Preclinical rodent models for human bone disease including a focus on cortical bone.

Preclinical models (typically the ovariectomized rat and genetically altered mice) have underpinned much of what we know about skeletal biology. They have been pivotal for developing therapies for osteoporosis and monogenic skeletal conditions, including osteogenesis imperfecta, achondroplasia, hypophosphatasia, and craniodysplasias. Further therapeutic advances, particularly to improve cortical strength, requires improved understanding and more rigorous use and reporting. We describe here how trabecular and cortical bone structure develop, are maintained, and degenerate with ageing in mice, rats, and humans, and how cortical bone structure is changed in preclinical models of endocrine conditions (e.g., postmenopausal osteoporosis, chronic kidney disease, hyperparathyroidism, diabetes). We provide examples of preclinical models used to identify and test current therapies for osteoporosis, and discuss common concerns raised when comparing rodent preclinical models to the human skeleton. We focus especially on cortical bone, because it differs between small and larger mammals in its organizational structure. We discuss mechanisms common to mouse and human controlling cortical bone strength and structure, including recent examples revealing genetic contributors to cortical porosity and osteocyte network configurations during growth, maturity, and ageing. We conclude with guidelines for clear reporting on mouse models with a goal for better consistency in the use and interpretation of these models.

Vitamin D status in post-medieval Northern England: Insights from dental histology and enamel peptide analysis at Coach Lane, North Shields (AD 1711-1857).

OBJECTIVES: The post-medieval period in Europe saw a dramatic increase in metabolic bone disease related to vitamin D deficiency (VDD). Recent paleopathological work has utilized interglobular dentin (IGD) as a proxy for poor vitamin D status during development, while enamel peptide analysis allows the identification of chromosomal sex in non-adult remains. Here we explore the relationship between sex, the presence of IGD, and macroscopic markers of VDD in an industrial era assemblage from Northeast England. MATERIALS AND METHODS: 25 individuals (9 females, 9 males, 9 unknown sex) from the cemetery site at Coach Lane, North Shields (1711-1857) were selected for paleopathological analysis, histological assessment of IGD, and enamel peptide determination of chromosomal sex. RESULTS: Ground tooth sections from 21 individuals were of suitable quality for detection of IGD, and enamel peptide analysis confirmed the chromosomal sex of ten individuals. Sixteen individuals (76.1%) exhibited ≥1 episode of IGD. Nine of these (42.8%) exhibited >1 episode and four (19%) exhibited ≥4 episodes in regular intervals. Male sex was significantly associated with the presence of IGD (p = 0.0351; 100% males vs. 54.5% females). Females were more likely to exhibit macroscopic evidence of VDD (45.5% females vs 30% males) but this was not statistically significant. DISCUSSION AND CONCLUSIONS: Periods of poor mineral metabolism during childhood appear much more prevalent at Coach Lane than macroscopic evidence suggests. Evidence of seasonal IGD episodes indicates that northern latitude played a major role in poor VD status in the Northeast of England. The significant association of IGD with male sex may be due to sex-related differences in dentinal mineralization or a higher risk of poor VD status in males aged <5 years. More work is needed to establish an evidence-based threshold for pathological levels of IGD before the presence of this feature can confidently be used as a biomarker for poor VD status.

Human femur morphology and histology variation with ancestry and behaviour in an ancient sample from Vietnam.

BACKGROUND: There is a genetic component to the minimum effective strain (MES)-a threshold which determines when bone will adapt to function-which suggests ancestry should play a role in bone (re)modelling. Further elucidating this is difficult in living human populations because of the high global genetic admixture. We examined femora from an anthropological skeletal assemblage (Mán Bac, Vietnam) representing distinct ancestral groups. We tested whether femur morphological and histological markers of modelling and remodelling differed between ancestries despite their similar lifestyles. METHODS: Static histomorphometry data collected from subperiosteal cortical bone of the femoral midshaft, and gross morphometric measures of femur robusticity, were studied in 17 individuals from the Mán Bac collection dated to 1906-1523 cal. BC. This assemblage represents agricultural migrants with affinity to East Asian groups, who integrated with the local hunter-gatherers with affinity to Australo-Papuan groups during the mid-Holocene. Femur robusticity and histology data were compared between groups of 'Migrant' (n = 8), 'Admixed' (n = 4), and 'Local' (n = 5). RESULTS: Local individuals had more robust femoral diaphyses with greater secondary osteon densities, and relatively large secondary osteon and Haversian canal parameters than the migrants. The Migrant group showed gracile femoral shafts with the least dense bone made up of small secondary osteons and Haversian canals. The Admixed individuals fell between the Migrant and Local categories in terms of their femoral data. However, we also found that measures of how densely bone is remodelled per unit area were in a tight range across all three ancestries. CONCLUSIONS: Bone modelling and remodelling markers varied with ancestral histories in our sample. This suggests that there is an ancestry related predisposition to bone optimising its metabolic expenditure likely in relation to the MES. Our results stress the need to incorporate population genetic history into hierarchical bone analyses. Understanding ancestry effects on bone morphology has implications for interpreting biomechanical loading history in past and modern human populations.

Female bone physiology resilience in a past Polynesian Outlier community.

Remodelling is a fundamental biological process involved in the maintenance of bone physiology and function. We know that a range of health and lifestyle factors can impact this process in living and past societies, but there is a notable gap in bone remodelling data for populations from the Pacific Islands. We conducted the first examination of femoral cortical histology in 69 individuals from ca. 440-150 BP Taumako in Solomon Islands, a remote 'Polynesian Outlier' island in Melanesia. We tested whether bone remodelling indicators differed between age groups, and biological sex validated using ancient DNA. Bone vascular canal and osteon size, vascular porosity, and localised osteon densities, corrected by femoral robusticity indices were examined. Females had statistically significantly higher vascular porosities when compared to males, but osteon densities and ratios of canal-osteon (~ 8%) did not differ between the sexes. Our results indicate that, compared to males, localised femoral bone tissue of the Taumako females did not drastically decline with age, contrary to what is often observed in modern populations. However, our results match findings in other archaeological samples-a testament to past female bone physiology resilience, also now observed in the Pacific region.

Secondary osteon variants and remodeling in human bone.

Histomorphometric analysis of human cortical bone has documented the occurrence of secondary osteon variants. These include drifting osteons which form tails as they move erratically through the cortex and Type II osteons which show partial resorption and redeposition within the cement line of the osteon. Little is known about the biological significance of these variants. Prior studies suggested correlations with age, biomechanics, diet, and mineral homeostasis. No study has yet tested for osteon variant associations with static measures of bone remodeling. In this study, thin sections (n = 112) of the posterior femur representing a late English Medieval adult human osteological collection, subdivided by age, sex, and socio-economic status, were examined to determine whether remodeling indicators reconstructed from osteon parameters (area, diameter, area ratios) and densities differed between categories of presence or absence of Type II and drifting osteon variants. Of the 112 sections, 33 presented with Type II osteons, and 38 had drifting osteons. Sporadic statistically significant results were identified. Haversian canal:osteon area ratio differed (p = 0.017) with Type II osteon presence, Type II osteons were more prevalent in males than females (p = 0.048), and drifting osteons were associated with smaller osteon (p = 0.049) and Haversian canal area (p = 0.05). These results may be explained through some biological (sex) and social (status) processes such as a period of physiological recovery (e.g., following lactation, malnutrition). However, the general lack of consistent relationships between osteon variants and remodeling indicators suggests they occur as a result of natural variation.

Growth of Neanderthal infants from Krapina (120-130 ka), Croatia.

Modern humans have a slow and extended period of childhood growth, but to what extent this ontogenetic pathway was present in Neanderthals is debated. Dental development, linked to the duration of somatic growth across modern primates, is the main source for information about growth and development in a variety of fossil primates, including humans. Studies of Neanderthal permanent teeth report a pace of development either similar to recent humans or relatively accelerated. Neanderthal milk teeth, which form and emerge before permanent teeth, provide an opportunity to determine which pattern was present at birth. Here we present a comparative study of the prenatal and early postnatal growth of five milk teeth from three Neanderthals (120 000-130 000 years ago) using virtual histology. Results reveal regions of their milk teeth formed quickly before birth and over a relatively short period of time after birth. Tooth emergence commenced towards the earliest end of the eruption schedules displayed by extant human children. Advanced dental development is consistent with expectations for Neanderthal infant feeding.

Forager and farmer evolutionary adaptations to malaria evidenced by 7000 years of thalassemia in Southeast Asia.

Thalassemias are inherited blood disorders that are found in high prevalences in the Mediterranean, Southeast Asia and the Pacific. These diseases provide varying levels of resistance to malaria and are proposed to have emerged as an adaptive response to malaria in these regions. The transition to agriculture in the Holocene has been suggested to have influenced the selection for thalassemia in the Mediterranean as land clearance for farming encouraged interaction between Anopheles mosquitos, the vectors for malaria, and human groups. Here we document macroscopic and microscopic skeletal evidence for the presence of thalassemia in both hunter-gatherer (Con Co Ngua) and early agricultural (Man Bac) populations in northern Vietnam. Firstly, our findings demonstrate that thalassemia emerged prior to the transition to agriculture in Mainland Southeast Asia, from at least the early seventh millennium BP, contradicting a long-held assumption that agriculture was the main driver for an increase in malaria in Southeast Asia. Secondly, we describe evidence for significant malarial burden in the region during early agriculture. We argue that the introduction of farming into the region was not the initial driver of the selection for thalassemia, as it may have been in other regions of the world.

Bone loss markers in the earliest Pacific Islanders.

Kingdom of Tonga in Polynesia is one of the most obese nations where metabolic conditions, sedentary lifestyles, and poor quality diet are widespread. These factors can lead to poor musculoskeletal health. However, whether metabolic abnormalities such as osteoporosis occurred in archaeological populations of Tonga is unknown. We employed a microscopic investigation of femur samples to establish whether bone loss afflicted humans in this Pacific region approximately 3000 years ago. Histology, laser confocal microscopy, and synchrotron Fourier-transform infrared microspectroscopy were used to measure bone vascular canal densities, bone porosity, and carbonate and phosphate content of bone composition in eight samples extracted from adult Talasiu males and females dated to 2650 BP. Compared to males, samples from females had fewer vascular canals, lower carbonate and phosphate content, and higher bone porosity. Although both sexes showed evidence of trabecularised cortical bone, it was more widespread in females (35.5%) than males (15.8%). Our data suggest experiences of advanced bone resorption, possibly as a result of osteoporosis. This provides first evidence for microscopic bone loss in a sample of archaeological humans from a Pacific population widely afflicted by metabolic conditions today.

Intra-skeletal vascular density in a bipedal hopping macropod with implications for analyses of rib histology.

Human ribs are thought to be less affected by mechanical strain at the microscopic level than limb bones, implying that rib remodelling better reflects bone physiological homeostasis. Here, we test the hypothesis that rib tissue will be well vascularized and thus enhance susceptibility to metabolic influence. An intra-skeletal comparison of bone vascular canal density was conducted using a macropod animal model adapted to bipedal habitual hopping. The right humerus, ulna, radius, femur, tibia, fibula, a mid-thoracic and upper-thoracic rib of an eastern grey kangaroo (Macropus giganteus) were sectioned at the midshaft, from which histological sections were prepared. Bone vascularity from a maximum of 12 mm2 of sub-periosteal parallel-fibred and lamellar bone was recorded, resulting in a total of 2047 counted vessels. Vascular canal density data were corrected by cortical width, maximum length, and midshaft circumference robusticity indices computed for each bone. The fibula consistently had the highest vascular canal density, even when corrected for maximum length, cortical width and midshaft circumference robusticities. This was followed by the mid- and upper-thoracic ribs. Vascularity differences between bones were relatively consistent whether vascular canal density was controlled for by cortical width or midshaft circumference robusticities. Vascular canal density and robusticity indices were also positively and negatively correlated (p < 0.05). Results confirm that the ribs are well vascularized, which facilitates bone metabolic processes such as remodelling, but the fibula also appears to be a well vascularized bone. Future research investigating human bone metabolism will benefit from examining thoracic rib or fibula samples.

An image analysis protocol for the quantification of interglobular dentine in anthropological tooth sections.

The histological identification of interglobular dentine (IGD) in archeological human remains with macroscopic evidence of rickets has opened a promising new avenue for the investigation of metabolic disease in the past. Recent paleopathological studies have shown that histological analysis of archeological human teeth may allow the identification of periods of vitamin D deficiency occurring within very narrow developmental windows, yielding new information on the seasonality or even maternal-fetal transmission of this disease. However, currently available techniques for recording IGD rely on subjective scoring systems or visual estimations, potentially leaving them open to inter and intra-observer error and rendering comparisons of datasets difficult. Here we describe a new imaging protocol that utilizes open access software and may yield more objective and quantitative data on the amount of IGD present within a dentinal region of interest. We demonstrate that grayscale histograms in FIJI®/ImageJ® might be used to provide less subjective estimates of the percentage of a region of interest affected by IGD. Application of this technique may enable more accurate comparison of datasets between researchers.

Asymmetric midshaft femur remodeling in an adult male with left sided hip joint ankylosis, Metal Period Nagsabaran, Philippines.

OBJECTIVE: This study investigated microstructural changes of the right and left midshaft femur in an archaeological individual afflicted with left-sided hip joint ankylosis to assess whether increased cortical porosity was present as a result of leg disuse. MATERIALS: The individual is a middle-aged adult male excavated from the Metal Period (∼2000 BP) Nagsabaran, Luzon Island, Philippines. METHODS: Following standard examination of femur gross anatomy and differential diagnosis of the hip joint fusion, ∼1 cm thick posterior midshaft femur samples were removed for microstructural examination. Using static histomorphometry, bone multi-cellular unit activity from Haversian canal (vascular pore) density, area, and circularity was reconstructed. Spatial positioning of Haversian canals was mapped using Geographic Information Systems software. Phosphate, carbonate, and carbonate:phosphate ratios were obtained using synchrotron-sourced Fourier transform infrared microspectroscopy. RESULTS: The left femur had greater cortical pore density, with smaller and rounder vascular canals, in addition to lower matrix levels of phosphate and carbonate, when compared to the right femur. CONCLUSIONS: Our data indicate compromised bone tissue in the left femur, and conform to expected bone functional adaptation paradigms of remodeling responses to pathological and biomechanical changes. SIGNIFICANCE: The preservation of this individual's hip abnormality created a unique opportunity to evaluate intra-skeletal bone health asymmetry, which may help other researchers evaluate the presence of limb disuse in archaeological samples. LIMITATIONS: A lack of lower limb data limits our interpretations to femur remodeling only. SUGGESTIONS FOR FURTHER RESEARCH: Future research efforts should aim to examine the presence of remodeling changes in all bones of the lower limb. LAYUNIN: Gamit ang buto ng magkabilang pemur ng isang taong natagpuan sa isang archaeological site na may sakit na ankylosis sa kaliwang balakang, pinag-aralan ang iba't-ibang microstructures galing sa gitnang bahagi o midshaft ng pemur upang malaman kung may makikitang mataas na cortical porosity ang buto dahil hindi ito malimit gamitin. GAMIT: Ang pinag-aaralang buto ay galing sa isang indibidwal na tinatayang middle-age na lalaki na namuhay noong Panahon ng Metal (∼2000 BP) sa Nagsabaran, Cagayan, Republika ng Pilipinas. PAMAMARAAN: Matapos ang unang pagkilatis sa femur at ang pagkilala ng sakit sa balakang, kumuha ng ∼1 sentimetro ng buto galing sa midshaft ng pemur upang lalong mapag-aralan ang kanyang microstructure. Gamit ang static histomorphometry, napag-aralan ang mga naiwang bakas ng multi-cellular unit activity ayon sa kapal, laki at pagkakabilog ng Haversian canal (vascular pore). Gumamit din ng Geographic Information Systems (GIS) software upang mapag-aralan ang kaugnayan ng posisyon ng Haversian canal. Panghuli, gumamit din ng synchroton-sourced Fourier transform infrared (sFTIR) microspectroscopy upang makuha ang bilang ng phosphate, carbonate, at carbonate:phosphate ratio. RESULTA: Napag-alaman na ang kaliwang pemur ay mayroong higit na maraming cortical pores, maliit at mabilog na vascular canals, at mababang bilang ng phosphate, carbonate kung ihahambing sa kanang pemur. KONKLUSYON: Ayon sa aming datos, ang kaliwang pemur ay umaayon sa mga katangian ng isang butong may sakit. Sumunod din ito sa inaasahang bone functional adaptation paradigms of remodeling ng buto dahil may sakit at hindi nagamit. KAHALAGAHAN: Dahil maganda ang pagkakalibing ng buto ng balakang, nagkaroon ng pagkakataong makilatis ang kalusugan ng sinaunang-tao sa pamamagitan ng pag-aaral ng kalusugan ng buto. Dagdag pa, makakatulong din ito upang malaman kung ibang mananaliksik ang pag-aaral ng ibang butong hindi nagagamit mula sa archaeological site. LIMITASYON: Dahil walang nakuhang ibang buto mula sa binti at paa, ang pemur lang ang naimbestigahan. MUNGKAHI PARA SA MGA SUSUNOD NA PAG-AARAL: Kung magkakaroon ng pagkakataon sa susunod, dapat maimbistigahan ang lahat ng buto ng binti (lower limb).

A structural biorhythm related to human sexual dimorphism.

Life on earth is regulated by biological rhythms, some of which oscillate with a circadian, monthly or lunar cycle. Recent research suggests that there is a near weekly biorhythm that may exert an influence on human skeletal growth. Evidence for the timing of this biorhythm is retained in tooth enamel as the periodicity of Retzius lines. Studies report that Retzius periodicity (RP) relates to adult human stature and enamel thickness. Adult human stature is sexually dimorphic, and so is enamel thickness of maxillary third molars (M3) but not mandibular M3. Yet, previous studies report sex differences in RP are apparent in some populations but not others, and it is unknown if dimorphism in enamel thickness relates to RP. To further our understanding of this biorhythm we analysed sex-related variation in RP and its relationship with enamel thickness in a sample of M3's (n = 94) from adults in Northern Britain. Results reveal RP was significantly higher in our sample of female molars compared to those of males, which is consistent with the previously reported correlation between the biorhythm and adult stature. The RP of maxillary M3 related to sex differences in enamel thickness, but this relationship was not present in mandibular M3. Our results support previous findings suggesting that this biorhythm is sexually dimorphic and provide the first evidence that RP may be one factor influencing sex differences in enamel thickness. Our study also shows that correlations between RP and enamel thickness appear to be most readily detected for tooth types with sufficiently wide ranges of enamel thickness variation, as is the case for maxillary but not mandibular M3. Achieving a sufficient sample size was critical for detecting a sex difference in periodicity.

Enamel neonatal line thickness in deciduous teeth of Australian children from known maternal health and pregnancy conditions.

BACKGROUND: Physiological disruptions to early human development have implications for health and disease in later life. Limited research has explored how prenatal factors influence dental development in children of mothers with known pregnancy conditions. Enamel in human deciduous teeth begins forming in utero and is highly susceptible to physiological upsets experienced perinatally. The moment of birth itself is marked in deciduous enamel by the Neonatal Line (NNL) as a baby transitions from the uterine to external environment. This study evaluates the effect of maternal health factors that include stress and alcohol consumption on NNL in teeth from Australian children. STUDY DESIGN AND SUBJECTS: Mothers (n = 53) were interviewed about their health during pregnancy and experience of birth. Sixty-five deciduous teeth (incisors, molars, one canine) from their children were donated for histological examination. Neonatal line thickness was measured from thin sections and evaluated against maternal and neonatal factors using statistical analyses, controlling for tooth type and birth number. RESULTS AND CONCLUSIONS: The only maternal factor of a statistically significant effect on NNL thickness was alcohol consumption. Children of mothers who drank occasionally during pregnancy had a thicker NNL when compared to children of mothers who abstained. These results suggest that maternal lifestyle factors influence NNL formation possibly due to physiological changes that disrupt calcium homeostasis during enamel deposition. We highlight large intra-specific variation in human NNL expression. The potential of dental sampling in identifying children with prenatal exposure to alcohol is suggested.

Microanatomical Record of Cortical Bone Remodeling and High Vascularity in a Fossil Giant Rat Midshaft Femur.

Rat cortical bone does not typically undergo secondary (Haversian) remodeling. Haversian organization of rat bone has been mainly observed in experimental settings following biomechanical or dietary manipulation. Here, we report an observation of cortical secondary osteons within a histological femur cross-section from an extinct (late Quaternary) form of Timorese giant rat (Murinae gen. et sp. indet). The medio-lateral midshaft diameter of its femur, used as a measure of bone size, is 6.15 mm and indicates a heavier than normal skeletal frame. We compare this sample to bone histology in a small rat's midshaft femur of 2.33 mm diameter. A complete lack of Haversian bone remodeling characteristics is noted for the smaller sample, which is dominated by radial vascular canals. The giant rat shows clear secondary osteons and diffuse vascularity mainly composed of tightly packed longitudinal canals across its cortex. It appears that rat cortical bone can undergo bone remodeling, and is organized in a highly vascularized manner, in insular giant cases. Our findings from Timor align with results reported in experimental rat model skeletal biology literature and other insular fossil rat material. Where macroanatomical examination is limited, histological observations on fossil rat limb bones have the potential to aid reconstructions of life history and skeletal growth aspects in these rodents. Anat Rec, 302:1934-1940, 2019. © 2019 American Association for Anatomy.

Microscopic markers of an infradian biorhythm in human juvenile ribs.

Recent studies have indicated that there may be an infradian systemic biorhythm that coordinates aspects of human hard tissue growth and influences adult body size. Here we investigate if evidence of this biorhythm retained in human teeth as the periodicity of Retzius lines (RP) corresponds with the microstructural growth of a non-weight bearing bone, the rib, in a sample of 50 human juvenile skeletons. Using static histomorphometric methods, the RP of one permanent tooth from each skeleton was calculated and combined with measures of bone remodeling in a rib from the same individual. Results provide the first evidence that the infradian biorhythm is linked to bone remodeling in children. Retzius periodicity was negatively correlated with relative osteon area (r = -0.563, p = 0.008) and positively related to Haversian canal area (r = 0.635, p = 0.002) and diameter (r = 0.671, p = 0.001) in children between the age of 8 to 12 years. There was also a negative correlation between RP and the relative cortical area of ribs (r = -0.500, p = 0.048). Relationships between bone remodeling and the biorhythm were much more variable in younger children. Results imply that as the biorhythm speeds up there is increased bone deposition during remodeling of the rib, leading to the larger osteonal lamellar bone areas and smaller Haversian canals in children between 8 and 12 years of age. Our results support the idea that there is an infradian biorhythm that coordinates aspects of human hard tissue growth.

The biorhythm of human skeletal growth.

Evidence of a periodic biorhythm is retained in tooth enamel in the form of Retzius lines. The periodicity of Retzius lines (RP) correlates with body mass and the scheduling of life history events when compared between some mammalian species. The correlation has led to the development of the inter-specific Havers-Halberg oscillation (HHO) hypothesis, which holds great potential for studying aspects of a fossil species biology from teeth. Yet, our understanding of if, or how, the HHO relates to human skeletal growth is limited. The goal here is to explore associations between the biorhythm and two hard tissues that form at different times during human ontogeny, within the context of the HHO. First, we investigate the relationship of RP to permanent molar enamel thickness and the underlying daily rate that ameloblasts secrete enamel during childhood. Following this, we develop preliminary research conducted on small samples of adult human bone by testing associations between RP, adult femoral length (as a proxy for attained adult stature) and cortical osteocyte lacunae density (as a proxy for the rate of osteocyte proliferation). Results reveal RP is positively correlated with enamel thickness, negatively correlated with femoral length, but weakly associated with the rate of enamel secretion and osteocyte proliferation. These new data imply that a slower biorhythm predicts thicker enamel for children but shorter stature for adults. Our results develop the intra-specific HHO hypothesis suggesting that there is a common underlying systemic biorhythm that has a role in the final products of human enamel and bone growth.

Cortical Histomorphometry of the Human Humerus During Ontogeny.

Modeling and remodeling are two key determinants of human skeletal growth though little is known about the histomorphometry of cortical bone during ontogeny. In this study, we examined the density and geometric properties of primary and secondary osteons (osteon area and diameter, vascular canal area and diameter) in subperiosteal cortical bone from the human humerus (n = 84) between birth and age 18 years. Sections were removed from the anterior midshaft aspect of humeri from skeletons. Age-at-death was reconstructed using standard osteological techniques. Analyses revealed significant correlation between the histomorphometric variables and age. Higher densities of primary osteons occurred between infancy and 7 years of age but were almost completely replaced by secondary osteons after 14 years of age. The geometry of primary osteons was less clearly related to age. Secondary osteons were visible after 2 years of age and reached their greatest densities in the oldest individuals. Osteon size was positively but weakly influenced by age. Our data imply that modeling and remodeling are age-dependent processes that vary markedly from birth to adulthood in the human humerus.

Enamel biorhythms of humans and great apes: the Havers-Halberg Oscillation hypothesis reconsidered.

The Havers-Halberg Oscillation (HHO) hypothesis links evidence for the timing of a biorhythm retained in permanent tooth enamel (Retzius periodicity) to adult body mass and life history traits across mammals. Potentially, these links provide a way to access life history of fossil species from teeth. Recently we assessed intra-specific predictions of the HHO on human children. We reported Retzius periodicity (RP) corresponded with enamel thickness, and cusp formation time, when calculated from isolated deciduous teeth. We proposed the biorhythm might not remain constant within an individual. Here, we test our findings. RP is compared between deciduous second and permanent first molars within the maxillae of four human children. Following this, we report the first RPs for deciduous teeth from modern great apes (n = 4), and compare these with new data for permanent teeth (n = 18) from these species, as well as with previously published values. We also explore RP in teeth that retain hypoplastic defects. Results show RP changed within the maxilla of each child, from thinner to thicker enameled molars, and from one side of a hypoplastic defect to the other. When considered alongside correlations between RP and cusp formation time, these observations provide further evidence that RP is associated with enamel growth processes and does not always remain constant within an individual. RP of 5 days for great ape deciduous teeth lay below the lowermost range of those from permanent teeth of modern orangutan and gorilla, and within the lowermost range of RPs from chimpanzee permanent teeth. Our data suggest associations between RP and enamel growth processes of humans might extend to great apes. These findings provide a new framework from which to develop the HHO hypothesis, which can incorporate enamel growth along with other physiological systems. Applications of the HHO to fossil teeth should avoid transferring RP between deciduous and permanent enamel, or including hypoplastic teeth.

Biorhythms, deciduous enamel thickness, and primary bone growth: a test of the Havers-Halberg Oscillation hypothesis.

Across mammalian species, the periodicity with which enamel layers form (Retzius periodicity) in permanent teeth corresponds with average body mass and the pace of life history. According to the Havers-Halberg Oscillation hypothesis (HHO), Retzius periodicity (RP) is a manifestation of a biorhythm that is also expressed in lamellar bone. Potentially, these links provide a basis for investigating aspects of a species' biology from fossilized teeth. Here, we tested intra-specific predictions of this hypothesis on skeletal samples of human juveniles. We measured daily enamel growth increments to calculate RP in deciduous molars (n = 25). Correlations were sought between RP, molar average and relative enamel thickness (AET, RET), and the average amount of primary bone growth (n = 7) in humeri of age-matched juveniles. Results show a previously undescribed relationship between RP and enamel thickness. Reduced major axis regression reveals RP is significantly and positively correlated with AET and RET, and scales isometrically. The direction of the correlation was opposite to HHO predictions as currently understood for human adults. Juveniles with higher RPs and thicker enamel had increased primary bone formation, which suggests a coordinating biorhythm. However, the direction of the correspondence was, again, opposite to predictions. Next, we compared RP from deciduous molars with new data for permanent molars, and with previously published values. The lowermost RP of 4 and 5 days in deciduous enamel extends below the lowermost RP of 6 days in permanent enamel. A lowered range of RP values in deciduous enamel implies that the underlying biorhythm might change with age. Our results develop the intra-specific HHO hypothesis.