Max Planck Institute for Evolutionary Anthropology

DEPARTMENT OF HUMAN EVOLUTION

DENTAL HARD TISSUE LABORATORY

Table of Contents
Tooth Histology and Human Evolution Dental Microstructure Principles of Incremental Development Histological Facilities Available Significance Contact Information 3 3 3 6 9 11

Tooth Histology and Human Evolution

The MPI-EVA Human Evolution Department has recently created state of the art facilities for the study of dental development in fossil hominids. Histological analysis of dental material facilitates understanding of the final functional products of the processes of development and growth, which may be understood in terms of enamel thickness (macrostructure) and enamel microstructure. Recent studies have provided information on age at death in hominids with developing dentitions, absolute and relative timing of dental development, age at first molar emergence, and differences in the developmental pathways of enamel formation. These studies have important implications for our understanding of hominid evolution and the origin of developmentally modern humans.

Dental Microstructure
Dental development in humans and great apes begins prior to birth and continues throughout adolescence. Like many biological systems, hard tissue formation is characterized by a circadian rhythm. Developmental rate and time are permanently recorded by incremental lines in enamel and dentine, which remain unchanged in these tissues for millions of years. Given that dental remains are the most common, well-preserved type of fossil evidence for extinct species of primates, examination of incremental growth processes may shed new light on the evolutionary developmental biology of early humans.

Principles of Incremental Development

Enamel is secreted by cells known as ameloblasts, which differentiate at the enamel-dentine junction and migrate outward towards what becomes the surface of the crown. The tracks left by these individual cells are known as enamel prisms (Figure 1). The prisms show cross-striations that result from the circadian rhythm of enamel secretion (Figures 1 & 2).

The successive positions of the advancing front of forming enamel are preserved as long-period incremental structures termed Retzius lines (Figures 1 & 3). These lines conform to the shape of the dentine horn in the cuspal enamel. In the lateral and cervical enamel, Retzius lines contact the enamel surface, forming circumferential rings known as perikymata (Figures 4 & 5). This region is referred to as lateral or imbricational enamel. The periodicity of Retzius lines may be determined in imbricational enamel by counting the number of cross-striations between Retzius lines (shown in the enlarged box in the upper right of Figure 1).

Figure 1. Schematic of enamel microstructure modified from Smith et al. (2003).

Dentine is produced by cells known as odontoblasts that generate dentine tubules (similar to enamel prisms), and show daily incremental lines known as von Ebners lines (equivalent to cross-striations) and long-period structures known as Andresens lines (equivalent to Retzius lines).

It has been demonstrated that both cross-striations and von Ebners lines show a 24-hour frequency, and because they are easier to image than von Ebners lines, cross-striations are used as a standard to determine the periodicity of long-period features in both tissues. Long-period features show a consistent periodicity within a single tooth and in all teeth belonging to the same individual, although this may vary within a taxon. Counts and measurements of these short- and long-period lines provide information on the rate and duration of enamel and dentine secretion, which may be combined to determine the total crown formation time and the rate and duration of root extension.

Figure 2. High magnification polarized light image of a Paranthropus boisei molar showing cross-striations. These light and dark bands run horizontally across the vertical enamel prisms, and are spaced approximately 5.5 microns apart. Fifty to 55 lines can be counted from the bottom to the top of the image, representing 50 - 55 days of tooth growth.

Histological Facilities Available

Hard tissue preparation and analysis laboratories are available for examination of dental material. Molding, casting, and replicating techniques may be used to study external features or naturally fractured surfaces. Standard refluxing and embedding techniques will be used to prepare samples, which may be sectioned with the use of a diamond wafering blade, ground, and polished to a fine sub-micron finish. For the preparation lab, we offer the use of two Buehler Isomet precision saws, a Buehler Ecomet Grinder-Polisher, and a large-diameter Logitech automated annular saw (APD1). Several forms of microscopy are available to study casts and histological sections, including transmitted and polarized light microscopy. We use the automated Olympus SZX9 Stereo microscope with the 12.5 mega-pixel DP 70 camera and SIS Imaging software for stereo microscopy, and the Olympus BX51 upright light microscope for transmitted and polarized light imaging, which can be coupled with the DP 70 and SIS software for a range of measurements, or used to capture images to 35 mm film. The microscopes are coupled to a high-speed computer with a 21-inch monitor and sufficient space to archive images to DVDs or for storage on our department server. A range of printing options is available, including local poster-sized image generation and high quality photo printing. Additional forms of analysis include scanning electron microscopy (SEM) and tandem scanning reflected light microscopy (confocal) available though collaborative arrangements with Stony Brook University and the University of Leipzig (Figures 6 & 7).

Figure 3. Polarized light photo montage of the lateral and cervical enamel of a Paranthropus boisei molar showing prominent accentuated growth lines and structural features known as Hunter-Schreger bands running from the lower border towards the tooth surface. The vibrant colors are partially due to post-mortem mineralization that is characteristic of fossil material.

Figures 4. (above) and 5. (below) illustrating perikymata (growth lines) on hominid teeth. These high-resolution replicas have been sputter coated with a metallic source to enhance the appearance of horizontally oriented long-period lines.

Significance
Early studies of fossil dentitions concentrated on aspects of crown size and morphology, root morphology, and the mechanics of occlusal wear. Additional studies have looked at the age of emergence and developmental chronology in fossil and extant taxa in an attempt to understand aspects of life history and phylogeny. Phylogenetic and functional importance has also been attached to tooth enamel thickness in analyses of Miocene hominoid fossils and interpretations of hominid origins. Examination of hard tissue development from a histological perspective is a relatively new field of odontological inquiry, particularly in relation to answering phylogenetic questions. Recently, there has been a dramatic increase in the number of studies on incremental dental development in hominoids. Studies on PlioPleistocene hominids and Neanderthals have indicated that the relatively slow developmental rate and prolonged duration of modern human crown formation is a fairly recent and unique development.

Figures 6. (above) and 7. (following page) illustrating prisms and incremental features on Moroccan hominid teeth. Figure 6 is a scanning electron micrograph of naturally fractured enamel (scale bar = 20 microns), and Figure 7 is a confocal micrograph of sub-surface enamel from the same tooth.

Members of the Department of Human Evolution are currently utilizing incremental structures preserved on the surface of teeth to study the growth processes of early humans (Figures 4 & 5). Current projects include a study of the development of Moroccan hominid dental material, as well as on-going work on Neanderthal growth processes. Additional projects include a study of fossil and extant Asian hominoid dental development, including a study of temporal and geographic developmental variation in Pongo, which may provide insight into the evolutionary developmental biology of the genus Homo. Over the next few years, we aim to build a substantial reference collection of histological sections of hominid material. This will yield additional developmental information that is not available from external surfaces or naturally fractured teeth, including developmental rate and total formation time. These types of data are illustrated here with images from histological sections of Paranthropus boisei (Figures 2 & 3). Fossil sections will be considered in the context of a large comparative collection of histological sections of extant hominoid material. We are open to considering proposals for collaborative work within all of these broad themes. Please do not hesitate to contact us for more information.

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Contact Information
Tanya M. Smith Max Planck Institute for Evolutionary Anthropology Department of Human Evolution Deutscher Platz 6 D-04103 Leipzig Germany Tel: + 49 (0) 341 - 35 50 362 Fax: + 49 (0) 341 - 35 50 399 E-mail: tsmith@eva.mpg.de

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2005 Department of Human Evolution

Max Planck Institute for Evolutionary Anthropology

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