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ARTICLES
Teeth are one of the best preserved and most commonly recovered elements ple, a recent study of a young captive
in primate fossil assemblages. Taxonomic, functional, and phylogenetic hypothe- gorilla demonstrated a strong corre-
ses often rely on dental characters, despite considerable evidence of homoplasy lation between accentuated lines in
in tooth form and large variation in tooth size within and among primates.1,2 tooth enamel and traumatic events,
Recent studies have led to new areas of research centered on incremental tooth including eye injury, subsequent hos-
development, chemical composition, and internal structure. Due to rapid techno- pital visits, and transfers to different
logical developments in imaging and elemental sampling, these new approaches enclosures.11 Other such studies have
have the potential to increase our understanding of developmental biology, suggested that patterns of develop-
including not only changes in the pace of growth and reproduction, but also our mental stress inside teeth may corre-
assessments of diets, migration patterns, environments, and taxonomy. The inte- late with social stress, weaning, eco-
gration of these temporal, chemical, and structural approaches heralds a bright logical variation, and menarche.12
future for the role of dental tissue research in evolutionary anthropology. Similar interpretations have been
based on patterns of hypoplasias on
external tooth surfaces,13–15 although
without knowledge of the precise
It is well established that dental in enamel and dentine microstruc- timing of tooth formation. Unfortu-
tissues preserve a permanent record ture.3–8 Counts and measurements of nately, relatively few data exist for
of their development through time, these incremental features have been individuals with corresponding be-
represented by incremental features used to determine the rate and dura- havioral, physiological, and ecologi-
tion of tooth formation, stress expe- cal records, a fact that prohibits con-
rienced during development, and the fident interpretations of accentuated
Tanya Smith is an Assistant Professor in age at death of juveniles. Incremen- lines or hypoplasias in fossil prima-
the Department of Anthropology at tal microstructure has traditionally
Harvard University, and an Associated tes or archeological material. This
Scientist in the Department of Human been assessed through physical sec- line of research would benefit from
Evolution at the Max Planck Institute for tioning and preparation of histologi- study of additional individuals with
Evolutionary Anthropology. Her primary cal sections, which reveal contrasting
research centers on the fundamental na- known histories, as was done by
ture of dental microstructure, including linear increments when viewed with Schwartz and colleagues.11 One
its variation in hominoid primates, as well light microscopy (Fig. 1). Develop- potentially complementary approach
as applications for understanding pri-
mate ontogeny and phylogeny. E-mail: mental features are often classified involves the assessment of changes
tsmith@fas.harvard.edu. as short- and long-period increments, in tooth chemistry, since dietary
Paul Tafforeau is a member of the imag- based on their rhythmic repeat inter-
ing group of the European Synchrotron changes associated with birth or
Radiation Facility (ESRF). His main vals (Box 1). Although histological weaning may be integrated with the
research is on fossil and modern primate investigations can yield highly accu- timing of tooth development.16–19
tooth structure, microstructure and de- rate estimates of tooth formation,
velopment. He is also in charge of the
development of synchrotron X-ray imag- until recently these types of studies
ing for paleontology at the ESRF. E-mail: have been limited to small samples INCREMENTAL TOOTH
paul.tafforeau@esrf.fr
due to their time-consuming and DEVELOPMENT
semi-destructive nature.6
Because tooth growth begins
Developmental Variation
Key words: tooth microstructure; incremental
feature; isotope; diagenesis; micro-computed before birth and continues through- Over the past 25 years, incremen-
tomography; micro-CT; synchrotron; phase out adolescence, assessment of incre- tal features have been investigated in
contrast; virtual histology
mental features may permit precise diverse primate taxa, particularly
reconstruction of an individual’s de- hominoid primates6 (Table 1). Dur-
V
C 2008 Wiley-Liss, Inc. velopmental history, including birth, ing the past decade, researchers have
DOI 10.1002/evan.20176
Published online in Wiley InterScience subsequent stress during develop- taken a rigorous comparative
(www.interscience.wiley.com). ment, and death (Fig. 2). For exam- approach through the use of large
214 Smith and Tafforeau ARTICLES
Accentuated line—a pronounced tine front; it relates to a stressor experi- neonatal line found in teeth developing
internal line corresponding to the posi- enced during tooth development (as during birth is the most common
tion of the developing enamel or den- opposed to an intrinsic rhythm). The example. See examples in Figure 2.
Figure B1. Incremental features in enamel (A,C) and dentine (B,D). Long-period lines are indicated by white arrows (Retzius lines
in A, Andresen lines in B) and blue dotted lines (perikymata in C, periradicular bands in D). Groups of four short-period lines are
indicated within each white bracket (cross-striations in A, von Ebner’s lines in B). Images are not to the same scale.
216 Smith and Tafforeau ARTICLES
Andresen lines—long-period which rapidly undergoes mineraliza- that are formed by Retzius lines as
(greater than circadian) incremental tion to form primary dentine. Den- they reach the enamel surface. This
features in dentine representing the tine formation begins at the dentine region of the crown is often called
successive positions of the dentine- horn underlying the future cusp tip the imbricational enamel. See exam-
forming front, and corresponding to and progresses inward through ples in Figure B1C.
periradicular bands on the root sur- secretion and downward through Periodicity—the number of days
face. They are temporally equivalent extension until it reaches the apex between long-period lines (Retzius
to Retzius lines and perikymata in of the root. lines or perikymata in enamel, Andre-
enamel. See examples in Figure B1B. Enamel development—enamel is sen lines or periradicular bands in
Cross-striations—short-period in- formed when ameloblasts secrete dentine). Periodicity typically is
cremental features in enamel running enamel matrix proteins that mineral- assessed by counting daily cross-
at right angles to enamel prisms. ize into long, thin bundles of hy- striations between pairs of Retzius
Cross-striations represent a 24-hour droxyapatite crystallites known as lines. The value for long-period line
cycle of ameloblast activity. Thus, enamel prisms. As the secretory periodicity is consistent within all teeth
the distance between adjacent cells progress outward toward the in an individual’s dentition, but may
cross-striations yields the daily rate future tooth surface, additional adja- vary among individuals in a taxon.
of enamel secretion. These striations cent cells are activated through Periradicular bands—external
are temporally equivalent to von extension until the forming front ridges and troughs encircling the
Ebner’s lines in dentine. See exam- reaches the cervix of the crown. tooth root that are temporally equiv-
ples in Figure B1A. Hypoplasia—feature formed when alent to the Andresen lines in the
Crown formation time—the dura- hard tissue formation has been dis- dentine (in addition to the long-pe-
tion of tooth crown secretion, either rupted by a stressor experienced dur- riod lines in enamel). See examples
for a single cusp or the entire crown ing development, which results in an in Figure B1D.
of multicusped teeth, which is typi- anomalous depression (furrow or pit) Retzius lines—long-period incre-
cally assessed through counts and on the surface of a tooth. These exter- mental features in enamel that repre-
measurements of cross-striations nal features are often associated with sent the position of the developing
and Retzius lines. accentuated lines internally. enamel front at successive points in
Dental development—the contin- Incremental features—micro- time. They are manifest on the tooth
uous process of tooth initiation, ma- scopic markings that represent surface as perikymata and are analo-
trix secretion, crown mineralization, intrinsic temporal rhythms in dental gous to the Andresen lines in dentine.
dental eruption, and root completion. hard tissue secretion. These can be The temporal repeat interval of Retzius
Primate dental development begins annual (cementum annulations), lines is known as their periodicity. See
before birth with initiation of the de- long-period (for example, Retzius examples in Figure B1A.
ciduous dentition, followed by initia- lines in enamel), or short-period (for von Ebner’s lines—short-period
tion of the permanent dentition. example, cross-striations in enamel). incremental features in dentine that
Dental eruption—the process of See examples in Figure B1. reflect a 24-hour cycle of dentine
tooth crown emergence; the tooth Laminations—daily incremental secretion and are temporally equiva-
must move past the bone margin features in enamel running parallel to lent to cross-striations and lamina-
(alveolar eruption) and the gum (gin- the developing enamel front and tions in enamel. See examples in
gival eruption) in order to emerge Retzius lines. Laminations are tempo- Figure B1B.
into the oral cavity and eventually rally equivalent to cross-striations but For additional information and
into functional occlusion. can be distinguished from them by illustrations, see Dean,3 FitzGerald,4
Dentine development—dentine is their relatively oblique orientation. Smith,5,6 Bromage,7 Tafforeau and
formed when odontoblasts secrete Perikymata—external ridges and colleagues,8 Dean and Scandrett,9
a collagenous matrix (predentine), troughs encircling the tooth crown and Smith and colleagues.10
debated topics in dental tissue logical age estimates of specific nyanzae, and a chimpanzee-like age
research. developmental stages, such as first at first molar eruption in A. turka-
molar eruption, which are correlated nensis.33–36 However, determination
Hominoid and Hominin with other developmental varia- of an ancestral great ape pattern has
bles.10,25,33,34 These studies suggest been complicated by recent docu-
Life History that, relative to other catarrhine mentation of substantial variation in
Dental microstructure analysis has primates, a prolonged hominoid life- extant great ape crown formation
frequently been employed in studies history pattern may have arisen in times and molar eruption ages,11,22,37
of the evolution of hominoid and the early Miocene, as shown by long as well as life-history parameters.38
hominin life histories.6,33 A key crown formation times in Proconsul Additional data on developmental
advantage of using incremental fea- nyanzae and Afropithecus turkanen- parameters in other early and middle
tures is their ability to yield chrono- sis, slow rates of root extension in P. Miocene taxa, as well as in addi-
ARTICLES New Visions of Dental Tissue Research 217
Figure 2. Illustration of histological reconstruction of an individual chimpanzee’s developmental history from birth (green line on left) to
death (blue arrow on right). The colored lines indicate unspecified postnatal developmental stress, with the corresponding age in days
given. The age at death determined from field notes was 1,372 days, or 3.76 years, suggesting that histologically derived times of stress
events are likely to be less than 2% different from the actual timing.
tional extant wild individuals, would tooth growth are conflicting. For greatly benefited from recent advan-
help in further clarifying the evolu- example, counts of perikymata have ces in technology. A synchrotron is a
tion of great ape life histories. been interpreted as suggesting machine that produces beams of
Dental microstructure studies of rapid development relative to recent light from accelerated electrons devi-
fossil hominins have almost com- humans42 or as indicating substan- ated by magnetic fields. Depending
pletely replaced determinations of tial overlap between Neanderthals on the energy, the light spectrum
dental eruption sequences or com- and modern humans.29,31 However, may range from radio frequencies to
parisons of relative degrees of devel- since perikymata numbers do not high-energy X-rays (hard X-rays).
opment. Dental eruption sequences directly yield formation times, infor- This type of light source facilitates
are known to be polymorphic within mation from internal features is criti- more efficient and diverse imaging
taxa,39 while comparisons of the cal to determine precise crown for- possibilities than do conventional lab-
degree of dental development mation and eruption ages. Recent oratory X-ray sources.45 This is a con-
requires an assumption of develop- histological studies of Neanderthal sequence of specific physical proper-
mental time or chronological age, permanent molar formation illustrate ties such as high flux, monochroma-
leading to circular reasoning.40 similarities in certain aspects of in- ticity, parallel geometry, and spatial
Bromage and Dean41 first applied ternal development, such as daily coherence (defined in Box 2). The first
dental microstructure analysis to rates of enamel secretion,43 but dif- three of these special properties pro-
estimate the age at first molar emer- ferences in cuspal enamel thickness duce high-quality absorption scans
gence in several juvenile Plio-Pleisto- and rates of crown extension, leading with gray levels that can be used to
cene hominins, which appeared to be to shorter molar formation times.10 quantify mineral densities.8
several years younger than modern Building on these differences, a A unique and critical aspect of
human children at a similar stage of study of the developmental status of synchrotron imaging for dental tis-
dental development. This and several the nearly complete dentition of a ju- sue research is based on phase-con-
subsequent studies have led to a gen- venile Belgian Neanderthal suggested trast techniques, which reveal subtle
eral consensus that the prolonged that Neanderthals did not share the variations in tissue structure that
childhood period and slow life his- prolonged life history of living and are invisible with other absorption
tory that are characteristic of living fossil Homo sapiens.10 Exactly where contrast imaging techniques such as
humans originated fairly recently, and when this transition took place radiography or conventional micro-
most likely within later members of within the genus Homo remains tomography. Over the past few
the genus Homo.6,33 unresolved. years, improvements in the micro-
Unfortunately, little is known tomography beamline ID19 at the
about incremental tooth growth in European Synchrotron Radiation
Middle Pleistocene hominins, save
Virtual Histology Facility in Grenoble, France, have
for counts of perikymata on anterior A promising new area of incremen- permitted rapid imaging of micro-
teeth of Homo antecessor and Homo tal feature research is the application structure in fully mineralized
heidelbergensis.42 As reviewed by of virtual histology via propagation- dental tissues at submicron resolu-
Smith and colleagues10 and Guatelli- phase contrast X-ray synchrotron tion.8,44–47 This nondestructive tech-
Steinberg,101 data on Neanderthal microtomography,44,45 which has nique yields images of incremental
218 Smith and Tafforeau ARTICLES
features in tooth enamel and den- can be used to characterize dental growth of a tooth at the cellular
tine, as well as the neonatal line, development and age at death in level.
which can be seen in highly miner- material that is unavailable for
alized fossils that are millions of traditional physical sectioning.47 It
years old.44 Highly precise assess- may also be used to provide novel DEVELOPMENTAL TIME AND
ments of tooth formation are now internal information from previously
TOOTH CHEMISTRY
possible though the nondestructive studied fossils. Beyond revealing de-
virtual determination of long-period velopmental information in fossil- Studies of tooth chemistry, partic-
line periodicity. In combination with ized dentitions nondestructively, it ularly the proportions of specific ele-
virtual data on cuspal enamel thick- is hoped that virtual histology will ments such as strontium and cal-
ness and the developmental status eventually facilitate precise under- cium or ratios of elemental isotopes
of unerupted teeth, this approach standing of the three-dimensional such as 13C/12C, 18O/16O, and
ARTICLES New Visions of Dental Tissue Research 219
Figure 3. Stable isotopes studied in dental tissues (reviewed in Mays49). *Enamel proteins
may one day yield isotopic data from individual amino acids.48 The Problem of Enamel
Mineralization
Isotopic studies of dental enamel
87
Sr/86Sr, have expanded over the ratios revealed dietary shifts and have recently underscored the impor-
past decade to integrate information migration events. Wright and tance of considering the complex
on the pattern and duration of Schwarcz18 examined changes in car- tooth mineralization process for
dental development. Initial analyses bon and oxygen isotopes across sev- accurate temporal interpretation
required bulk samples taken from a eral teeth of the same individuals of chemical signatures.8,16,46,51,55–58
large proportion of the tooth crown spanning the time from birth to ado- Mineralization proceeds in a series
or root to infer an organism’s diet lescence. Their results suggested that of stages as apatite crystallites grow
geographic location, or climate of infant diets in their archeological in length and thickness, with the ma-
the region in which the individual population were supplemented with jority of mineral incorporation
lived during tooth formation48,49 solid food by two years of age, occurring some time after the end of
(Fig. 3). In contrast to bone, dental although breastfeeding appeared to secretion.8,16,55 Isotopic studies of
tissues mineralize over a relatively continue for an extended period. intratooth variation often sample
short time without subsequent Building on this bulk sampling enamel serially from one end of the
remodeling,50 providing a narrower approach, Fuller, Richards, and crown to the other, assuming a rate
window of recorded time. Isotopic Mays19 used the internal spatial pat- of constant growth and mineraliza-
studies of dental tissues may provide tern of tooth formation to track tion, which allows isotopic values to
information about young individuals; changes in nitrogen and carbon iso- be regressed against sampling posi-
once the tooth roots are complete, topes in serial sections of succes- tion as a proxy for a specific point in
relatively little tissue is added later sively forming tooth crowns and developmental time (see, for exam-
in life (in the form of small amounts roots, demonstrating differences ple, Fig. 3, p. 208, in Balasse and col-
of secondary/tertiary dentine and between earlier- and later-formed leagues59). However, the final miner-
cementum). regions within a tooth, which were alization process does not follow the
interpreted as pre- and postweaning pattern of matrix secretion, which is
dietary signals. represented by incremental fea-
Variation Within Individuals Most recently, Humphrey and col- tures.8,60,61 This is particularly criti-
Several recent studies have used leagues16,102 used laser ablation cal for studies that attempt to relate
information on tooth growth to look inductively coupled plasma mass isotopic information to local incre-
at migration patterns and dietary spectrometry to sample strontium/ mental features.8,16,17,54 It is still
changes in human archeological calcium ratios of pre- and postnatal unclear when particular elements are
contexts.51 Sealy, Armstrong, and enamel in histological thin sections incorporated into enamel, and thus
Schrire52 examined skeletal and den- of teeth from modern humans. This if the recovered signal has been
tal samples from five unknown technique involves focusing a thin ‘‘dampened’’ (shifted in time or modi-
individuals, including pairs of ear- laser beam on a small area of a tooth fied in degree) by subsequent miner-
lier- and later-forming teeth, and surface or thin section that trans- alization during maturation.16,51 Fur-
concluded that differences in carbon, forms (ablates) the solid into a ther complicating the situation,
nitrogen, and strontium isotope plasma, which is then analyzed with many studies of tooth mineralization
220 Smith and Tafforeau ARTICLES
have been performed on nonprimate minor for particular elements.16 This ology were used to suggest that the
mammals,8,55,57–59 and thus may not study demonstrated a consistent individual had migrated from a
clarify how primate teeth mature, change in ratios sampled before the region at least 20 kilometers away.
since tooth mineralization and matu- neonatal (birth) line and after this An advantage of using tooth
ration processes vary among mam- line, suggesting that physiological enamel for isotopic studies is its
mals.50,62 differences in strontium discrimina- greater resistance than bone to dia-
Passey and Cerling55 attempted to tion were recorded at the time of genetic change.67,68 Diagenesis, or
model the dampening effect that sub- matrix secretion and were not lost the process of chemical change after
sequent mineralization may have on during subsequent mineralization. death (including fossilization) is
the original ‘‘input signal’’ recorded Humphrey and colleagues17 also highly variable and still not com-
as a tooth develops. By measuring recently applied laser ablation to pletely understood in hard tis-
phosphorous, an indicator of hy- developmentally overlapping teeth in sues.51,54,66 However, diagenesis can
droxyapatite content, they found that two baboon individuals and found strongly affect the assessment of
tooth mineralization occurred in a some correspondence in ratio tooth chemistry in archeological or
spatially diffuse pattern that differed changes between simultaneously fully fossilized tissues. Because
among teeth and continued after ma- forming enamel (between teeth). enamel is approximately 95% miner-
trix secretion was finished. Based on This provides additional evidence alized by the end of formation, in
this information, they proposed a that subsequent mineralization does contrast to dentine or bone at
model to predict the original input not completely dampen the original approximately 70%–75%, the chemi-
signal from knowledge of the (biogenetic) signal. cal composition of mature enamel is
‘‘mature (measured) signal’’ and the believed to provide a more faithful
process of amelogenesis for teeth record of the original mineralization
growing at a constant rate through- Fossil Hominins and Tooth process. Furthermore, given the
out the life of the animal, with the highly inorganic composition of
aim of interpreting temporal infor-
Diagenesis enamel, it has been suggested to ap-
mation present in the pattern of Isotopic analyses of fossil hominin proximate a ‘‘closed system’’ that is
chemical variation. However, this dental tissues have primarily cen- more impermeable to alteration from
model appears to be of limited utility tered on elucidating southern African the postmortem burial environment
for hominoid teeth, as rates of Plio-Pleistocene hominin diets54,63,64 (but see evidence of enamel diagene-
enamel secretion and extension vary and potential environmental shifts,65 sis below and in Wang and Cerling,68
from the beginning to the end of and these studies are beginning to Kohn, Schoeninger, and Barker,69
crown formation (reviewed in provide insight into hominin mobil- and Schoeninger and colleagues70).
Smith6). Additional study is of criti- ity.53 Dental tissues are of particular However, one limitation of inorganic
cal importance to document the tim- importance for older time periods component analyses is the lack of in-
ing and relative degree of incorpora- because little organic material (col- ternal verification of results; in cer-
tion of particular elements in lagen) remains in fossil bone and tain cases it is difficult to exclude
primate tooth enamel. diagenetic processes are highly likely environmental sources of particular
Tafforeau and colleagues8 applied to have modified the inorganic com- elements that may bias results.
X-ray synchrotron microtomography ponent.66 Although most dietary In a study of modern and fossil
to the study of tooth mineralization, studies have used bulk sampling, a southern African fauna, Sponheimer
finding that rhinoceros enamel recent approach by Sponheimer and and colleagues64 argued that similar-
shows a 20-micron thick zone near colleagues54 related sequential iso- ity in strontium/calcium ratios
the enamel-dentine junction that topic laser ablation samples from the between modern and fossil animals
appears to mineralize rapidly after tooth surface to counts of periky- demonstrated that diagenesis was
secretion. This suggests that isotopic mata in four Paranthropus teeth. unlikely to have a major impact on
analyses of intratooth variation may Their results showed a high degree these ratios in their sample of fossil-
recover a near-primary signal from of variation within and between ized dental enamel. However, others
sampling along this zone (also see teeth. This, based on estimates of the have abandoned the use of trace-ele-
Balasse56 and Zazzo, Balasse, and time represented by perikymata, was ment ratios such as that of strontium
Patterson58). Even when the spatial interpreted as seasonal and interan- to calcium, in part because it is diffi-
pattern of mineralization is taken nual variation, implying that Para- cult to detect when elements have
into account, it is unlikely that a sin- nthropus was not a dietary specialist. leeched in or out of a tooth or
gle isotopic sample will represent a Less research has been done on bone.71,72 Because even slight
discontinuous or instantaneous point hominin migration. Richards and amounts of particular elements or iso-
in time because some degree of time colleagues53 recently presented iso- topes will be detected in most cases, a
averaging will occur during minerali- topic evidence of Neanderthal mobil- ratio is always obtained, but it is often
zation.8,56,57 However, a recent study ity based on strontium isotopes in quite difficult to verify that it repre-
of strontium/calcium ratios in dental enamel. Differences in the sents the original signal. Hydroxyapa-
human molar enamel suggests that strontium detected in the internal tite, the major mineralized compo-
the effect of time averaging may be tooth enamel relative to the local ge- nent of enamel, dentine, and bone,
ARTICLES New Visions of Dental Tissue Research 221
Figure 5. Developing chimpanzee tooth (shown in Figs. 1 and 2) imaged with an isotropic voxel size of 13.8 lm using a conventional
micro-CT before sectioning. A) 3D rendering of the occlusal surface. B) Virtual vertical cut through the mesial cusps. C) 3D model of the
tooth after segmentation showing the enamel cap (transparent yellow), EDJ (red), and dentine (transparent blue). D) Isolated enamel
cap used for volume measurement (178.6 mm3). E) EDJ used for surface area measurement (188 mm2). F) Dentine used for volume
measurement (293.3 mm3). The volume of dentine and pulp in the enamel cap delimited by the basal plane (following Olejniczak and
colleagues86) is 239 mm3, yielding a 3D average enamel thickness of 0.95 mm and a 3D relative enamel thickness of 15.31 (following
Tafforeau,46 Olejniczak and colleagues,78 and Kono.84)
ment over medical CT imaging teeth (for example, cases A to C in utility of micro-CT for assessment of
because fine anatomical details can Figure 4), there is little advantage in three-dimensional (3D) molar enamel
be resolved (Fig. 5) and accurate using a synchrotron. However, prop- thickness and enamel distribution.
metric assessment can be done on agation phase-contrast X-ray syn- Three-dimensional enamel thickness
relatively small dental samples.46,77,79 chrotron microtomography often data are now available for a broad
This is particularly important for reveals the enamel-dentine junction sample of extant primates,46,78,84 the
studies of enamel thickness, given in strongly modified teeth even when fossil ape Gigantopithecus blacki,85
the limitations of conventional radi- there is little or no absorption con- Neanderthals,43,86 australopiths,103
ography or tomography,79 as well as trast.45,46 This is a particularly valua- and Middle Stone Age Homo sapi-
limitations in physically sectioning ble tool for imaging diagenetically ens.87 New 3D data have confirmed
invaluable fossil material. Aspects of altered fossil teeth, especially tissue broad trends in primate enamel
microscopic dental tissue structure structure in older hominins and ear- thickness derived from earlier two-
can also be investigated nondestruc- lier primate fossils. dimensional studies78 and have also
tively with high precision; this is one demonstrated that thick molar
of the main features of synchrotron enamel is not found in all fossil hom-
imaging. Despite these advantages,
Tooth Crown and Root Structure inin taxa.86 Future applications of
more commonly available conven- Most anthropological applications micro-CT are needed to clarify phylo-
tional microtomographs may yield of micro-CT techniques have exam- genetic or functional aspects of hom-
absorption data quite similar to syn- ined internal tooth structure from inin enamel thickness, particularly
chrotron data for voxel sizes between virtual two-dimensional planes of given the diverse range of measure-
5 and 50 microns. For standard sections.46,80–83 Reiko Kono’s land- ment techniques employed in previ-
absorption imaging of well-preserved mark study84 firmly established the ous studies. In particular, such data
ARTICLES New Visions of Dental Tissue Research 223
tructive 3D perspective, demonstrat- of dietary or environmental change secretion determined along the prism
ing that dental traits commonly iden- while providing the potential for length from the cross-striation spac-
tified on the outer enamel surface identifying life history events such as ing. These examples of research inte-
may result from different EDJ con- birth or weaning.8,16,17 gration represent a few promising
figurations. It appears that in some Other examples of this synergistic means to advance our understanding
cases these traditional ‘‘discrete approach include studies of hominin of primate developmental biology,
traits’’ may not be developmentally dental development that have used and to initiate the formulation of
homologous.94 This study also dem- micro-CT imaging to characterize more refined areas of dental tissue
onstrated another potential advant- aspects of internal structure or the research over the coming years.
age of micro-CT studies: Teeth that developmental status of unerupted
show marked attrition that obscures teeth in situ.10,47,87 Structural fea-
or obliterates external morphology tures of developmental significance, ACKNOWLEDGMENTS
can still be included in analyses of such as the thickness of cuspal Special thanks to John Fleagle for
EDJ shape, underscoring the value of enamel, the length of the EDJ, and patience with this manuscript, and to
nondestructive studies of internal the length of tooth roots, can be Jean-Jacques Hublin for support of
tooth structure. Similar research on accurately characterized nondestruc- this research. Comments on the
external dental tissue structure may tively. Data obtained from conven- manuscript were kindly provided by
one day allow worn teeth to be rebuilt tional or virtual histological investi- Ben Fuller, Vaughan Grimes, Donald
volumetrically to better understand gations can then be used to estimate Reid, Chris Dean, Mary Maas, Allison
their original form, as well as the the duration of formation along Cleveland, and one anonymous
complex process of tooth attrition. these virtual growth axes, leading to reviewer. We thank Ben Fuller,
more precise estimations of tooth Vaughan Grimes, and Mike Richards
growth and age at death in rare fos- for illuminating discussions of iso-
A WAY FORWARD: INTEGRATION sil juveniles. topes; Louise Humphrey and Wendy
OF TEMPORAL, CHEMICAL, AND A final example of research inte- Dirks for access to unpublished data;
gration is the quantification of volu- Bernard Wood for assistance with an
STRUCTURAL INFORMATION
metric growth rates, which can be earlier version of Box 1; Jose Baruchel
One challenge of having new tools calculated as overall crown averages with Box 2; and Christophe Boesch,
is learning how to best apply them. using knowledge of incremental de- Lawrence Martin, and Michel Tous-
It is clear that decades of studies on velopment and 3D dental tissue met- saint, who kindly provided material.
tooth development, chemistry, and rics. For example, a chimpanzee first The Max Planck Society and Euro-
structure have provided valuable molar tooth was subjected to micro- pean Synchrotron Radiation Facility
insight into the evolution of primate CT scanning before histological sec- provided financial support.
life history, paleodiets, and taxon- tioning and analysis (Figs. 1, 2, 4),
omy. Future studies of dental tissues yielding the volume of enamel (178.6
will undoubtedly benefit from mm3) and dentine (293.3 mm3) and
REFERENCES
increased technological efficiency in the maximum formation time of
chemical sampling, tomographic each tissue (722 and 1427 days, 1 Collard M, Wood B. 2000. How reliable are
human phylogenetic hypotheses? Proc Natl
scanning resolution and sensitivity, respectively). This information was Acad Sci USA 97:5003–5006.
and increased computing power for used to estimate minimum average 2 Hooijer DA. 1948. Prehistoric teeth of man
data processing. We suggest that volumetric crown growth rates and of the orang-utan from central Sumatra,
with notes on the fossil orang-utan from Java
additional advancements may be (247.4 lm3/day for enamel and 205.5 and southern China. Zool Mededeelingen
found through the synergy of these lm3/day for dentine), along with the 29:175–301.
seemingly disparate research areas. minimum average coronal extension 3 Dean MC. 1995. The nature and periodicity
For example, studies of tooth rate (260.4 lm2/day, using a mea- of incremental lines in primate dentine and
their relationship to periradicular bands in OH
chemistry may be better informed sured EDJ surface area of 188 mm2). 16 (Homo habilis). In: Moggi-Cecchi J, editor.
by complementary applications of Volumetric rates of primate tooth Aspects of dental biology: paleontology, an-
thropology and evolution. Florence: Interna-
nondestructive microtomography to growth can be compared to rates of tional Institute for the Study of Man. p 239–
characterize mineral density and the brain growth100 or other aspects of 265.
duration of mineralization8 (Fig. 6). somatic development, as well as to 4 FitzGerald CM. 1998. Do enamel microstruc-
Furthermore, absorption micro-CT characterize volumetric increases in tures have regular time dependency? Conclu-
sions from the literature and a large-scale
imaging of teeth before sampling the formation of the deciduous denti- study. J Hum Evol 35:371–386.
may yield information about internal tion or the successive formation of 5 Smith TM. 2006. Experimental determination
diagenetic modification, leading to the permanent dentition. This analy- of the periodicity of incremental features in
enamel. J Anat 208:99–114.
more efficient selection of material sis can also be extended to volumet-
6 Smith TM. 2008 Incremental dental develop-
for chemical analyses. Similarly, the ric quantification at the microstruc- ment: methods and applications in hominoid
integration of incremental develop- tural level with propagation phase evolutionary studies. J Hum Evol 54:205–224.
ment, maturation patterns, and contrast X-ray synchrotron microto- 7 Bromage TG. 1991. Enamel incremental peri-
odicity in the pig-tailed macaque: a polychrome
isotopic sampling strategies may mography, as prisms may be virtu- fluorescent labeling study of dental hard tis-
facilitate more precise assessments ally extracted in 3D and the rate of sues. Am J Phys Anthropol 86:205–214.
ARTICLES New Visions of Dental Tissue Research 225
8 Tafforeau P, Bentaleb I, Jaeger J-J, Martin C. 24 Macho GA. 2001. Primate molar crown for- 45 Tafforeau P, Boistel R, Boller E, Bravin A,
2007. Nature of laminations and mineralization mation times and life history evolution revis- Brunet M, Chaimanee Y, Cloetens P, Feist M,
in rhinoceros enamel using histology and X-ray ited. Am J Primatol 55:189–201. Hoszowska J, Jaeger J-J, Kay RF, Lazzari V,
synchrotron microtomography: potential impli- 25 Dirks W, Bowman JE. 2007. Life history Marivaux L, Nel A, Nemoz C, Thibault X,
cations for palaeoenvironmental isotopic stud- theory and dental development in four species Vignaud P, Zabler S. 2006. Applications of
ies. Palaeogeogr Palaeoclimatol Palaeoecol of catarrhine primates. J Hum Evol 53:309– X-ray synchrotron microtomography for non-
246:206–227. 320. destructive 3D studies of paleontological speci-
9 Dean MC, Scandrett AE. 1996. The relation mens. Appl Phys A 83:195–202.
26 Godfrey LR, Schwartz GT, Samonds KE,
between long-period incremental markings in Jungers W, Catlett KK. 2006. The secrets of 46 Tafforeau P. 2004. Aspects phyloǵnétiques et
dentine and daily cross-striations in enamel in lemur teeth. Evol Anthropol 15:142–154. fonctionnels de la microstructure de l’émail
human teeth. Arch Oral Biol 41:233–241. dentaire et de la structure tridimensionnelle des
27 Reid DJ, Dean MC. 2000. Brief communica-
10 Smith TM, Toussaint M, Reid DJ, Olejniczak molaires chez les primates fossiles et actuels:
tion: the timing of linear hypoplasias on human
AJ, Hublin J-J. 2007. Rapid dental development apports de la microtomographie à rayonnement
anterior teeth. Am J Phys Anthropol 113:135–139.
in a Middle Paleolithic Belgian Neanderthal. X synchrotron. PhD Thesis, Université de Mont-
28 Reid DJ, Dean MC. 2006. Variation in mod- pellier II, France. http://www.paleoanthro.org/
Proc Natl Acad Sci USA 104:20220–20225.
ern human enamel formation times. J Hum dissertation_list.htm
11 Schwartz GT, Reid DJ, Dean MC, Zihlman Evol 50:329–346.
AL. 2006. A faithful record of stressful life 47 Smith TM, Tafforeau PT, Reid DJ, Grün R,
29 Reid DJ, Guatelli-Steinberg D, Walton P. Eggins S, Boutakiout M, Hublin J-J. 2007. Ear-
events preserved in the dental developmental
2008. Variation in modern human premolar liest evidence of modern human life history in
record of a juvenile gorilla. Int J Primatol
enamel formation times: implications for Nean- North African early Homo sapiens. Proc Natl
22:837–860.
dertals. J Hum Evol 54:225–235. Acad Sci USA 104:6128–6133.
12 Dirks W, Reid DJ, Jolly CJ, Phillips-Conroy
30 Mann A, Monge JM, Lampl M. 1991. Investi- 48 Schoeninger MJ. 1995. Stable isotope stud-
JE, Brett FL. 2002. Out of the mouths of
gation into the relationship between perikymata ies in human evolution. Evol Anthropol 4:83–
baboons: stress, life history, and dental develop-
counts and crown formation times. Am J Phys 98.
ment in the Awash National Park hybrid zone,
Anthropol 86:175–188.
Ethiopia. Am J Phys Anthropol 118:239–252. 49 Mays S. 2000. New directions in the analysis
31 Guatelli-Steinberg D, Reid DJ. 2008. What of stable isotopes in excavated bones and teeth.
13 Guatelli-Steinberg D. 2001. What can devel-
molars contribute to an emerging understand- In: Cox M, Mays S, editors. Human osteology
opmental defects of enamel reveal about physi-
ological stress in nonhuman primates? Evol ing of lateral enamel formation in Neandertals in archaeology and forensic science. Cam-
Anthropol 10:138–151. vs. modern humans. J Hum Evol 54:236–250. bridge: Cambridge University Press. p 425–438.
14 Skinner MF, Hopwood D. 2004. Hypothesis 32 Reid DJ, Ferrell RJ. 2006. The relationship 50 Boyde A. 1997. Microstructure of enamel.
for the causes and periodicity of repetitive lin- between number of striae of Retzius and their In: Chadwick DJ, Cardew G, editors. Dental
ear enamel hypoplasia in large, wild African periodicity in imbricational enamel formation. enamel. Chichester: Wiley. p 18–31.
(Pan troglodytes and Gorilla gorilla) and Asian J Hum Evol 50:195–202. 51 Montgomery J, Evans JA. 2006. Immigrants
(Pongo pygmaeus) apes. Am J Phys Anthropol 33 Dean MC. 2006. Tooth microstructure tracks on the Isle of Lewis: combining traditional
123:216–235. the pace of human life-history evolution. Proc funerary and modern isotope evidence to inves-
15 Katzenberg MA, Herring DA, Saunders SR. R Soc B 273:2799–2808. tigate social differentiation, migration and die-
1996. Weaning and infant mortality: evaluating 34 Kelley J, Smith TM. 2003. Age at first molar tary change in the Outer Hebrides of Scotland.
the skeletal evidence. Yearbook Phys Anthropol emergence in early Miocene Afropithecus turka- In: Gowland R, Kn̈sel C, editors. Social archae-
39:177–199. nensis and life-history evolution in the Homi- ology of funerary remains. Oxford: Oxbow
16 Humphrey LT, Dean MC, Jeffries TE. 2007. noidea. J Hum Evol 44:307–329. Books. p 122–142.
An evaluation of changes in strontium/calcium 35 Beynon AD, Dean MC, Leakey MG, Reid DJ, 52 Sealy J, Armstrong R, Schrire C. 1995.
ratios across the neonatal line in human decid- Walker A. 1998. Comparative dental develop- Beyond lifetime averages: tracing life histories
uous teeth. In: Bailey SE, Hublin J-J, editors. ment and microstructure of Proconsul teeth through isotopic analysis of different calcified
Dental perspectives on human evolution: state from Rusinga Island, Kenya. J Hum Evol tissues from archaeological human skeletons.
of the art research in dental paleoanthropology. 35:163–209. Antiquity 69:290–300.
Dordrecht: Springer. p 301–317. 36 Smith TM, Martin LB, Leakey MG. 2003. 53 Richards M, Harvati K, Grimes V, Smith C,
17 Humphrey LT, Dirks W, Dean MC, Jeffries Enamel thickness, microstructure and develop- Smith T, Hublin J-J, Karkanas P, Panagopoulou
TE. 2008. Tracking dietary transitions in wean- ment in Afropithecus turkanensis. J Hum Evol E. 2008. Isotope evidence of Neanderthal mobil-
ling baboons (Papio hamadryas anubis) using 44:283–306. ity. J Archeol Sci 35:1251–1256.
strontium/calcium ratios in enamel. Folia Pri- 37 Zihlman A, Bolter D, Boesch C. 2004. Wild 54 Sponheimer M, Passey BH, de Ruiter DJ,
matol 79:197–212. chimpanzee dentition and its implications for Guatelli-Steinberg D, Cerling TE, Lee-Thorp JA.
18 Wright LE, Schwarcz HP. 1998. Stable car- assessing life history in immature hominin fos- 2006. Isotopic evidence for dietary variability
bon and oxygen isotopes in human tooth sils. Proc Natl Acad Sci USA 101:10541–10543. in the early hominin Paranthropus robustus.
enamel: identifying breastfeeding and weaning 38 Robson SL, van Schaik CP, Hawkes K. 2006. Science 314:980–982.
in prehistory. Am J Phys Anthropol 106:1–18. The derived features of human life history. In: 55 Passey BH, Cerling TE. 2002. Tooth enamel
19 Fuller BT, Richards MP, Mays SA. 2003. Sta- Hawkes K, Paine RP, editors. The evolution of mineralization in ungulates: implications for
ble carbon and nitrogen isotope variations in human life history. Santa Fe: School of Ameri- recovering a primary isotopic time-series. Geo-
tooth dentine serial sections from Wharram can Research Press. p 17–44. chim Cosmochim Acta 66:3225–3234.
Percy. J Archeol Sci 30:1673–1684. 39 Smith BH, Garn SM. 1987. Polymorphisms 56 Balasse M. 2003. Potential biases in sam-
20 Schwartz GT, Reid DJ, Dean C. 2001. Devel- in eruption sequence of permanent teeth in pling design and interpretation of intra-tooth
opmental aspects of sexual dimorphism in American children. Am J Phys Anthropol isotope analysis. Int J Osteoarcheol 13:3–10.
hominoid canines. Int J Primatol 22:837–860. 74:289–303. 57 Hoppe KA, Stover SM, Pascoe JR, Amund-
21 Guatelli-Steinberg D, Reid DJ, Bishop TA, 40 Smith BH. 2004. The paleontology of growth son R. 2004. Tooth enamel biomineralization in
Larsen CS. 2005. Anterior tooth growth periods and development. Evol Anthropol 13:239–241. extant horses: implications for isotopic micro-
in Neanderthals were comparable to those of 41 Bromage TG, Dean MC. 1985. Re-evaluation sampling. Palaeogeogr Palaeoclimatol Palaeoe-
modern humans. Proc Natl Acad Sci USA of the age at death of immature fossil homi- col 206:355–365.
102:14197–14202. nids. Nature 317:525–527. 58 Zazzo A, Balasse M, Patterson WP. 2005.
22 Smith TM, Reid DJ, Dean MC, Olejniczak 42 Ramirez Rozzi FV, Bermudez de Castro JM. High-resolution d13C intratooth profiles in bo-
AJ, Martin LB. 2007. Molar development in 2004. Surprisingly rapid growth in Neander- vine enamel: implications for mineralization
common chimpanzees (Pan troglodytes). J Hum thals. Nature 428:936–939. pattern and isotopic attenuation. Geochim Cos-
Evol 52:201–216. mochim Acta 69:3631–3642.
43 Macchiarelli R, Bondioli L, Debénath A,
23 Smith TM, Reid DJ, Dean MC, Olejniczak Mazurier A, Tournepiche J-F, Birch W, Dean C. 59 Balasse M, Smith AB, Ambrose SH, Leigh
AJ, Ferrell RJ, Martin LB. 2007. New perspec- 2006. How Neanderthal molar teeth grew. SR. 2003. Determining sheep birth seasonality
tives on chimpanzee and human dental devel- Nature 444:748–751. by analysis of tooth enamel oxygen isotope
ratios: the Late Stone Age site of Kasteelberg
opment. In: Bailey SE, Hublin J-J, editors. Den- 44 Tafforeau P, Smith TM. 2008. Nondestruc-
tal perspectives on human evolution: state of (South Africa). J Archeol Sci 30:205–215.
tive imaging of hominoid dental microstructure
the art research in dental paleoanthropology. using phase contrast X-ray synchrotron micro- 60 Suga S. 1983. Comparative histology of the
Dordrecht: Springer. p 177–192. tomography. J Hum Evol 54:272–278. progressive mineralization pattern of develop-
226 Smith and Tafforeau ARTICLES
ing enamel. In: Suga S, editor. Mechanisms of 75 Fisher DC, Fox DL. 1998. Oxygen isotopes in tocene hominids. IV. Mandibular postcanine
tooth enamel formation. Tokyo: Quintessence mammoth teeth: sample design, mineralization root morphology. J Anat 156:107–139.
Publishing. p 167–203. patterns, and enamel-dentine comparisons. J 89 Chaimanee Y, Yamee C, Tian P, Khaowiset
61 Wilson PR, Beynon AD. 1989. Mineraliza- Vert Paleo 18:41A–42A. K, Marandat B, Tafforeau P, Nemoz C, Jaeger
tion differences between human deciduous 76 Hayakawa T, Mishima H, Yokota I, Sakae T, J-J. 2006. Khoratpithecus piriyai, a late Miocene
and permanent enamel measured by quantita- Kozawa Y, Nemoto K. 2000. Application of hominoid of Thailand. Am J Phys Anthropol
tive microradiography. Arch Oral Biol 34:85– high resolution microfocus X-ray CT for the 131:311–323.
88. observation of human tooth. Dent Mat J 19:87–
90 Kupczik K, Dean MC. 2008. Comparative
62 Kohn MJ. 2004. Comment: tooth enamel 95.
observations on the tooth root morphology of
mineralization in ungulates: implications for 77 Olejniczak AJ, Grine FE, Martin LB. 2007. Gigantopithecus blacki. J Hum Evol 54:196–
recovering a primary isotopic time-series, by Micro-computed tomography of the post-canine 204.
B.H. Passey and T.E. Cerling (2002). Geochim dentition: methodological aspects of three-
Cosmochim Acta 68:403–405. dimensional data collection. In: Bailey SE, 91 Bailey SE. 2005. Diagnostic dental differen-
Hublin J-J, editors. Dental perspectives on ces between Neandertals and Upper Paleolithic
63 Lee-Thorp JA, van der Merwe NJ, Brain CK. modern humans: getting to the root of the mat-
1994. Diet of Australopithecus robustus at human evolution: state of the art research in
dental paleoanthropology. Dordrecht: Springer. ter. In: Zadzinska E, editor. Current trends in
Swartkrans from stable carbon isotopic analy-
p 103–116. dental morphology research. Lodz: University
sis. J Hum Evol 27:361–372.
78 Olejniczak AJ, Tafforeau P, Feeney RNM, Mar- of Lodz Press. p 201–210.
64 Sponheimer M, de Ruiter D, Lee-Thorp J,
tin LB. 2008. Three-dimensional primate molar 92 Kupczik K. 2003. Tooth root morphology in
Späth A. 2005. Sr/Ca and early hominin diets
enamel thickness. J Hum Evol 54:187–195. primates and carnivores. Ph.D. Thesis, Univer-
revisited: new data from modern and fossil
79 Olejniczak AJ, Grine FE. 2006. Assessment sity of London.
tooth enamel. J Hum Evol 48:147–156.
of the accuracy of dental enamel thickness 93 Avishai G, Muller R, Gabet Y, Bab I, Zilber-
65 Lee-Thorp JA, Sponheimer M, Luyt J. 2007.
measurements using micro-focal X-ray com- man U, Smith P. 2004. New approach to quan-
Tracking changing environments using stable
puted tomography. Anat Rec 288A:263–275. tifying developmental variation in the dentition
carbon isotopes in fossil tooth enamel: an
80 Chaimanee Y, Jolly D, Benammi M, Taffor- using serial microtomographic imaging.
example from the South African hominin sites.
eau P, Duzer D, Moussa I, Jaeger J-J. 2003. Microsc Res Tech 65:263–269.
J Hum Evol 53:595–601.
Middle Miocene hominoid from Thailand and 94 Skinner MM, Wood B, Boesch C, Olejniczak
66 Nielsen-Marsh C, Gernaey A, Turner-Walker
orangutan origins. Nature 422:61–65. AJ, Rosas A, Smith TM, Hublin J-J. 2008. Den-
G, Hedges R, Pike A, Collins M. 2000. The
81 Brunet M, Guy F, Pilbeam D, Lieberman tal trait expression at the enamel-dentine junc-
chemical degradation of bone. In: Cox M, Mays
DE, Likius A, Mackaye HT, Ponce de León MS, tion of lower molars in extant and fossil homi-
S, editors. Human osteology in archaeology and noids. J Hum Evol 54:173–186.
forensic science. Cambridge: Cambridge Uni- Zollikofer CPE, Vignaud P. 2005. New material
versity Press. p 439–454. of the earliest hominid from the upper Miocene 95 Olejniczak AJ, Gilbert CC, Martin LB, Smith
of Chad. Nature 434:752–755. TM, Ulhaas L, Grine FE. 2007. Morphology of
67 Lee-Thorp JA, van der Merwe NJ. 1991. the enamel-dentine junction in sections of
Aspects of the chemistry of modern and fossil 82 Suwa G, Kono RT, Katoh S, Asfaw B,
Beyene Y. 2007. A new species of great ape anthropoid primate maxillary molars. J Hum
biological apatites. J Arch Sci 18:343–354. Evol 53:292–301.
from the late Miocene epoch in Ethiopia.
68 Wang Y, Cerling TE. 1994. A model of fossil Nature 448:921–924. 96 Olejniczak AJ, Martin LB, Ulhaas L. 2004.
tooth and bone diagenesis: implications for Quantification of dentine shape in anthropoid
paleodiet reconstruction from stable isotopes. 83 Kunimatsu Y, Nakatsukasa M, Sawada Y,
Sakai T, Hyodo M, Hyodo H, Itaya T, Nakaya primates. Ann Anat 186:479–485.
Palaeogeogr Palaeoclimatol Palaeoecol 107:281–
H, Saegusa H, Mazurier A, Saneyoshi M, Tsuji- 97 Smith TM, Olejniczak AJ, Reid DJ, Ferrell
289.
kawak H, Yamamoto A, Emma Mbua. 2007. A RJ, Hublin J-J. 2006. Modern human molar
69 Kohn MJ, Schoeninger MJ, Barker WW. new Late Miocene great ape from Kenya and enamel thickness and enamel-dentine junction
1999. Altered states: effects of diagenesis on its implications for the origins of African great shape. Arch Oral Biol 51:974–995.
fossil tooth chemistry. Geochim Cosmochim apes and humans. Proc Natl Acad Sci USA 98 Korenhof CAW. 1961. The enamel-dentine
Acta 63:2737–2747. 104:19220–19225. border: a new morphological factor in the study
70 Schoeninger MJ, Hallin K, Reeser H, Valley 84 Kono R. 2004. Molar enamel thickness and of the (human) molar pattern. Proc Koninkl
JW, Fournelle J. 2003. Isotopic alteration of distribution patterns in extant great apes and Nederl Acad Wetensch 64B:639–664.
mammalian tooth enamel. Int J Osteoarch humans: new insights based on a 3-dimensional 99 Kono RT, Suwa G, Tanijiri T. 2002. A three-
13:11–19. whole crown perspective. Anthropol Sci dimensional analysis of enamel distribution
71 Burton JH, Price TD. 2000. The use and 112:121–146. patterns in human permanent first molars.
abuse of trace elements for paleodietary 85 Olejniczak AJ, Smith TM, Wei W, Potts R, Arch Oral Biol 47:867–875.
research. In: Ambrose SH, Katzenberg MA, edi- Ciochon R, Kullmer O, Schrenk F, Hublin J-J. 100 Leigh S. 2004. Brain growth, life history,
tors. Biogeochemical approaches to paleodiet- 2008. Molar enamel thickness and dentine horn and cognition in primate and human evolution.
ary analysis. New York: Kluwer Academic/ height in Gigantopithecus blacki. Am J Phys Am J Phys Anthropol 62:139–164.
Plenum Publishers. p 159–171. Anthropol 135:85–91. 101 Guatelli-Steinberg D. n.d. Recent studies of
72 Fabig A, Herrmann B. 2002. Trace 86 Olejniczak AJ, Smith TM, Feeney RNM, dental development in Neandertals: Implica-
elements in buried human bones: intra-popula- Macchiarelli R, Mazurier A, Bondioli L, Rosas tions for Neandertal life histories. Evol Anthro-
tion variability of Sr/Ca and Ba/Ca ratios— A, Fortea J, de la Rasilla M, Garcı́a-Tabernero pol In Press.
diet or diagenesis? Naturwissenschaften 89: A, Radovčić J, Skinner MM, Toussaint M,
115–119. 102 Humphrey LT, Dean MC, Jeffries TE, Penn
Hublin J-J. 2008. Dental tissue proportions and
M. 2008. Unlocking evidence of early diet from
73 Zazzo A, Lécuyer C, Sheppard SMF, Grand- enamel thickness in Neandertal and modern
tooth enamel. Proc Natl Acad Sci USA 105:
jean P, Mariotti A. 2004. Diagenesis and the human molars. J Hum Evol 55:12–23. 6834–6839.
reconstruction of paleoenvironments: a method 87 Smith TM, Olejniczak AJ, Tafforeau P, Reid 103 Olejniczak AJ, Smith TM, Skinner MM,
to restore original d18O values of carbonate and DJ, Grine FE, Hublin J-J. 2006. Molar crown Grine FE, Feeney RNM, Thackeray FJ, Hublin
phosphate from fossil tooth enamel. Geochim thickness, volume, and development in South J-J. 2008. Three-dimensional patterning and
Cosmochim Acta 68:2245–2258. African Middle Stone Age humans. S Afr J Sci thickness of molar enamel in Australopithecus
74 Fejerskov O, Larsen MJ, Richards A, Baelum 102:513–517. and Paranthropus. Biol Lett 4:406–410.
V. 1994. Dental tissue effects of fluoride. Adv 88 Wood BA, Abbott SA, Uytterschaut H. 1988.
Dent Res 8:15–31. Analysis of the dental morphology of Plio-Pleis- V
C 2008 Wiley-Liss, Inc.