Histomorphometric analysis of osteocyte lacunae in human and pig: exploring its potential for species discrimination

In recent years, several studies have focused on species discrimination of bone fragments by histological analysis. According to literature, the most consistent distinguishing features are Haversian canal and Haversian system areas. Nonetheless, there is a consistent overlap between human and non-human secondary osteon dimensions. One of the features that have never been analyzed for the purpose of species discrimination is the osteocyte lacuna, a small oblong cavity in which the osteocyte is locked in. The aim of this study is to verify whether there are significant quantitative differences between human and pig lacunae within secondary osteons with similar areas. Study sample comprises the midshaft of long bones (humerus, radius, ulna, femur, tibia, and fibula) of a medieval human adult and a juvenile pig. Sixty-eight secondary osteons with similar areas have been selected for each species and a total of 1224 osteocyte lacunae have been measured. For each osteon, the total number of lacunae was counted, and the following measurements were taken: minimum and maximum diameter, area, perimeter, and circularity of nine lacunae divided between inner, intermediate, and outer lacunae. Statistical analysis showed minimal differences between human and pig in the number of lacunae per osteons and in the minimum diameter (P > 0.05). On the contrary, a significant difference (P < 0.001) has been observed in the maximum diameter, perimeter, area, and circularity. Although there is the need for further research on different species and larger sample, these results highlighted the potential for the use of osteocyte lacunae as an additional parameter for species discrimination. Concerning the difference between the dimensions of osteocyte lacunae based on their position within the osteon (inner, intermediate, and outer lacunae), results showed that their size decreases from the cement line towards the Haversian canal both in human and pig

Swine cortical and cancellous bone: histomorphometric and densitometric characterisation

Introduction: Swine bone morphology, composition and remodelling are similar to humans\u2019, therefore they are considered good models in bone-related research. They have been used for several studies involving bone growth, bone and cartilage fractures and femoral head osteonecrosis. Nevertheless, the literature about pig normal bone features is incomplete. This work aims to fill the literature gaps on the microarchitecture and Bone Mineral Density (BMD) of swine femoral diaphysis and distal epiphysis and tibial plateau and diaphysis. Materials and methods: Five hind limbs were collected from slaughtered 80-100 kg pigs. Microscopic analysis of cortical and cancellous bone from middle/distal femur and proximal/middle tibia was performed to determine basic histomorphometric parameters at different sites. Dual-energy X-Rays Absorptiometry was also employed to evaluate BMD. ANOVA and correlation between BMD, bone area (BA) and cortical thickness were performed. Results and discussion: Diaphyseal cortical bone was mostly plexiform both in the tibia and the femur; primary/secondary osteons without clear organization were also found. Mean values for bone area, bone perimeter, trabecular width, number and separation and BMD at different anatomical sites were defined. No significant difference was found for these values at different anatomical sites. BMD proved to be positively correlated with cortical thickness (r=0,80; p<0,01). Despite the small sample size, these results seem homogeneous. They could therefore represent reference values for normal bone parameters in pigs. Applied anatomy and regenerative medicine, in fact, demand very precise information about bone micromorphology, composition and density to provide reliable indication in bone substitutes building. Moreover, since the interpretation of bone abnormalities is based on mastering normal bone characteristics, the definition of reference parameters is mandatory to avoid misinterpretation and allow comparative evaluation. Conclusion: The results of this study, although preliminary, may be considered a dependable starting point for the definition of normal bone features in pigs

The Issue of Age Estimation in a Modern Skeletal Population : Are Even the More Modern Current Aging Methods Satisfactory for the Elderly?

The main idea behind age assessment in adults is related to the analysis of the physiological degeneration of particular skeletal structures with age. The main issues with these procedures are due to the fact that they have not been tested on different modern populations and in different taphonomic contexts and that they tend to underestimate the age of older individuals. The purpose of this study was to test the applicability and the reliability of these methods on a contemporary population of skeletal remains of 145 elderly individuals of known sex and age. The results show that, due to taphonomic influences, some skeletal sites showed a lower survival. Therefore, the methods with the highest percentage of applicability were Lovejoy (89.6%) and Roug\ue9-Maillart (81.3%), followed by Suchey-Brooks (59.3%), and those with the lowest percentage of applicability were Beauthier (26.2%) and Iscan (22.7%). In addition, this research has shown how for older adults the study of both acetabulum and auricular surface may be more reliable for aging. This is also in accordance with the fact that auricular surface and the acetabulum are the areas more frequently surviving taphonomic insult

The Issue of Age Estimation in a Modern Skeletal Population : Are Even the More Modern Current Aging Methods Satisfactory for the Elderly?

The main idea behind age assessment in adults is related to the analysis of the physiological degeneration of particular skeletal structures with age. The main issues with these procedures are due to the fact that they have not been tested on different modern populations and in different taphonomic contexts and that they tend to underestimate the age of older individuals. The purpose of this study was to test the applicability and the reliability of these methods on a contemporary population of skeletal remains of 145 elderly individuals of known sex and age. The results show that, due to taphonomic influences, some skeletal sites showed a lower survival. Therefore, the methods with the highest percentage of applicability were Lovejoy (89.6%) and Rougé-Maillart (81.3%), followed by Suchey-Brooks (59.3%), and those with the lowest percentage of applicability were Beauthier (26.2%) and Iscan (22.7%). In addition, this research has shown how for older adults the study of both acetabulum and auricular surface may be more reliable for aging. This is also in accordance with the fact that auricular surface and the acetabulum are the areas more frequently surviving taphonomic insult

The “forensic paradox” of aging unaccompanied minors in the migration crisis: Why medicine and forensics are a must

only between 2014 and 2017 the European Migration Network (EMN) recorded 268,348 unaccompanied minors entering Europe1, hence in the past years the European Union has intervened in order to guarantee special protective measures for minors. Yet only a few weeks ago news concerning incidents in Greece have once again drawn attention to the need to protect migrant children2. Particularly when dealing with adolescents, protection derives, for those without documents, from properly ascertaining that they are minors. Adequate verification of age is therefore fundamental, yet in Europe it is still badly managed. Italy is a perfect example of such mismanagement, but not the only one. Our country is one of the main doors to Western Europe and on May 6th 2017, the Italian Parliament enacted the “Provision on protective measures for unaccompanied foreign minors”, known as the “Zampa law” (n°47/17), which aims at guaranteeing the protection of unaccompanied minors. However, as reflected in the whole of Europe, within the lines of this law, the paradox of the lacking, restricted and somewhat maligned role of medical and forensic sciences in age assessment emerges, which is dangerously leading to a sloppy administration of the rights of minors. Proof of this is the amount of resources spent in appeals which follow inadequate aging procedures which in turn can lead to minors being judged as adults (the most alarming risk since what is most at heart is the protection of true minors), or to judging adults as minors, leading to an excessive burden on administrations which can severely challenge the system in the long run. According to principles of medical ethics and human rights assessed in the practical guide recommended by the European Asylum Support Office (EASO) as well as other sources, the age assessment procedure, when necessary, recommends that a multidisciplinary holistic approach be applied. This approach involves an interview with a social worker, a neuropsychiatric or psychological evaluation, and a pediatric auxological examination, (all in this order), preferably performed together with an intercultural-linguistics mediator; medical procedures should come last, favour those methods that are less intrusive for the minor leaving the most invasive ones (including radiological examination) as a last resort (though one wonders which reliable medical age assessment for this purpose is at the moment non radiological). Here the double paradox emerges: first that of the secondary role of medical evaluation, then that of the omitted role of forensics. Age assessments by psychosocial tests and non-medical methods can measure behavior and cognitive ability but they are highly influenced by environmental and cultural factors. Furthermore child and adolescent interviews and psychological evaluations do not provide any validated margin of error (let alone referred to specific ethnic groups) as requested by most administrative or judicial agencies, and the assessment of mental maturity is very difficult in individuals who have little or no education, come from different cultural backgrounds and who may have experienced trauma. On the other hand, medical radiological methods (which are far less invasive than usually proclaimed8) have fairly large error ranges but these are better known and quantifiable than those of psychosocial assessments.9 Hence it is unreasonable and counterproductive to proceed to verify age in adolescents without medical and radiological tests. Furthermore, one wonders why the world of forensic experts is frequently left aside in guidelines and laws, since these are the scientists best trained to discuss “error ranges” in administrative and judicial (not “only - clinical”) scenarios. Age assessment based on dental and/or skeletal maturity has since decades been the object of research by forensic experts. In the last years several forensic groups and societies10 have dealt with this issue and provided a plethora of publications relating to this topic as well as appropriate scientific methodology with which to present the strength of evidence. It is our opinion that the recommended holistic approach in the age estimation process must always include, when dealing with minors (particularly post pubertal minors) with no valid documents, medical and radiological evaluation, and that it cannot be considered multidisciplinary and complete without the involvement of forensic expertise

Age estimation in unaccompained foreign minors: Comparative analysis of some Italian protocols. [STIMA DELL'ETÀ NEI MINORI STRANIERI NON ACCOMPAGNATI: ANALISI COMPARATIVA DI ALCUNI PROTOCOLLI ITALIANI]

Secondo le normative nazionali ed internazionali, la determinazione dell’età anagrafica assume un’importanza fondamentale, stante le correlazioni esistenti tra minore età ed il riconoscimento di alcuni diritti fondamentali. In caso di fondati dubbi in merito all’età dei rifugiati e richiedenti asilo, il procedimento di stima dell’età dovrebbe essere esperito mediante un approccio olistico e multidisciplinare caratterizzato dal graduale e progressivo utilizzo di metodi non-medici e medici. Tuttavia, in ambito forense non vi è consenso ed uniformità in merito alle procedure ed alle metodologie da utilizzare e ciò anche in relazione alle controversie sui limiti ed i rischi di errore correlati ad una valutazione effettuata con metodi invasivi e facendo ricorso all’utilizzo di radiazioni ionizzanti per finalità giuridico-amministrative e non diagnostico-terapeutiche. In relazione a tali criticità ed alle evidenze che suggeriscono un’applicazione disomogenea dell’attuazione della norma che prescrive il carattere multidisciplinare dell’accertamento, con il presente contributo si mira ad intraprendere un’analisi comparativa di alcuni protocolli per la stima dell’età cronologica vigenti sul territorio italiano

Three-dimensional facial anatomy evaluation: Reliability of laser scanner consecutive scans procedure in comparison with stereophotogrammetry

Laser scanner devices are acquiring a growing importance in facial anatomy. Most studies have analysed facial scans obtained through two simultaneous captures, whereas the same result can be obtained by consecutive three-dimensional (3D) scans. However, this latter procedure has not yet been validated. Fourteen volunteers underwent face image capture through stereophotogrammetry (VECTRA M3) and three consecutive facial scans through a laser scanner (Konica Minolta Vi910). The concordance between 14 linear distances, 12 angles, facial surface area and volume measurements was verified by the Bland\u2013Altman test and calculation of absolute and relative technical errors of measurement (TEM/rTEM). The two facial images obtained by the different devices were then registered to calculate point-to-point distance. Most of linear distances and angles showed a high agreement, with \u201cvery good\u201d or \u201cgood\u201d rTEMs, ranging between 1.1% and 6.4%. Surface area measurements agreed well between the devices (rTEM: 6.3%), while volumes were poorly comparable (rTEM: 25.8%); the root mean square point-to-point distance was 0.80 mm (SD: 0.41). This study first tested the concordance of measurements on facial images obtained by stereophotogrammetry and consecutive laser scans. Results highlight the reliability of linear distances, angles and surface areas measurements, but discourage volume assessment and registration of surfaces acquired through different devices

3D quantitative analysis of early decomposition changes of the human face

Decomposition of the human body and human face is influenced, among other things, by environmental conditions.The early decomposition changes that modify the appearance of the face may hamper the recognition and identification of the deceased. Quantitative assessment of those changes may provide important information for forensic identification. This report presents a pilot 3D quantitative approach of tracking early decomposition changes of a single cadaver in controlled environmental conditions by summarizing the change with weekly morphological descriptions. The root mean square (RMS) value was used to evaluate the changes of the face after death. The results showed a high correlation (r = 0.863) between the measured RMS and the time since death. RMS values of each scan are presented, as well as the average weekly RMS values. The quantification of decomposition changes could improve the accuracy of antemortem facial approximation and potentially could allow the direct comparisons of antemortem and postmortem 3D scans

Histological determination of the human origin from dry bone: a cautionary note for subadults

Anthropologists are frequently required to confirm or exclude the human origin of skeletal remains; DNA and protein radioimmunoassays are useful in confirming the human origin of bone fragments but are not always successful. Histology may be the solution, but the young subadult structure could create misinterpretation. Histological tests were conducted on femur and skull of 31 human subjects. Each sample was observed focusing on presence or absence of fibrous bone, lamellar bone, radial lamellar bone, plexiform bone, reticular pattern, osteon banding, Haversian bone, primary osteons, secondary osteon and osteon fragments. Samples were divided into five age classes; 1 (&lt;1&nbsp;year), 2 (1\u20135&nbsp;years), 3 (6\u201310&nbsp;years), 4 (11\u201315&nbsp;years) and 5 (16\u201320&nbsp;years). Regarding femurs, class 1 presented the following: 87.5&nbsp;% fibrous bone, 37.5&nbsp;% plexiform bone, 12.5&nbsp;% reticular pattern and 12.5&nbsp;% lamellar bone radially oriented. Class 2 showed 37.5&nbsp;% of fibrous bone, 12.5&nbsp;% of reticular pattern and 37.5&nbsp;% of osteon banding. In the higher age classes, the classical human structures, lamellar bone and osteons were frequently visible, except for one case of reticular pattern, generally considered a distinctive non-human structure. The situation appeared different for the skull, where there was a lack of similar information, both in human and non-human. An analysis of the percentage of lamellar bone and osteons was conducted on femur and skull fragments. A trend of increase of primary osteon number and a decrease of the lamellar bone area has been detected in the femur. The present study has therefore shed some light on further pitfalls in species determination of subadult bone