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.     

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.     

胎儿牙齿硬组织发育的临床应用解剖

目的：探讨国人胎儿牙齿硬组织发育状况。方法：对正常胎龄10-38周的胎儿上、下颌骨作连续切片,观察乳、恒牙胚及其相关结构发生规律。结果：牙体硬组织形成于14-20周,牙齿的增殖、钙化、萌出同时进行,牙釉质、牙本质厚度随胎龄增长而增加,各乳磨牙牙尖总是近颊尖最先发牛,恒牙板总是位于乳牙胚舌侧。结论：国人牙体硬组织发育成熟较低,牙釉质、牙本质厚度与胎龄呈正相关。     

Megadontia, striae periodicity and patterns of enamel secretion in Plio-Pleistocene fossil hominins

Early hominins formed large and thick-enamelled cheek-teeth within relatively short growth periods as compared with modern humans. To understand better the developmental basis of this process, we measured daily enamel increments, or cross striations, in 17 molars of Plio-Pleistocene hominins representing seven different species, including specimens attributed to early Homo. Our results show considerable variation across species, although all specimens conformed to the known pattern characterised by greater values in outer than inner enamel, and greater cuspal than cervical values. We then compared our results with the megadontia index, which represents tooth size in relation to body mass, for each species to assess the effect of daily growth rates on tooth size. Our results indicate that larger toothed (megadont) taxa display higher rates or faster forming enamel than smaller toothed hominins. By forming enamel quickly, large tooth crowns were able to develop within the constraints of shorter growth periods. Besides daily increments, many animals express long-period markings (striae of Retzius) in their enamel. We report periodicity values (number of cross striations between adjacent striae) in 14 new specimens of Australopithecus afarensis, Paranthropus aethiopicus, Paranthropus boisei, Homo habilis, Homo rudolfensis and Homo erectus, and show that long-period striae express a strong association with male and average male–female body mass. Our results for Plio-Pleistocene hominins show that the biological rhythms that give rise to long-period striae are encompassed within the range of variation known for modern humans, but show a lower mean and modal value of 7 days in australopithecines. In our sample of early Homo, mean and modal periodicity values were 8 days, and therefore similar to modern humans. These new data on daily rates of enamel formation and periodicity provide a better framework to interpret surface manifestations of internal growth markings on fossil hominin tooth crowns. Importantly, our data on early hominin cross striation variation may now contribute towards solving difficult taxonomic diagnoses where much may depend on fragmentary molar remains and enamel structure.     

Characterization of short-period and long-period incremental markings in porcine enamel and dentine—Results of a fluorochrome labelling study in wild boar and domestic pigs

Mammalian dental hard tissues exhibit incremental markings that reflect the periodic variation of appositional growth rates. In order to use these markings to characterize dental growth processes and to infer life-history traits, an unequivocal identification of their periodicities is required. We performed a fluorochrome labelling study on forming enamel and dentine in molar teeth of wild boar and domestic pigs to establish the periodicity and temporal correspondence of incremental markings in enamel and dentine. The dominant incremental markings in enamel (laminations) and dentine (von Ebner lines) recorded in the pig teeth are of a daily nature. In addition, long-period incremental markings with a periodicity of 2 days were recorded in enamel (striae of Retzius) and dentine (Andresen lines). The 2-day growth rhythm was also expressed at the lateral crown surface, as evidenced by the pattern of perikymata. In enamel, also markings with a sub-daily periodicity, representing an ultradian growth rhythm, were observed. Our study provides experimental evidence for the periodicity of incremental markings in porcine enamel and dentine. The findings correct previous misconceptions on incremental markings in dental hard tissues of pigs and other ungulates that had led to erroneous conclusions regarding crown formation parameters.     

Experimental determination of the periodicity of incremental features in enamel

Vital labelling of hard tissues was used to examine the periodicity of features of dental enamel microstructure. Fluorescent labels were administered pre- and postnatally to developing macaques (Macaca nemestrina), which were identified histologically in dentine and related to accentuated lines in enamel, allowing for counts of features within known-period intervals. This study demonstrates that cross-striations represent a daily rhythm in enamel secretion, and suggests that intradian lines are the result of a similar 12-h rhythm. Retzius lines were found to have a regular periodicity within individual dentitions, and laminations appear to represent a daily rhythm that also shows 12-h subdivisions. The inclusion of intradian lines and laminations represents the first empirical evidence for their periodicities in primates; these features frequently complicate precise measurements of secretion rate and Retzius line periodicity, which are necessary for determination of crown formation time. The biological basis of incremental feature formation is not completely understood; long-period features may result from interactions between short-period rhythms, although this does not explain the known range of Retzius line periodicities within humans or among primates. Studies of the genetic, neurological and hormonal basis of incremental feature formation are needed to provide more insight into their physiological and structural basis.     

Disturbed enamel formation in wild boars (Sus scrofa L.) from fluoride polluted areas in Central Europe

The pathological alterations of enamel structure in the teeth of wild boars from fluoride polluted areas in N-Bohemia (Czech Republic) and S-Saxony (Germany) were studied on a macroscopic and a microscopic level. Mandibular bone fluoride concentration (mg F(-)/kg, dry wt; mean +/-SD, individuals <24 months of age) in the specimens from N-Bohemia (754.3+/-149.6) and S-Saxony (490.8+/-135.1) was significantly higher than that of controls (free of dental fluorosis), originating from the western part of Germany (304.7+/-91.0). Fluoride content in bulk enamel (mg F(-)/kg, ash wt) of fluorotic permanent teeth from N-Bohemia (382.1+/-165.2) and S-Saxony (125.0+/-38.3) was likewise significantly increased over that of non-fluorotic control teeth from W-Germany (33.6+/-26.7). Macroscopically, fluorosed wild boar enamel exhibited opacity and discoloration of varying extent, accentuated perikymata as well as hypoplastic and posteruptive surface defects. Microradiographic and scanning electron microscopic analyses revealed enamel subsurface hypomineralization, accentuated Retzius lines and occurrence of broad, hypomineralized incremental bands of abnormal structure underlying hypoplastic enamel surface defects. The presence of zones of aprismatic enamel was associated with these bands. Incremental bands with altered enamel structure and enamel surface hypoplasias, both denoting a severe disturbance during the secretory stage of amelogenesis, have previously been observed in rodents following acute parenteral fluoride dosing. It is concluded that in the chronically fluoride exposed wild boars periods of especially elevated plasma fluoride levels exerted an acute toxic effect on the secretory ameloblasts. A feature not previously reported from fluorosed enamel was the occurrence of canal-like structures that originated at the broad incremental bands and extended into the external enamel. The presence of these canals presumably results from a delay in the resumption of secretory activity by groups of ameloblasts following a fluoride insult. Based on experimental evidence in domestic pigs and in sheep, the overall subsurface hypomineralization of fluorosed wild boar enamel is attributed to a disturbance of enamel maturation. The distribution of fluorotic enamel changes within the dentition of the wild boars could be related to the developmental sequence of tooth formation in the species. Teeth whose crown formation took place prenatally (deciduous teeth) or largely pre-weaning (permanent first molars) exhibited no or only moderate fluorotic enamel alterations. Based on the extension of enamel surface hypoplasias along the coronoapical axes of the tooth crowns, the timing of excess fluoride exposure that caused a marked disruption of enamel matrix secretion was estimated in specimens with a known date of death. The results indicate that the wild boars had been exposed to a particularly severe fluoride impact during autumn and winter of their first year of life.     

Light and electron microscopic studies of retzius lines in human cervical enamel

Unaltered mature enamel and partially demineralized mature enamel from the cervical third of human permanent teeth were examined by means of transmission electron microscopy, microradiography, phase contrast and Nomarski differential interference microscopy. The Retzius lines in the outer third of the cervical enamel had a staircase configuration. The steps in the stair-case pattern formed the bases of triangular regions which were radiolucent and ultrastructurally contained fewer crystallites than the adjacent enamel. In the outer prismless layer, the Retzius lines had a curvilinear configuration and were also crystallite deficient. The structure of the Retzius lines in the inner portion of the cervical enamel was not nearly as well defined.     

In Vivo Microstructural Analysis of Enamel in Permanent and Deciduous Teeth

Confocal microscope was used to analyze human enamel from 10 deciduous and 10 permanent teeth. Optically sectioned images were obtained. A more intense autofluorescence was found in primary teeth. This finding might be due to the greater presence of organic substances in deciduous enamel. The mean prism diameter measurement in permanent teeth enamel was 3.150?µm and 2.602?µm in deciduous teeth. The mean prism diameter in deciduous teeth was statistically least. The results indicate that a confocal microscope may be of help in analyzing and defining the microscopic features of human enamel.     

Comparison between deciduous and permanent incisor teeth in morphology of bovine enamel.

Functional incisor teeth (deciduous and permanent teeth) from Bovidae (14 species) were prepared for scanning electron microscopic observation. Ultrastructural patterns of the enamel layer of deciduous and permanent incisor teeth varied (ex. prisms, arrangement pattern of matrices, and in thickness of enamel layer) in each species. The ultrastructures of prisms in longitudinal sections were classified into three types; A, radial, B, tangential, and C, mix of A and B arrangement enamel; modified Koenigswald's method (1982) in examined species. Type A was found in a large part of permanent and a small part of deciduous incisor teeth, while types B and C were mainly found in the deciduous teeth. These morphological features show the remarkable correlation between permanent and deciduous teeth.     

Effect of Down syndrome on the dimensions of dental crowns and tissues

Abnormal growth in Down syndrome (DS) is reflected by variable reduction in size and simplification in form of many physical traits. This study aimed to compare the thickness of enamel and dentine in deciduous and permanent mandibular incisor teeth between DS and non‐DS individuals and to clarify how these tissues contribute to altered tooth size in DS. Sample groups comprised 61 mandibular incisors (29 permanent and 32 deciduous) from DS individuals and 55 mandibular incisors (29 permanent and 26 deciduous) from non‐DS individuals. Maximum mesiodistal and labiolingual crown dimensions were measured initially, then the crowns were sectioned midsagittally and photographed using a stereomicroscope. Linear measurements of enamel and dentine thickness were obtained on the labial and lingual surfaces of the crowns, together with enamel and dentine–pulp areas and lengths of the dentino‐enamel junction. Reduced permanent crown size in DS was associated with a reduction in both enamel and dentine thickness. After adjustments were made for tooth size, DS permanent incisors had significantly thinner enamel than non‐DS permanent teeth. The DS permanent teeth also exhibited significant differences in shape and greater variability in dimensions than the non‐DS permanent teeth. Crown dimensions of deciduous incisors were similar in size or larger in DS compared with non‐DS deciduous teeth. Enamel and dentine thicknesses of the deciduous teeth were similar in DS and non‐DS individuals. The findings indicate that growth retardation in DS reduces both enamel and dentine deposition in the permanent incisors but not in the earlier‐forming deciduous predecessors. The results are also consistent with the concept of amplified developmental instability for dental traits in DS. Am. J. Hum. Biol. 13:690–698, 2001. © 2001 Wiley‐Liss, Inc.     

Structure of retzius lines in partially demineralized human enamel

Human third molars were partially demineralized in an acid-alcohol solution and embedded in Epon 812. Six-micron sections were cut from regions of the cervical enamel exhibiting prominent Retzius lines. The plastic was removed from the specimens by microincineration and were examined with the scanning electron microscope. The most prominent structural feature seen along the Retzius lines was the cervical translocation of some of the prisms. The scanning electron microscopic images also suggested that prisms were translocating in the transverse plane of the tooth. A series of pores, which appeared to be empty, were observed in association with the translocations occurring along the Retzius lines.     

Structure of Retzius lines in partially demineralized human enamel.

Human third molars were partially demineralized in an acid-alcohol solution and embedded in Epon 812. Six-micron sections were cut from regions of the cervical enamel exhibiting prominent Retzius lines. The plastic was removed from the specimens by microincineration and were examined with the scanning electron microscope. The most prominent structural feature seen along the Retzius lines was the cervical translocation of some of the prisms. The scanning electron microscopic images also suggested that prisms were translocating in the transverse plane of the tooth. A series of pores, which appeared to be empty, were observed in association with the translocations occurring along the Retzius lines.     

Enamel and dentine development and the life history profile of Victoriapithecus macinnesi from Maboko Island, Kenya

Two permanent lower second molar teeth (KNM-MB 19841 and KNM-MB 27844) attributed to Victoriapithecus macinnesi were prepared for histological analysis. A further five male and three female canine teeth were replicated with a silicone impression material and perikymata counts subsequently made on epoxy casts over the whole crown surface. Daily enamel cross striations averaged 6 μm apart in both cuspal and lateral enamel of the molars and the total crown formation times were 1.24 years in the molars and, using the same periodicity determined from the molars, 1.84 and 1.36 years respectively in the male and female canines. Rates of dentine formation matched those known for extant macaques and were used to calculate root extension rates, which averaged 11.5 μm per day over the whole 8 mm root length of KNM-MB 19841. The period between M2 initiation and gingival emergence was estimated to be 1.95 years in Victoriapithecus which is greater than estimates for Cebus albifrons and Chlorocebus aethiops, (which are similar in body mass to Victoriapithecus), but less than estimates made here for several macaque species. A speculative picture of dental development in Victoriapithecus emerges that is slower than that known for modern vervet monkeys and may have been more similar to that in some smaller modern macaque species.     

The presence of accessory cusps in chimpanzee lower molars is consistent with a patterning cascade model of development

Tooth crown morphology is of primary importance in fossil primate systematics and understanding the developmental basis of its variation facilitates phenotypic analyses of fossil teeth. Lower molars of species in the chimp/human clade (including fossil hominins) possess between four and seven cusps and this variability has been implicated in alpha taxonomy and phylogenetic systematics. What is known about the developmental basis of variation in cusp number – based primarily on experimental studies of rodent molars – suggests that cusps form under a morphodynamic, patterning cascade model involving the iterative formation of enamel knots. In this study we test whether variation in cusp 6 (C6) presence in common chimpanzee and bonobo lower molars (n = 55) is consistent with predictions derived from the patterning cascade model. Using microcomputed tomography we imaged the enamel‐dentine junction of lower molars and used geometric morphometrics to examine shape variation in the molar crown correlated with variation in C6 presence (in particular the size and spacing of the dentine horns). Results indicate that C6 presence is consistent with predictions of a patterning cascade model, with larger molars exhibiting a higher frequency of C6 and with the location and size of later‐forming cusps correlated with C6 variation. These results demonstrate that a patterning cascade model is appropriate for interpreting cusp variation in Pan and have implications for cusp nomenclature and the use of accessory cusp morphology in primate systematics.     

Deciduous teeth in tuberous sclerosis

Shed deciduous teeth from patients with tuberous sclerosis, cerebral palsy, Down syndrome, phenylketonuria and healthy persons were examined with a surface microscope. We found enamel pits in all 87 deciduous teeth from the 20 patients with tuberous sclerosis, but in none of the 253 deciduous teeth from 142 controls constituting patients with cerebral palsy, phenylketonuria and Down syndrome as well as healthy persons. Enamel pits always occurred in the facial surface of the central incisor, lateral incisor and canine, while the number of enamel pits in the other surfaces of the deciduous teeth varied from none to nine. Ground sections examined microscopically revealed an undisturbed pattern of incremental lines (Retzius striae) surrounding the pits. In five dental sacs from patients with tuberous sclerosis, microscopic examination showed that the inner surface of the operculum was remarkably more irregular than in control patients.     

Evolution and development of the mammalian dentition: Insights from the marsupial Monodelphis domestica

To understand developmental mechanisms of evolutionary change, we must first know how different morphologies form. The vast majority of our knowledge on the developmental genetics of tooth formation derives from studies in mice, which have relatively derived mammalian dentitions. The marsupial Monodelphis domestica has a more plesiomorphic heterodont dentition with incisors, canines, premolars, and molars on both the upper and the lower jaws, and a deciduous premolar. The complexity of the M. domestica dentition ranges from simple, unicusped incisors to conical, sharp canines to multicusped molars. We examine the development of the teeth in M. domestica, with a specific focus on the enamel knot, a signaling center in the embryonic tooth that controls shape. We show that the tooth germs of M. domestica express fibroblast growth factor (FGF) genes and Sprouty genes in a manner similar to wild‐type mouse molar germs, but with a few key differences. Developmental Dynamics, 2011. © 2010 Wiley‐Liss, Inc.     

Development and size of the teeth of Macaca mulatta

A cross-sectional sample of 151 skulls from Macaca mulatta of known age and similar rearing in U.S. Primate Centers was analyzed to determine age-related “norms” of stages of development and size of teeth. The stages of development from the follicle of a deciduous incisor in the fetus to completion of the root with apex closed of the permanent third molar were related to age. The age range observed for eruption of each tooth was noted and related to its stage of development. The crown of each erupted tooth was found to be completely developed, but growth of its root continued for a longer, indeterminate period. When a deciduous tooth was exfoliated, the crown of the permanent successor was found to be completed and root growth had begun. Measurements of both mesiodistal and faciolingual diameters and of crown length of the teeth in situ and of total length and root length on roentgenograms were examined for sexual dimorphism. The faciolingual diameter of the deciduous mandibular second incisor and of both second molars showed the greatest sexual dimorphism among both diameters of all deciduous teeth. The mesiodistal and faciolingual diameters of the mandibular premolars were found to be the best dimensions in discriminant functions for identifying sex in the absence of permanent canines.     

Dental Defects as a Potential Indicator of Chronic Malnutrition in a Population of Fallow Deer (Dama dama) from Northwestern Germany

We studied enamel defects in mandibular cheek teeth of fallow deer from an enclosed population that had grown far beyond the carrying capacity of its habitat. Macroscopic inspection revealed a high frequency of pathological enamel alterations in the permanent premolars and the third molar, which form late during dental development, while earlier forming teeth (deciduous premolars and first molar) were generally not affected. Macroscopic enamel alterations comprised opacity and posteruptive discoloration, loss of occlusal enamel ridges, and presence of enamel surface lesions. Backscattered electron imaging in the SEM revealed a marked hypomineralization and related increased porosity of the outer compared to the central and inner enamel portions in the affected teeth. In contrast, the enamel of the unaffected first molars showed a homogeneous high degree of mineralization. Microindentation hardness testing demonstrated a significantly reduced and highly variable hardness of the outer enamel of the affected teeth. The recorded enamel alterations are indicative of a disturbance of enamel maturation. Based on findings of experimental studies by other authors that explored the effects of dietary deficiencies on the mineralization of dental hard tissues in different mammal species, we suggest a dietary calcium deficiency as the cause of the observed pathological enamel changes in the fallow deer. The supposed calcium deficiency only affected teeth whose crown mineralization takes place after weaning, while the deciduous premolars and the first molar, whose crown formation occurs prenatally or during the early postnatal period of intense milk feeding, were unaffected. Anat Rec, 299:1409–1423, 2016. © 2016 Wiley Periodicals, Inc.     

Mechanical properties and microstructure of hypomineralised enamel of permanent teeth

Isolated enamel defects are commonly seen in first permanent molar teeth but there has been little work on the physical and morphological composition of affected molars. The aim of this study was to determine the mechanical and morphological properties of hypomineralised first permanent molar teeth, utilising the Ultra-Micro-Indentation System (UMIS) and scanning electron microscope, respectively. Further investigations using Energy Dispersive X-ray Spectrometery (EDS), Back Scatter Electron (BSE) Imaging, and X-ray diffraction were employed to attempt to determine the chemical composition, mineral content and crystalline structure of the hypomineralised tissue, respectively, of eight first permanent molars with severe enamel hypomineralisation. The hardness and modulus of elasticity were found to be statistically significantly lower (0.53+/-0.31 and 14.49+/-7.56 GPa, respectively) than normal enamel (3.66+/-0.75 and 75.57+/-9.98 GPa, respectively). Although the fractured surface of the hypomineralised enamel was significantly more disorganised and the relative mineral content was reduced by approximately 5% in comparison to sound enamel, the mineral phase and Ca/P ratio was similar in hypomineralised and sound enamel. The dramatic reduction in the mechanical properties of first permanent molar teeth has ramifications when clinicians are choosing restorative materials to restore the defects. The reason for the dramatic reduction in mechanical properties of hypomineralised first permanent molar teeth is at present unknown.