Different Preparation Techniques – Similar Results? On the Quality of Thin‐Ground Sections of Archaeological Bone

Palaeohistology as a valuable diagnostic instrument is dependent on the production of high‐quality thin‐ground sections from dry bone. The objective of this study was to consider technical differences and assess the qualitative outcomes of five techniques for preparing thin‐ground sections from dry archaeological bone. Established techniques with long follow‐up times and excellently documented results were compared with simpler and cheaper time‐saving techniques. Evaluations were made of the quality of thin sections obtained by one classical machine‐based embedding technique, two revised versions of the same technique, one manual moulding technique based on Frost's rapid technique and one manual hybrid technique. Five osteological specimens of differing quality were prepared following the manuals for these five techniques and examined microscopically with respect to a list of standardised histological and diagenetic parameters. Alterations in the specimens attributable to preparation effects were recorded, and observations were scored with reference to three criteria: section quality, technical quality and staining. The results show that embedding techniques are to prefer. Superglue should not be used as a mounting or embedding medium. Manual grinding comes with several limitations, and machine cutting and grinding are preferred. Haematoxylin staining can be successfully applied to embedded specimens, giving more information on microscopic diagenetic processes. A stepwise manual for a revision of the classical embedding technique is presented. The time required for producing sections using classical embedding techniques is shortened from 6 weeks to 3.7 days by refining the preparation/polymerization processes involved with no loss of osteological data. Copyright © 2014 John Wiley & Sons, Ltd.     

Staining ground sections of natural dry bone tissue for microscopy

The histological staining of bone tissue is of great value in identifying structural changes in human bone tissue when examined microscopically, and is therefore routinely used by clinical pathologists. However, for archaeological and other types of undecalcified dry bone tissue, histochemical staining is currently not widely used. The purpose of this paper is to describe a new method developed for the staining of undecalcified archaeological bone which may be a valuable addition to the palaeopathological toolkit. Sections were ground by hand and stained with haematoxylin and eosin. The microstructure and architecture of the stained sections become clearly visible without compromising total tissue envisioning. In addition, staining enhances differentiation between taphonomically affected and non‐affected bone tissue. This paper accordingly describes a reproducible stepwise method for the production of ground stained sections. An additional troubleshooting paragraph discusses the most often encountered problems and provides solutions. Copyright © 2010 John Wiley & Sons, Ltd.     

Assessing Post‐traumatic Time Interval in Human Dry Bone

The post‐traumatic status of antemortem fractures in human dry bone remains is currently defined as being either ‘healing’ or ‘healed’. However, detailed ‘dating’ of the related post‐traumatic time interval would be desirable, since it would aid in assessing individual medical status and care at the time of death. Within forensic pathology practice, fresh tissue healing phases are routinely used as an intrinsic parameter for the length of the post‐traumatic time interval. Unfortunately, the direct application of such a method is hampered when applied to dry bone skeletal material. This study explores the possibility of applying a fracture dating system, drawn forth from the traditional forensic pathology method, on dry bone remains. More specifically, the aim is to establish the extent to which various histomorphological features indicative of specific time intervals of healing are consistently detectable. Human dry bones with fractures and amputations in various phases of healing were studied. Results show that the complementary use of radiological and histological investigation techniques improves the differentiation between various healing phases and thus allow for a more detailed dating of lesions. For future use, healing features that have proven to be consistently detectable and their related post traumatic time intervals are listed. The system aids in demarcating a considerably more ‘narrow’ post‐traumatic time interval than usual. Copyright © 2012 John Wiley & Sons, Ltd.     

Ossified haematomas and infectious bone changes on the anterior tibia: histomorphological features as an aid for accurate diagnosis

Examination of the histological structure of bone not only helps investigators to estimate age at death but can also aid in the diagnosis of palaeopathological lesions. The purpose of this paper is to assess whether histological features as described in the literature can confirm the macroscopic diagnoses of ossified subperiosteal haematomas associated with healed scurvy and syphilitic bone changes observed on the anterior tibiae of individuals from a 19th century mining community from Kimberley South Africa. The frequent occurrence of these two diseases amongst the deceased was well established in related hospital documents and governmental documents. A section of bone was removed from lesions on the tibiae of 14 individuals. These bone changes were macroscopically diagnosed as being indicative of either treponematosis ossified subperiosteal haematomas or non‐specific periostitis. Cross‐sections were prepared for microscopic investigation using a manual ground section technique. Ossified haematomas were histologically identified in seven individuals. These sections were characterised by normal cortical bone an intact original periosteal surface and newly formed radiating trabecular bone apposing it. Three phases of ossified subperiosteal haematoma formation and remodelling could be distinguished. Infectious bone changes most likely associated with treponematosis were observed in one individual. These were characterised by lysis and numerous resorption holes/channels. No clear distinction could be made between the internal spongy cortical or newly formed bone. Histological features described by some authors as characteristic of this condition could not be identified. In addition three individuals presented with microscopic features indicative of both the aforementioned bone affections and three did not show any pathological changes on microscopic level. It was concluded that although specific pathological conditions can most likely not be diagnosed purely on the basis of histomorphological observations broad distinctions could be made between lesions caused by the ossification of subperiosteal haematomas and bone changes due to infectious diseases. Copyright © 2009 John Wiley & Sons Ltd.     

Bone preservation in human remains from the Terme del Sarno at Pompeii using light microscopy and scanning electron microscopy

Archaeological bone can show marked and complex alterations depending on the environment in which it was buried. In this study, the state of preservation of 27 femurs recovered from the archaeological site of Pompeii was evaluated by light microscopy and scanning electron microscopy. Most of the bone samples, prepared by the grinding method, showed good histological preservation, although they were characterized by microfissures (microcracking). Nine bone samples showed different states of histological preservation, including worst preservation (two femurs), due to diagenetic processes. Cryostat bone sections stained with thionin or 4′,6′-diamidino-phenylindole (DAPI) revealed the persistence of DNA within some osteocyte lacunae. Scanning electron microscopy analysis showed that ultrastructural characters, such as lamellae and collagen fibres, are recognizable only in unaltered bone. Our results reveal that most Pompeian samples are well preserved since they have a bone microstructure virtually indistinguishable from that of fresh bone. In methodological terms, although each of the various morphological methods used contributes information, histological and histochemical analyses are the most informative for studying the preservation state of bone and allow for rapid essential screening of archaeological bone.     

Maize and Bone: Residues of Grinding in Northwestern Argentina*

Archaeometric and archaeobotanical analyses of grinding residues inside an archaeological milling stone tool from the Cueva de los Corrales 1 site in northwestern Argentina are shown. Multiple analytical techniques are used to identify ground substances, and their results are compared and complemented. They give direct and reliable evidence of maize (Zea mays L.) and burned bone grinding in the pre‐hispanic Argentine north‐west, although the latter is little related to milling tasks.     

An investigation of historical and archaeological evidence for age‐related bone loss and osteoporosis

A growing awareness of the problems caused by osteoporosis‐related fractures has led to increased investigation of the condition in past populations. The present study seeks to examine the ways in which osteoporosis‐related fractures may be explored through different approaches; modern clinical information, historical records of fractures and analysis of archaeological bone may provide fuller information on the condition. The three types of fractures upon which this study focuses are fractures of the femoral neck, Colles' fractures and vertebral compression fractures. The results of the investigation clearly demonstrate the way in which the various sources produce a slightly different picture of the condition. The archaeological significance of the different types of fractures was found to be the reverse of that which would be suspected in the light of information derived from historical sources and the clinical literature. To date vertebral crush fractures have been the most commonly found osteoporosis‐related fracture in archaeological material, but their significance to people in the past is likely to have been minimal. In contrast hip fractures have rarely been reported from archaeological bone material, but historical information makes it clear they were well known, and knowledge derived from clinical sources emphasizes the serious consequences of such a fracture for the affected individuals. The results indicate the value of undertaking a multi‐disciplinary approach where possible. Copyright © 2002 John Wiley & Sons, Ltd.     

A distinct region of microarchitectural variation in femoral compact bone: Histomorphology of the endosteal lamellar pocket

Bone is a dynamic tissue, responding locally to differential mechanical loading and systemically to hormonal stimuli. Although the tendency is to consider compact bone a homogenous tissue, regional variations in microarchitecture are known to have quite different properties relating to processes of growth and mechanical loading. Specifically, we investigated an example of microstructural variation of the human, adult femoral endocortex, referred to here as the endosteal lamellar pocket (ELP). The femoral ELP is characterised as a histomorphological meta‐feature, made distinct from general circumferential lamellae by its hemicircumferential lamellar orientation, medial positioning, radially oriented Volkmann's canals and decreased presence of Haversian systems. Our preliminary observations of mid‐diaphyseal transverse thin‐ground sections demonstrate the ELP as a regular meta‐feature of adult human femora unearthed from the Maya archaeological site of Xcambó, Mexico. ELPs were easily identified in 38 of 45 examined individuals. Their size, shape and degree of secondary remodelling were variable. In comparison, ELP lamellae and those originating from the periosteum are divergent, or non‐concentric in orientation. This divergence, along with the ELP's medial position, suggest it forms during growth through endosteal modelling drift. Possible considerations for ELP formation and persistence with age are discussed as well as its implications for the study of bone dynamics during growth and activity. Further research is necessary to quantitatively measure the morphology of ELPs and to identify any co‐variance between ELP characteristics and age, sex or indicators of mechanical loading. Copyright © 2010 John Wiley & Sons, Ltd.     

A comparative experiment in the consolidation of cremated bone

This experiment assessed whether consolidation of cremated remains facilitated cross‐sectioning and grinding, which often are required for microstructural analyses such as the histomorphometric estimation of age and the examination of pathological lesions. Green, bovine femoral diaphyses were cremated at three temperature ranges (346–357°C; 610–755°C; 684–838°C) in an electric furnace and then consolidated with Acryloid B‐72 or Butvar B‐98 using simple brushing and immersion techniques. After consolidation was complete, bone pieces were cross‐sectioned and ground to a thickness of 100–120 μm. Results indicate that specimens treated with Acryloid B‐72 are harder and stronger than Butvar treated and untreated control specimens. Acryloid B‐72 is, therefore, recommended for the consolidation of cremated remains in preparation for microstructural analysis. Copyright © 2004 John Wiley & Sons, Ltd.     

High Resolution Stratigraphic Distribution of Coprolites within Eneolithic Middens,a Case Study: Hârsova-Tell (Constanta County,Southeast Romania)

Abstract

Quantitative distribution of coprolites, fish bones and mammal bones recovered by wet screening from an Eneolithic midden at Hârsova-tell (southeast Romania) is surveyed within a high resolution stratigraphic framework. The coprolites, all containing bone fragments, are attributed to carnivores/omnivores: dog, pig, man. Highly significant correlation (Spearman's coefficient of rank correlation r_s = 0.46, P < 0.0001) between coprolite distribution and the seasonality-related fish bone distribution suggests that seasonality controls the stratigraphic distribution of coprolites recovered by wet screening within middens. Periods of high fish bone input into the midden, correlated with frequent coprolite occurrences, correspond to the warm season (late spring to early autumn). Assessment of potential factors controlling coprolite distribution within the midden suggests that sections of high coprolite concentration reflect preservation of faeces until embedding in sediments, and/or rapid embedding of faeces. Preservation of faeces is favored by dry periods, characteristic of the summer season in the highly continental climate of the region. Rain, freeze-thaw cycles or other wet season processes favor faeces disintegration. Our results suggest that the stratigraphic distribution of coprolites obtained by wet screening may represent an indicator of seasonality in middens. Additional studies are needed to better characterise this relationship and refine interpretations.     

脆弱纸张加固材料研究

作为一种天然的有机材料,纸张在环境因素的影响下很容易老化。对于脆弱纸张,可通过加固来提高纸张的耐久性。本研究对羟丙基甲基纤维素、有机硅改性丙烯酸脂、有机氟树脂在加固脆弱纸张方面的应用进行了研究。结果表明,羟丙基甲基纤维素(HPMC)能有效改善纸张强度,不改变纸张的外表,不发粘不发黄,耐老化性好,不影响以后的再处理。     

The risk of harm to archaeological artefacts in soil from dynamic subsurface pressures generated by agricultural operations: experimental studies

An investigation was made of the fracture risk to buried ceramic pots and bones from the subsurface pressures generated by agricultural operations. A method for generating subsurface pressures at 0.25 m depth in a soil bin was developed and used to explore fracture failure of modern replicates of ceramic pots corresponding to different archaeological periods and aged medieval human bone. Application of conductive traces to the objects’ surfaces allowed detection of fracturing in real‐time without excavation. Predictive models relating fracture of objects to subsurface pressure application were developed by relating the probability of fracture to observed survival statistics at increasing subsurface pressures. Subsurface pressures of above 100 kPa were sufficient to fracture the more fragile pots at a probability level of 90%. The lowest subsurface pressure causing bone fracture was 280 kPa; however, no relationship was observed between subsurface pressure and bone fracture, probably due to variation in bone material. The subsurface pressures generated and applied are within the range reported from field measurements during typical agricultural operations. They indicate that measures to control the generation of subsurface pressures would be necessary to mitigate risk of harm from agriculture to buried archaeology; for example, via low‐pressure tyres, tracks and appropriate tillage.     

Accessory navicular: A heritable accessory bone of the human foot

The accessory navicular is a supernumerary bone of the human foot located medial to the navicular tuberosity and represents a secondary center of ossification that has failed to fuse to the main body of the navicular. Three forms of the accessory bone have been identified: Type I is an independent ossicle that is often embedded within the tibialis posterior tendon; Type II is a triangular accessory bone that attaches to the navicular tuberosity by means of a cartilaginous or fibrocartilaginous bridge; and Type III represents a fused Type II, which forms a hook‐like protuberance extending from the tuberosity. The Type II accessory navicular is the most common of the three forms and is the most readily identifiable in skeletal material since it causes the navicular tuberosity to become abnormally flattened and porous. The purpose of this study was to describe the various manifestations of the accessory navicular in dry bone, to present and compare frequency data for several skeletal populations, and to consider questions of laterality and sex bias in trait expression. In total, the skeletons of 497 Danes, 460 Euro‐Americans, 300 African Americans, 100 Japanese, and 205 Europeans were examined for the presence of the Type II accessory navicular. Overall frequencies for the five groups ranged from 2% in the African American sample to 5% in the Japanese sample. Since several family pedigrees have documented the accessory navicular as being an inherited skeletal defect, the relatively low frequency found in the present study makes this trait a potentially useful indicator of genetic relatedness within archaeological cemeteries. In addition, the trait was found to occur more often unilaterally than bilaterally and there was nearly equal incidence among males and females. Copyright © 2010 John Wiley & Sons, Ltd.     

MULTIANALYTICAL STUDY OF PALAEOLITHIC REINDEER ANTLER. DISCOVERY OF ANTLER TRACES IN LASCAUX PIGMENTS BY TEM*

Palaeolithic and modern reindeer antler were analysed using complementary techniques to evaluate the conservation state of bone material. X‐ray diffraction, FT‐IR spectroscopy, scanning electron microscopy (SEM–EDX) and transmission electron microscopy (TEM–EDX) as well as proton induced X‐ray and γ‐ray emission (PIXE–PIGE) at the particle accelerator AGLAE, C2RMF were applied. This study enabled us to define the chemical and structural features of ancient antler at the micro‐ and nanometre scale and compare them to those of other bone materials such as bone and ivory. Antler is richer in its organic fraction compared to bone and ivory. Its mineral fraction, carbonated hydroxylapatite, shows very specific crystal shapes at the nanometre scale. This specific property allowed us to recognize antler traces in pigment samples originating from rock art paintings in the Lascaux cave, Dordogne, southwestern France, Magdalenian period. Therefore, TEM–EDX is the technique of choice for characterizing antler traces in complex mixtures. In the paint material of Lascaux, antler seems most likely to be a pollutant introduced either by stirring the pigments in water with a piece of antler or by carving antler artefacts next to the preparation of paint material. However, it could eventually be used as a marker of paintings that were created contemporaneously.     

Magdalenian Children: Projectile Points,Portable Art and Playthings

Children, no doubt, were a significant component of Upper Palaeolithic societies. Despite this fact, however, serious identification and consideration of material culture which may have belonged to children – at least at one time during their use‐life – have not been undertaken. This situation extends to the best represented and most intensively studied of the European Palaeolithic techno‐complexes, the Magdalenian (c.21,000–14,000 cal BP), and consequently, we know very little about the children of this enigmatic people. As play, including object play, is a ‘true cultural universal’, we can be certain that Magdalenian children integrated objects into their games, with these playthings later incorporated into the archaeological record. Through examining ethnographic accounts of recent hunter‐gatherer children and reconsidering archaeological assemblages in light of these data, this paper suggests that Magdalenian playthings probably included full‐sized adult weapon tips and – more significantly – pieces of what archaeologists term ‘art mobilier’.     

SYNCHROTRON FTIR MICRO‐SPECTROSCOPY APPLIED TO THE STUDY OF POLISHED SERPENTINITE ARTEFACTS: A NON‐DESTRUCTIVE ANALYTICAL APPROACH

In prehistory, serpentinite was one of the most frequently used raw materials to produce polished stone artefacts. Several conventional analytical techniques can be applied to identify the serpentine minerals, but their application generally requires a powdered sample. This implies that the artefacts to be analysed must be damaged, and the possibility of analysing a mixture of different serpentine polytypes is high. The use of spatially resolved techniques is therefore a necessity to overcome this problem. Several thin sections of serpentinitic rocks and prehistoric axes have been analysed by synchrotron radiation Fourier transform infrared micro‐spectroscopy. The spectra were acquired directly on specific points of polished stone artefacts and this allowed the recognition of the different polytypes of serpentine minerals without causing any damage to the objects. The results show the infrared micro‐spectroscopy technique to be a useful tool for the characterization of archaeological lithic material.     

Was He Murdered or Was He Not?—Part II: Multi‐Elemental Analyses of Hair and Bone Samples from Tycho Brahe and Histopathology of His Bones

Hair and bone samples procured from the remains of Tycho Brahe were analysed by several analytical techniques. In segmented hair samples, concentrations of Fe, As, Ag and Au at the tips exceeded values for the contemporary population; however, they decreased towards the hair bulbs, similarly to Hg, indicating that recent exposure that was discontinued ~2 months prior to Brahe's death. Several other elements did not follow this pattern. Analyses of bones revealed signs of long‐term exposure to Au, while many other elements were within expected ranges. Histopathological examination of bone sections yielded no signs of severe bone metabolic disorders.     

The application of 3D‐microprofilometry as a tool in the surface diagnosis of fossil and sub‐fossil vertebrate hard tissue. An example from the Pliocene Upper Laetolil Beds,Tanzania

Non‐contact optical 3D‐profiling instruments are often used in the study of surface modifications on fossil mammalian bone. The advantage of optical laser scanning for the study of fossil and sub‐fossil bone is its non‐contact nature, allowing the investigation of fragile and poorly preserved surfaces. The high resolution and fast measuring rate of this method make it an alternative to scanning electron microscopy (SEM) investigation if topography is to be visualized. This study analyses clusters of incisions in a suid humeral fragment from the Pliocene Upper Laetolil Beds. The marks show a characteristic crest structure that is also frequently found in mandible marks produced by the Australian termite species Mastotermes darwiniensis. The marks from the Laetolil Beds are, therefore, interpreted as also being caused by insect mandible action. An as yet unknown large insect species capable of modifying bone with their mandibles is thus postulated in the palaeohabitat represented by the hominid‐bearing Upper Laetolil Beds. Copyright © 2001 John Wiley & Sons, Ltd.