Mineral metabolism and microstructural defects in primate teeth.

There are numerous structural defects that occur in primate teeth due to variable calcification during certain growth stages. These interruptions are usually areas of hypomineralization in enamel and dentin which are referred to as Striae of Retzius and Hunter Shreger bands in the enamel or Incremental Lines of von Ebner and Contour Lines of Owen in the case of the dentin. The frequency of occurrence of these variations in mineralization can be related to such factors as growth tempo, dietary quality, state of health, and past disease episodes. Another structure that appears in the dentin is a region that fails to calcify and is referred to as Inter-globular Dentin. Our studies have shown that the microstructural quality of different species' dentition varies widely. Samples obtained from certain fre-ranging cercopithcoids show that these species have the least hypomineralizations while man has the most. Other primate species range between these two extremems with the pongids nearer to man in these characteristics, as previously noted by Sognnaes. Additionally, out initial study shows a great deal of diversity between prehistoric human populations in the microstructural quality of their teeth. We offer the tentative explanation that this variation is due to differences in the calcitying properties of the diet and hence a difference in the general state of their health.     

Dental Phenotype in Jalili Syndrome Due to a c.1312 dupC Homozygous Mutation in the CNNM4 Gene

Jalili syndrome denotes a recessively inherited combination of an eye disease (cone-rod dystrophy) and a dental disorder (amelogenesis imperfecta), which is caused by mutations in the CNNM4 gene. Whereas the ophthalmic consequences of these mutations have been studied comprehensively, the dental phenotype has obtained less attention. A defective transport of magnesium ions by the photoreceptors of the retina is assumed to account for the progressive visual impairment. Since magnesium is also incorporated in the mineral of dental hard tissues, we hypothesized that magnesium concentrations in defective enamel resulting from mutations in CNNM4 would be abnormal, if a similar deficiency of magnesium transport also accounted for the amelogenesis imperfecta. Thus, a detailed analysis of the dental hard tissues was performed in two boys of Kosovan origin affected by Jalili syndrome. Retinal dystrophy of the patients was diagnosed by a comprehensive eye examination and full-field electroretinography. A mutational analysis revealed a c.1312 dupC homozygous mutation in CNNM4, a genetic defect which had already been identified in other Kosovan families and putatively results in loss-of-function of the protein. The evaluation of six primary teeth using light and scanning electron microscopy as well as energy-dispersive X-ray spectroscopy showed that dental enamel was thin and deficient in mineral, suggesting a hypoplastic/hypomineralized type of amelogenesis imperfecta. The reduced mineral density of enamel was accompanied by decreased amounts of calcium, but significantly elevated levels of magnesium. In dentin, however, a similar mineral deficiency was associated with reduced magnesium and normal calcium levels. It is concluded that the c.1312 dupC mutation of CNNM4 results in mineralization defects of both enamel and dentin, which are associated with significantly abnormal magnesium concentrations. Thus, we could not disprove the hypothesis that a disrupted magnesium transport is involved in the development of the dental abnormalities observed in Jalili syndrome.     

Developmental and Post-Eruptive Defects in Molar Enamel of Free-Ranging Eastern Grey Kangaroos (Macropus giganteus) Exposed to High Environmental Levels of Fluoride

Dental fluorosis has recently been diagnosed in wild marsupials inhabiting a high-fluoride area in Victoria, Australia. Information on the histopathology of fluorotic marsupial enamel has thus far not been available. This study analyzed the developmental and post-eruptive defects in fluorotic molar enamel of eastern grey kangaroos (Macropus giganteus) from the same high-fluoride area using light microscopy and backscattered electron imaging in the scanning electron microscope. The fluorotic enamel exhibited a brownish to blackish discolouration due to post-eruptive infiltration of stains from the oral cavity and was less resistant to wear than normally mineralized enamel of kangaroos from low-fluoride areas. Developmental defects of enamel included enamel hypoplasia and a pronounced hypomineralization of the outer (sub-surface) enamel underneath a thin rim of well-mineralized surface enamel. While the hypoplastic defects denote a disturbance of ameloblast function during the secretory stage of amelogenesis, the hypomineralization is attributed to an impairment of enamel maturation. In addition to hypoplastic defects, the fluorotic molars also exhibited numerous post-eruptive enamel defects due to the flaking-off of portions of the outer, hypomineralized enamel layer during mastication. The macroscopic and histopathological lesions in fluorotic enamel of M. giganteus match those previously described for placental mammals. It is therefore concluded that there exist no principal differences in the pathogenic mechanisms of dental fluorosis between marsupial and placental mammals. The regular occurrence of hypomineralized, opaque outer enamel in the teeth of M. giganteus and other macropodids must be considered in the differential diagnosis of dental fluorosis in these species.     

Incisor Schmelzmuster Diversity in South America's Oldest Rodent Fauna and Early Caviomorph History

Investigation of the enamel microstructure of 20 isolated rodent incisors from the ?Eocene Santa Rosa local fauna (Peru) yielded exclusively schmelzmuster with multiserial Hunter–Schreger bands (HSB). All three subtypes of multiserialHSB with parallel, acute angular, and rectangular interprismatic matrix (IPM) that were previously reported for caviomorph rodents are present. Two lower incisors with rectangular IPM can be attributed to the Octodontoidea, a caviomorph superfamily exhibiting this highly derived enamel type. The plesiomorphic pauciserial condition that characterizes early Paleogene rodents such as North American Ischyromyoidea (including “Franimorpha”) has not been detected. It is therefore probable that the founder populations of South American Caviomorpha already possessed a derived incisor schmelzmuster with multiserial HSB that is shared with African Thryonomyoidea. Because on the North American continent a possible stem-lineage representative of Caviomorpha with multiserial HSB has never been detected, incisor enamel microstructure supports the hypothesis of an African origin of Caviomorpha from a common ancestor shared with Thryonomyoidea.     

Deciduous enamel defects and caries susceptibility in a prehistoric Ohio population

Clinical studies of the relationship between developmental enamel defects and caries susceptibility have often produced conflicting results. This has been due in part to a failure to distinguish between different types of defects. Studies of this association in prehistoric populations have been rare. The complete deciduous dentitions of 57 subadults from the Libben site, a large Late Woodland cemetery in Ottawa County, Ohio, were selected for analysis. Defects were classified as either hypoplasias (deficiencies in matrix apposition) or hypocalcifications (deficiencies in mineralization) and were graded for severity. The presence or absence of carious lesions was recorded for each tooth. Results indicate a strong positive relationship between hypocalcifications and caries susceptibility. The elevated caries susceptibility of hypocalcified teeth may be related to high levels of magnesium or altered enamel microcrystallite orientation within these teeth. Variations in the frequency of hypocalcifications may partially explain differences in caries rates that have been observed in different prehistoric populations.     

Cyclical phenomena occurring during the maturation of the enamel of rat incisor teeth

Summary A pattern of obliquely oriented bands has been demonstrated at the surface of the maturation zone enamel of freshly dissected rat incisor teeth as they dry. This pattern, of which there is no evidence in the fresh, wet, or completely dry teeth, consists of up to 4 or 5 pale grey, translucent linesseparated by wider, whiter, more opaque bands and has been shown to correlate directly with a similar pattern seen on the same teeth after staining with toluidine blue and previously described as the maturation cycle banding pattern (Boyde and Reith 1982). A second pattern comprised of much more closely spaced bands is also described. In this pattern, which again correlates with a similar pattern seen after toluidine blue staining, translucent and opaque bands cross the maturation zone more transversely and have a width of about 200 microns, approximating to 8h tooth growth. It is postulated that these banding patterns reflect alternately different drying rates of the maturation zone enamel and that they may correspond to cyclical changes in the hydrophobicity and hydrophilicity of enamel matrix both on a daily basis and on a larger time scale.     

The Incidence of Enamel Hypoplasia among the Krapina Neandertals

Developmental defects of teeth often appear as a record of metabolic disturbances during growth. One defect in particular, enamel hypoplasia, has been described among recent prehistoric human populations, but few analyses have been made of fossil hominids. This study describes the enamel quality of the dental remains of the Krapina Neandertals. Of the 18 individuals represented by dental arch fragments, 13 showed evidence of one or more teeth with enamel defects described as hypoplasia. This was a high frequency of occurrence when compared to Bronze and Copper Age populations in Central Europe or Woodland and Mississippian populations in North America. The reason for the higher incidence among the Neandertals is not fully explainable given the data available. However, there is a suggestion that fluorosis could be a factor that reduced the expression of enamel hypoplasia in the European populations.     

Cyclical phenomena occurring during the maturation of the enamel of rat incisor teeth. Their manifestation during drying

A pattern of obliquely oriented bands has been demonstrated at the surface of the maturation zone enamel of freshly dissected rat incisor teeth as they dry. This pattern, of which there is no evidence in the fresh, wet, or completely dry teeth, consists of up to 4 or 5 pale grey, translucent lines separated by wider, whiter, more opaque bands and has been shown to correlate directly with a similar pattern seen on the same teeth after staining with toluidine blue and previously described as the maturation cycle banding pattern (Boyde and Reith 1982). A second pattern comprised of much more closely spaced bands is also described. In this pattern, which again correlates with a similar pattern seen after toluidine blue staining, translucent and opaque bands cross the maturation zone more transversely and have a width of about 200 microns, approximating to 8 h tooth growth. It is postulated that these banding patterns reflect alternately different drying rates of the maturation zone enamel and that they may correspond to cyclical changes in the hydrophobicity and hydrophilicity of enamel matrix both on a daily basis and on a larger time scale.     

Prevalence and the duration of linear enamel hypoplasia: a comparative study of Neandertals and Inuit foragers

As a dental indicator of generalized physiological stress, enamel hypoplasia has been the subject of several Neandertal studies. While previous studies generally have found high frequencies of enamel hypoplasia in Neandertals, the significance of this finding varies with frequencies of enamel hypoplasia in comparative samples. The present investigation was undertaken to ascertain if the enamel hypoplasia evidence in Neandertals suggests a high level of physiological stress relative to a modern human foraging group, represented here by an archaeological sample of Inuit from Point Hope, Alaska. Unlike previous studies, this study focused specifically on linear enamel hypoplasia (LEH), emphasizing systemic over localized causes of this defect by considering LEH to be present in an individual only if LEH defects occur on two anterior teeth with overlapping crown formation periods. Moreover, this study is the first to evaluate the average growth disruption duration represented by these defects in Neandertals and a comparative foraging group. In the prevalence analysis, 7/18 Neandertal individuals (from Krapina and southern France) and 21/56 Neandertal anterior teeth were affected by LEH, or 38.9% and 37.5% respectively. These values do not differ significantly from those of the Inuit sample in which 8/21, or 38.1% of individuals, and 32/111, or 28.8% of anterior teeth were affected. For the growth disruption duration analysis, 22 defects representing separate episodes of growth disruption in Neandertals were compared with 22 defects in the Inuit group using three indicators of duration: the number of perikymata (growth increments) in the occlusal walls of LEH defects, the total number of perikymata within them, and defect width. Only one indicator, the total number of perikymata within defects, differed significantly between the Inuit and Neandertal groups (an average of 13.4 vs. 7.3 perikymata), suggesting that if there is any difference between them, the Inuit defects may actually represent longer growth disruptions than the Neandertal defects. Thus, while stress indicators other than linear enamel hypoplasia may eventually show that Neandertal populations were more stressed than those of modern foragers, the evidence from linear enamel hypoplasia does not lend support to this idea.     

Scanning electron microscope study of cat and dog enamel structure

Scanning electron microscopy revealed several similarities as well as significant differences in the enamel structure between cat and dog teeth. Three enamel layers were present in both species; a surface rodless (aprismatic) layer, an outer layer of parallel rods (only at some sites), and an inner layer with prominent Hunter-Schreger bands. In the inner layer of both carnivores, the diameter of individual rods varied significantly and frequently their course changed abruptly with respect to neighboring rods. In dog teeth the cross-sectional shape of inner enamel rods was pleomorphic, but hexagonal in outer enamel. In contrast, cat enamel rods were rounded in both inner and outer enamel layers. Hunter-Schreger bands of cats circumscribed the teeth in relatively straight segments, but these bands showed pronounced waviness in dog teeth. In cats and dogs the surface rodless layer was structurally continuous with subjacent interrod enamel and covered all tooth surfaces with the exception of the cervical areas. The data show that the structure of inner and outer enamel layers differ between these two carnivore species and that the enamel structure of the cat was most similar to that described in humans. One principal difference between carnivore and human teeth is that the growth lines of carnivores do not terminate at perikymata on the tooth surface.     

Health of infants in an Imperial Roman skeletal sample: perspective from dental microstructure

This study examines general health in the first year of life of a population of 127 subadults from the Imperial Roman necropolis of Isola Sacra (2nd-3rd century ACE). Health status was determined by analyzing 274 deciduous teeth from these children for Wilson bands (also known as accentuated striae), microscopic defects caused by a disruption to normal enamel development arising from some generalized external stressor. While macroscopic enamel defects, or hypoplasias, have long been used as proxies of general population health, we believe that this is the first population-wide study of microscopic defects in deciduous teeth. We used microstructural markers of enamel to attach very precise chronologies to Wilson band formation that allowed us to calculate maximum prevalence (MAP) and smoothed maximum prevalence (SMAP) distributions to portray what we believe to be a realistic risk profile for a past population of children. There appear to be two periods of high prevalence, the first beginning around age 2 months and continuing through month 5, and the second higher period beginning around month 6 and continuing through month 9. These results are discussed in light of historical records of Roman childhood rearing practices.     

Histological enamel indicator of childhood stress in prehistoric skeletal samples.

Although previous paleopathological studies have used disturbances in enamel formation as indicators of childhood stress, the full potential of this technique has not been realized. This paper presents a test case which demonstrates that the frequency of disturbed enamel formation (i.e., Wilson bands) is associated with other stress indicators (i.e., probability of dying and infectious lesions) in three prehistoric skeletal samples representing the Middle Woodland (10.3%), Mississippian Acculturated Late Woodland (21.4%), and the Middle Mississippian (40.0%). Additionally, the mean ages at death of individuals with at least one Wilson band are lower than those without bands.These results suggest that Wilson bands are an indicator of the relative proportion of individuals who are high susceptibles in prehistoric populations. The data also corroborate the hypothesis that the adoption of maize agriculture in the prehistoric American Midwest is associated with increased stress.     

Microstructural defects and enamel hypoplasia in teeth of Toxodon Owen, 1837 from the Pleistocene of Southern Brazil

Enamel hypoplasia is characterized by reduction in the enamel thickness, resulting from a disruption of ameloblast activity due to systemic physiological stress. The euhypsodont teeth of Toxodon, a notoungulate from the Pleistocene of South America, often exhibit signs of enamel hypoplasia, in the form of continuous grooves or a series of pits where the enamel is thinner than in normal areas. These defects alternate with areas of normal enamel, and sometimes more than one form of enamel hypoplasia is present on the same tooth. This study analysed teeth of Toxodon from the Pleistocene Touro Passo Formation and the coastal plain of State of Rio Grande do Sul, Southern Brazil. Six types of enamel hypoplasia were observed. Upper teeth present mainly superficial grooves on the buccal surface, and the defects are less severe than those observed in the lower teeth. In the lower incisors, deep grooves with mesiodistal rows of pits were observed, showing clearly cyclical changes, which to a lesser degree, exist in all teeth. These changes are likely related to the continuous growth of euhypsodont teeth. Seven specimens were analysed under scanning electron microscopy and optical microscopy, which showed the occurrence of microstructural changes associated with the macroscopic enamel defects. Enamel underlying in the vicinity of hypoplastic defects was aprismatic and associated with prominent pathologic striae. These pathological findings might indicate that toxodonts were exposed to some stressing conditions or that their teeth were more easily abraded due to a change in diet items, related to shifting climatic conditions.     

Dental indicators of stress and reduced age at death in prehistoric Native Americans

Considerable evidence supports the hypothesis that developmental enamel defects represent stress-induced growth disruptions. In this investigation, the relationship between different kinds of enamel defects and age at death is examined in the prehistoric Libben population from Ottawa County, Ohio. The sample consisted of the permanent dentitions of 143 individuals. Defects were classified based on the criteria of the Developmental Defects of Enamel (DDE) Index. The multifactorial age at death determinations of Lovejoy and coworkers (1977) were used in this analysis. Results reveal a significantly lower mean age at death for individuals with enamel defects vs. individuals with normal teeth. This pattern was clearly present for all defect types examined. No significant differences by sex were detected. The age-at-death distribution for individuals with normal teeth approximated the normal curve. The modal value was reached in the 35–40 year age class. The age-at-death distribution for individuals with enamel defects showed two peaks. The mode occurred in the 15–20 year age class, and the second, lower peak occurred in the 30–35 year age class. The early mortality of individuals with enamel defects may be related to biological damage to the immune system during prenatal or postnatal development. © 1996 Wiley-Liss, Inc.     

Test of histological methods of determining chronology of accentuated striae in deciduous teeth

One of the difficulties that has prevented Wilson bands, internal defects of enamel growth, from achieving maximum usefulness as indicators of stress in bioarchaeological studies is the inaccuracy of the methods for determining ages of defect occurrence. This study tests a technique that uses microstructural growth markers of enamel to establish the chronology of Wilson bands in deciduous teeth. A sample of 274 teeth from 127 subadults from an Imperial Roman necropolis was sectioned and examined under a microscope. The sample ranged in age at death from birth to 13 years. Sixty-four teeth from 50 individuals were found to have 447 Wilson bands in total. Of those contributing multiple teeth to the sample, 13 individuals had at least one Wilson band in each of two teeth that could be identified as having been formed coevally. These provided the basis for testing the methodology. Paired t-tests found no significant differences between the chronologies of matched pairs in the two teeth, with mean ages from each differing by less than 1 day. The authors propose a hypothesis that explains the development of Wilson bands, and classifies them within the context of other features of enamel. The most important implications arising from this paper are: 1) aging methods using microstructural growth markers can be applied to deciduous teeth; 2) physiological stressors leave different traces in enamel, depending on severity and time of occurrence relative to total crown development; 3) no threshold level exists; all physiological stress will leave a record; 4) therefore, no minimal definition of a Wilson band can be delineated that recognizes all such events; 5) implying that studies using them will only identify minimum rates of occurrence.     

Intraflagellar transport is essential for endochondral bone formation

While cilia are present on most cells in the mammalian body, their functional importance has only recently been discovered. Cilia formation requires intraflagellar transport (IFT), and mutations disrupting the IFT process result in loss of cilia and mid-gestation lethality with developmental defects that include polydactyly and abnormal neural tube patterning. The early lethality in IFT mutants has hindered research efforts to study the role of this organelle at later developmental stages. Thus, to investigate the role of cilia during limb development, we generated a conditional allele of the IFT protein Ift88 (polaris). Using the Cre-lox system, we disrupted cilia on different cell populations within the developing limb. While deleting cilia in regions of the limb ectoderm had no overt effect on patterning, disruption in the mesenchyme resulted in extensive polydactyly with loss of anteroposterior digit patterning and shortening of the proximodistal axis. The digit patterning abnormalities were associated with aberrant Shh pathway activity, whereas defects in limb outgrowth were due in part to disruption of Ihh signaling during endochondral bone formation. In addition, the limbs of mesenchymal cilia mutants have ectopic domains of cells that resemble chondrocytes derived from the perichondrium, which is not typical of Indian hedgehog mutants. Overall these data provide evidence that IFT is essential for normal formation of the appendicular skeleton through disruption of multiple signaling pathways.     

Health and differential survival in prehistoric populations: prenatal dental defects.

Linear hypoplasia of the deciduous teeth is rare in most human populations, but common where nutritional status is poor. Deciduous enamel hypoplasia, hypocalcification, and hypoplasia-related caries are described in Middle and Late Woodland skeletal series from the Lower Illinois Valley. Gross enamel defects that can be referred to pre-natal development are found in 83 of 170 children under six years of age at death. Circular caries secondary to hypoplasia is significantly more common in the Late Woodland series, reflecting the apparent higher cariogenicity of Late Woodland diets. There is a significant association between prenatal dental defects and bony evidence for anemia and infectious disease. Children with enamel defects show relatively higher weaning age mortality than those without. These relationships suggest that at least moderate levels of malnutrition existed in Illinois Woodland populations.     

Insights from the inside: histological analysis of abnormal enamel microstructure associated with hypoplastic enamel defects in human teeth

We studied the abnormalities in enamel microstructure associated with enamel hypoplasia in human teeth from the early medieval (5th-7th century AD) cemetery of Barbing, Germany, using light and scanning electron microscopy. The main aim of the study was to test the hypothesis that by analyzing the microstructure of fully formed enamel it is possible to reconstruct the reaction pattern of secretory ameloblasts to stress events leading to enamel hypoplasia. From the histological findings, a sequence of increasing impairment of secretory ameloblast function involving three thresholds was deduced. Surpassing of each of these thresholds is assumed to result in characteristic changes in enamel microstructure attributable to specific functional/morphological alterations of secretory ameloblasts. Based on our results we propose a model identifying the principal factors influencing the reaction of secretory ameloblasts to stress. The present study demonstrates that by including microscopic analysis in the study of enamel hypoplasia, it is possible to obtain a more complete picture of the formation of these developmental defects than is possible by inspection of crown surface features alone, and to draw more substantiated conclusions about the possible nature of developmental defects of enamel.