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Incremental lines of dental cementum in biological age estimation

Article  in  HOMO - Journal of Comparative Human Biology · February 2001

DOI: 10.1078/0018-442X-00020 · Source: PubMed

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HOMO Vol. 52/1, pp. 59–71
© 2001 Urban & Fischer Verlag HOMO
http://www.urbanfischer.de/journals/homo

Incremental lines of dental cementum

in biological age estimation

R. JANKAUSKAS, S. BARAKAUSKAS, R. BOJARUN

Department of Anatomy, Histology and Anthropology, Faculty of Medicine, Vilnius University,

Čiurlionio 21, Vilnius LT 2009, Lithuania

Summary
The aim of this study is to evaluate the use of counting incremental lines of dental root cementum for
biological age determination, and to compare it with alternative methods. Two samples were taken: 51
teeth from 49 individuals of known age obtained at the Stomatological Clinic, Vilnius University, as
well as the canine teeth from the remains of 48 individuals from the mass graves of Tuskul ė˙ nai in Vil-
nius (inhumed 1944–47). In the latter sample, the chronological age of 43 individuals was known
through personal identification. Undecalcified teeth were sectioned with the Leica SP 1600 microtome
diamond saw, and incremental line count as a blind test was made on sections of 35 to 100 µm thick-
ness. Incremental line count was possible in 82–86 percent of cases. The results of three independent
counts showed that intra-observer bias has no significant impact. Biological age was estimated by
adding incremental line number to the average age of tooth eruption. It was found that mean absolute
error was 6.46 years for the 1st sample, 6.27 years for the 2nd sample, and in some cases exceeded 10
years. For the 2nd sample, the results were compared to those of other methods such as endocranial
suture ossification, pubic symphysis morphology and the «combined» method of Nemeskéri. All four
methods yield a similar correlation in regard to an individual’s chronological age. The highest correla-
tion was found for the combined method, and the lowest one for pubic symphysis morphology. All cor-
relations had a similar standard error. Thus our assessment is less enthusiastic than in some past stud-
ies; it is suggested that the incremental lines rather have a similar use as other methods.

Introduction
Accurate determinations of the biological age of an individual remains a problem
in physical anthropology. In bioarchaeology, the age of an individual is the basis
for further analysis of the demographic and adaptive characteristics of skeletal
populations (ACSÁDI & NEMESKÉRI 1970; UBELAKER 1989). In forensic sciences, a
precise age determination is a necessary requirement for individual identification
(GARMUS 1996). An entire set of age determination methods has been elaborated –
both macroscopic (for summary, see SZILVÁSSY 1988) and microscopic (UYTTER-
SCHAUT 1985). However, both anthropologists and forensic experts judge these
methods to be far from perfect (see RÖSING & KVAAL 1998). The search for new
methods thus continues.

0018-442X/01/52/01–59/$ 15.00/0
60 R. Jankauskas et al.

Dentition is the most durable and often a very informative system for vertebrate
organisms, and many age determination methods are based on teeth. The colour
and fluorescence, although correlated with age, are of limited use in bioarchaeol-
ogy as teeth are prone to discoloration due to soil conditions (RÖSING & KVAAL
1998). Attrition of the masticatory surface, often used in bioarchaeology, depends
on dietary patterns (MILES 1978; UBELAKER 1989), and, unless the calibration for a
particular population is performed (MAYS 1998), provides only approximate
results. Use of other traits such as periodontal resorption or thickening of the
cement of the root (LAMENDIN et al. 1992) also may result in large errors. Dental
X-rays are technically complicated, and the standard error is substantial (KVAAL &
SOLHEIM 1994). The root cement thickening only moderately correlates with age
and depends on an individual’s sex (SOLHEIM 1990), as well as periodontal disease
(see HÜRZELLER & ZANDER 1959, but disputed by GROSSKOPF et al. 1996). Since
the 1950s, undecalcified thin sections of teeth have been used (GUSTAFSON 1950,
1966; LAMPE & RÖTZSCHER 1994) for the evaluation of six traits on longitudinal
sections – crown attrition, secondary dentine, periodontal change, dental root
cement apposition, resorption of the root, and root translucency. According to
KILIAN (1975), this method has a low error of only 3.6–1.8 years, although other
specialists are more sceptical (MAPLES & RICE 1979; LUCY & POLLARD 1995). Sev-
eral attempts have been made to improve and/or to modify Gustafson’s method
(MAPLES 1978; KASHYAP & KOTESWARA RAO 1990; XU et al. 1991; LUCY et al.
1996; for others see RÖSING & KVAAL 1998).
Layer structures in teeth have been known for many years (Malpighi noted lay-
ers in cementum in the 17th c., Retzius and Owen in dentine in the 19th c.) and
studied in various mammals (e.g., SCHEFFER 1950; SERGEANT & PIMLOTT 1959;
RAUSCH 1961; SAXON & HIGHAM 1968; STOTT et al. 1981, ZVJAGIN & STANCHEV
1986). Sometimes these three-dimensional, cone-like layers are called «rings»,
because in cross-section they do appear as concentric annulations. Although the
precise mechanism of their appearance is still disputed, it is thought that they are
related to ontogenetic growth, thus the term «incremental lines» is frequently
used. As layers of secondary dentine are deposited in the dental pulp cavity, the
process is restricted by the limited space, which later complicates the attempt to
count the lines. Thus, dental root cementum seems more attractive, as layers are
deposited on the external surface, and there are no restrictions for the incremental
line number increase. Annual deposition lines are composed from a wide and light
«summer» layer rich in cementocytes, and a narrow and darker «winter» layer,
constituted almost exclusively of calcium salts. Investigations on animals revealed
that the regularities of deposition are influenced by the environment (climatic
changes, quantity and quality of food), as well as metabolic fluctuations including
reproduction cycle and feeding habits (GRUE & JENSEN 1979; STALLIBRASS 1982;
GORDON 1993). Thus, the annulations are extremely pronounced for animals with
marked seasonal differences in activity and feeding (KLEVEZAL 1970; STOTT et al.
1981). The method was employed for age estimation of various wild mammals
(SERGEANT & PIMLOTT 1959; MCEWAN 1963; WADA et al. 1975; KLEVEZAL &
KLEINEBERG 1976; MORRIS 1978). In such age determinations, annual rings are
counted and average tooth eruption ages added (ZUHRT et al. 1978); the addition
of the age attained at a specific root length (RÖSING & KVAAL 1998) might be more
 Incremental lines of dental cementum in biological age estimation 61

precise but has not been tested so far. In human age estimation both demineralised
and mineralised, stained and unstained teeth sections can be used. A majority of
researchers found a high correlation between the number of incremental lines and
chronological age (STOTT et al. 1981; CONDON et al. 1986; KVAAL & SOLHEIM
1995). Undecalcified ground sections of teeth are recommended (NAYLOR et al.
1985), especially when the quantity of organic components is diminished due to
various factors such as cremation and soil conditions (GROSSKOPF 1989, 1990).
Some studies, however, have failed to find relations between the number of cemen-
tal annulations and the age of the individual and consider the use of the method
impossible (MILLER et al. 1988).
The purpose of this paper is to count incremental lines of dental root cementum
in two samples of teeth of individuals of known and of diagnosed chronological
age and to compare the results with other methods of biological age determina-
tion.

Materials and methods

Two samples were chosen for analysis:
Sample 1. 51 teeth of 49 individuals that were extracted at the Stomatological
Clinic, Vilnius University (table 1). The patients’ ages were known but the reasons
for extraction were not known for all cases.

Table 1: Incremental line counts in the sample of clinically extracted teeth (Sample 1).

Refer. Age Tooth Line Line Line Average** Age according Difference from
Case (years) (FDI count – count – count – average line chronological
No. No.) 1* 2* 3* count age

1 56 43 41 48 57 51.00 61.30 +5.30

2 33 42 24 30 26 26.67 34.07 +1.07
3 54 13 30 27 26 27.67 38.97 –15.03
4 20 15 8 7 7 7.33 18.43 –1.57
5 26 18 6 4 8 6.00 23.00 –3.00
6 36 28 9 12 8 9.67 26.67 –9.33
7 39 18 15 14 13 14.00 31.00 –8.00
8 24 18 17 24 15 18.67 35.67 +11.67
9 24 38 7 11 10 9.33 26.33 +2.33
10 36 48 12 14 12 12.67 29.67 –6.33
11 41 38 15 18 20 17.67 34.67 –6.33
12 30 41 22 16 20 19.33 25.83 –4.17
13 16 32 12 14 13 13.00 20.40 +4.40
14 65 26 54 62 60 58.67 65.17 +0.17
15 53 17 50 45 52 49.00 61.60 +8.60
16 27 46 24 18 23 21.67 28.07 +1.07
17 26 47 Count failure
18 29 48 Count failure
62 R. Jankauskas et al.

Table 1: (Continued).

Refer. Age Tooth Line Line Line Average** Age according Difference from
Case (years) (FDI count – count – count – average line chronological
No. No.) 1* 2* 3* count age

19 66 48 Count failure
20 12 24 7 9 10 8.67 19.77 +7.77
21 31 36 21 25 26 24.00 30.40 –0.60
22 30 38 24 16 26 22.00 39.00 +9.00
23 48 14 39 38 42 39.67 49.97 +1.97
24 12 24 8 4 7 6.33 16.63 +4.63
25 17 36 10 14 9 11.00 17.40 +0.40
26 46 38 Count failure
27 23 36 21 17 23 20.33 26.73 +3.73
28 72 31 51 52 43 48.67 55.17 –16.83
29 61 21 70 76 58 66.75 74.05 +13.05
30 57 47 38 37 39 38.00 50.10 –6.90
31 51 17 32 36 38 35.33 47.93 –3.07
32 41 25 58 52 42 50.67 61.77 +20.77
33 69 48 Count failure
34 68 46 81 87 82 83.33 89.73 +21.73
35 17 35 9 10 12 10.33 21.73 +4.73
36 17 41 8 7 8 7.67 14.17 –2.83
37 35 47 23 28 27 26.00 38.10 +3.10
38 35 28 34 35 35 34.67 51.67 +16.67
39 38 48 22 21 20 21.00 38.00 0.00
40 63 47 Count failure
41 23 23 15 13 13 13.67 24.97 +1.97
42 30 18 Count failure
43 45 46 36 40 41 39.00 45.40 +0.40
44 54 12 39 31 36 35.33 43.63 –10.37
45 52 44 44 46 41 43.67 54.07 +2.07
46 32 38 Count failure
47 62 37 52 49 55 52.00 64.10 +2.10
48 30 48 26 24 23 24.33 41.33 +11.33
49 44 37 42 43 44 43.00 55.10 +11.10
50 50 23 44 43 40 42.33 53.63 +3.63
51 45 43 45 44 41 43.33 53.63 +8.63

Total (N): 42 42 42 42 42 42
Average difference (and standard deviation) from chronological age 6.46 (5.63)
Maximal difference from chronological age 21.73

* – results of the first, second and third blind count

**– average from three counts

Sample 2. Canine teeth of 48 individuals from the Stalin era mass grave of
Tuskul ė˙ nai in Vilnius. The calendar age of 43 of these individuals had been previ-
ously established after personal identification. The canines were preferred as there
was no postmortal loss during exhumation and no antemortal stomatological
interventions. In this sample, biological age was determined in the majority of indi-
viduals by means of endocranial suture ossification (ZVJAGIN 1982), morphology
of the pubic symphysis (GARMUS 1996) and a «combined» method (ACSÁDI &
NEMESKÉRI 1970, SJØVOLD 1975) (table 2).
 Incremental lines of dental cementum in biological age estimation 63

Table 2: Age determination on individuals from the Tuskul ė˙ nai mass graves in Vilnius (four methods)
(Sample 2).

Forensic Age by Age Age Age according the incremental Chrono-

case No. endocranial according according lines of cementum logical
suture pubic «combined» age
closing symphysis method Tooth Incremental Age
(deviation (deviation (deviation FDI line count (deviation
from from from No. from
calendar calendar calendar calendar
age) age) age) age)

44 26.10 29.41 30.50 23 16 29.00 27.00

(–1.10) (+2.41) (+3.50) (+2.00)
56 22.68 29.63 34.00 23 23 36.00 20.00
(+22.68) (+9.63) (+14.00) (+16.00)
80 35.61 – 37.00 13, 23 22 35.00 30.00
(+5.61) (+7.00) (+5.00)
86 31.70 – 33.00 13 17 30.00 41.00
(–9.30) (–7.00) (–11.00)
161 22.68 – 23.00 13 8 21.00 23.00
(–0.32) (0.00) (–2.00)
204 38.45 – 33.00 23 16 29.00 42.00
(–3.55) (–9.00) (–13.00)
214 23.39 30.12 31.00 13 14 27.00 27.00
(–3.61) (+3.12) (+4.00) (0.00)
337 44.32 47.10 53.75 23 36 49.00 57.00
(–12.68) (–9.90) (–3.25) (–8.00)
481 40.60 – 34.00 23 23 36.00 38.00
(+2.60) (–4.00) (–2.00)
484 22.70 – 27.50 13 13 26.00 28.00
(–5.30) (–.050) (–2.00)
488 – 24.50 27.50 13 10 23.00 22.00
(+2.50) (+5.50) (+1.00)
491 43.40 47.10 51.75 13, 23 22 35.00 52.00
(–8.60) (–4.90) (–0.25) (–17.00)
492 44.00 39.39 32.00 23 18 31.00 31.00
(+13.00 (+8.39) (+1.00) (0.00)
493 35.43 47.10 47.75 23 21 34.00 34.00
(+1.43) (+13.10) (+13.75) (0.00)
495 39.87 47.10 47.57 23 16 29.00 31.00
(+8.87) (+16.10) (+16.57) (–2.00)
507 45.50 49.00 55.75 43 31 44.00 63.00
(–17.50) (–14.00) (–7.25) (–19.00)
510 37.80 – 34.00 23 22 35.00 24.00
(+13.80) (+10.00) (+11.00)
511 44.70 44.42 44.25 33 35 48.00 59.00
(–14.30) (–14.58) (–14.75) (–11.00)
514 48.94 38.61 43.25 23, 33 28 41.00 36.00
(+12.94) (+2.61) (+7.25) (+5.00)
515 45.40 – 51.75 33 28 41.00 43.00
(+2.40) (+8.75) (–2.00)
518 34.20 – 31.00 43 10 23.00 25.00
(+9.20) (+6.00) (–2.00)
64 R. Jankauskas et al.

Table 2: (Continued).

Forensic Age by Age Age Age according the incremental Chrono-

case No. endocranial according according lines of cementum logical
suture pubic «combined» age
closing symphysis method Tooth Incremental Age
(deviation (deviation (deviation FDI line count (deviation
from from from No. from
calendar calendar calendar calendar
age) age) age) age)

520 24.58 57.52 52.75 23 – – 31.00

(–6.42) (+26.52) (+21.75)
538 33.30 – 34.00 15 23 36.00 34.00
(–0.70) (0.00) (+2.00)
543 22.68 28.22 26.50 13, 23 15 28.00 26.00
(–3.32) (+2.22) (+0.50) (+2.00)
552 31.19 47.10 41.50 23 – – 37.00
(–5.81) (+10.10) (+4.50)
553 39.60 – 34.00 33 – – 28.00
(+11.60) (+6.00)
554 – – 34.00 23 20 33.00 26.00
(+8.00) (+7.00)
559 22.70 19.89 27.50 33 19 32.00 20.00
(+2.70) (–0.11) (+7.50) (+12.00)
560 41.07 – 47.25 23 23 36.00 48.00
(–6.93) (–0.25) (–12.00)
561 42.60 – 50.00 23 38 51.00 50.00
(–7.40) (0.00) (+1.00)
563 22.70 – 23.00 33 10 23.00 19.00
(+3.70) (+4.00) (+4.00)
564 28.90 – 23.00 23 16 29.00 22.00
(+6.90) (+1.00) (+7.00)
566 – – 27.50 23 8 21.00 19.00
(+8.50) (+2.00)
568 22.70 – 26.00 33 25 38.00 23.00
(–0.30) (+3.00) (+15.00)
569 42.80 – 50.00 13 – – 40.00
(+2.80) (+10.00)
572 45.30 – 61.00 23 27 40.00 56.00
(–10.70) (+5.00) (–16.00)
573 28.30 – 23.00 23 20 33.00 23.00
(+5.30) (0.00) (+10.00)
574 22.70 – 30.50 23 21 34.00 27.00
(–4.30) (+3.50) (+7.00)
576 26.10 – 30.50 23 13 26.00 26.00
(+0.10) (+4.50) (0.00)
577 22.68 19.89 27.50 23 12 25.00 22.00
(+0.68) (–2.11) (+5.50) (+3.00)
579 44.32 – 63.00 13 – – 62.00
(–17.68) (+1.00)
580 28.50 – 30.50 13 14 27.00 26.00
(+2.50) (+4.50) (+1.00)
666 39.50 – 31.00 23 – – 35.00
(+4.50) (–4.00)
353 44.80 40.00 38.50 13 29 42.00 –
 Incremental lines of dental cementum in biological age estimation 65

Table 2: (Continued).

Forensic Age by Age Age Age according the incremental Chrono-

case No. endocranial according according lines of cementum logical
suture pubic «combined» age
closing symphysis method Tooth Incremental Age
(deviation (deviation (deviation FDI line count (deviation
from from from No. from
calendar calendar calendar calendar
age) age) age) age)

360 34.89 47.10 47.75 13 20 33.00 –

423 47.20 51.00 49.00 23 – – –
485 40.17 47.10 51.25 13 30 43.00 –
706 52.43 47.60 49.25 23 37 50.00
–
Total (N): 45 22 48 52 41 41 43

Absolute difference from chronological age (years):

Average 6.32 8.37 5.90 – – 6.27 0.00

Maximal 17.68 26.52 21.75 – – 19.00 0.00

Before sectioning, the teeth were embedded in special forms in polymere Medim
K-Plast™ and left for hardening in room temperature for 2–3 days. The blocks
were fixed in the support of the microtome. Undecalcified teeth were sectioned
with the Leica SP 1600 microtome diamond saw. Three to four transverse cuts
were made of each tooth root at the mid-upper region. Prior data (ZVJAGIN &
STANCHEV 1986) and our own experience indicated that the use of 3–4 sections
was sufficient – independent incremental line counts on different sections of the
same tooth were highly correlated. Thickness of sections were 35 to 100 µm. Each
section was washed under a weak stream of tap water, dried on filter paper for
15–20 minutes and mounted on pre-marked objective glass with the glue E®. Sec-
tions were covered with mounting glass and left for final fixation for 8–12 hours.
Using a light microscope on each section where the lines were best distinguishable,
the direct count of incremental lines was performed as a blind test, three times for
each section at one day intervals. Results of all three calculations are presented in
table 1, while table 2 shows the average number. Dentine/cementum junction line
was not taken into account. Biological age was determined summing up the aver-
age number of incremental lines and the average age of tooth eruption (ZUHRT et
al. 1978; STALLIBRASS 1982; GROSSKOPF 1990). Regretfully, the method of so-called
«gliding correction» (adding the developmental age of exact section position)
(RÖSING & KVAAL 1998) appeared to be hardly applicable, as the exact location
(distance from enamel-cementum junction) of each section was not fixed.
66 R. Jankauskas et al.

Results and discussion

Results of Sample 1 analysis
An incremental line count was possible in 42 teeth, or 82.3% of the sample (see
table 1), which is less than CONDON et al. (1986) found in their study. The average
difference from the chronological (calendar) age, the mean absolute error was 6.46
years, and in some cases exceeded 10 years. The results of three independent
counts are highly correlated (r > 0.95; table 3). This means that intra-observer bias
has no significant impact on the results. Most other studies also note that intra-
observer error is minimal. The incremental line number is highly correlated to

Table 3: Pearson’s correlation coefficients between chronological age, incremental lines and estimated
age (complete Sample 1). Confidence of all coefficients p < 0.01.

Chronological Line count Line count Line count Average

age –1 –2 –3

Line count – 1 0.854

Line count – 2 0.852 0.976
Line count – 3 0.861 0.963 0.963
Average 0.866 0.989 0.990 0.988
Estimated age 0.879 0.966 0.966 0.965 0.977

Figure 1: Scatterplot of chronological age versus estimated age (complete sample 1)

 Incremental lines of dental cementum in biological age estimation 67

Table 4: Pearson’s correlation coefficients between chronological age, incremental lines and estimated
age (Sample 1, third molars excluded). Confidence of all coefficients p < 0.01.

Chronological Line count Line count Line count Average

age –1 –2 –3

Line count – 1 0.891

Line count – 2 0.882 0.978
Line count – 3 0.900 0.956 0.965
Average 0.902 0.987 0.991 0.987
Estimated age 0.905 0.980 0.979 0.982 0.992

chronological age. The correlation between chronological and estimated age is also
high (r = 0.879). The scatterplot of estimated versus chronological age revealed a
significant dispersion without detectable regularities of under- or overestimation,
increasing slightly with increasing calendar age (figure 1). The most significant lin-
ear regression model (with F = 105.915, at 41 degrees of freedom) was:
Y = 0.770 x + 6.686,
where Y = chronological age, x = estimated age, and the standard error of esti-
mates is 8.633 years.
As 11 teeth in this sample were third molars (18, 28, 38, 48 according to FDI),
whose eruption age varies significantly, we made an attempt to recalculate the
data. Table 4 shows that the results were only slightly better. The average differ-
ence from the chronological age was 6.05 years (standard deviation 5.91 years). In
some cases, the difference from chronological age again exceeded 10 years. The
linear regression model (F = 94.692, at 30 degrees of freedom) in this case is:
Y = 0.801 x + 5.501.
Exclusion of third molars in this regression, however, leads to an even higher
error of estimate (8.995 years). Exclusion of the third molars therefore only
slightly improves the result.

Results of Sample 2 analysis

Canine teeth were the focus of this sample. An incremental line count was made in
37 cases, or 86.0% (table 2). Average difference from the chronological age was
6.27 (standard deviation 5.69) years, so it does not differ substantially from other
methods. In some cases, the difference from actual age exceeds 10 years, and the
maximal difference was 19 years. The scatterplot of incremental line number versus
chronological age also revealed significant dispersal of data around the linear regres-
sion line (figure 2). The linear regression (F = 41.756 at 36 degrees of freedom):
Y = 1.214 x + 8.656
gives a standard error of estimate of 8.612 years. This does not differ from results
obtained during the Sample 1 analysis, which included teeth with different erup-
tion ages.
68 R. Jankauskas et al.

Figure 2: Scatterplot of incremental lines versus chronological age (sample 2)

Table 5: Pearson’s correlation coefficients between age estimations in the Tuskule·nai mass grave sam-
ple.

Chronological age Biological age by incremental lines

Biological age according 0.773 0.741

endocranial suture closure (p <0.01) (p <0.01)
Biological age according 0.671 0.732
pubic symphysis (p <0.01) (p <0.01)
Biological age according 0.840 0.738
«combined» method (p <0.01) (p < 0.01)
Biological age according 0.765 1.000
incremental lines (p <0.01)

In an attempt to compare all four age estimation methods, we performed a Pear-

son correlation analysis (table 5). All four methods show a reliable correlation
with the individual’s chronological age. The highest correlation was represented by
the «combined» method, and the lowest correlation with the pubic symphysis
morphology.
 Incremental lines of dental cementum in biological age estimation 69

Conclusions
This study revealed both positive and negative aspects of the incremental line
count method. First, incremental line count is possible in about 80–85 percent of
examined cases (a frequency that is lower than in previous studies). A probable
factor is pathological changes of the cementum (i.e. massive and numerous cemen-
tocytes obscuring the lines). Such a large number of count failures could also be
related to the simple technique employed, which was a direct count using light
microscopy. One might expect that the use of more sophisticated techniques, e.g.,
phase contrast with digital image analysis, could improve the results. The experi-
ence of other analysts (BUBA 2000; KAGERER & GRUPE 2000) points to substantial
difficulties in the application of the method in case of pathological periodontal
processes. However, minor pathologies do not affect results substantially, as
shown by our analysis of both samples. It was found that the correlation to
chronological age and average errors of biological age estimation are similar to the
errors established by the majority of other studies (see RÖSING & KVAAL 1998 for
discussion) and the errors of other methods. Only GROSSKOPF (1990) and KAGERER
& GRUPE (2000) obtained substantially lower errors. Differences in opinion about
the reliability of the method reflect the use of different techniques (undecalcified or
decalcified sections; stained or unstained slices; longitudinal or transverse sections;
simple light or phase contrast microscopy). One might speculate that seasonal fluc-
tuation in metabolism rate for humans born in the 20th c. is less pronounced than
that of archaeological populations, and therefore the incremental lines of dental
cementum do not demonstrate expected simple regularity. As RÖSING & KVAAL
(1998) have mentioned, there are possibilities that some humans appose more than
one layer every year, and in older individuals some layers could be skipped. The
hypothesis that a larger sample and calculation of multiple regression equations
for each tooth (ZVJAGIN & STANCHEV 1986) could increase the reliability of the
method still needs verification. We conclude that the incremental line count
method can be used as an independent verification of conventional methods, and,
in the case of very fragmentary skeletons, it is the method of choice.

Acknowledgements
We thank PD Dr. Ursula Wittwer-Backofen, Justus-Liebig-Universität Gießen, for practical training and
advice. We are also grateful to the reviewers Prof. Dr. G. Grupe and PD Dr. I. Schröder, for their thor-
ough analysis of the manuscript and useful suggestions. Credits should be given also to Raymond
Sidrys for the editing of the English version of the paper. This research was supported by grants from
DAAD and Lithuanian State Science Foundation (grant no. 344/98).

References
ACSÁDI G, NEMESKÉRI J (1970) History of Human Life Span and Mortality. Akadémiai Kiado, Budapest.
BUBA H (2000) Berücksichtigung der Parodontose bei der Methode der Zahnzementannulation. Homo
51/Suppl.: 13.
CONDON K, CHARLES DK, CHEVERUD JM, BUIKSTRA JE (1986) Cementum annulation and age determi-
nation in Homo sapiens II. Estimates of accuracy. Am J Phys Anthrop 71: 321–330.
70 R. Jankauskas et al.

GARMUS A (1996) Lithuanian Forensic Osteology. Baltic Medico-Legal Association, Vilnius.

GORDON BC (1993) Archaeological tooth and bone seasonal increments: the need for standardised
terms and techniques. Archaeozoologia V/2: 9–16.
GROSSKOPF B (1989) Incremental lines in prehistoric cremated teeth. A technical note. Z Morph
Anthrop 77: 309–311.
GROSSKOPF B (1990) Individualaltersbestimmung mit Hilfe von Zuwachsringen im Zement bodenge-
lagerter menschlicher Zähne. Z Rechtsmed 103: 351–359.
GROSSKOPF B, DEUDEN JM, KRÜGER W (1996) Untersuchung zur Zementapposition bei Parodontitis
marginalis profunda. Dt Zahnärztl Z 51: 295–297.
GRUE H, JENSEN B (1979) Review of the formation of incremental lines in tooth cementum of terrestrial
mammals. Danish Review of Game Biology 117: 2–49.
GUSTAFSON G (1950) Age determination on teeth. J Am Dent Ass 41: 45–54.
GUSTAFSON G (1966) Forensic Odontology. Staples Press, London.
HÜRZELER B, ZANDER HA (1959) Cementum apposition in periodontally diseased teeth. Helv. Odont
Acta 3: 1–3.
KAGERER P, GRUPE G (2000) Zahnzementzuwachsringe: Dechiffrierbare Annalen in der Paläopathologie,
Paläodemographie und Rechtsmedizin. Homo 51/Suppl.: 62.
KASHYAP VK, KOTESWARA RAO NR (1990) A modified Gustafson method of age estimation from teeth.
Forensic Sci Int 47: 237–247.
KILIAN J (1975) Age determination on teeth by means of Gustafson’s method. Scripta Medica (Brno) 48:
197–201.
KLEVEZAL GA (1970) A retrospective evaluation of the individual features of mammal growth based on
the structure of dentine and bone layers. Ontogenez 1: 362–372.
KLEVEZAL GA, KLEINEBERG SE (1976) Opredelenije vozrasta mlekopitajuš čich po sloistym strukturam
zubov i kosti. Nauka, Moskva.
KVAAL S, SOLHEIM T (1994) A non-destructive dental method for age estimation. J For Odonto-Stomatol
12: 6–11.
KVAAL S, SOLHEIM T (1995) Incremental lines in human cementum in relation to age. Europ J Oral Sci
103: 225–230.
LAMENDIN H, BACCINO E, HUMBERT JF, TAVERNIER JC, NOSSINTCHOUK RM, ZERILLI A (1992) A simple
technique for age estimation in adult corpses: the two criteria dental method. J Forensic Sci 37:
1373–1379.
LAMPE H, RÖTZSCHER K (1994) Forensic odontology: age determination from adult human teeth. Med
Law 13: 623–628.
LUCY D, AYKROYD RG, POLLARD AM, SOLHEIM T (1996) A Bayesian approach to adult human age esti-
mation from dental observations by Johanson’s age changes. J For Sci 41: 189–194.
LUCY D, POLLARD AM (1995) Further comments on the estimation of error associated with the
Gustafson dental age estimation method. J For Sci 40: 222–227.
MAYS S (1998) The archaeology of human bones. Routledge, London and New York.
MAPLES WR (1978) An improved technique using dental histology for estimation of adult age. J For Sci
23: 764–770.
MAPLES WR, RICE PM (1979) Some difficulties in the Gustafson dental age estimations. J For Sci 24:
168–172.
MCEWAN EH (1963) Seasonal annuli in the cementum of the teeth of Barren Ground caribou. Can J
Zool 41: 111–113.
MILES AEW (1978) Teeth as an indicator of age in man. In BUTLER PM, JOYSEY KA (eds) Development,
function and evolution of teeth. Academic Press, London, 455–464.
MILLER CS, DOVE SB, COTTONE JA (1988) Failure of use of cemental annulations in teeth to determine
the age of humans. J For Sci 33: 137–143.
MORRIS P (1978) The use of teeth for estimating the age of wild mammals. In BUTLER PM, JOYSEY KA
(eds) Development, function and evolution of teeth. Academic Press, London, 483–494.
NAYLOR JW, MILLER WG, STOKES GN, STOTT GG (1985) Cemental annulation enhancement: a tech-
nique for age determination in man. Am J Phys Anthrop 69: 197–200.
RAUSCH RL (1961) Notes on the black bear Ursus americanus pallus, in Alaska, with particular refer-
ence to dentition and growth. Z Säugetierkde 26: 77–107.
RÖSING FW, KVAAL SI (1998) Dental age in adults. A review of estimation methods. In ALT KW, RÖSING
FW, TESCHLER-NICOLA M (eds) Dental Anthropology. Fundamentals, Limits, and Prospects. Springer,
Wien, 443–468.
 Incremental lines of dental cementum in biological age estimation 71

SAXON A, HIGHAM CFW (1968) Identification and interpretation of growth rings in the secondary den-
tal cementum of Ovis aries L. Nature 219: 634–635.
SCHEFFER VB (1950) Growth layers on the teeth of Pinnipedia as an indication of age. Science 112:
309–311.
SERGEANT DE, PIMLOTT DH (1959) Age determination in moose from sectioned incisor teeth. J. of
Wildlife Management 23: 315–321.
SJØVOLD T (1975) Tables of the combined method for determination of age at death given by
Nemeskéri, Harsányi and Acsádi. Anthrop Közl 19: 9–22.
SOLHEIM T (1990) Dental cementum apposition as an indicator of age. Scand. J Dent Res 98: 510–519.
STALLIBRASS S (1982) The use of cementum layers for absolute ageing of mammalian teeth: a selective
review of the literature with suggestions for further studies and alternative applications. In WILSON
B, GRIGSON C, PAYNE S (eds) Aging and sexing animal bones from archaeological sites. BAR, BS 109,
Oxford, 109–126.
STOTT GG, SIS RF, LEVY BM (1981) Cemental annulation as an age criterion in forensic dentistry J Dent
Res 61: 814–817.
SZILVÁSSY J (1988) Altersdiagnose am Skelett. In KNUSSMAN R (ed) Anthropologie: Handbuch der ver-
gleichenden Biologie des Menschen. Gustav Fischer Verlag, Stuttgart, 421–443.
UBELAKER DH (1989) Human Skeletal Remains: Excavation, Analysis, Interpretation. 2nd ed. Tarax-
acum, Washington DC.
UYTTERSCHAUT HT (1985) Determination of skeletal age by histological methods. Z Morph Anthrop
75: 331–340.
WADA K, OHTAISHI N, HACHIYA N (1975) Determination of age in the Japanese monkey from growth
layers in the dental cementum. Primates 19: 775–784.
XU XH, PHILIPSEN HP, JABLONSKI NG, WEATHERHEAD B, PANG KM, ZHU JZ (1991) Preliminary report
on a new method of human age estimation from single adult teeth. For Sci Int 51: 281–288.
ZUHRT T R, ROTTSTOCK F, WINTERFELD RI (1978) Möglichkeiten und Methoden der Stomatologie bei
der Identifizierung. In HUNGER H, LEOPOLD D (Hrg) Identifikation. Springer-Verlag, Berlin, p.
287–340.
ZVJAGIN VN (1982) Individuelle Altersbestimmung des Menschen an Hand des Schädels unter Anwen-
dung von Gleichungen der multiplen Regression. Kriminalistik und forensische Wissenschaften 44:
33–44.
ZVJAGIN VN, STANCHEV NA (1986) Diagnostika vozrasta cheloveka po mikrostrukturnym osobennos-
tiam tviordykh tkanej zubov. Sudebno-medicinskaja ekspertiza 29: 54–57.

Authors address: Prof Dr RIMANTAS JANKAUSKAS, Department of Anatomy, Histology and Anthropol-
ogy, Faculty of Medicine, Vilnius University, Čiurlionio 21, Vilnius LT2009, Lithuania, tel. (+370 2)
651764, fax (+370 2) 263167, e-mail: rimantas.jankauskas@mf.vu.lt

Received 4 Sept 2000, accepted 31 Oct 2000, resubmitted 20 Nov 2000

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