Three-dimensional reconstruction of Haversian systems in human cortical bone using synchrotron radiation-based micro-CT: morphology and quantification of branching and transverse connections across age.

This study uses synchrotron radiation-based micro-computed tomography (CT) scans to reconstruct three-dimensional networks of Haversian systems in human cortical bone in order to observe and analyse interconnectivity of Haversian systems and the development of total Haversian networks across different ages. A better knowledge of how Haversian systems interact with each other is essential to improve understanding of remodeling mechanisms and bone maintenance; however, previous methodological approaches (e.g. serial sections) did not reveal enough detail to follow the specific morphology of Haversian branching, for example. Accordingly, the aim of the present study was to identify the morphological diversity of branching patterns and transverse connections, and to understand how they change with age. Two types of branching morphologies were identified: lateral branching, resulting in small osteon branches bifurcating off of larger Haversian canals; and dichotomous branching, the formation of two new osteonal branches from one. The reconstructions in this study also suggest that Haversian systems frequently target previously existing systems as a path for their course, resulting in a cross-sectional morphology frequently referred to as 'type II osteons'. Transverse connections were diverse in their course from linear to oblique to curvy. Quantitative assessment of age-related trends indicates that while in younger human individuals transverse connections were most common, in older individuals more evidence of connections resulting from Haversian systems growing inside previously existing systems was found. Despite these changes in morphological characteristics, a relatively constant degree of overall interconnectivity is maintained throughout life. Altogether, the present study reveals important details about Haversian systems and their relation to each other that can be used towards a better understanding of cortical bone remodeling as well as a more accurate interpretation of morphological variants of osteons in cross-sectional microscopy. Permitting visibility of reversal lines, synchrotron radiation-based micro-CT is a valuable tool for the reconstruction of Haversian systems, and future analyses have the potential to further improve understanding of various important aspects of bone growth, maintenance and health.

Does 3D orientation account for variation in osteon morphology assessed by 2D histology?

The primary microstructural unit of cortical bone, the secondary osteon or Haversian system, is widely assumed to have a cylindrical shape. It is generally accepted that osteons are roughly circular in cross-section and deviations from circularity have been attributed to deviations from longitudinal orientation. To our knowledge this idealized geometric relationship, which assumes osteons are perfect cylinders, has not been rigorously explored. As such, we sought to explore two research questions: (i) Does the orientation of osteons in 3D explain variation in shapes visualized in 2D? (ii) Can differences in osteon 3D orientation explain previously reported age-related differences observed in their 2D cross-sectional shape (e.g. more circular shape and decreased area with age)? To address these questions we utilized a combination of 2D histology to identify osteon shape and superimposed micro-computed tomography data to assess osteon orientation in 3D based upon the osteonal canal. Shape was assessed by the inverse of Aspect Ratio (On.AspR(-1), based on a fitted ellipse) - which ranged from 0 (infinitely elongated shape) to 1 (perfectly circular). A sample (n = 27) of human female anterior femoral cortical bone samples from across the human lifespan (20-87 years) were included in the analysis, which involved 1418 osteons. The overall mean measure of On.AspR(-1) was 0.703 (1.42 Aspect Ratio). Mean osteon orientation was 79.1° (90° being longitudinal). While we anticipated a positive relation between orientation and On.AspR(-1), we found the opposite - a weak negative correlation (with more oblique 3D osteon alignment, the 2D shape became more circular as reflected by increased On.AspR(-1)). When analysis of covariance was performed with age and orientation as covariates, the negative relation with orientation was replaced by a significant relation with age alone. This relation with age accounted for 41% of the variation of On.AspR(-1). The results revealed that osteons, on average, are not circular in cross-section and that 3D orientation cannot account for deviation from circular shape. Osteons thus are strictly speaking not cylinders, as they tend to have elliptical cross-sections. We observed that osteons did become less elliptical in cross-section with age independent of orientation - suggesting this is a real change in morphology.

Normal variation in cortical osteocyte lacunar parameters in healthy young males.

The most abundant cell in bone, osteocytes form an interconnected system upon which the regulation of healthy bone relies. Although the complete nature of the role of osteocytes has yet to be defined, they are generally accepted to play a part in the sensing of load and the initiation of damage repair. A previous study conducted by our group identified variation of up to 30% in osteocyte lacunar density and morphological parameters between regions of a single cross-section of human femoral shaft; that study, however, was limited to a single individual. The aim of the current study was to determine whether this pattern consistently occurs in healthy young male femora. Anterior, posterior, medial and lateral blocks were prepared from the proximal femoral shaft of seven males and synchrotron radiation micro-CT imaged. Average lacunar densities (± SD) from the anterior, posterior, medial and lateral regions were 23 394 ± 1705, 30 180 ± 4860, 35 946 ± 5990 and 29 678 ± 6081 lacunae per mm(3) of bone tissue, respectively. These values were significantly different between the anterior and both the medial and posterior regions (P < 0.05). The density of the combined anterior and posterior regions was also significantly lower (P = 0.006) than the density of the combined medial and lateral regions. Although no difference was found in predominant orientation, shape differences were found; with the combined anterior-posterior regions having lacunae that were significantly more elongated and less flat than the combined medial-lateral values (P < 0.001). As expected, in this larger study, there was a dramatic difference in lacunar density between the medial and anterior region (up to ~ 54%). The study clearly demonstrates that the high variation seen in osteocyte lacunar density as well as other lacunar parameters, noted in a number of biomechanical, age and pathology studies, are well within the range of normal variation; however, the reasons for and consequences of this variation remain unclear. Lacunar parameters including abundance and shape are being increasingly incorporated into computational modeling of bone biology and this paper represents a more comprehensive description of normal healthy lacunae.

Endocortical bone loss in osteoporosis: the role of bone surface availability.

Age-related bone loss and postmenopausal osteoporosis are due to a dysregulation of bone remodelling in which less bone is reformed than resorbed. This dysregulation of bone remodelling does not occur with equal strength in all bone regions. Loss of bone is more pronounced near the endocortical surface. This leads to thinning of the cortical wall proceeding from the endosteum, a process sometimes called 'trabecularisation'. In this paper, we investigate the influence of the nonuniform distribution of bone surface within bone tissue for osteoporotic bone losses. We use a spatio-temporal computational model of bone remodelling in which microstructural changes of bone tissue are represented by a phenomenological relationship between bone specific surface and bone porosity. The simulation of an osteoporotic condition by our model shows that the evolution of bone porosity within a bone cross section is significantly influenced by the nonuniform availability of bone surface. Greater bone loss occurs near the endocortical wall, leading to cortical wall thinning and to an expansion of the medullary cavity similar to cross-sectional observations from human femur midshafts. Our model suggests that the rate of cortical wall thinning is fast/slow in the presence/absence of an adjacent trabecular or trabecularised bone compartment.

Femoral osteocyte lacunar density, volume and morphology in women across the lifespan.

Osteocytes are believed to be the primary agents of mechanosensing in bone. Due to this important role in the structure-function relationship of bone, osteocytes and the spaces they occupy (lacunae) are of increasing interest. Changes in lacunae with age are of particular interest in women since they are more susceptible to bone loss and fragility associated with senescent diseases including osteoporosis. This study's purpose was to test whether differences exist in lacunar density (lacunae/mm(3) of bone), orientation and morphology in the cortex of adult women spanning the human lifespan. Anterior blocks from the femoral shaft from 30 women aged 20-86years were imaged by synchrotron-radiation micro-CT. No significant relation between lacunar density and age was detected. A significant reduction in lacunar volume with age (p<0.001) was observed, alongside changes in lacunar morphology. When divided into two groups (<50 and >50years) the younger group's lacunae were ∼30% larger and were flatter (p<0.001) and less equant (spherical) (p<0.001). To our knowledge the observation that lacunar volume and morphology change over the human lifespan is novel, potentially resulting from preferential surface infilling within the extracellular space. The functional impact of this infilling is unclear but such a change in scale likely impacts the mechanosensing function of the osteocyte network. Limitations in resolution prevented us from assessing if this infilling is associated with disruption of the canaliculi. This hypothesis warrants further investigation as, if confirmed, it would represent a profound negative impact on the osteocyte network and may provide new insights into age-related bone loss.

Variation in osteocyte lacunar morphology and density in the human femur--a synchrotron radiation micro-CT study.

In recent years there has been growing interest in the spatial properties of osteocytes (including density and morphology) and how these potentially relate to adaptation, disease and aging. This interest has, in part, arisen from the availability of increasingly high-resolution 3D imaging modalities such as synchrotron radiation (SR) micro-CT. As resolution increases, field of view generally decreases. Thus, while increasingly detailed spatial information is obtained, it is unclear how representative this information is of the skeleton or even the isolated bone. The purpose of this research was to describe the variation in osteocyte lacunar density, morphology and orientation within the femur from a healthy young male human. Multiple anterior, posterior, medial and lateral blocks (2 mm × 2 mm) were prepared from the proximal femoral shaft and SR micro-CT imaged at the Advanced Photon Source. Average lacunar densities (± standard deviation) from the anterior, posterior, medial and lateral regions were 27,169 ± 1935, 26,3643 ± 1262, 37,521 ± 6416 and 33,972 ± 2513 lacunae per mm(3) of bone tissue, respectively. These values were significantly different between the medial and both the anterior and posterior regions (p<0.05). The density of the combined anterior and posterior regions was also significantly lower (p=0.001) than the density of the combined medial and lateral regions. Although no difference was found in predominant orientation, shape differences were found; with the combined anterior and posterior regions having more elongated (p=0.004) and flattened (p=0.045) lacunae, than those of the medial and lateral regions. This study reveals variation in osteocyte lacunar density and morphology within the cross-section of a single bone and that this variation can be considerable (up to 30% difference in density between regions). The underlying functional significance of the observed variation in lacunar density likely relates to localized variations in loading conditions as the pattern corresponds well with mechanical axes. Lower density and more elongate shapes being associated with the antero-posterior oriented neutral axis. Our findings demonstrate that the functional and pathological interpretations that are increasingly being drawn from high resolution imaging of osteocyte lacunae need to be better situated within the broader context of normal variation, including that which occurs even within a single skeletal element.

Trabecular Plate Loss and Deteriorating Elastic Modulus of Femoral Trabecular Bone in Intertrochanteric Hip Fractures.

Osteoporotic hip fracture is associated with significant trabecular bone loss, which is typically characterized as low bone density by dual-energy X-ray absorptiometry (DXA) and altered microstructure by micro-computed tomography (µCT). Emerging morphological analysis techniques, e.g. individual trabecula segmentation (ITS), can provide additional insights into changes in plate-like and rod-like trabeculae, two major microstructural types serving different roles in determining bone strength. Using ITS, we evaluated trabecular microstructure of intertrochanteric bone cores obtained from 23 patients undergoing hip replacement surgery for intertrochanteric fracture and 22 cadaveric controls. Micro-finite element (µFE) analyses were performed to further understand how the abnormalities seen by ITS might translate into effects on bone strength. ITS analyses revealed that, near fracture site, plate-like trabeculae were seriously depleted in fracture patients, but trabecular rod volume was maintained. Besides, decreased plate area and rod length were observed in fracture patients. Fracture patients also showed decreased elastic moduli and shear moduli of trabecular bone. These results provided evidence that in intertrochanteric hip fracture, preferential loss of plate-like trabeculae led to more rod-like microstructure and deteriorated mechanical competence adjacent to the fracture site, which increased our understanding of the biomechanical pathogenesis of hip fracture in osteoporosis.

Femoral neck trabecular bone: loss with aging and role in preventing fracture.

Hip fracture risk rises 100- to 1000-fold over six decades of age, but only a minor part of this increase is explained by declining BMD. A potentially independent cause of fragility is cortical thinning predisposing to local crushing, in which bone tissue's material disintegrates at the microscopic level when compressed beyond its capacity to maintain integrity. Elastic instability or buckling of a much thinned cortex might alternatively occur under compression. In a buckle, the cortex moves approximately at right angles to the direction of load, thereby distorting its microstructure, eventually to the point of disintegration. By resisting buckling movement, trabecular buttressing would protect the femoral neck cortex against this type of failure but not against crushing. We quantified the effect of aging on trabecular BMD in the femoral neck and assessed its contribution to cortical elastic stability, which determines resistance to buckling. Using CT, we measured ex vivo the distribution of bone in the midfemoral necks of 35 female and 33 male proximal femurs from cases of sudden death in those 20-95 yr of age. We calculated the critical stress sigma(cr), at which the cortex was predicted to buckle locally, from the geometric properties and density of the cortical zone most highly loaded in a sideways fall. Using long-established engineering principles, we estimated the amount by which stability or buckling resistance was increased by the trabecular bone supporting the most stressed cortical sector in each femoral neck. We repeated these measurements and calculations in an age- and sex-matched series of femoral necks donated by women who had suffered intracapsular hip fracture and controls, using histological measurements of cortical thickness to improve accuracy. With normal aging, trabecular BMD declined asymmetrically, fastest in the supero-lateral one-half (in antero-posterior projection) of the trabecular compartment. When viewed axially with respect to the femoral neck, the most rapid loss of trabecular bone occurred in the posterior part of this region (supero-posterior [S-P]), amounting to a 42% reduction in women (34% in men) over five decades of adult age. Because local cortical bone thickness declined comparably, age had no significant effect on the relative contributions of cortical and trabecular bone to elastic stability, and trabecular bone was calculated to contribute 40% (in men) and 43% (in women) to the S-P cortex of its overall elastic stability. Hip fracture cases had reduced elastic stability compared with age-matched controls, with a median reduction of 49% or 37%, depending on whether thickness was measured histologically or by CT (pQCT; p < 0.002 for both). This effect was because of reduced cortical thickness and density. Trabecular BMD was similar in hip fracture cases and controls. The capacity of the femur to resist fracture in a sideways fall becomes compromised with normal aging because cortical thickness and trabecular BMD in the most compressed part of the femoral neck both decline substantially. This decline is relatively more rapid than that of femoral neck areal BMD. If elastic instability rather than cortical crushing initiates the fracture event, interventions that increase trabecular bone in the proximal femur have great potential to reduce fracture risk because the gradient defining the increase in elastic stability with increasing trabecular BMD is steep, and most hip fracture cases have sufficient trabecular bone for anabolic therapies to build on.

The relation of femoral osteon geometry to age, sex, height and weight.

As computational modeling becomes an increasingly common tool for probing the regulation of bone remodeling, the need for experimental data to refine and validate such models also grows. For example, van Oers et al. (R.F. van Oers, R. Ruimerman, B. van Rietbergen, P.A. Hilbers, R. Huiskes, Relating osteon diameter to strain. Bone 2008;43: 476-482.) recently described a mechanism by which osteon size may be regulated (inversely) by strain. Empirical data supporting this relation, particularly in humans, are sparse. Therefore, we sought to determine if there is a link between body weight (the only measure related to loading available for a cadaveric population) and osteon geometry in human bone. We hypothesized that after controlling for age, sex and height, weight would be inversely related to femoral osteon size (area, On.Ar; diameter, On.Dm). Secondarily we sought to describe the relation between osteon circularity (On.Cr) and these parameters. Osteons (n=12,690) were mapped within microradiographs of femoral mid-diaphyseal specimens (n=88; 45 male, 43 female; 17-97 yrs). Univariate analysis of covariance was conducted (n=87; 1 outlier) with sex as a fixed factor and height, weight and log-transformed age as covariates. Weight was negatively related to On.Ar and On.Dm (p=0.006 and p=0.004, respectively). Age was significantly related to osteon and, it was also significantly related to circularity (all p<0.001). This relation was negative for On.Ar and On.Dm and positive for On.Cr (increasing circularity with age). On.Ar and On.Dm were found to be significantly different between the sexes (p=0.021 and p=0.019, respectively), with females having smaller osteons. No relation between sex and On.Cr was detected (p=0.449). Height was not significantly related to any of the geometric parameters. Partial eta-squared values revealed that age accounted for the largest proportion (On.Ar: 28%, On.Dm: 18%, On.Cr: 30%), weight accounted for the second largest (On.Ar: 9%, On.Dm: 10%) and sex accounted for the smallest proportion (On.Ar: 6%, On.Dm: 7%) of the variance in geometry. While previous studies have reported relations between osteon size and sex/age, we believe that our findings are the first to demonstrate a link with weight. We believe that this negative relation with weight is most probably mechanical in nature; however, alternative (endocrine) links between bone and adipose tissue cannot be ruled out by our design.

Collagen turnover in the adult femoral mid-shaft: modeled from anthropogenic radiocarbon tracer measurements.

We have measured the (14)C content of human femoral mid-shaft collagen to determine the dynamics of adult collagen turnover, using the sudden doubling and subsequent slow relaxation of global atmospheric (14)C content due to nuclear bomb testing in the 1960s and 1970s as a tracer. (14)C measurements were made on bone collagen from 67 individuals of both sexes who died in Australia in 1990-1993, spanning a range of ages at death from 40 to 97, and these measurements were compared with values predicted by an age-dependent turnover model. We found that the dataset could constrain models of collagen turnover, with the following outcomes: 1) Collagen turnover rate of females decreases, on average, from 4%/yr to 3%/yr from 20 to 80 years. Male collagen turnover rates average 1.5-3%/yr over the same period. 2) For both sexes the collagen turnover rate during adolescent growth is much higher (5-15%/yr at age 10-15 years), with males having a significantly higher turnover rate than have females, by up to a factor of 2. 3) Much of the variation in residual bomb (14)C in a person's bone can be attributed to individual variation in turnover rate, but of no more than about 30% of the average values for adults. 4) Human femoral bone collagen isotopically reflects an individual's diet over a much longer period of time than 10 years, including a substantial portion of collagen synthesised during adolescence.

Age-dependent change in the 3D structure of cortical porosity at the human femoral midshaft.

Microstructural change associated with cortical bone remodeling has been extensively explored with 2D techniques. However, relatively little is known regarding the 3D dynamic microstructure of cortical bone. Therefore, we employed micro-CT imaging to investigate 3D remodeling-related change in the structure of cortical bone porosity across the human lifespan. Anterior femoral midshaft specimens (n=51 male, 28 female) spanning 18 to 92 years of age were scanned with 7 mum nominal isotropic resolution. Canal volume fraction (Ca.V/TV), mean diameter (Ca.Dm), mean separation (Ca.Sp), degree of anisotropy (DA), connectivity density (Ca.ConnD), and number (Ca.N) were calculated for subperiosteal cylindrical regions of interest. Ca.N was calculated in 2D (Ca.N(2D)) and 3D (Ca.N(3D)). Regression was used to examine the relation between the structural parameters and age. Additionally, the impact of sex, height, and weight were investigated collectively (MANCOVA) and individually (ANCOVA). For all analyses, Ca.V/TV and Ca.Dm were inverted (Ca.V/TV(-1), Ca.Dm(-1)) to establish normality and linear relations with age. Ca.N values (2D and 3D) were non-linearly (quadratic) related to age, increasing until the 6th decade then decreasing. This relation was only significant for the pooled sexes Ca.N(3D) values (p=0.012). Ca.ConnD was positively related to age (p<0.05), while all remaining 3D parameters, except DA for males (p=0.070), were negatively related (p<0.05). In all cases, the relation with age was strongest for females. MANCOVA revealed that age was the only significant (p<0.001) covariate overall. Univariate ANCOVA indicated significant differences between the sexes for Ca.V/TV(-1) and Ca.Dm(-1) (p=0.018 and 0.010, respectively). Relative to males, females had lower values for these parameters, translating into larger mean canal diameter and overall porosity. Body weight had a significant (p=0.043) positive relation with Ca.Dm(-1), indicating lower weight was also associated with increased mean canal diameter. Therefore, while age was the most important factor, sex and body size were found to play a role in parameters related to canal size and the overall level of porosity. This study is unique in that changes in cortical bone microstructure were examined across the adult human lifespan in three rather than two dimensions.

Increase in pore area, and not pore density, is the main determinant in the development of porosity in human cortical bone.

This study investigated the relative contributions of pore size and pore density (number of pores per mm2) to porosity in the midshaft of the human femur. Cross-sections were obtained from 168 individuals from a modern Australian population (mostly Anglo-Celtic). The study group comprised 73 females and 95 males, aged from 20 to 97 years. Microradiographs were made of 100-microm sections and porosity, pore areas and pore densities determined using image processing software. The cortex was divided into three rings radially and into octants circumferentially, and the porosity, pore area and pore density of each segment were calculated. Results show that 81% of the variance in porosity can be explained by changes in mean pore area with only a further 12-16% explained by changes in pore density. These effects were found to be constant across all areas of the cortex and in both sexes. These results are significant in their consistency and ordered gradation and indicate a well-regulated and systematic process of bone removal with ageing. The results show a regular progression from less porous to more porous bone; this is a uniform process that occurs in all individuals, and factors such as sex and rate of ageing determine where on this continuum any individual is at a particular time.

Three-dimensional microcomputed tomography imaging of basic multicellular unit-related resorption spaces in human cortical bone.

This study employed microcomputed tomography (micro-CT) as a novel means for visualizing the morphology and quantifying the range (length) of basic multicellular unit (BMU)-related resorption spaces in human cortical bone. We tested the hypotheses that the density and range of spaces vary with age and sex. The sample included 82 human (18-92 years) anterior femoral midshaft samples. The morphology of the spaces (n = 99) was varied, including unidirectional, bidirectional, branched, and even highly clustered forms. The density of resorption spaces was negatively correlated with age for the combined sexes and females, with Spearman's rho values of -0.355 (P < 0.001) and -0.522 (P = 0.002), respectively. The density of spaces did not differ significantly between the sexes (P = 0.735). Mean range +/- SD for the combined sexes, females, and males was 2,706 +/- 1,177, 2,681 +/- 1,247, and 2,718 +/- 1,150 microm, respectively. Numerical simulation of the effect of the 7,000 microm scan field of view suggested that the actual mean range of the spaces for the pooled sample was actually on the order of 3,770 microm. Range did not correlate significantly with age for the combined sexes (P = 0.587) or females (P = 0.345) and males (P = 0.896) considered separately and was not significantly different (P = 0.883) between the sexes. These results suggest that the range of BMUs is not affected by age. The age-dependent decrease in resorption space density for the females and pooled sexes was most likely a consequence of cortical rarefaction, leading to difficulty detecting resorption spaces with micro-CT, rather than a decrease in overall remodeling activity.

Effects of tensioning the lumbar fasciae on segmental stiffness during flexion and extension: Young Investigator Award winner.

STUDY DESIGN: Biomechanical study of unembalmed human lumbar segments. OBJECTIVE: To investigate the effects of tensioning the lumbar fasciae (transversus abdominis [TrA]) aponeurosis) on segment stiffness during flexion and extension. SUMMARY OF BACKGROUND DATA: Animal and human studies suggest that TrA may influence intersegmental movement via tension in the middle and posterior layers of lumbar fasciae (MLF, PLF). METHODS: Compressive flexion and extension moments were applied to 17 lumbar segments from 9 unembalmed cadavers with 20 N lateral tension of the TrA aponeurosis during: 1) "static" tests: load was compared when fascial tension was applied during static compressive loads into flexion-extension; 2) "cyclic loading" tests: load, axial displacement, and stiffness were compared during repeated compressive loading cycles into flexion-extension. After testing, the PLF was incised to determine the tension transmitted by each layer. RESULTS: At all segments and loads (<200 N), fascial tension increased resistance to flexion loads by approximately 9.5 N. In 15 of 17, fascial tension decreased resistance to extension by approximately 6.6 N. Fascial tension during cyclic flexion loading decreased axial displacement by 26% at the onset of loading (0-2 N) and 2% at 450 N (13 of 17). During extension loading, fascial tension increased displacement at the onset of loading (10 of 17) by approximately 23% and slightly (1%) decreased displacement at 450 N. Segment stiffness was increased by 6 N/mm in flexion (44% at 25 N) and decreased by 2 N/mm (8% at 25 N) in extension. More than 85% of tension was transmitted through the MLF. CONCLUSIONS: Tension on the lumbar fasciae simulating moderate contraction of TrA affects segmental stiffness, particularly toward the neutral zone.

Regional variation of intracortical porosity in the midshaft of the human femur: age and sex differences.

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.

Delineation of facial archetypes by 3d averaging.

The objective of this study was to investigate the feasibility of creating archetypal 3D faces through computerized 3D facial averaging. A 3D surface scanner Fiore and its software were used to acquire the 3D scans of the faces while 3D Rugle3 and locally-developed software generated the holistic facial averages. 3D facial averages were created from two ethnic groups; European and Japanese and from children with three previous genetic disorders; Williams syndrome, achondroplasia and Sotos syndrome as well as the normal control group. The method included averaging the corresponding depth (z) coordinates of the 3D facial scans. Compared with other face averaging techniques there was not any warping or filling in the spaces by interpolation; however, this facial average lacked colour information. The results showed that as few as 14 faces were sufficient to create an archetypal facial average. In turn this would make it practical to use face averaging as an identification tool in cases where it would be difficult to recruit a larger number of participants. In generating the average, correcting for size differences among faces was shown to adjust the average outlines of the facial features. It is assumed that 3D facial averaging would help in the identification of the ethnic status of persons whose identity may not be known with certainty. In clinical medicine, it would have a great potential for the diagnosis of syndromes with distinctive facial features. The system would also assist in the education of clinicians in the recognition and identification of such syndromes.

A Japanese computer-assisted facial identification system successfully identifies non-Japanese faces.

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.

Assessing growth and development of the facial profile.

The study of growth and development of the facial profile is of interest to clinicians and researchers in the fields of pediatric dentistry, orthodontics, and craniofacial surgery, enabling diagnosis, planning, and evaluation of treatment. Until recently, craniofacial studies addressed facial growth, facial asymmetry, and gender differences by examining changes in size. However, size changes alone do not represent fully the complicated process of craniofacial growth which also involves changes in shape. The shape of the facial profile can now be quantified with Fourier analysis, contributing to a better understanding of growth. A combination of recently developed methods, such as 3-dimensional facial morphometry and Fourier analysis, should allow a more comprehensive knowledge of growth and development of the craniofacial structures, including the facial profile. This article examines various methods for assessing facial growth and development currently available with particular reference to the facial profile, and addresses the value of Fourier analysis in assessing shape changes.

Lateral facial soft-tissue prediction model: analysis using Fourier shape descriptors and traditional cephalometric methods.

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.

Preferred collagen fiber orientation in the human mid-shaft femur.

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.