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J Forensic Sci, May 2014, Vol. 59, No.

3
doi: 10.1111/1556-4029.12385
TECHNICAL NOTE Available online at: onlinelibrary.wiley.com

ODONTOLOGY

Ashith B. Acharya,1 B.D.S., G.D.F.O.

Forensic Dental Age Estimation by Measuring


Root Dentin Translucency Area Using a New
Digital Technique*

ABSTRACT: Dentin translucency measurement is an easy yet relatively accurate approach to postmortem age estimation. Translucency area
represents a two-dimensional change and may reflect age variations better than length. Manually measuring area is challenging and this paper
proposes a new digital method using commercially available computer hardware and software. Area and length were measured on 100 tooth
sections (age range, 19–82 years) of 250 lm thickness. Regression analysis revealed lower standard error of estimate and higher correlation
with age for length than for area (R = 0.62 vs. 0.60). However, test of regression formulae on a control sample (n = 33, 21–85 years) showed
smaller mean absolute difference (8.3 vs. 8.8 years) and greater frequency of smaller errors (73% vs. 67% age estimates ≤ 10 years) for area
than for length. These suggest that digital area measurements of root translucency may be used as an alternative to length in forensic age
estimation.

KEYWORDS: forensic science, forensic odontology, age prediction, dentin transparency, area measurement, Adobe Photoshopâ, regression
analysis

Postmortem age estimation is an important contributor to contributed more often to stepwise multiple regression formulae
establishing identity of the deceased in forensic casework. Age than length measurements. Based on their results, they recom-
estimation, in conjunction with sex assessment, stature predic- mended measuring translucency area for age estimation.
tion, and population designation, assists in reconstructive identi- Translucency area has previously been measured manually
fication; alternatively, when law enforcers already have a (4,5), although this may not be an ideal approach—while manual
putative age of the deceased, a forensic age estimate that is close measurements of translucency length are easily taken using cali-
to the presumed age provides the police clarity in their line of pers, the same is not true for area because it depends on an
investigation. Since it was first proposed by Gustafson (1) as approximation of a sq. mm grid placed over the translucent zone
one of six parameters for adult age estimation, and backed by (e.g., Ref. [5]). Hence, a method that records area more objec-
Johanson (2) as the most strongly correlated with age among the tively is desirable and digital approaches may offer a solution.
six, root dentinal translucency has gained immense interest and While few papers have proposed digital approaches to capture
used as a single parameter in age estimation (3–7). Apart from translucency area (9,10,14), they either lack a detailed descrip-
its relative accuracy, the simplicity of its evaluation is an added tion of the method (9,14) or require capturing tooth images on a
attraction to assessing it. video camera, converting the analog signal to digital format, and
Conventionally, dentinal translucency has been examined subsequent image processing using customized software program
using calipers to measure its length on unsectioned or sectioned (10)—steps that, today, may be perceived as complex and out-
teeth (4,5). In the last two decades, digital methods have also dated. With advances in computing technology, a simpler
been proposed to quantify dentinal translucency (8–11) and have method is now feasible using commercially available computer
been reported as producing more accurate age estimates when hardware and software. Using these, this paper proposes a new
compared to caliper-based measurement (11,12). Most of these, technique to measure the area of root dentinal translucency and
however, have focused on measuring the length of translucency, compares age estimates obtained from it with those of length
and a simple method to capture translucency area is still lacking. measurements.
Lorentsen and Solheim (13) found that translucency area had a
better correlation than length with chronological age, and also
Methods
1
Department of Forensic Odontology, S.D.M. College of Dental Sciences Materials
and Hospital, Sattur, Dharwad, 580009 Karnataka, India.
*Funded by an institutional grant from the S.D.M. College of Dental
The material consisted of teeth collected from 133 subjects
Sciences and Hospital, Karnataka, India. aged 19–85 years (mean age = 48.7 years) encompassing a het-
Received 6 Nov. 2012; and in revised form 15 Feb. 2013; accepted 23 erogeneous sample of females and males of different age groups
Feb. 2013. in relatively equal numbers (Table 1). Six subjects had multiple

© 2014 American Academy of Forensic Sciences 763


764 JOURNAL OF FORENSIC SCIENCES

TABLE 1––Sample distribution across age groups, the sexes, and tooth clas- Measurement of Translucency Area
ses.
For measuring translucency area, the scanned images were
Sex Tooth Type opened in Adobe Photoshopâ 7.0.1 image-editing software pro-
Age group gram (Adobe Systems Inc., Mountain View, CA) installed in a
(years) Subjects Sections M F Incisors Canines Premolars Lenovo ThinkCentre desktop computer (Intelâ CoreTM i3-2100
19–30 22 22 6 16 11 – 11 Processor; 3.10-GHz CPU, 4.00 GB RAM) (Lenovo Group Ltd.,
31–40 23 24 13 10 10 7 7 Hong Kong, China). The root translucency area was selected
41–50 27 27 12 15 8 7 12 using the following steps:
51–60 25 29 15 10 7 7 15
>60 36 45 20 16 16 10 19 • The Zoom Tool in the Toolbox of Photoshopâ was selected
Total 133 147 66 67 52 31 64 to magnify the scanned image to a zoom setting of 66.7%
(about 94 magnification).
teeth (ranging from 2 to 7) contributing to a total of 147 teeth. • The Magic Wand Tool in the Toolbox was selected and the
The teeth were collected from the Department of Oral and Max- tolerance level set to 18; the Anti-aliased and Contiguous
illofacial Surgery of two dental schools and two private clinics boxes were checked (both options visible on the menu bar).
of this region, spread across a radius of c. 80 km. Only fully The Magic Wand Tool cursor was placed within the translu-
erupted permanent teeth extracted for valid clinical reasons such cent zone visible in the section’s root and clicked. (Note: The
as periodontal disease, malocclusion/orthodontic treatment, and tool selects areas of similar color or grayscale pixel values on
caries were included. Carious teeth were included only when the the image [15]. Manually setting the tolerance level allows
root, or root dentinal translucency, was unaffected macroscopi- one to specify the pixel range. The tolerance for both gray-
cally by the disease. scale and color has a range from 0 to 255 [15]. A lower tol-
erance level allows selecting zones with similar pixels values,
while a higher level selects a broader range of color.)
Tooth Processing and Digitization
• Ideally, the entire root translucency area should get selected
The extracted teeth were thoroughly cleaned and soft tissue with this action (indicated by the dynamic black and white
remnants removed from the root surface with a scalpel. Follow- lines, also referred to as the “marching ants”) (Fig. 1). How-
ing preservation in 10% formalin, teeth were mounted in autopo- ever, it is likely that either more or less than the desired area
lymerizing acrylic for sectioning by a hard-tissue microtome of root dentin translucency gets selected because the translu-
(Leica SP 1600, Leica Microsystems GmßH, Wetzlar, Germany). cency zones may be of slightly different pixel values or
The mounted teeth were sectioned longitudinally in the buccolin- because zones of opaque root dentin separate them (Fig. 2).
gual plane to 250 lm, as close as was possible to the central • If more, the selected translucent zone is deselected (Ctrl + D,
axis of the tooth. This thickness has, repeatedly, been found to or Command + D for Macintosh computers), and the toler-
be the best for assessing translucency in terms of clarity (2,6). ance reduced from 18 to, say, 14. The Magic Wand Tool
The sections were coded to ensure blind analysis. cursor is clicked again within the translucent zone. The action
Tooth sections were digitized using a previously described is repeated, with a still lower tolerance (12 or 8 usually
method (11). Briefly, the sections were placed adjacent to an suffices) if necessary, until the desired translucent area is
ABFO No. 2 scale (Lightning Powder Co. Inc., Jacksonville, selected.
FL) on a flatbed scanner (HP Scanjet G3010, Hewlett-Packard • If less, the tolerance of 18 is left unchanged; the shift key on
Co., Palo Alto, CA) and scanned at a resolution of 600 dpi. the keyboard is held down which allows adding to, or

FIG. 1––The entire area of root dentinal translucency has been selected, which is indicated by the black and white line, or “marching ants;” also, note that
the crown and root dentin have been separated by a line using the Line Tool in the Toolbox.
ACHARYA . DIGITAL TRANSLUCENCY AREA MEASUREMENT 765

FIG. 2––Multiple selections of root dentin translucency is necessary when it is separated by regions of opaque dentin. Note the Rectangle Tool, beneath
which the Line Tool is hidden.

extending, the selected translucent zone. A tiny “+” appears


next to the Magic Wand Tool cursor (indicating the additive
mode), and the selection of the desired translucency area is
continued (Fig. 2). The additive/multiple selections may also
be obtained with varying degrees of tolerance, as desired by
the examiner.
• If translucency extends into the crown, the Line Tool in the
Toolbox (Fig. 1), which is hidden below the Rectangle Tool
(Fig. 2), may be used to separate the crown from the root by
drawing a line at the cemento-enamel junction. This facili-
tates selecting only the root dentin translucency. However,
because drawing the line creates a new layer in the image, go
to Layer in the menu bar, click on Flatten Image and proceed
from the first step.
• While selecting the translucent area, however, any visibly
thin bands of peripheral translucency (c. <0.5 mm thick)
(Fig. 3) are excluded because this has been attributed to
periodontal inflammation and a pathologic origin rather than
age-related physiologic alteration (4).
FIG. 3––Thin bands of peripheral root dentin translucency (arrowed), just
Calculation of translucent area in sq. mm was undertaken adjacent to the root surface/cementum, was not included in the selected area
using the following steps: (outlined in white).

• On the menu bar, click on Image, and click on Histogram in


the drop-down menu. In the displayed Histogram dialog box,
“Pixels” reveal the pixel value of the selected translucency Translucency area (in sq. mm) = ([image area in sq. mm]/
area. [image area in pixels]) 9 (translucency area in pixels).
• Once this value is noted, the dialog box is closed. The selected
translucent area is deselected and the above step repeated. Repeat Observations
Because, now, no translucent area is selected, the pixel value
seen is that of the entire image. This is also noted. To assess potential intra-observer variation, translucency area
• Again, click on Image in the menu bar, followed by clicking was measured again on 20 randomly selected tooth sections after
on Image Size in the drop-down menu. This action displays 2 weeks, and the translucency area was re-calculated.
the image width and height. After ensuring that the width
and height units are in mm, their values are noted to calculate Measurement of Translucency Length
the area of the entire image (in sq. mm).
• The aforementioned three steps have produced three values— Using the same software program, translucency length was
the translucency area in pixels and the image area in pixels derived applying two previously published methods (11,16).
and sq. mm. To calculate the area of root dentinal translu- These papers describe digital techniques to obtain the maximum
cency in sq. mm, a simple arithmetic equation is applied: length (distance between the apical and coronal extent of root
766 JOURNAL OF FORENSIC SCIENCES

dentinal translucency) and the average translucency length (as based on the standard deviation. Instead, he encourages calculat-
described by Bang and Ramm [4]) of root dentin translucency. ing mean differences, which is more appropriate in real-life situ-
ations, produces lower error rates, and is easier to understand
(19). Gorard believes that this argument can be extended to SEE
Statistical Analysis
versus MAD (S. Gorard, personal communication). Hence, the
To evaluate the degree of intra-observer congruity, the repeat MAD was used because it has the potential to represent the error
measurements were compared to the primary measurements by more objectively; the SEE was also calculated separately for the
applying the paired t-test using the SPSS 10.0 program (SPSS test dataset to compare it with the MAD.
Inc., Chicago, IL; now IBM Corp., Armonk, NY). In addition, the number (and percentage) of estimates with
Following calculation of translucency area for the 133 sub- errors ≤ 10 years and ≥ 15 years was also calculated because
jects, the sections were decoded to reveal the actual age and Solheim and Sundnes (20) have categorized the former as
other demographic data. The sample was divided into a reference “acceptable” and designated the latter as “unsatisfactory” in
(or training) dataset of 100 subjects and a control (or test) data- forensic age estimation (20; pg. 11). All arithmetic calculations
set comprising sections from 33 individuals. All six subjects were undertaken on an Excel spreadsheet (Office 2011; Micro-
with multiple teeth described under materials were included as soft Corp., Redmond, WA).
part of the test dataset so as to imitate cases wherein multiple
teeth are available for postmortem age estimation. The distribu-
Results
tion of tooth classes, sex, and age groups in the test dataset was
similar to the training dataset, except for younger age groups The difference between the primary and repeat measurements
(≤40 years), who were proportionately less in the former of translucency area was statistically insignificant (t-value =
(Table 2). The training dataset was used to develop first- and 0.615; p > 0.05). The regression analysis revealed that maximum
second-degree polynomial functions (i.e., linear and quadratic translucency length had the highest correlation with age, fol-
regression equations, respectively), also using the SPSS 10.0 lowed by average translucency length and translucency area
program (SPSS Inc.). Linear and quadratic functions were devel- (Table 3). The correlation coefficients for the quadratic functions
oped because two studies (4,16) showed that the latter had a were greater than those of their linear counterparts. The refer-
better age–translucency correlation for length measurements, and ence sample SEE was lowest for the quadratic function derived
similar results for area measurements were expected here. for maximum translucency length, and quadratic functions in
These functions, or formulae, were applied on the test dataset. general (Table 3). However, test of functions on the control
In each of the six cases with multiple teeth, age was estimated dataset (n = 33) revealed that the quadratic function derived for
separately for every section and averaged, which was taken as translucency area produced the smallest SEE, MAD, and the
the age for that case. The difference between the estimated and highest number of “acceptable” age estimates (Table 4). The
actual age was determined for each case in the test dataset, and quadratic functions, overall, produced lower SEEs, MADs, and
the mean absolute difference (MAD) was calculated. The abso- higher “acceptable” age estimates than their linear counterparts
lute difference is the difference between the estimated and actual (Table 4), the exception being maximum translucency length
age devoid of a positive or negative sign (the “unsigned” value). wherein linear and quadratic functions produced the same level
The MAD therefore is the average magnitude of difference in a of “acceptable” age estimates. On the other hand, the linear
set of estimates and has been used as a measure of accuracy of functions produced fewer “unacceptable” age estimates than the
age estimation methods (16,17). Although the standard error of quadratic functions, with the exception of translucency area,
the estimate (SEE) produced from regression analyses gives an where it was the same for both (Table 4).
indication of the expected error rate, there is an inherent bias
because it is based on the training dataset (18) and does not
Discussion
reflect the error in an independent test sample. Moreover, Gorard
(19) has convincingly criticized the use of statistical indicators The assessment of translucency is an easy, practical, and rela-
tively accurate method of age estimation, which is usable by
experts and novices alike. The 250-lm-thick tooth sections
TABLE 2––Comparison of the proportion of age groups, sexes, and tooth
ensured the best possible visualization of translucency. While
classes in the training and test dataset. this study obtained sections using a hard-tissue microtome, simi-
lar sections may also be obtained through manual grinding and
Group Training Dataset (%) Test Dataset (%) their thickness verified using a micrometer (e.g., Digimatic
Age group
Micrometer, Mitutoyo Corp., Kawasaki, Japan). While manual
19–30 19.0 9.1 grinding would not require mounting the section in autopolymer-
31–40 20.0 9.1 izing acrylic, the process of grinding per se would take a consid-
41–50 18.0 27.3 erably longer time.
51–60 18.0 21.2 Nevertheless, dentin translucency correlates most closely with
>60 25.0 33.3
Total 100 100 chronological age when compared to other microscopically visi-
Subjects ble age changes on the dental tissues (2,9). Hence, large-scale
Males 49.0 51.5 interest in assessing this parameter resulted in several published
Females 51.0 48.5 papers that evaluated both length and area measurements
Total 100 100
Sections
(5,6,9,14,21). However, in contrast to Lorentsen and Solheim’s
Incisors 37.0 31.9 findings (13), these studies reported higher age correlations for
Canines 21.0 21.3 length than for area, which is reflected in the results of this
Premolars 42.0 46.8 paper. The smaller diameter and less number per unit area (5) of
Total 100 100 dentinal tubules toward the root apex and root surface have been
ACHARYA . DIGITAL TRANSLUCENCY AREA MEASUREMENT 767

TABLE 3––Correlation coefficients (R), standard errors of the estimate (SEE), and regression formulae derived for translucency length and area measure-
ments.

Variables and Regression Function R SEE (Years) Regression Formulae


Avg. T length (ATL) linear 0.55 13.3 Age = 35.56 + (3.48 9 ATL)
Avg. T length (ATL) quadratic 0.60 12.8 Age = 29.91 + (7.45 9 ATL) + ( 0.44 9 (ATL 9 ATL))
Maximum T length (MTL) linear 0.60 12.8 Age = 33.39 + (2.81 9 MTL)
Maximum T length (MTL) quadratic 0.62 12.5 Age = 28.61 + (5.50 9 MTL) + ( 0.24 9 (MTL 9 MTL))
T area (TA) linear 0.52 13.6 Age = 38.36 + (1.06 9 TA)
T area (TA) quadratic 0.60 12.7 Age = 31.75 + (2.97 9 TA) + ( 0.07 9 (TA 9TA))
T, translucency.

TABLE 4––Comparison of accuracy of the formulae derived for the measurements of translucency length and area.

Function ≤ 5 Years (%) ≤ 10 Years (%) ≥ 15 Years (%) MAD (Years) SEE (Years)
Avg. T linear 11/33 (33.3) 17/33 (51.5) 6/33 (18.2) 9.45 12.04
Avg. T quadratic 9/33 (27.3) 22/33 (66.7) 7/33 (21.2) 8.92 11.40
Maximum T linear 14/33 (42.4) 21/33 (63.6) 6/33 (18.2) 8.91 11.82
Maximum T quadratic 12/33 (36.4) 21/33 (63.6) 8/33 (24.2) 8.84 11.80
T area linear 15/33 (45.5) 17/33 (51.5) 6/33 (18.2) 8.94 11.73
T area quadratic 14/33 (42.4) 24/33 (72.7) 6/33 (18.2) 8.27 10.97
T, translucency.

considered as reasons for the commencement of translucency


from the root tip and periphery. As an extension, it may be
suggested that the narrower tubules closer to the root surface
vis-a-vis those toward the root canal become translucent more
consistently as age progresses, thereby contributing to a vertical
linear (i.e., length) increase in root dentinal translucency that is
more regular with an increase in age, than vertical and horizontal
increase combined (i.e., area).
According to Thomas et al. (14), because translucent dentin is
not deposited uniformly, its three-dimensional volume may be
more accurate in age estimation. However, they concede that the
problem of computing three-dimensional volume may preclude
its usage. In fact, Lorentsen and Solheim (13) have stated that a
method previously developed for measuring translucency volume
(viz., Ref. [22]) is a “difficult technique requiring expensive
equipment and the time-consuming measurement of a sufficient
number of teeth” (pg. 8). Therefore, they believe that measuring
area of translucent dentin on tooth sections may be a simpler FIG. 4––Scatter plot for the training dataset (n = 100) showing correla-
procedure for estimating age (13). The results herein show that it tion (linear and quadratic regression lines) of translucency area with age.
The quadratic regression line begins to curve just after the age of 60 years.
is repeatable too. Moreover, the higher accuracy of the quadratic
formula derived for area measurements in the present study’s test
dataset may be an added justification for its usage.
The higher correlation and lower error rates of the quadratic accuracy of methods and variables. However, Giles and Klepin-
functions derived for area and length measurements here confirm ger (18) have stated that it is merely “the square root of the
those obtained previously for length measurements (4,16). This average of the squared errors for the sample” (pg. 1218). Fur-
suggests that the rate of increase in the dimensions of translu- ther, Gorard (19) points out that the relative efficiency of mea-
cency slows down or plateaus in old age, probably because most sures of dispersion such as the SEE depends “on there being no
of the root dentin has become translucent, impeding its further errors at all in the observations” (19; pg. 421). But for normal
increase (4). In the present sample, this appears to occur just distributions with small “contaminations” in the data, the relative
after 60 years of age, as seen by the curve of the quadratic func- advantages which sample deviations (e.g., SEE) may have in
tion (Fig. 4). uncontaminated situations over mean differences (e.g., MAD)
No paper that previously assessed and derived regression for- are radically reversed. For example, observed differences tend to
mulae for translucency length and area had tested the same on a be longer-tailed with more extreme values than would be
control dataset. The intention of such an approach was to gauge expected under ideal conditions. Unlike in MAD, the errors are
the possible accuracy and applicability of these formulae in squared in SEE, which makes it exponentially (rather than addi-
forensic casework, and not just stop at an inference based on tively) greater, and the act of square-rooting its average “does
correlation coefficients (R) and SEEs. The SEE, which is inver- not completely eliminate this bias” (19; pg. 421). Therefore, the
sely related to R, has been used regularly in age estimation stud- SEE may give misleading answers in these cases, and measures
ies—including translucency assessment (8,12,23)—to depict the of difference such as the MAD are more appropriate. Hence, the
768 JOURNAL OF FORENSIC SCIENCES

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Acknowledgments sic Sci Int 2007;165:178–81.
The author expresses his gratitude to Professor Srinath L. Tha- 25. Jeevan MB, Kale AD, Angadi PV, Hallikerimath S. Age estimation by
pulp/tooth area ratio in canines: Cameriere’s method assessed in an
kur, Principal of this institution, for his continued support to Indian sample using radiovisiography. Forensic Sci Int 2011;204:209.
research in forensic odontology. Thanks are also owed to Profes- e1–5.
sor C. Bahsker Rao, former Principal, for approving an institu-
tional grant for the study and his support. The author also Additional Information and reprint requests:
thanks Professor Sadashiva Shetty, Principal, Bapuji Dental Col- Ashith B. Acharya, B.D.S., G.D.F.O.
Associate Professor and Head
lege, Davangere, India, for providing subsidized access to the Department of Forensic Odontology
hard-tissue microtome at his institution and Professor Mandana S.D.M. College of Dental Sciences and Hospital
Donoghue, Head of Oral Pathology at the College of Dental Sci- Sattur
ences, Davangere, for making available sections from 14 sub- Dharwad 580009
jects. Many thanks are owed to Profs. Anand Tavargeri and Karnataka
India
Kiran Radder for providing some of the extracted teeth from E-mail: ashithacharya@hotmail.com
their private practices.