Dietary health does affect histological age assessment: an evaluation of the Stout and Paine (1992) age estimation equation using secondary osteons from the rib

An age at death estimation equation that uses rib histological variables presented by Stout and Paine was used to evaluate a skeletal population of individuals with a known age at death and cause of death from either malnutrition or the niacin deficiency disease pellagra. The sample was comprised of 26 autopsied black South Africans. Histological analysis of mounted thin sections involved the microscopic measurement of cortical area and a count of the number of intact and fragmentary secondary osteons for the entire cross-section of the rib. Rib osteon population density values were then calculated for each case. It was found that this equation under-aged individuals on average by 29.2 years. Overall, secondary osteon size and Haversian canals tended to be larger than expected, while cortical bone area was less when compared with a control population. The implications of these findings are critical given that many of the skeletal remains examined by forensic anthropologists come from marginalized backgrounds, including malnutrition. This research suggests that measurements based on healthy cases may not be useful in an analysis of individuals with poor diet and health. It is argued that new standards for histological age assessment methods need to be created that account for variation in the health status of individuals examined by forensic anthropologists.     

A Four‐Stage Method of Age at Death Estimation for Use in the Subadult Rib Cortex*,†

Abstract: Age estimation in the subadult skeleton can be rather precise when the epiphyses and dentition are present, but incomplete or commingled remains still present a challenge. Histomorphometric age‐at‐death estimation methods developed for use on adults are based on the age‐associated accumulation of osteons. In the growing skeleton, there is a poor correlation between osteon numbers and age until the latter half of the second decade. As a result, there has been no histological aging method for use in subadults. The analysis of the rib cortex of 72 subadults ranging in age from 2 to 21 years has identified a series of developmental changes in the bone microstructure that can be used to estimate age. This qualitative method utilizes the systematic changes in rib cortical morphology to classify ribs into one of four age phases. This method can be applied to immature skeletons in forensic, archaeological, and paleontological contexts.     

The use of bone histomorphometry in skeletal identification: the case of Francisco Pizarro

Two lines of approach are utilized in a histomorphometric analysis of a rib sample from the postcranial remains believed to be those of Francisco Pizarro. Using a newly developed age predicting formula, age at death is estimated to be 62 years. This age agrees with several ages reported in historical documents. The histomorphometry of cortical area/total area (C/T) ratio, mean osteonal cross-sectional area, and mean annual Haversian bone formation rate conform to those of a healthy individual in his early sixties.     

Analysis of histomorphometric variables: Proposal and validation of osteon definitions

Histological analysis of bone tissue has been used to explore a variety of questions relating to age-at-death, habitual behaviors, health, and nutritional stress. Identification of intact and fragmentary osteons is of key interest to many researchers in these studies, yet the definitions of these features vary between researchers making cross-study comparisons problematic. Furthermore, histological variable definitions are often ambiguous or require subjective classifications by the observer. As a result, and as indicated by previous studies, observer error and misclassification of certain variables, namely intact and fragmentary osteons, can be significant. This study proposes new definitions for intact and fragmentary osteons that are designed to limit observer subjectivity and also explore efficacy of combining osteon types into one variable. A sample of 30 6th rib cross-sections from a modern forensic population was used to test the validity of the proposed definitions. Observations of intact osteon population density (OPD(I)) and fragmentary osteon population density (OPD(F)) were made by three observers for each cross-section. These observations were used to explore the interobserver error associated with the proposed definitions and determine if combining variables into one variable (OPD) mitigates persisting classification difficulties. Results indicate that the proposed definitions significantly reduce interobserver error and misclassification of intact and fragmentary osteons. However, the interobserver error associated with fragmentary osteons is still high. Evaluation of the variables independently indicates that combining variables has potential to reduce the predictive strength of an age estimation model and the ability to interpret age-related bone remodeling.     

Assessment of intra- and intercostal variation in rib histomorphometry: its impact on evidentiary examination

Rib histological age estimation requires the evaluation of the middle third of the sixth rib. Human ribs have thin cortices and, when recovered, are often fragmented or absent, making it difficult to identify a specific midthoracic rib. This research explores the amount of microstructure variation in the middle third of the midthoracic ribs and determines whether the sixth rib age prediction equation can be applied to non-sixth ribs with similar accuracy. The amount of variability must be evaluated in order to meet the criterion for evidentiary examination. The sample consists of 120 cortical bone cross-sections from the middle third of ribs 3-8 removed from 20 cadavers. For each rib, osteon population densities (OPDs) and associated age estimates were calculated. The results demonstrate that non-sixth ribs can provide similar OPD values compared with those of the sixth ribs; however, individual variation proved to be significantly associated with bias, suggesting that individual factors influence the magnitude and direction of bias in non-sixth rib OPD values. This demonstrates the importance of evaluating multiple cross-sections (both intra- and inter-rib) to estimate age due to the normal remodeling variation within individuals.     

Estimation of age at death by tibial osteon remodeling in an autopsy series

Estimation of age at death by histological methods in tibiae has been found to be inaccurate in individuals less than 55 years of age. This study describes reasons for the inaccurate age estimates and provides a new regression equation for age estimation. Bone cores were removed from tibiae in 53 individuals at autopsy ranging in age from 17 years to 53 years of age; 48 males and 5 females. The sample contained 41 whites and 12 blacks. Variables ascertained from each core were: cortical thickness, cortical bone density, secondary osteon number, area, and perimeter, and Haversian canal area and perimeter. The results showed significant differences in secondary osteon number, area, and perimeter between groups aged less than and greater than 35 years of age. Multiple linear regression analysis indicated osteon number to be the best predictor of age in this sample aged less than 55 years of age. Age at death was accurately estimated for 11 additional forensic cases using osteon number in tibiae. No significant histological differences were observed between males and females and between blacks and caucasians.     

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.     

Rib Variation at Multiple Locations and Implications for Histological Age Estimation*

Existing histological age estimation methods using the rib were developed mainly from the midshaft; however, in forensic practice, uncertainty of sampling location often arises due to fragmented or previously sampled ribs. The potential for error increases when sampling location is uncertain and utilizing a section beyond the midshaft (either anterior or posterior) may result in erroneous age estimates. Additionally, there is debate within the field regarding the minimum number of sections needed for accurate age estimation. The aim of this research is to determine the importance of the midshaft distinction for age-at-death assessment and the necessity of analyzing serial sections by evaluating histological variables at sampling locations along the length of the rib. Three seriated histological sections at three sampling locations (anterior, midshaft, and posterior) were obtained from sixth ribs of ten postmortem human subjects. Cortical area (Ct.Ar) and osteon population density (OPD) were collected from each section (n = 90). Significant differences were determined in Ct.Ar between sampling locations, demonstrating the variation present along the length of the rib. A comparison of OPD at sampling locations revealed significant differences, suggesting that sampling site is critical to accurate age estimates. When sampling location is uncertain, a more anterior section should be taken. Analysis of serial sections within locations revealed no significant differences in OPD or Ct.Ar, supporting the practice of collecting data from one section for age estimation. While an age estimate can be achieved through the analysis of one section, best practice suggests reading two sections to capture intraindividual variation.     

X线片的骨骼年龄判定

目前,对儿童青少年骨骼年龄评价以手腕部及膝部较好,方法也较多,但主观性强,随机误差大。随着计算机图像识别及智能化技术的应用,将使得骨骼年龄评价更为客观、准确。对于成年人X线片骨龄推断,以胸骨、耻骨及锁骨等方法具有一定的应用价值,尚需更广泛的方法学研究以增加推断年龄的方法及准确性。本文介绍了X线片的骨骼年龄判定方法的现状及研究进展。     

Assessment of histomorphological features of the sternal end of the fourth rib for age estimation in Koreans

The aim of this study was to assess the histomorphological features of the fourth rib and to develop age-predicting equations for Koreans. Sixty-four rib samples (36 males and 28 females) obtained from forensic cases were used for developing equations. Two thin sections (<100-microm thick) per sample were prepared by manual grinding. Multivariate analysis of covariance revealed statistically significant differences in age-adjusted histomorphological variables between sexes. Using stepwise regression analysis, osteon population density and average osteon area were correlated with unknown sex (r2 = 0.826), and sex plus two histomorphological variables provided the best results for an age-predicting equation given the assumption of knowing the sex of a specimen (r2 = 0.839). Average Haversian canal area had little influence on age estimation for male or female samples, and relative cortical area was not significantly related to age for any specimen.     

A Test of Hartnett's Revisions to the Pubic Symphysis and Fourth Rib Methods on a Modern Sample

Estimating age at death is one of the most important aspects of creating a biological profile. Most adult age estimation methods were developed on North American skeletal collections from the early to mid‐20th century, and their applicability to modern populations has been questioned. In 2010, Hartnett used a modern skeletal collection from the Maricopia County Forensic Science Centre to revise the Suchey‐Brooks pubic symphysis method and the ??can et al. fourth rib methods. The current study tests Hartnett's revised methods as well as the original Suchey‐Brooks and ??can et al. methods on a modern sample from the William Bass Skeletal Collection (N = 313, mean age = 58.5, range 19–92). Results show that the Suchey‐Brooks and ??can et al. methods assign individuals to the correct phase 70.8% and 57.5% of the time compared with Hartnett's revised methods at 58.1% and 29.7%, respectively, with correctness scores based on one standard deviation of the mean rather than the entire age range. Accuracy and bias scores are significantly improved for Hartnett's revised pubic symphysis method and marginally better for Hartnett's revised fourth rib method, suggesting that the revised mean ages at death of Hartnett's phases better reflect this modern population. Overall, both Hartnett's revised methods are reliable age estimation methods. For the pubic symphysis, there are significant improvements in accuracy and bias scores, especially for older individuals; however, for the fourth rib, the results are comparable to the original ??can et al. methods, with some improvement for older individuals.     

Age estimation from the rib by phase analysis: white females

Metamorphosis at the sternal extremity of the rib has already been established as a reliable indicator of age at death. Using a sample of white males, it was shown that an accurate estimation of age can be made by direct examination of the bone itself. However, because of sexual differences in hormonal production and dimorphism in the skeleton, the present study was carried out to develop an appropriate standard for age determination from the sternal rib in white females. The sample consisted of 86 ribs of known age, sex, and race. Observations were made at the costochondral junction with special attention to pit formation (its shape and depth), changes in the walls and rim surrounding it, and overall bone density and texture. Based on changes in these areas, the ribs were separated into nine phases (0 through 8). The most rapid metamorphosis occurred in Phases 1 through 4 (mean ages 14-28) with changes noticeable at 3 to 4 year intervals. After mean age 28, this process slowed, considerably expanding the interval between phases to 10 to 15 years. The female ribs showed both earlier initial pit formation and a different morphologic pattern of aging as compared with males. Statistical analysis revealed that the features chosen to delineate the phases are valid predictors of age. This study has shown that the sternal rib can provide an accurate estimation of age in females spanning a mean age of 14 to 76 years.     

14岁青少年手腕骨发育的研究I．男生骨龄标准的制定

目的研究我国当前青少年骨发育状况,制订专用于法医学推断男性刑事责任年龄14岁的骨龄标准。方法以湖南冷水江地区青少年为对象,按照纳入标准,选取14岁±3个月的健康男性中学生103名,拍摄其左手腕部后前位X线片。选择手腕骨15个部位,按照统一标准进行观察、测量,采用最大百分比法确定发育等级标准,建立骨龄评估方法,以单盲法进行验证。结果除第一掌骨、近节第四指骨骨骺发育等级为部分闭合外,其他均为未闭合;对于第四、五掌骨和近三指骨,不足14岁组以未闭合为主,已满14岁组以部分闭合为主。各个观察部位骨骺横径宽于干骺横径。30例的验证结果准确率为80.0%。结论根据发育等级特征及测量学方法建立骨龄评估标准可行,可望得到推广。本文以刑事责任年龄14周岁为对象,制订法医学专业骨龄推断标准,并引入测量学方法,辅助推断年龄。     

Internal Morphological Analysis for Age Estimation of Blow Fly Pupae (Diptera: Calliphoridae) in Postmortem Interval Estimation*

Abstract: Larvae and pupae of blow fly species are frequently used in postmortem interval estimation, their age indicating minimum time since death. Most studies have considered age estimation of larvae, neglecting study of pupae. Relative development of external pupal features is useful, but there are also internal changes during metamorphosis that may be indicators of age, utilizing histological techniques. This study aimed to optimize preservation and histological analysis of blow fly pupae, specifically Calliphora vicina Robineau‐Desvoidy and Lucilia sericata (Meigen), and to examine internal features with potential for age estimation. Effect of hot‐water‐killing and different preservatives were examined. It was determined that blow fly pupae should be pierced through the three tagma, hot‐water‐killed, and preserved in 80% ethanol as the optimal preservation for subsequent analyses. Hematoxylin and eosin stained pupal sections revealed differences in brain and thoracic muscle development throughout the pupal stage with potential for age estimation.     

Age estimation by measuring the racemization of aspartic acid from total amino acid content of several types of bone and rib cartilage: a preliminary account

Age estimation has been often performed based on the rate of aspartic acid racemization using bones. In this study, we investigated various kinds of bone and a cartilage for suitability to estimate age by racemization. Ten male cadavers aged 22 to 77 years at death were selected, and bone specimens and cartilage were taken from seven sites in each individual. The rate of racemization of aspartic acid among total amino acid contained in each specimen was analyzed by gas chromatography. The correlation coefficient between the rate of racemization and chronological age was relatively high in the sternum, skull, and femur. The rates of aspartic acid racemization were high in the costal cartilage, femur, and skull. In addition, we found that the rate of racemization was only slightly lower after the second irrigation than after the first irrigation in femur and skull bone specimens, but those of others were significantly lower. These findings showed that among the six different bones and rib cartilage, the skull and femur might be used most effectively for age estimation using total amino acid fraction.     

Estimation of Age Using Alveolar Bone Loss: Forensic and Anthropological Applications

The objective of this study was to utilize a new odontological methodological approach based on radiographic for age estimation. The study was comprised of 397 participants aged between 9 and 87 years. A clinical examination and a radiographic assessment of alveolar bone loss were performed. Direct measures of alveolar bone level were recorded using CT scans. A medical examination report was attached to the investigation file. Because of the link between alveolar bone loss and age, a model was proposed to enable simple, reliable, and quick age estimation. This work added new arguments for age estimation. This study aimed to develop a simple, standardized, and reproducible technique for age estimation of adults of actual populations in forensic medicine and ancient populations in funeral anthropology.