Human Bone Collagen Research Articles

Light Stable Isotope Analysis of Diet in Corded Ware Culture Communities: Święte, Jarosław District, South-Eastern Poland

Abstract The presented study was based on isotopic analysis of δ13C and δ15N in human bone collagen samples from graves of the Corded Ware culture in Święte, south-east Poland. Isotopic values demonstrate a relatively narrow variation, ranging from -20.4‰ to -19.8‰ and 10.6‰ to 12.0‰ for δ13C and δ15N values, respectively. The diet was likely C3 plant-based with a substantial animal protein component, including predominantly terrestrial and possibly riverine resources.

Variability along the frontier: stable carbon and nitrogen isotope ratio analysis of human remains from the Late Roman–Early Byzantine cemetery site of Joan Planells, Ibiza, Spain

Carbon (δ13C) and nitrogen (δ15N) stable isotope analysis of human bone collagen from 38 individuals was undertaken to assess diet at the Late Roman–Early Byzantine (AD 300–700) cemetery site, Joan Planells, in Ibiza, Spain. The results (δ13C = − 18.7 ± 0.5‰ and δ15N = 10.1 ± 1.3‰) show that the diet of this population was derived predominantly from C3 terrestrial resources; plant foods were likely dietary staples along with meat and/or dairy produce comprising an important component of diet. Variation in stable isotope ratio values suggests individual differences in diet. Two individuals, both males, are statistical outliers with distinctive δ15N values (14.4 and 14.8‰) that point to significant consumption of marine resources. Females, on average, have higher δ13C values than males. The parsimonious explanation for this observation is the greater inclusion of C4 resources such as millet in the diets of females. Comparison of the diet of the Joan Planells population with other Late Roman period sites on the Hispanic mainland and other parts of the Mediterranean region suggests that populations may have been responding to a combination of socio-political and environmental factors that could have included Roman influence of food consumptive practices in some of these distant locales.

Mobility in the Atacama Desert, northern Chile, in the Late Intermediate Period (AD 900–1450): A re-evaluation using stable isotope analysis

Research on the Late Intermediate Period (AD 900–1450) in northern Chile has been strongly influenced by two mobility models: John Murra's classic vertical ecological archipelago model, with highland colonies in the lowlands; and the gyratory mobility model, with pastoralists and their llama caravans controlling trade. The widespread application of these two models, however, suffers from a lack of supporting archaeological evidence. Stable isotope analysis provides a more direct approach for studying both diet and mobility patterns, complementing material culture studies. The aim of this paper is to evaluate the presence of colonies and/or specialised highland caravans in the lowlands of northern Chile through the application of δ13C, δ15N and δ18O to human tooth enamel, bone collagen and bone apatite. The results show strong local traditions in terms of diet and mobility associated with the Loa-San Pedro, Antofagasta and Pica-Tarapacá cultural groups. Our results suggest a non-local origin for some individuals in Pica-Tarapacá and Loa-San Pedro, but provide little evidence to support either the presence of ‘colonies’ or of specialised ‘caravans’. Mobility patterns were, instead, diverse and flexible, including female and male individuals that moved at different moments of their lives.

Relationships between human cortical bone toughness and collagen cross-links on paired anatomical locations.

Human cortical bone fracture processes depend on the internal porosity network down to the lacunar length scale. Recent results show that at the collagen scale, the maturation of collagen cross-links may have a negative influence on bone mechanical behavior. While the effect of pentosidine on human cortical bone toughness has been studied, the influence of mature and immature enzymatic cross-links has only been studied in relation to strength and work of fracture. Moreover, these relationships have not been studied on different paired anatomical locations. Thus, the aim of the current study was to assess the relationships between both enzymatic and non-enzymatic collagen cross-links and human cortical bone toughness, on four human paired anatomical locations. Single Edge Notched Bending toughness tests were performed for two loading conditions: a quasi-static standard condition, and a condition representative of a fall. These tests were done with 32 paired femoral diaphyses, femoral necks and radial diaphyses (18 women, age 81 ± 12 y.o.; 14 men, age 79 ± 8 y.o.). Collagen enzymatic and non-enzymatic crosslinks were measured on the same bones. Maturation of collagen was defined as the ratio between immature and mature cross-links (CX). The results show that there was a significant correlation between collagen cross-link maturation and bone toughness when gathering femoral and radial diaphyses, but not when considering each anatomical location individually. These results show that the influence of collagen enzymatic and non-enzymatic cross-links is minor when considering human cortical bone crack propagation mechanisms.

Did military orders influence the general population diet? Stable isotope analysis from Medieval Tomar, Portugal

This study integrates bone collagen stable isotope data (carbon, nitrogen and sulphur) from 33 human adult tibiae (15 females; 18 males) and 13 faunal remains from Tomar, while it was under the Military Orders domain (eleventh–seventeenth centuries). Historical literature indicates that the amount of meat consumption amongst Templars was lower than in individuals with similar social status. In Medieval times, these Military Orders had total control of towns and angling and fishing rights, but their influence on the general population diet remains unknown. While no statistically significant differences (p > 0.05) were found between sexes, social status, or for bone collagen δ13C and δ34Sbetween age groups, δ15N did differ significantly with age, which may be related to tooth loss in old individuals. Additionally, the human samples have higher stable isotope differences, in comparison with faunal samples, than would be expected within the food web, particularly for δ13C. This human bone collagen δ13C enrichment may reflect a diet rich in aquatic protein intake, which is also supported by δ34S archived in human and faunal samples, and the presence of oysters and cockles shells at the excavation. The religious diet restrictions might have led to a higher intake of aquatic protein when meat consumption was not allowed.

Estimating C4 plant consumption in Bronze Age Northeastern Italy through stable carbon and nitrogen isotopes in bone collagen

AbstractThe application of biomolecular techniques for the study of food practices in the Italian Bronze Age has revealed an interesting complexity. This is particularly true for the Po plain, in northern Italy, where the use of “alternative” grains (i.e., the millets) has been assessed isotopically through the measurement of stable carbon (δ13C) and nitrogen (δ15N) isotope ratios in human and animal bone collagen at the site of Olmo di Nogara (Verona). This work provides new isotopic data from 12 Early, Middle, and Late Bronze Age sites from western Veneto and Friuli. Data obtained contribute to the understanding of mode and tempo of the spread of new crops in northeastern Italy, which appears to be a hotspot for the study of Bronze Age farming economies in Europe. We have successfully analyzed 146 specimens to find that δ13C values are higher, in a way that we interpret as indicative of C4 plant consumption. Four of the sites analyzed, with a specific chronological indication, show this evidence. We explain this phenomenon as the result of a discontinuous spread of new crops in northeastern Italy at a very specific moment of the Bronze Age. The data presented might call for a reconsideration of food production and consumption among Bronze Age groups of southern Europe.

Marine resource reliance in the human populations of the Atacama Desert, northern Chile – A view from prehistory

The Atacama Desert is one of the most inhospitable terrestrial environments on Earth, yet the upwelling of the Humboldt Current off the coast has resulted in the presence of a rich marine biota. It is this marine environment which first enabled the human settlement of the northern Atacama Desert, and continues to form the basis of regional economies today. In this paper we explore how the desert has shaped human dietary choices throughout prehistory, using carbon and nitrogen isotope analysis of human bone collagen (n = 80) to reconstruct the diets of the inhabitants of the Arica region of the northern Atacama. This area is one of the driest parts of the desert, but has been generally understudied in terms of dietary adaptation. Statistical analysis using FRUITS has allowed deconvolution of isotopic signals to create dietary reconstructions and highlight the continued importance of marine resources throughout the archaeological sequence. Location also appears to have played a role in dietary choices, with inland sites having 10–20% less calories from marine foods than coastal sites. We also highlight evidence for the increasing importance of maize consumption, coinciding with contact with highland polities. In all periods apart from the earliest Archaic, however, there is significant variability between individuals in terms of dietary resource use. We conclude that marine resource use, and broad-spectrum economies persisted throughout prehistory. We interpret these results as reflecting a deliberate choice to retain dietary diversity as a buffer against resource instability.

Palaeodietary Reconstruction of 6th Century Naju Yeongdong‐ri People Recorded in Stable Carbon and Nitrogen Isotope Analysis of Human Bone Collagen

뼈에서 추출된 탄소와 질소 ì•ˆì •ë™ìœ„ì›ì†Œ ì •ë³´ëŠ” 과거 사람들의 식생활, 영양상태, ìƒê³„ê²½ì œì™€ ê³ í™˜ê²½ 등을 연구하는데 매우 중요한 자료이다. 본 연구에서는 6세기대 나주 영동리 ê³ ë¶„êµ° ì œ1호분 돌방무덤과 돌덧널무덤에서 ì¶œí† ëœ 옛사람 뼈 9개체에서 추출된 콜라겐의 ì•ˆì •ë™ìœ„ì›ì†Œ 분석을 실시하였다. 탄소와 질소 ì•ˆì •ë™ìœ„ì›ì†Œ 결과는 다음과 같다(δ13C= -19.5±0.7‰, δ15N= 9.6±0.7‰, (n=9)). 탄소 ì•ˆì •ë™ìœ„ì›ì†Œ 분석 ê²°ê³¼ 나주 영동리 ê³ ë¶„êµ° 피장자 집단은 ë²¼, 보리, 콩 등 C3 작물 위주의 섭취를 í•˜ì˜€ë‹¤ê³ ì¶”ì •ë˜ë©°, 질소 ì•ˆì •ë™ìœ„ì›ì†Œ 값으로부터 동물성 단백질은 주로 육상 동물로부터 ì–»ì—ˆë‹¤ê³ ì¶”ì •ëœë‹¤. 이 결과로부터 ë°±ì œì‹ 무덤 양식을 ê°–ê³ ì˜ì‚°ê°•ìœ ì—­ í† ì°© ì„¸ë ¥ì˜ 매장 양식이 보이는 피장자 집단의 식생활 양상을 í™•ì¸í• ìˆ˜ 있었다. 중심어: 사람뼈, 뼈 콜라겐, 탄소 ·질소 ì•ˆì •ë™ìœ„ì›ì†Œ, 식생활, 나주 영동리 ê³ ë¶„êµ°

Early indicators to C4 plant consumption in central Kazakhstan during the Final Bronze Age and Early Iron Age based on stable isotope analysis of human and animal bone collagen

In this paper, we present new stable isotope data from central, southern and eastern Kazakhstan (KZ) that date to the Early Iron Age. Our primary data together with results from previously published studies demonstrate that the consumption of C4 plants, possibly millet, started in the Final Bronze Age in central KZ and continued into the Early Iron Age. Data from southern KZ, however, demonstrate that over half the human population consumed C4 crops in the Early Iron Age as opposed to the central regions of KZ, where just a few individuals within the population, often males buried in elite kurgans, have high δ13C values indicative of C4 plant consumption. In this paper we aim first to understand if any dietary changes can be seen in the central KZ population during the transitional period between the Bronze and Iron Ages; secondly, we investigate the extent of C4 plant consumption in central KZ during these time periods. Here we present new human isotopic data from nine central sites of the Tasmola culture (n=11), two eastern KZ sites (n=3) and two southern KZ sites (n=26).

Social status in late medieval and early modern Brittany: insights from stable isotope analysis

We document for the first time the diet of a privileged French population from Brittany, a region that was the center of battles between the Kingdoms of England and France until the end of the fifteenth century. We present here the results of stable isotope analyses of carbon, nitrogen, and sulfur of human and animal bone and tooth collagen for a late medieval to early modern Breton population. The isotopic values observed for the Dominican convent of Rennes, Brittany, are very similar to those reported for medieval archaeological populations in Great Britain, namely they have enriched δ15N values combined with almost entirely terrestrial carbon signals. We discuss the consumption of young animals in a diet made up of terrestrial, marine, and freshwater resources. We report dietary differences between socio-economic groups and gender, with women and nobles (male and female) showing patterns consistent with high animal product consumption and lower mobility. The S isotope ratios of both humans and fauna are very homogeneous and generally have coastal δ34S values. The convent is known to have been an interregional pilgrimage site during the early modern period, but the isotope values indicate that the identified migrants were not pilgrims. Stable isotope analysis therefore complements the available historical information on human diets and mobility.

The effect of ribose pre-treatment of cortical bone on γ-irradiation sterilization effectiveness.

Reconstruction of large skeletal defects is a significant and challenging issue. Tissue banks often use γ-irradiation (15-35kGy) to sterilize bone allografts, which, however, drastically impairs the post-yield mechanical properties. In previous studies, we reported the development of a method that protects human bone collagen connectivity through ribose crosslinking while still undergoing γ-irradiation. Given these promising results, the next step was to determine if the presence of ribose within the bone tissue would interfere with the effectiveness of the γ-irradiation sterilization against bacteria. This study had two stages. The aim of the first stage was to assess the protective effect of ribose in solution using a Bacillus pumilus spore strip model. The aim of the second stage was to assess the protective effect of ribose (R) on spores within a human cortical bone model in comparison to conventionally irradiated bone (I). Treatment of B. pumilus spore strips with ribose in solution led to temperature-dependent effects on spore viability versus spore strips treated with PBS alone. Ribose solution at 60°C led to a notable two logs decrease in spore count relative to PBS at 60°C. In the human bone model, the number of spores in the I and R groups were greatly decreased in comparison to the non-irradiated N group. No spore colonies were detected in the R group (n=4) whereas two of the four plates of group I formed colonies. This study provides evidence that the method of pre-treating bone with ribose crosslinking prior to irradiation sterilization, while improving irradiation sterilized bone allograft quality, also may improve the effectiveness of the sterilization process.

Using Stable Isotopes and a Bayesian Mixing Model (FRUITS) to Investigate Diet at the Early Neolithic Site of Carding Mill Bay, Scotland

AbstractWe present δ13C, δ15N, and δ34S measurements on archaeological human and animal bone collagen samples from a shell midden dating to the Neolithic ca. 4000–3500 cal BC, together with measurements on modern fish and shellfish. These data were used in conjunction with the Bayesian mixing model, Food Reconstruction Using Isotopic Transferred Signals (FRUITS), to reconstruct human diet at the site. We demonstrate the importance of using a geographically appropriate faunal baseline in stable isotope paleodietary studies, and suggest that Neolithic individuals at this site consumed up to ca. 21% of dietary protein from marine resources, despite stable isotope ratios that imply a wholly terrestrial diet. This marine resource consumption does not significantly shift the radiocarbon (14C) dates of these individuals, so although we must consider the use of marine resources at the site, the chronology that has previously been established is secure. The δ13C and δ15N measurements from the archaeological herbivore bone collagen indicate that it is unlikely they ate plants enriched with fertilisers such as manure or seaweed. The δ34S values reveal a sea-spray effect; therefore, in this instance, δ34S cannot be used as a dietary indicator but can be used to demonstrate the likely locality of the fauna.

Diet uniformity at an early farming community in northwest Anatolia (Turkey): carbon and nitrogen isotope studies of bone collagen at Aktoprakl\u0131k

Aktopraklık is a settlement site composed of three areas (A–C) in the Marmara region of northwest Anatolia, with phases of occupation that date to the Late Neolithic and Early Chalcolithic periods, mid-seventh to mid-sixth millennium bc (ca. 6400–5600 cal. bc). Here, we present 54 human and fauna bone collagen stable isotope results from the site, alongside five modern fish bone collagen isotope results, to examine the nature of human diet. The stable isotope analysis shows that human diet comprised the consumption of select C3 terrestrial resources, with a preference for domestic animal proteins over plant proteins. The evidence to date suggests that animal husbandry was at the forefront of Late Neolithic and Early Chalcolithic subsistence practices. No isotopic difference in humans is observed between biological sex or between areas B and C at the settlement.

Development of a novel method for the strengthening and toughening of irradiation-sterilized bone allografts.

Reconstruction of large skeletal defects is a significant and challenging issue. Bone allografts are often used for such reconstructions. However, sterilizing bone allografts by using γ-irradiation, damages collagen and causes the bone to become weak, brittle and less fatigue resistant. In a previous study, we successfully protected the mechanical properties of human cortical bone by conducting a pre-treatment with ribose, a natural and biocompatible agent. This study focuses on examining possible mechanisms by which ribose might protect the bone. We examined the mechanical properties, crosslinking, connectivity and free radical scavenging potentials of the ribose treatment. Human cortical bone beams were treated with varying concentration of ribose (0.06-1.2M) and γ-irradiation before testing them in 3-point bending. The connectivity and amounts of crosslinking were determined with Hydrothermal-Isometric-Tension testing and High-Performance-Liquid-Chromatography, respectively. The free radical content was measured using Electron Paramagnetic Resonance. Ribose pre-treatment improved the mechanical properties of irradiation sterilized human bone in a pre-treatment concentration-dependent manner. The 1.2M pre-treatment provided >100% of ultimate strength of normal controls and protected 76% of the work-to-fracture (toughness) lost in the irradiated controls. Similarly, the ribose pre-treatment improved the thermo-mechanical properties of irradiation-sterilized human bone collagen in a concentration-dependent manner. Greater free radical content and pentosidine content were modified in the ribose treated bone. This study shows that the mechanical properties of irradiation-sterilized cortical bone allografts can be protected by incubating the bone in a ribose solution prior to irradiation.

Fabrication and Characterization of Three-Dimensional (3D) Core-Shell Structure Nanofibers Designed for 3D Dynamic Cell Culture.

Three-dimensional elastic nanofibers (3D eNFs) can offer a suitable 3D dynamic microenvironment and sufficient flexibility to regulate cellular behavior and functional protein expression. In this study, we report a novel approach to prepare 3D nanofibers with excellent mechanical properties by solution-assisted electrospinning technology and in situ polymerization. The obtained 3D eNFs demonstrated excellent biocompatible properties to meet cell culture requirements under a dynamic environment in vitro. Moreover, these 3D eNFs also promoted human bone marrow mesenchymal stem cells (hMSCs) adhesion and collagen expression under biomechanical stimulation. The results demonstrated that this dynamic cell culture system could positively impact cellular collagen but has no significant effect on the proliferation of hMSCs grown in the 3D eNFs. This work may give rise to a new approach for constructing a 3D cell culture for tissue engineering.

Reconstructing diet at the Neolithic stalled cairn of the Knowe of Rowiegar, Rousay, Orkney, using stable isotope analysis

In this study, human remains from the Neolithic stalled cairn of the Knowe of Rowiegar, Rousay, Orkney (3620–2880cal BC, 95.4% probability), were analysed for bone collagen stable carbon (δ13C) and nitrogen (δ15N) isotope ratios in order to determine the dietary adaptations of individuals buried at the site, particularly the contribution of marine protein in the diet. Collagen was extracted from bone from 13 individuals (11 males, 1 female, and 1 sub-adult), and stable isotope data generated were compared with previously-published Neolithic Orcadian faunal data, and with human and animal bone collagen isotope data from other published British Neolithic sites. The results from the Knowe of Rowiegar suggest that the dietary protein of those buried at the site was largely terrestrial in origin, which is similar to other British Neolithic bone collagen datasets, albeit with the possible minor inclusion of marine protein. Intra-group comparison highlights the potentially different dietary habits of the single female and sub-adult individuals sampled from the site compared to the interred males. Geographical variations in both humans and animals (particularly in nitrogen isotope ratios) across Britain are examined, and the consumption of marine fish and the influence of herbivore baseline variability in the study of Neolithic human diet are explored.

Modeling middle Neolithic funnel beaker diet on Falbygden, Sweden

In this paper, a quantitative reconstruction of Middle Neolithic diet in the area of Falbygden, Sweden, is attempted. The reconstruction is based on carbon and nitrogen isotope values for human bone collagen in the passage grave at Frälsegården as well as on a series of animal and cereal isotope measurements from the same area and period. The Bayesian software package FRUITS is employed, resulting in estimates of the caloric contribution of plants, terrestrial animals, and fish to the diet.Two models were developed, one based on carbon and nitrogen isotope values in adult bone collagen and one based on carbon and nitrogen isotopes in tooth enamel and collagen.The estimates from both models are similar, suggesting a high caloric contribution from plants and a smaller one from terrestrial animals while the contribution from freshwater fish is marginal. These results contradict some earlier views regarding Neolithic economy on Falbygden and suggests that Funnel Beaker society in this region was heavily reliant on cultivation, rather than a pastoral economy with only marginal importance of cereals.

Dietary reconstruction of the Iron Age population at the Fantzuyuan site, Taiwan, revealed by isotopic analysis on human and faunal bone collagen

In this study, we analyzed the carbon and nitrogen stable isotopic composition of human bone collagen in 33 individuals found at the Fantzuyuan site in Taiwan in order to investigate the dietary patterns of this Iron Age group. Forty-three faunal collagen samples were also analyzed to ascertain the variability of baseline isotopic signatures in the area. Mean δ13C and δ15N values of −12.5±0.7‰ and 8.1±0.5‰, respectively, were found in 26 human individuals. In conjunction with archaeological evidence, this study showed that human diet at this site derived mainly from terrestrial animals, with a minor component derived from marine shellfish. No significant difference in isotopic compositions was detected between male and female adults or between adults and juveniles. However, six individuals had dietary patterns that were different from others, which probably reveals that they had special social status and/or non-local origins.

Assessment of collagen quality associated with non-enzymatic cross-links in human bone using Fourier-transform infrared imaging

Aging and many disease conditions, most notably diabetes, are associated with the accumulation of non-enzymatic cross-links in the bone matrix. The non-enzymatic cross-links, also known as advanced glycation end products (AGEs), occur at the collagen tissue level, where they are associated with reduced plasticity and increased fracture risk. In this study, Fourier-transform infrared (FTIR) imaging was used to detect spectroscopic changes associated with the formation of non-enzymatic cross-links in human bone collagen. Here, the non-enzymatic cross-link profile was investigated in one cohort with an in vitro ribose treatment as well as another cohort with an in vivo bisphosphonate treatment. With FTIR imaging, the two-dimensional (2D) spatial distribution of collagen quality associated with non-enzymatic cross-links was measured through the area ratio of the 1678/1692cm−1 subbands within the amide I peak, termed the non-enzymatic crosslink-ratio (NE-xLR). The NE-xLR increased by 35% in the ribation treatment group in comparison to controls (p<0.005), with interstitial bone tissue being more susceptible to the formation of non-enzymatic cross-links. Ultra high-performance liquid chromatography, fluorescence microscopy, and fluorometric assay confirm a correlation between the non-enzymatic cross-link content and the NE-xLR ratio in the control and ribated groups. High resolution FTIR imaging of the 2D bone microstructure revealed enhanced accumulation of non-enzymatic cross-links in bone regions with higher tissue age (i.e., interstitial bone). This non-enzymatic cross-link ratio (NE-xLR) enables researchers to study not only the overall content of AGEs in the bone but also its spatial distribution, which varies with skeletal aging and diabetes mellitus and provides an additional measure of bone's propensity to fracture.

Dietary patterns in human populations from northwest Patagonia during Holocene: an approach using Binford’s frames of reference and Bayesian isotope mixing models

The goal of this paper is to assess the variation in the proportional contribution of diverse resources to the diet of human populations from northwest Patagonia (Argentina) throughout the Middle-Late Holocene. Particularly, we assessed the variation among three geographic areas and two periods. We first estimated the expected proportions of terrestrial animals and plants and aquatic resources for each area according to the Binford’s frames of references approach. A Bayesian mixing method was then applied to calculate the proportion of plants and animals in the diets from stable isotopes (δ13C and δ15N) of human bone collagen. The isotope values suggest that the composition of diets differed spatially and temporally. Diets of South Mendoza were mainly composed of terrestrial animals (Rhea-Lama and rodents) with a greater incorporation of C3 plants towards the later Late Holocene; in North Neuquen, Rhea and Lama represent a proportion of 0.84 of the diet consumed; and finally, the sample of Center Neuquen is the only one with high values of Araucaria in the diet. The isotopic values obtained for the three studied areas did not fit to the expectations of Binford’s model, North Neuquen being the area that departs most from the predicted proportions of terrestrial animals and plants and aquatic organisms in the diet. These findings open up new questions about the local conditions that influenced regional variation in the diet of prehistoric hunter-gatherers.