Variation in cortical bone histology within the human femur and its impact on estimating age at death

Histological methods for the estimation of age at death using cortical bone are based on the evaluation of microstructural changes over time. Since histological analysis is a destructive method, most techniques attempt to limit the amount of cortical bone needed for analysis. Sample location, however, can have a significant effect on the accuracy of these methods. Furthermore, research demonstrates that both intersection and intrasection variation is present at the midshaft of the femur, which is the primary location for estimating age at death in humans. This research determines the extent of regional variation within the adult human femur and its effect on age estimation. Secondary osteon lamellae and Haversian canal ratio and cortical thickness were quantified. Thompson's All Males Left Femur regression equation was used to estimate age. Results show that significant regional variation occurs in the estimated ages derived from the posterior aspect of the femoral shaft and significant intrasection variation occurs in age estimates from the mid and mid-distal cross-sections. Thus, the inter and intrasection variation that occurs in bone remodeling within the femoral cortex has the potential to produce significant differences amongst age estimates taken from various femoral diaphyseal locations compared to the age estimated from the standard location used in Thompson's core method (1978). The results indicate that the use of this histological method is dependant on the ability to correctly identify the four anatomical locations, but the extracted core used for age estimation is not necessarily confined to the anterior midshaft.     

Cortical thickness measurements and photon absorptiometry for determination of bone quantity.

Reproducibility of estimates of cortical bone quantity in the appendicular skeleton by two methods was studied in healthy individuals and patients undergoing long-term hemodialysis. Repeated measurements of cortical thickness (CT) at the midpoint of the second metacarpal were taken from single radiographs of both hands by two independent observers. Repeated measurements by the same observer were more reproducible and the degree of reproducibility was far greater in healthy subjects than in dialysis patients. Repeated measurements were made of bone mineral content (BMC) and bone width (W) of the distal radius by photon absorptiometry. Repeated BMC/W determinations were highly reproducible in both healthy subjects and dialysis patients. High correlation was found between BMC and cross-sectional cortical area and between both simple cortical thickness and cortical area/width. Thus the photon absorptiometric technique is superior for the serial monitoring of bone quantity, particularly in patients with uremic osteodystrophy, but results obtained by the two methods in group studies should be comparable.     

A meta-analysis of cigarette smoking,bone mineral density and risk of hip fracture: recognition of a major effect.

OBJECTIVE: To determine the magnitude and importance of the relation between smoking, bone mineral density, and risk of hip fracture according to age. DESIGN: Meta-analysis of 29 published cross sectional studies reporting the difference in bone density in 2156 smokers and 9705 non-smokers according to age, and of 19 cohort and case-control studies recording 3889 hip fractures reporting risk in smokers relative to non-smokers. RESULTS: In premenopausal women bone density was similar in smokers and non-smokers. Postmenopausal bone loss was greater in current smokers than non-smokers, bone density diminishing by about an additional 2% for every 10 year increase in age, with a difference of 6% at age 80. In current smokers relative to non-smokers the risk of hip fracture was similar at age 50 but greater thereafter by an estimated 17% at age 60, 41% at 70, 71% at 80, and 108% at 90. These estimates of relative risk by age, derived directly from a regression analysis of the studies of smoking and hip fracture, were close to estimates using the difference in bone density between smokers and non-smokers and the association between bone density and risk of hip fracture. The estimated cumulative risk of hip fracture in women in England was 19% in smokers and 12% in non-smokers to age 85; 37% and 22% to age 90. Among all women, one hip fracture in eight is attributable to smoking. Limited data in men suggest a similar proportionate effect of smoking as in women. The association was not explained by smokers being thinner, younger at menopause, and exercising less nor by actions of smoking on oestrogen, but smoking may have a direct action on bone. CONCLUSIONS: Hip fracture in old age is a major adverse effect of smoking after the menopause. The cumulative excess bone loss over decades is substantial, increasing the lifetime risk of hip fracture by about half.     

In vivo tibial stiffness is maintained by whole bone morphology and cross-sectional geometry in growing female mice

Whole bone morphology, cortical geometry, and tissue material properties modulate skeletal stresses and strains that in turn influence skeletal physiology and remodeling. Understanding how bone stiffness, the relationship between applied load and tissue strain, is regulated by developmental changes in bone structure and tissue material properties is important in implementing biophysical strategies for promoting healthy bone growth and preventing bone loss. The goal of this study was to relate developmental patterns of in vivo whole bone stiffness to whole bone morphology, cross-sectional geometry, and tissue properties using a mouse axial loading model. We measured in vivo tibial stiffness in three age groups (6, 10, 16 wk old) of female C57Bl/6 mice during cyclic tibial compression. Tibial stiffness was then related to cortical geometry, longitudinal bone curvature, and tissue mineral density using microcomputed tomography (microCT). Tibial stiffness and the stresses induced by axial compression were generally maintained from 6 to 16 wks of age. Growth-related increases in cortical cross-sectional geometry and longitudinal bone curvature had counteracting effects on induced bone stresses and, therefore, maintained tibial stiffness similarly with growth. Tissue mineral density increased slightly from 6 to 16 wks of age, and although the effects of this increase on tibial stiffness were not directly measured, its role in the modulation of whole bone stiffness was likely minor over the age range examined. Thus, whole bone morphology, as characterized by longitudinal curvature, along with cortical geometry, plays an important role in modulating bone stiffness during development and should be considered when evaluating and designing in vivo loading studies and biophysical skeletal therapies.     

Machine vision photogrammetry: a technique for measurement of microstructural strain in cortical bone.

Understanding local microstructural deformations and strains in cortical bone may lead to a better understanding of cortical bone damage development, fracture, and remodeling. Traditional experimental techniques for measuring deformation and strain do not allow characterization of these quantities at the microstructural level in cortical bone. This study describes a technique based on digital stereoimaging used to measure the microstructural strain fields in cortical bone. The technique allows the measurement of material surface displacements and strains by comparing images acquired from a specimen at two distinct stress states. The accuracy of the system is investigated by analyzing an undeformed image set; the test image is identical to the reference image but translated by a known pixel amount. An increase in the correlation sub-image train parameter results in an increase in displacement measurement accuracy from 0.049 to 0.012 pixels. Errors in strain calculated from the measured displacement field were between 39 and 564 microstrain depending upon the sub-image train size and applied image displacement. The presence of a microcrack in cortical bone results in local strain at the crack tip reaching 0.030 (30,000 microstrain) and 0.010 (10,000 microstrain) near osteocyte lacunae. It is expected that the use of this technique will allow a greater understanding of bone strength and fracture as well as bone mechanotransduction.     

Decreased body weight in young Osterix-Cre transgenic mice results in delayed cortical bone expansion and accrual

Conditional gene inactivation using the Cre/loxP system has lead to significant advances in our understanding of the function of genes in a wide range of disciplines. It is becoming increasingly apparent in the literature, that Cre transgenic mice may themselves have a phenotype. In the following study we describe the bone phenotype of a commonly used Cre transgenic mouse line to study osteoblasts, the Osx-GFP::Cre (Osx-Cre) mice. Cortical and trabecular bone parameters were determined in the femurs of Osx-Cre mice at 6 and 12?weeks of age by microtomography (μCT). At 6?weeks of age, Osx-Cre mice had reduced body weight by 22% (P?相似文献   

Bone lamina thickness,bone apposition rates,and age estimates in sauropod humeri and femora

Laminar fibrolamellär bone from the mid-shaft region of sauropod humeri and femora was analyzed quantitatively to understand Variation in lamina thickness and apposition rate. The samples were derived from five taxa (Apatosaurus, Barosaurus, an indeterminate gracile diplodocid,Brachiosaurus, Janenschia) and two localities (Tendaguru beds, Tanzania, and Howe-Stephens Quarry, Morrison Formation, Wyoming, USA). Lamina thickness is rather constant at 4–6 laminae/mm (mean 4.8), independent of locality, taxon, bone, and ontogenetic age, suggesting structural constraints as a cause. Bone apposition rate was calculated in bones with annual growth lines (polish lines) by two methods to serve in age estimates of individuals without growth lines and unsampled individuals. Apposition rates are rather variable (1–20 (jm/day), resulting in a wide bracket for the ages estimated. Our age estimates are compatible with recent estimates derived by other methods, however.     

Estimation of age at death from second metacarpals.

This study examined the estimation of age at death from the second metacarpal in 227 individuals aged 30-98 years. Variables ascertained from each bone were: cortical thickness and microdensitometric cortical bone density measured on radiographs of the bone and total osteon count and density recorded on microradiographs of the complete cross section at its midshaft. Based on the latter two variables, two age groups were formed; a middle age group representing those individuals aged 30-65 years, and an older group aged 65+. Stepwise regression analysis of the four variables produced a series of regression equations for age estimation for the middle, old and combined age groups for each sex and sexes combined. Sex-specific equations provided better results than nonspecific ones, especially in females. Total osteon density and combined cortical thickness were found to be the most useful estimators in the middle and the old age group, respectively. The standard error of estimate was 6.71 and 6.90 years in each age group for the sexes combined. In the combined age group, age could be estimated accurately from total osteon count, cortical thickness and MD cortical bone density with the standard error of estimate of 11.10 years. The relative error of estimate ranged within +/- 30% in almost all individuals aged above 60 years.     

Roentgenographic and direct measurement of femoral cortical involution in a prehistoric Mississippian population

Recent roentgenographic studies suggest that periosteal apposition adds femoral cortical bone faster than it is lost by endosteal involution, so that cortical bone area increases with age. A comparison was made of direct and roentgenographic measurements of femoral cortical bone using femora from 23 females and 20 males of a prehistoric Mississippian population dated at A.D. 1540–1700. The comparison revealed that significant errors existed in the technique of radiographic measurement (p<0.000001), in estimating male and female cortical thickness (p<0.000001), and the changes in thickness with age (p<0.000001). There was no significant interaction of the variables: age, sex, and method. The errors arose partly from the inability of the roentgenograph to reveal clearly endosteal porosity, and partly in the assumption that the medial and lateral cortical wall thickness would be representative of the mean cortical thickness. The reduction in cortical thickness (11.3% males and 29.3% females) with age as measured directly is comparable with loss in modern populations.     

Estimation of stature from the skeletal reconstruction of an immature Neandertal from Dederiyeh cave, Syria

Skeletal reconstruction of a child Neandertal unearthed at Dederiyeh Cave, Syria in 1993, is undertaken and the acquired stature discussed. Although the skeletal remains were well preserved, the reconstruction required several assumptions to be made because of the immature status of the specimen. The assumptions were mainly concerned with distances between bones in the inter-vertebral spaces and in the joints of the hip, knee, and ankle. These were estimated from X-ray films of modern children and data from previous studies. Stature was directly measured on the reconstruction, and found to be 79.2 cm. After corrections for soft tissue thickness and shrinkage of the casts, the stature became 81.7 cm. This estimate is consistent with estimates based on regression equations of long bone lengths, especially from those of the lower extremity. In comparison with longitudinal data for white American boys, the assessment of stature for Dederiyeh varied according to the estimated age. For a younger estimated age, the stature falls in the lower half of the white American range of variation, but with an older estimated age, it falls below the lower limit of the range of variation. Other immature Neandertals including two European specimens, Roc de Marsal and La Ferrassie 6, fall below the lower limit of the 5th to 95th percentile range based on the estimated statures from their long bone lengths. More comprehensive age assessment covering both fossil and modern humans is required before accurate conclusions in relation to Neandertal growth can be drawn.     

The role of estrogen receptor-beta, in the early age-related bone gain and later age-related bone loss in female mice

The molecular and cellular mechanism of estrogen action in skeletal tissue remains unclear. The purpose of this study was to understand the role of estrogen receptor-beta, (ERbeta) on cortical and cancellous bone during growth and aging by comparing the bone phenotype of 6- and 13-month-old female mice with or without ERbeta. Groups of 11-14 wild-type (WT) controls and ERbeta knockout (BERKO) female mice were necropsied at 6 and 13 months of age. At both ages, BERKO mice did not differ significantly from WT controls in uterine weight and uterine epithelial thickness, indicating that ERbeta does not regulate the growth of uterine tissue. Femoral length increased significantly by 5.5% at 6 months of age in BERKO mice compared with WT controls. At 6 months of age, peripheral quantitative computerized tomography (pQCT) analysis of the distal femoral metaphysis (DFM) and femoral shafts showed that BERKO mice had significantly higher cortical bone content and periosteal circumference as compared with WT controls at both sites. In contrast to the findings in cortical bone, at 6 months of age, there was no difference between BERKO and WT mice in trabecular density, trabecular bone volume (TBV), or formation and resorption indices at the DFM. In 13-month-old WT mice, TBV (-41%), trabecular density (-27%) and cortical thickness decreased significantly. while marrow cavity and endocortical circumference increased significantly compared with 6-month-old WT mice. These age-related decreases in cancellous and endocortical bone did not occur in BERKO mice. At 13 months of age, BERKO mice had significantly higher total, trabecular and cortical bone, while having significantly lower bone resorption, bone formation and bone turnover in DFM compared with WT mice. These results indicate that deleting ERbeta protected against age-related bone loss in both the cancellous and endocortical compartments by decreasing bone resorption and bone turnover in aged female mice. These data demonstrate that in female mice, ERbeta plays a role in inhibiting periosteal bone formation, longitudinal and radial bone growth during the growth period, while it plays a role in stimulating bone resorption, bone turnover and bone loss on cancellous and endocortical bone surfaces during the aging process.     

Quantitative histology of changes with age in rat bone cortex

Changes with age in bone cortex of the rat were investigated by establishing histological parameters which could be quantitated to estimate age at death. Decalcified cross sections of mandible, femur and tibia were prepared from rats two to 120 days old, and measurements were made of: (1) number of osteons, (2) average number of lamellae per osteon, (3) average Haversian canal diameter, and (4) number of non-Haversian (primary) canals. Multiple regression techniques were used to estimate age at death from several combinations of these variables. With age, the number of osteons per unit area of bone and the number of lamellae per osteon increased, but Haversian canal diameter and the number of primary canals decreased. Multiple regression analyses indicated that age at death could be estimated to ± 3 days of the true value in 95% of the cases. Nomographs based on histological measurements of each bone were prepared which can provide accurate estimates of age between two and 120 days in the Sprague-Dawley female rat. It was concluded that microstructure of bone cortex can not only be quantitated to provide accurate estimates of age but it may also constitute a sensitive measure of the metabolic state of the organism. The techniques utilized should prove useful in anthropology as well in studies of bone aging.     

Nanoindentation and whole-bone bending estimates of material properties in bones from the senescence accelerated mouse SAMP6

The senescence accelerated mouse, strain P6 (SAMP6) has been described as a model of senile osteoporosis. Recent results from whole-bone bending tests indicate that, despite having increased moments of inertia, SAMP6 long bones are weak and brittle compared to SAMR1 controls. In the current study we determined material properties of cortical bone from SAMP6 and SAMR1 femora and tibiae by two methods-nanoindentation and whole-bone bending tests combined with simple beam theory. We hypothesized that: (1) SAMP6 mice have reduced cortical bone material properties compared to SAMR1 controls; and (2) modulus estimated from whole-bone bending tests correlates well with modulus determined by nanoindentation. Results from nanoindentation indicated that modulus and hardness are approximately 10% higher in SAMP6 mice compared to SAMR1 controls (p<0.001), a finding consistent with slightly higher mineralization in SAMP6 bones. Despite their superior elastic and hardness properties, the bending failure properties of SAMP6 bones were markedly inferior--ultimate stress and toughness were reduced by 40% and 60%, respectively (p<0.001). Comparisons between the two testing methods for determining modulus showed poor agreement. Modulus estimated from whole-bone bending tests was not correlated with modulus determined by nanoindentation (p=0.054; r2=0.03) and the absolute values differed by a factor of five between the two methods (bending [wet], 6GPa; nanoindentation [dry], 31GPa). Moreover, relative differences between groups were inconsistent between the two methods. We conclude: (1) cortical bone from the SAMP6 mouse has increased modulus and hardness but poor material strength and toughness, which underscores the relevance of the SAMP6 mouse for studies of skeletal fragility, and (2) values of elastic modulus of bone tissue estimated using simple beam theory and bending tests of mouse femora and tibiae are inaccurate and should be interpreted with caution.     

Multi-scale characterization of swine femoral cortical bone

Multi-scale experimental work was carried out to characterize cortical bone as a heterogeneous material with hierarchical structure, which spans from nanoscale (mineralized collagen fibril), sub-microscale (single lamella), microscale (lamellar structures), to mesoscale (cortical bone) levels. Sections from femoral cortical bone from 6, 12, and 42 months old swine were studied to quantify the age-related changes in bone structure, chemical composition, and mechanical properties. The structural changes with age from sub-microscale to mesoscale levels were investigated with scanning electron microscopy and micro-computed tomography. The chemical compositions at mesoscale were studied by ash content method and dual energy X-ray absorptiometry, and at microscale by Fourier transform infrared microspectroscopy. The mechanical properties at mesoscale were measured by tensile testing, and elastic modulus and hardness at sub-microscale were obtained using nanoindentation. The experimental results showed age-related changes in the structure and chemical composition of cortical bone. Lamellar bone was a prevalent structure in 6 months and 12 months old animals, resorption sites were most pronounced in 6 months old animals, while secondary osteons were the dominant features in 42 months old animals. Mineral content and mineral-to-organic ratio increased with age. The structural and chemical changes with age corresponded to an increase in local elastic modulus, and overall elastic modulus and ultimate tensile strength as bone matured.     

Effects of age and occupation on cortical bone in a group of 18th-19th century British men

The effects of age and occupation on cortical bone in a group of adult males from the 18th-19th century AD skeletal collection from Christ Church Spitalfields, London, were investigated. Cortical bone was monitored using metacarpal radiogrammetry. Individual age at death was known exactly from coffin plates. Occupation for individuals was known from historical sources. Results showed that continued periosteal apposition was evident throughout adult life, but from middle age onwards this was outstripped by about 2:1 by endosteal resorption, so that there was net thinning of cortical bone. The rate of cortical thinning resembled that seen in modern European males. Cross-sectional properties, as measured by second moments of area, bore no relationship to occupation. The results may suggest that, firstly, patterns of loss of cortical bone have remained unchanged in males for at least two centuries in Britain, and secondly, that biomechanical analyses of metacarpal cortical bone may be rather insensitive indicators of intensity of manual activity.     

Osteosclerosis in F344/DuCrj rats

Osteosclerosis was observed in the tibia and sternum in F344/DuCrj rats of both sexes at 6, 18 and 30 months of age. The lesion first seen was a proliferation of osteogenic tissues on the marrow surface of the cortical bone and bone trabeculae, resulting in replacement of the marrow cavity by lamellar bone. Most of the affected rats had associated degenerative osteoarthrosis and regressive changes of the growth plate. Osteosclerosis was considered to be an aging change, lesions were observed at 6 months and increased in frequency with age.     

Tooth cementum annulation for age estimation: results from a large known-age validation study

Recent research indicates that tooth-cementum annulations (TCA) may be used more reliably than other morphological or histological traits of the adult skeleton to estimate age. Until now, however, confidence intervals for age estimated by this method have not been available for paleodemographic and forensic applications. The present study addresses this problem. Based on a large known-age sample, age estimates by TCA were conducted in a blind study involving 363 teeth. Tooth-root cross sections were made using a refined preparation technique. Improved digital graphic procedures and enhancement strategies were used to produce digital images with a specially adapted software package. This resulted in high concordance between the TCA age estimates and chronological age. Assessment of the method's accuracy, as expressed by 95% confidence intervals, showed that error bounds for age estimates do not exceed 2.5 years. Sex differences, intraindividual correlations, and the effects of periodontal disease were studied. None of these indicators had a quantitative effect on the number of TCA bands when the proposed methodological standard was followed. We conclude that the TCA technique is a reliable method for estimating a subject's age from cementum annulations.     

Response of cortical and cancellous bones to mild calcium deficiency in young growing female rats: a bone histomorphometry study.

The purpose of the present study was to clarify the differences in the alterations of cellular activities of osteoblasts and osteoclasts, mineralization, and bone mass in cortical and cancellous bones of young growing rats with mild calcium deficiency. Twenty female Sprague-Dawley rats, 6 weeks of age, were randomized by the stratified method into two groups with 10 rats in each group: 0.5% (normal) calcium diet group and 0.1% (low) calcium diet group. After 10 weeks of feeding, bone histomorphometric analysis was performed on cancellous bone of the proximal tibia as well as cortical bone of the tibial shaft. Calcium deficiency increased eroded surface (ES/bone surface [BS]) and the number of osteoclast (N.Oc/BS) with an increase in osteoblast surface (ObS/BS), but decreased bone formation rate (BFR/BS) in cancellous bone. However, cancellous bone volume was preserved, while cortical bone area was decreased as a result of decreased periosteal bone gain and enlargement of the marrow cavity. These results suggest that short-term mild calcium deficiency in young growing female rats increased bone resorption by increasing osteoclastic recruitment, and suppressed mineralization followed by increased osteoblastic recruitment in cancellous bone, but cancellous bone loss was counteracted through redistribution of calcium from cortical bone to cancellous bone.     

Comparison of the egg flotation and egg candling techniques for estimating incubation day of Canada Goose nests

ABSTRACT Both egg flotation and egg candling have been used to estimate incubation day (often termed nest age) in nesting birds, but little is known about the relative accuracy of these two techniques. We used both egg flotation and egg candling to estimate incubation day for Canada Geese (Branta canadensis interior) nesting near Cape Churchill, Manitoba, from 2000 to 2007. We modeled variation in the difference between estimates of incubation day using each technique as a function of true incubation day, as well as, variation in error rates with each technique as a function of the true incubation day. We also evaluated the effect of error in the estimated incubation day on estimates of daily survival rate (DSR) and nest success using simulations. The mean difference between concurrent estimates of incubation day based on egg flotation minus egg candling at the same nest was 0.85 ± 0.06 (SE) days. The positive difference in favor of egg flotation and the magnitude of the difference in estimates of incubation day did not vary as a function of true incubation day. Overall, both egg flotation and egg candling overestimated incubation day early in incubation and underestimated incubation day later in incubation. The average difference between true hatch date and estimated hatch date did not differ from zero ( days) for egg flotation, but egg candling overestimated true hatch date by about 1 d (true – estimated; days). Our simulations suggested that error associated with estimating the incubation day of nests and subsequently exposure days using either egg candling or egg flotation would have minimal effects on estimates of DSR and nest success. Although egg flotation was slightly less biased, both methods provided comparable and accurate estimates of incubation day and subsequent estimates of hatch date and nest success throughout the entire incubation period.     

A comparison of cortical elastic properties in the craniofacial skeletons of three primate species and its relevance to the study of human evolution

When a force is applied to an object, the resulting pattern of strain is a function of both the object's geometry and its elastic properties. Thus, knowledge of elastic properties in craniofacial cortical bone is indispensable for exploring the biomechanics and adaptation of primate skulls. However, elastic properties, such as density and stiffness, cannot be measured in all species, particularly extinct species known only from fossils. In order for advanced engineering techniques such as finite element analysis (FEA) to be applied to questions of primate and hominid craniofacial functional morphology, it is important to understand interspecific patterns of variation in elastic properties. We hypothesized that closely related species would have similar patterns of bone elastic properties, and that similarities with extant species should allow reasonable predictions of elastic properties in the skeletons of extinct primate species. In this study, we tested this hypothesis by measuring elastic properties in five areas of the external cortex of the baboon craniofacial skeleton using an ultrasonic technique, and by comparing the results to existing data from macaque and human crania. Results showed that cortical density, thickness, elastic and shear moduli, and anisotropy varied among areas in the baboon cranium. Similar variation had previously been found in rhesus and human crania, suggesting area-specific elastic patterns in the skulls of each species. Comparison among species showed differences, suggesting species-specific patterns. These patterns were more similar between macaques and baboons for density, maximum elastic and shear stiffness, and anisotropy than between either of these and humans. This finding demonstrates that patterns of cortical elastic properties are generally similar in closely related primate species with similar craniofacial morphology. Thus, reasonable estimates of cortical bone elastic properties should be possible for extinct species through the study of phylogenetically related and functionally similar modern forms. For example, reasonable elastic property estimates of cortical bone from fossil hominid skulls should be possible once adequate information about such properties in extant great apes is added to our current data from humans, macaques, and baboons. Such data should eventually allow FEA of craniofacial function in fossil hominids.