One of the fundamental questions in forensic medicine and anthropology is whether or not a bone or bone fragment is human. Surprisingly at times for the extreme degradation of the bone (charred, old), DNA cannot be successfully performed and one must turn to other methods. Histological analysis at times can be proposed. However, the variability of a single human skeleton has never been tested. Forty-nine thin sections of long, flat, irregular and short bones were obtained from a well-preserved medieval adult human skeleton. A qualitative histomorphological analysis was performed in order to assess the presence of primary and secondary bone and the presence, absence and orientation of vascular canals. No histological sections exhibited woven or fibro-lamellar bone. Long bones showed a higher variability with an alternation within the same section of areas characterized by tightly packed secondary osteons and areas with scattered secondary osteons immersed in a lamellar matrix. Flat and irregular bones appeared to be characterized by a greater uniformity with scattered osteons in abundant interstitial lamellae. Some cases of "osteon banding" and "drifting osteons" were observed. Although Haversian bone represent the most frequent pattern, a histomorphological variability between different bones of the same individual, in different portions of the same bone, and in different parts of the same section has been observed. Therefore, the present study has highlighted the importance of extending research to whole skeletons without focusing only on single bones, in order to have a better understanding of the histological variability of both human and non-human bone.

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.

In the last decades, the histomorphometric analysis of bone tissue has been utilized to develop equations for species discrimination of fragmentary bone. Although this technique showed promising results, its main limitation concerns the lack of knowledge on the histomorphometric variability which may exist between different bones of the skeleton. In a previous study, we demonstrated a significant histomorphological variability in different bones of the same individual and even in different sections of the same bone. The present study aimed at investigating the extent of intraindividual variability in bone histomorphometry throughout the human adult skeleton and areas of a single bone.Samples were taken along an entire medieval male adult human skeleton (aged between 26-45 years), including long, flat, irregular and sesamoid bones for a total of 49 cross-sections.The histomorphometric analysis revealed that the size of both Haversian systems and Haversian canals were statistically significantly larger in long and irregular bones compared to flat bones. Moreover, osteons were generally bigger in the diaphysis compared to the proximal and distal metaphyses, whereas Haversian canals showed a higher uniformity in the different portions of each bone.The present study has highlighted the importance of conducting similar studies on both human and nonhuman skeletons at different stages of skeletal maturity in order to shed light on the extent of variability in the size of osteons and Haversian canals. This, in fact, represents an important prerequisite to develop reliable histological methods for species discrimination of fragmented bone.

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