). Raw and height-normalized values were analyzed with SPSS using t-tests, analysis of variance, and Tukey's honestly significant difference (HSD) tests. The results show that with age the femoral mid-shaft in both sexes becomes larger and more circular, with a slight shift towards the anterior. Apposition is least on the posterior and resorption greatest on the anterior, the latter being particularly evident in postmenopausal females. The greatest sex differences are seen in the middle years, lessening again in the old. We conclude that differential circumferential modeling in response to functional and postural changes occurs in both sexes with age.

Twins studies provide a powerful approach to determining the relative contribution of genetics and environment to observed variation. Such studies assume trait differences in monozygous (MZ) twins are due to environmental factors and those in dizygous (DZ) twins are due to both genetic and environmental factors. This study quantitated facial profiles of twins using Fourier equations, determining their value in profile analysis and the assessment of the genetic contribution to facial shape. Standardized profile slide photographs of 79 pairs of 4-6 year-old twins (37 MZ pairs, 42 DZ pairs) were scanned and x and y coordinates were extracted from each profile using sellion and Camper's plane as references. The coordinates were subjected to Fourier analysis and the normalised vertex projection coefficients were studied. The means of the differences between coefficients for MZ co-twins did not differ significantly from that of DZ co-twins, although the DZ group showed higher mean differences in the higher harmonics. Subjective examination of superimposed reconstructions showed wider variation between DZ co-twins than MZ co-twins. Correct classification of twins by discriminant function analysis using Fourier coefficients was similar for both groups (MZ: 70.3%; DZ: 73.8%). Fourier analysis could quantitate facial profiles of young children and differentiate some details, but was unable to discriminate between genetic and environmental influences, and any possible interactions between these influences, on their overall facial profiles at this developmental stage.

The matrix of human cortical bone is arranged around a network of vascular spaces (hereafter referred to as "pores"). Our aim was to investigate age-related differences in human cortical porosity (total pore area divided by cortical bone area), pore size and number, and surface to volume ratios, while adjusting for sex, height, and weight. Ninety-six specimens of entire transverse sections of human femoral diaphysis, from subjects aged 21-92 years, were examined. We used our established automated image acquisition and analysis system which measures pores from entire sections of multiple specimens of bone. Over 400,000 pores were recorded. Results showed a greater porosity in older bone (p < 0.01) but marked variation in porosity for any given age. The cohort median, of the specimen medians, of pore cross-sectional area was 2050 microns 2. Older specimens did not have more pores than younger specimens but had a greater proportion of larger pores (p < 0.05) and greater intraspecimen variation in pore size (p < 0.001). The pore surface to bone matrix volume ratio was a median 2.3 mm2/mm3. This varied more than 4-fold between individuals but did not relate to age. No simple relationships were found between any of the measured parameters and either sex, height, or weight, even after adjustment for age. We conclude that the greater porosity in older specimens is due to greater pore size rather than a larger number of pores. Age, however, explains little of the inter-individual variation in the parameters studied.

Characteristics of microscopic structures in bone cross sections carry essential clues in age determination in forensic science and in the study of age-related bone developments and bone diseases. Analysis of bone cross sections represents a major area of research in bone biology. However, traditional approaches in bone biology have relied primarily on manual processes with very limited number of bone samples. As a consequence, it is difficult to reach reliable and consistent conclusions. In this paper we present an image processing system that uses microstructural and relational knowledge present in the bone cross section for bone image segmentation. This system automates the bone image analysis process and is able to produce reliable results based on quantitative measurements from a large number of bone images. As a result, using large databases of bone images to study the correlation between bone structural features and age-related bone developments becomes feasible.