Ontogenetic Patterning of Human Subchondral Bone Microarchitecture in the Proximal Tibia.

High-resolution computed tomography images were acquired for 31 proximal human tibiae, age 8 to 37.5 years, from Norris Farms #36 cemetery site (A.D. 1300). Morphometric analysis of subchondral cortical and trabecular bone architecture was performed between and within the tibial condyles. Kruskal−Wallis and Wilcoxon signed-rank tests were used to examine the association between region, age, body mass, and each morphometric parameter. The findings indicate that age-related changes in mechanical loading have varied effects on subchondral bone morphology. With age, trabecular microstructure increased in bone volume fraction (p = 0.033) and degree of anisotropy (p = 0.012), and decreased in connectivity density (p = 0.001). In the subchondral cortical plate, there was an increase in thickness (p < 0.001). When comparing condylar regions, only degree of anisotropy differed (p = 0.004) between the medial and lateral condyles. Trabeculae in the medial condyle were more anisotropic than in the lateral region. This research represents an innovative approach to quantifying both cortical and trabecular subchondral bone microarchitecture in archaeological remains.

Body size and social status in medieval Alba (Cuneo), Italy.

OBJECTIVES: Previous work by Vercellotti et al. in 2011 found significant status-related differences in body size in males but not in females from the Italian bioarchaeological assemblage of San Michele di Trino (8th-14th centuries CE). The purpose of the present work is twofold: (a) to determine if status-related body size differences could be observed in the nearby collection of San Lorenzo di Alba (7th-15th centuries CE) and (b) to add to the emerging narrative of medieval Italians. MATERIALS AND METHODS: Osteometric data (maximum length for the humerus, radius, ulna, femur, tibia, and fibula; bicondylar length of the femur, condylo-malleolar length of the tibia, foot height, maximum vertebral heights, and basion-bregma height) were collected for 50 (20 female, 30 male) individuals from Alba, and Monte Carlo analysis was used to assess differences in skeletal element size, skeletal height, living stature, and body mass across sex and status. RESULTS: Significant differences were detected between high status and low status males in Alba for radial maximum length (p = 0.013), tibial maximum length (p = 0.011), tibial condylo-malleolar length (p = 0.012), skeletal height estimated from condylo-malleolar tibial length (p = 0.002), and stature estimated from condyle-malleolar tibial length with the age component (p = 0.003). In contrast, no significant status-based differences were observed between female subsamples (p > 0.05). DISCUSSION: The patterns of intrapopulation variation observed at Alba are similar but not as pronounced as those observed at Trino, suggesting that overall life conditions experienced by the two groups were comparable.

Variability of in vivo linear microcrack accumulation in the cortex of elderly human ribs.

Excessive accumulation of microdamage in the skeleton in vivo is believed to contribute to fragility and risk of fracture, particularly in the elderly. Current knowledge of how much in vivo damage accrual varies between individuals, if at all, is lacking. In this study, paired sixth ribs from five male and five female elderly individuals (76-92 years, mean age = 84.7 years) were examined using en bloc staining and fluorescent microcopy to quantify linear microcracks present at the time of death (i.e. in vivo microdamage). Crack number, crack length, crack density, and crack surface density were measured for each complete cross-section, with densities calculated using the variable of bone area (which accounts for the influence of porosity on the cortex, unlike the more frequently used cortical area), and analyzed using a two-way mixed model analysis of variance. Results indicate that while microcracks between individuals differ significantly, differences between the left and right corresponding pairs within individuals and the pleural and cutaneous cortices within each rib did not. These results suggest that systemic influences, such as differential metabolic activity, affect the accumulation of linear microcracks. Furthermore, variation in remodeling rates between individuals may be a major factor contributing to differential fracture risk in the elderly. Future work should expand to include a wider age range to examine differences in in vivo microdamage accumulation across the lifespan, as well as considering the influence of bisphosphonates on microdamage accumulation in the context of compromised remodeling rates in the elderly.

Variation in osteon histomorphometrics and their impact on age-at-death estimation in older individuals.

Histomorphometric studies have reported relations between osteon size and age; however, data focused on the shape of osteons is sparse. The purpose of this study was to determine how osteon circularity (On.Cr) varies with age in different skeletal elements. Regression analysis was used to evaluate the relationship between age and osteon shape and size. We hypothesized that age would be negatively related to osteon size (area, On.Ar) and positively related to osteon shape (On.Cr). On.Cr and On.Ar were determined for the ribs and femora of 27 cadaveric specimens with known age-at-death. As predicted, age was significantly related to osteon size and shape for both the femur and rib. With age, there was a decrease in size and an increase in circularity. No relationship between sex and On.Cr was detected. An age predicting model, including On.Cr, On.Ar and OPD, is proposed to improve our ability to estimate age-at-death, especially for older individuals.

Methods and theory in bone modeling drift: comparing spatial analyses of primary bone distributions in the human humerus.

This study compares two novel methods quantifying bone shaft tissue distributions, and relates observations on human humeral growth patterns for applications in anthropological and anatomical research. Microstructural variation in compact bone occurs due to developmental and mechanically adaptive circumstances that are 'recorded' by forming bone and are important for interpretations of growth, health, physical activity, adaptation, and identity in the past and present. Those interpretations hinge on a detailed understanding of the modeling process by which bones achieve their diametric shape, diaphyseal curvature, and general position relative to other elements. Bone modeling is a complex aspect of growth, potentially causing the shaft to drift transversely through formation and resorption on opposing cortices. Unfortunately, the specifics of modeling drift are largely unknown for most skeletal elements. Moreover, bone modeling has seen little quantitative methodological development compared with secondary bone processes, such as intracortical remodeling. The techniques proposed here, starburst point-count and 45° cross-polarization hand-drawn histomorphometry, permit the statistical and populational analysis of human primary tissue distributions and provide similar results despite being suitable for different applications. This analysis of a pooled archaeological and modern skeletal sample confirms the importance of extreme asymmetry in bone modeling as a major determinant of microstructural variation in diaphyses. Specifically, humeral drift is posteromedial in the human humerus, accompanied by a significant rotational trend. In general, results encourage the usage of endocortical primary bone distributions as an indicator and summary of bone modeling drift, enabling quantitative analysis by direction and proportion in other elements and populations.

Drifting Diaphyses: Asymmetry in Diametric Growth and Adaptation Along the Humeral and Femoral Length.

This study quantifies regional histomorphological variation along the human humeral and femoral diaphysis in order to gain information on diaphyseal growth and modeling drift patterns. Three thin sections at 40, 50, and 60% bone length were prepared from a modern Mexican skeletal sample with known age and sex to give a longitudinal perspective on the drifting cortex (12 adults and juveniles total, 7 male and 5 female). Point-count techniques were applied across eight cross-sectional regions of interest using the starburst sampling pattern to quantify percent periosteal and endosteal primary lamellar bone at each diaphyseal level. The results of this study show a posterio-medial drift pattern in the humerus with a posterior rotational trend along the diaphysis. In the femur, we observed a consistent lateral to anteriolateral drift and an increase in primary lamellar bone area of both, periosteal and endosteal origin, towards the distal part of the diaphysis. These observations characterize drifting diaphyses in greater detail, raising important questions about how to resolve microscopic and macroscopic cross-sectional analysis towards a more complete understanding of bone growth and mechanical adaptation. Accounting for modeling drift has the potential to positively impact age and physical activity estimation, and explain some of the significant regional variation in bone histomorphology seen within (and between) bone cross-sections due to differing ages of tissue formation. More study is necessary, however, to discern between possible drift scenarios and characterize populational variation.

Intraskeletal Variability of Relative Cortical Area in Humans.

Histomorphometric and cross-sectional geometric studies of bone have provided valuable information about age at death, behavioral and activity patterns, and pathological conditions for past and present human populations. While a considerable amount of exploratory and applied research has been completed using histomorphometric and cross-sectional geometric properties, the effects of intraskeletal variability on interpreting observed histomorphometric data have not been fully explored. The purpose of this study is to quantify intraskeletal variability in the relative cortical area of long bones and ribs from modern humans. To examine intraskeletal variability, cross-sections of the femur, tibia, fibula, humerus, radius, ulna, and rib when present, were examined within individuals from a cadaveric collection (N = 34). Relative cortical area was compared within individuals using a repeated measurements General Linear Model, which shows significant differences between bones, particularly between the rib and the remaining long bones. Complementarily, correlations between bones' relative cortical area values suggest an important allometric component affecting this aspect of long bones, but not of the rib. This study highlights the magnitude of intraskeletal variability in relative cortical area in the human skeleton, and because the relative cortical area of any particular bone is affected by a series of confounding factors, extrapolation of relative cortical area values to infer load history for other skeletal elements can be misleading.

Exploring the multidimensionality of stature variation in the past through comparisons of archaeological and living populations.

Adult stature variation is commonly attributed to differential stress-levels during development. However, due to selective mortality and heterogeneous frailty, a population's tall stature may be more indicative of high selective pressures than of positive life conditions. This article examines stature in a biocultural context and draws parallels between bioarchaeological and living populations to explore the multidimensionality of stature variation in the past. This study investigates: 1) stature differences between archaeological populations exposed to low or high stress (inferred from skeletal indicators); 2) similarities in growth retardation patterns between archaeological and living groups; and 3) the apportionment of variance in growth outcomes at the regional level in archaeological and living populations. Anatomical stature estimates were examined in relation to skeletal stress indicators (cribra orbitalia, porotic hyperostosis, linear enamel hypoplasia) in two medieval bioarchaeological populations. Stature and biocultural information were gathered for comparative living samples from South America. Results indicate 1) significant (P < 0.01) differences in stature between groups exposed to different levels of skeletal stress; 2) greater prevalence of stunting among living groups, with similar patterns in socially stratified archaeological and modern groups; and 3) a degree of regional variance in growth outcomes consistent with that observed for highly selected traits. The relationship between early stress and growth is confounded by several factors-including catch-up growth, cultural buffering, and social inequality. The interpretations of early life conditions based on the relationship between stress and stature should be advanced with caution.

A comparison of histomorphometric data collection methods.

Although many variables that skeletal biologists examine have been standardized, the actual techniques used to collect these data from bone thin sections vary. This project compares different methods of obtaining data (relative cortical area values) for histomorphometric research. One visual and three digital methods of histomorphometric data collection are compared: (i) Merz microscopic eyepiece counting reticule, (ii) flatbed scanner, (iii) overlaying multiple images of a thin section, and (iv) digital SLR camera with macro settings. The discussion includes a comparison of usability factors such as cost, time, user-experience, and ease-of-use, which vary for each method. Values from the different methods are compared using ANOVA tests to evaluate inter-method, inter-observer, and intra-observer variability. Intra-observer error was greater for the microscopic method, although the error values are concomitant with experience. We found no statistically significant differences between the four methods examined, but certain caveats must be addressed when these methods are used.

Brief communication: the effects of disuse on the mechanical properties of bone: what unloading tells us about the adaptive nature of skeletal tissue.

The intricate link between load environment and skeletal health is exemplified by the severe osteopenia that accompanies prolonged periods of immobilization, frequently referred to as disuse osteoporosis. Investigating the effects disuse has on the structural properties of bone provides a unique opportunity to better understand how mechanical loads influence the adaptation and maintenance of skeletal tissue. Here, we report results from an examination of multiple indicators of bone metabolism (e.g., mean osteon density, mean osteon size, bone mass, and bone area distribution) within the major long bones of individuals with distinct activity level differences. Results are based on a sample comprising two subjects that suffered from long-term quadriplegia and 28 individuals of comparable age that had full limb mobility. Although limited in sample size, our findings suggest bones associated with long-term disuse have lower osteon densities and larger osteon areas compared to individuals of normal mobility, reflecting dramatically lower remodeling rates potentially related to reduced strain levels. Moreover, immobilized skeletal elements demonstrate a reduced percentage of cortical area present resulting from endosteal resorption. Differences between mobility groups in the percentage of cortical area present and bone distribution of all skeletal elements, suggests bone modeling activity is negligible in the unloaded adult skeleton. Additional histomorphometric comparisons reveal potential intraskeletal differences in bone turnover rates suggesting remodeling rates are highest within the humeri and femora. Addition of more immobilized individuals in the future will allow for quantitative statistical analyses and greater consideration of human variation within and between individuals.

Rib histomorphometry for adult age estimation.

Estimating the age at death in the adult skeleton is problematic owing to the biological variability in morphological age indicators and the differential response to environmental factors over an individual's life. It is becoming increasingly important for anthropologists to improve age estimates through the use of multiple age indicators and various modalities of assessment (e.g., macroscopic, microscopic, and radiological). Lack of instructional texts describing how to prepare histological samples and evaluate bone under the microscope has been a restricting factor in the widespread use of current histological methods within the field of forensic anthropology. The limited use of histological methods for age estimation often lies in the misunderstanding that the preparation and evaluation of cortical bone thin sections is a highly technical and an expensive endeavor. Like any method of age estimation, the researcher/practitioner must be guided through the analytical process to ensure reliable and repeatable results. This chapter provides a step-by-step instructional guide in the preparation and evaluation of histological samples removed from the sixth rib for histological age estimation.

Technical note: The use of geographical information systems software for the spatial analysis of bone microstructure.

Geographic information systems (GIS) software is typically used for analyzing geographically distributed data, allowing users to annotate points or areas on a map and attach data for spatial analyses. While traditional GIS-based research involves geo-referenced data (points tied to geographic locations), the use of this technology has other constructive applications for physical anthropologists. The use of GIS software for the study of bone histology offers a novel opportunity to analyze the distribution of bone nano- and microstructures, relative to macrostructure and in comparison to other variables of interest, such as biomechanical loading history. This approach allows for the examination of characteristics of single histological features while considering their role at the macroscopic level. Such research has immediate promise in examining the load history of bone by surveying the functional relationship between collagen fiber orientation (CFO) and strain mode. The diversity of GIS applications that may be utilized in bone histology research is just beginning to be explored. The goal of this study is to introduce a reliable methodology for such investigation and our objective is to quantify the heterogeneity of bone microstructure over an entire cross-section of bone using ArcGIS v 9.3 (ESRI). This was accomplished by identifying the distribution of remodeling units in a human metatarsal relative to bending axes. One biomechanical hypothesis suggests that CFO, manifested by patterns of birefringence, is indicative of mode of strain during formation. This study demonstrates that GIS can be used to investigate, describe, and compare such patterns through histological mapping.

Brief communication: Reevaluating osteoporosis in human ribs: the role of intracortical porosity.

Osteoporosis is a major health concern in modern society and is continually being evaluated in past populations by quantifying bone loss. Cortical area measures are commonly used in anthropological analyses to assess bone loss in the ribs, but these values are typically based on endosteal expansion and do not account for intracortical bone loss. The objective of this study is to evaluate the effectiveness of using absolute cortical area, compared to traditional cortical area measures to describe global bone loss in elderly ribs. Transverse sections were prepared from sixth ribs of ten elderly subjects, and bone area measurements were made from 100× magnification composites of each rib for calculation of cortical area (Ct.Ar) and percent cortical area (% C/T). In addition, all areas of intracortical porosity were measured and percent porosity area (% Po.Ar) calculated. Absolute cortical area (Ct.Ar(A)) was calculated by subtracting porosity area from cortical area, and a percent absolute cortical area (% C(A)/T) calculated. ANOVA results reveal significant interindividual variation in percent porosity area (% Po.Ar). Percent cortical area and percent absolute cortical area values were compared and results show a mean difference of 4.08% exists across all subjects, with a range of 1.19-11.73%. This suggests that intracortical porosity is variable and does play a role in age-associated bone loss in the rib. All future investigations of osteoporosis should account for intracortical porosity in bone loss.

Differentiating fragmented human and nonhuman long bone using osteon circularity.

Distinguishing between human and nonhuman bone is important in forensic anthropology and archeology when remains are fragmentary and DNA cannot be obtained. Histological examination of bone is affordable and practical in such situations. This study suggests using osteon circularity to distinguish human bone fragments and hypothesizes that osteons will more closely resemble a perfect circle in nonhumans than in humans. Standard histological methods were used, and circularity was determined using an image analysis program, where circularity was controlled for by Haversian canal measurements. Homogeneity was first tested for multiple variables within human and nonhuman samples. No significant differences were found between human sexes (p = 0.657) or among nonhuman species (p = 0.553). Significant differences were found among intraskeletal elements of both humans (p = 0.016) and nonhumans (p = 0.013) and between pooled samples of humans and nonhumans (p < 0.001). Results of this study indicate that osteon circularity can be used to distinguish between fragmented human and nonhuman long bone.

Intrapopulation variation in stature and body proportions: social status and sex differences in an Italian medieval population (Trino Vercellese, VC).

The phenotypic expression of adult body size and shape results from synergistic interactions between hereditary factors and environmental conditions experienced during growth. Variation in body size and shape occurs even in genetically relatively homogeneous groups, due to different occurrence, duration, and timing of growth insults. Understanding the causes and patterns of intrapopulation variation can foster meaningful information on early life conditions in living and past populations. This study assesses the pattern of biological variation in body size and shape attributable to sex and social status in a medieval Italian population. The sample includes 52 (20 female, 32 male) adult individuals from the medieval population of Trino Vercellese, Italy. Differences in element size and overall body size (skeletal height and body mass) were assessed through Monte Carlo methods, while univariate non-parametric tests and Principal Component Analysis (PCA) were employed to examine segmental and overall body proportions. Discriminant Analysis was employed to determine the predictive value of individual skeletal elements for social status in the population. Our results highlight a distinct pattern in body size and shape variation in relation to status and sex. Male subsamples exhibit significant postcranial variation in body size, while female subsamples express smaller, nonsignificant differences. The analysis of segmental proportions highlighted differences in trunk/lower limb proportions between different status samples, and PCA indicated that in terms of purely morphological variation high status males were distinct from all other groups. The pattern observed likely resulted from a combination of biological factors and cultural practices.

The effects of total hip arthroplasty on the structural and biomechanical properties of adult bone.

The responsiveness of bone to mechanical stimuli changes throughout life, with adaptive potential generally declining after skeletal maturity is reached. This has led some to question the importance of bone functional adaptation in the determination of the structural and material properties of the adult skeleton. A better understanding of age-specific differences in bone response to mechanical loads is essential to interpretations of long bone adaptation. The purpose of this study is to examine how the altered mechanical loading environment and cortical bone loss associated with total hip arthroplasty affects the structural and biomechanical properties of adult bone at the mid-shaft femur. Femoral cross sections from seven individuals who had undergone unilateral total hip arthroplasty were analyzed, with intact, contralateral femora serving as an approximate internal control. A comparative sample of individuals without hip prostheses was also included in the analysis. Results showed a decrease in cortical area in femora with prostheses, primarily through bone loss at the endosteal envelope; however, an increase in total cross-sectional area and maintenance of the parameters of bone strength, I(x), I(y), and J, were observed. No detectable differences were found between femora of individuals without prostheses. We interpret these findings as an adaptive response to increased strains caused by loading a bone previously diminished in mass due to insertion of femoral prosthesis. These results suggest that bone accrued through periosteal apposition may serve as an important means by which adult bone can functional adapt to changes in mechanical loading despite limitations associated with senescence.

Intraskeletal variability in bone mass.

For methodological or other reasons, a variety of skeletal elements are analyzed and subsequently used as a basis for describing general bone loss and mass. However, bone loss and mass may not be uniform within and among skeletal elements of the same individual because of biomechanical factors. We test the hypothesis that a homogeneity in bone mass exists among skeletal elements of the same individual. Measures indicative of bone mass were calculated from the midshafts of six skeletal elements from the same individuals (N = 41). The extent of intraskeletal variability in bone mass (relative cortical area) was then examined for the entire sample, according to age, sex, and pathological status. The results of the analysis showed that all measures reflect a heterogeneity in bone mass (P

Cortical bone remodeling rates in a sample of African American and European American descent groups from the American Midwest: comparisons of age and sex in ribs.

This study employs regression analysis to explore population and sex differences in the pattern of age-associated bone loss, as reflected by histomorphometric variables that are measures of intracortical and endocortical bone remodeling. A comparison of an African American sample from the Washington Park Cemetery in St. Louis, Missouri, and a European American rib sample composed of cadavers, autopsies, and forensic cases from Missouri reveals the existence of complex age-associated patterns for differences in measures of intracortical remodeling and cortical area. Females from the two samples express similar bone dimensions and dynamics. The African American females appear to lose more bone than their male counterparts, but this difference is absent in the European American sample. When age-associated patterns are considered, it is in the younger cohorts that African Americans exhibit greater relative cortical area than European Americans, but this is reversed in the older ages, when the latter group manifests greater bone mass. The European American males consistently differ in the slopes and intercepts for the variables compared to the other groups, and differences are highly significant with African American females, with the former group maintaining bone mass while the latter exhibit a more rapid bone loss. Achieving larger relative cortical area due to smaller endosteal area, coupled with better bone quality due to lower intracortical porosity early in life, may be a mechanism by which African Americans, especially females, maintain adequate bone mass in older ages, which buffers them from bone loss and related fragility fractures despite higher rates of intracortical remodeling and endosteal expansion later in life. These results suggest that both genetic and environmental factors are responsible for the differences in bone remodeling and bone mass observed between these samples.

Population-specific histological age-estimating method: a model for known African-American and European-American skeletal remains.

Previously developed histological age-estimating methods have been based on samples lacking interpopulation variability. A comparison of age-associated rib histomorphometrics between an European-American sample and an African-American sample indicates that ethnicity can have a significant effect on osteon population density (OPD), osteon cross-sectional area (On.Ar), and relative cortical area (Ct.Ar/Tt.Ar). Based upon these findings, new histological age-predicting formulae are presented that are recommended when estimating age for African-American or European-American skeletal remains. A general formula that is applicable to remains of unknown ethnicity is also provided.