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Forensic Science International 277 (2017) 2129

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Forensic age estimation by morphometric analysis of the manubrium

from 3D MR images
Naira P. Martnez Veraa,b,* , Johannes Hllera , Thomas Wideka , Bernhard Neumayera,b ,
Thomas Ehammera , Martin Urschlera,b,c
Ludwig Boltzmann Institute for Clinical Forensic Imaging, Universittsplatz 4, 8010 Graz, Austria
BioTechMed-Graz, Graz, Austria
Institute for Computer Graphics and Vision, Graz University of Technology, Inffeldgasse 16, 8010 Graz, Austria


Article history:
Received 9 December 2016 Forensic age estimation research based on skeletal structures focuses on patterns of growth and
Received in revised form 12 April 2017 development using different bones. In this work, our aim was to study growth-related evolution of the
Accepted 2 May 2017 manubrium in living adolescents and young adults using magnetic resonance imaging (MRI), which is an
Available online 12 May 2017 image acquisition modality that does not involve ionizing radiation. In a rst step, individual manubrium
and subject features were correlated with age, which conrmed a statistically signicant change of
manubrium volume (Mvol : p < 0.01, R2 0:50) and surface area (Msur : p < 0.01, R2 0:53) for the
Forensic age estimation
MRI studied age range. Additionally, shapes of the manubria were for the rst time investigated using
Manubrium principal component analysis. The decomposition of the data in principal components allowed to analyse
Morphometry the contribution of each component to total shape variation. With 13 principal components, 96% of
Principal component analysis shape variation could be described (Mshp : p < 0.01, R2 0:60). Multiple linear regression analysis
modelled the relationship between the statistically best correlated variables and age. Models including
manubrium shape, volume or surface area divided by the height of the subject (Y  MshpMsur/
Sh : p < 0.01, R2 0:71; Y  MshpMvol/Sh : p < 0.01, R2 0:72) presented a standard error of estimate of
two years. In order to estimate the accuracy of these two manubrium-based age estimation models, cross
validation experiments predicting age on held-out test sets were performed. Median absolute difference
of predicted and known chronological age was 1.18 years for the best performing model (Y  MshpMsur/
Sh : p < 0.01, R2p 0:67). In conclusion, despite limitations in determining legal majority age, manubrium
morphometry analysis presented statistically signicant results for skeletal age estimation, which
indicates that this bone structure may be considered as a new candidate in multi-factorial MRI-based age
2017 Elsevier B.V. All rights reserved.

1. Introduction most frequently used biological features for estimating CA [7],

with skeletal and dental development predominantly being
Forensic age estimation is an important tool in anthropology investigated by radiographic imaging methods [810]. Deriving
for identication of human remains [1], however, recently it has CA from biological development is challenging due to the
gained a lot of interest especially for age estimation of living variation of human development [11,12] as well as uncertainties
adolescents and young adults to verify chronological age (CA) in introduced by both limited study populations [13] and used
legal proceedings [24] or sports applications [5,6]. For this radiological staging methods [8,9,14]. Therefore, there is an
purpose, skeletal, sexual and dental maturity indicators are the increased interest in identication of novel indicators of
biological development to support accurate multi-factorial age
estimation approaches [15].
Skeletal age (SA) estimation is a core component in forensic age
* Corresponding author at: Ludwig Boltzmann Institute for Clinical Forensic estimation of the living from childhood into late adolescence. It is
Imaging, Universittsplatz 4, 8010 Graz, Austria.
based on skeletal maturation and may be followed until
E-mail addresses: (N.P. Martnez Vera), (M. Urschler).
development ceases to progress and maturity of the specic bone
0379-0738/ 2017 Elsevier B.V. All rights reserved.
22 N.P. Martnez Vera et al. / Forensic Science International 277 (2017) 2129

is attained [16]. To increase the potential age range after the changes across age [32]. They standardized the sternum according
completion of hand bone ossication, which occurs around the age to size and shape by applying morphometric analysis using indices
of 18 years in males [8,9,1719], research has been performed on of manubrium breadth and length proportion, thickness, area and
other patterns of growth and development using different bones. volume. Based on the model for rib cage shape by Gayzik et al. [34],
The medial clavicular epiphysis (MCE) has been intensively studied a complete quantication of size and shape variation was
in plain chest radiography [20] and thin-slice computed tomogra- characterized for ages between 0 and 100 years from landmarks
phy (CT) of the thorax [21,22]. However, use of ionizing radiation of CT scans of the sternum by Weaver et al. [35]. In summary, all
without medical indication for examining healthy children and these works suggest a signicant, reproducible change in
adolescents is problematic with these imaging modalities. morphometry of the sternum during growth. Thus, the aim of
Therefore, magnetic resonance imaging (MRI) is nowadays seeing our study was to verify the hypothesis that growth-related
increasing research interest for bone age estimation using hand morphometric changes in the manubrium can be used to predict
and wrist bones [5,23,24] and especially MCE [2528]. A very chronological age in a forensically relevant data set of male
obvious extension to MRI-based investigation of the clavicular adolescents and young adults around majority age.
ossication is to additionally study the manubrium, i.e. the upper
part of the sternum, since it is well visible on MR images of the 2. Material and methods
upper thorax and therefore requires no extra image acquisition.
Age-dependent morphometric variability of the manubrium has 2.1. Study design
already been shown in the literature in relation to growth and
maturation [29]. These changes are due to ossication in the We retrospectively analysed the manubrium based on an
cartilaginous sternum that begins during fetal development and is available dataset of upper thorax MR images that was acquired
completely nished around the age of 25 except for the for studying age estimation using the clavicle (see Fig. 1). The
manubriosternal joint [30]. Previous studies have analysed accompanying study investigating the clavicle was performed in
standard linear measurements such as manubrium length, area accordance with the Declaration of Helsinki and was approved
and volume changes with age using radiographs or osteological by the ethical committee of the local medical university. All
collections [31,32]. In [33], CT data of the sternum was analysed eligible volunteering participants of the accompanying study
and the results showed a volume increase in a population of 48 provided written informed consent and from underage subjects
children between 4 months and 15 years which was modelled by written consent signed by the parents was additionally
an exponential function. Selthofer et al. have further studied shape obtained.

Fig. 1. T1-weighted coronal MRI slices showing clavicles (C) and manubrium (M) from a 13.02- (left) and a 23.71-year-old (right) male subject including semi-automatically
derived segmentations in red (top row). 3D point clouds visualizing the respective 3D shapes of the manubria at the different ages (bottom row). (For interpretation of the
references to colour in this gure legend, the reader is referred to the web version of this article.)
N.P. Martnez Vera et al. / Forensic Science International 277 (2017) 2129 23

Marching Cubes algorithm [40]. The exported 3D triangle mesh

was then pre-processed with the MeshLab software.3 This pre-
processing consisted of Poisson reconstruction to achieve a
consistent, hole-free surface, followed by mesh simplication
according to a quadric surface error measure [41] that reduced the
number of vertices to 5000 (bottom row in Fig. 1). From these 3D
meshes, volume (Mvol) and surface area (Msur) of the manubria
were computed. After roughly aligning all meshes by shifting each
to its centre of gravity, a base mesh was randomly selected and all
other meshes were non-rigidly registered to the base mesh with
the method from [42]. As a result, correspondence of 3D points for
all 3D meshes of the manubria could be established. Dened in
analogy to [32], the determined correspondences allowed to
extract four conventional linear measurements (upper and lower
manubrium width Muw, Mlw, manubrium height Mh and
manubrium thickness Mt) by identifying eight homologous
Fig. 2. Distribution per year among the study population of 130 subjects. Each landmarks (3D points). The input for principal component analysis
histogram bin represents the illustrated age 0.5 years. (PCA) based shape analysis was provided by the full set of
corresponding 3D points for all 3D meshes of the manubria. PCA is
2.2. Subjects a statistical procedure that transforms observations, i.e. the 3D
points for all 3D meshes, into a set of coefcients called principal
In this study morphometric changes [36] were analysed in components [43]. The rst principal component encodes the
relation to CA on MRI data from 130 male Caucasian subjects with largest variance seen in the dataset, thus describing the principal
ages ranging from 13.01 to 24.98 years (mean age 19.00 years, mode of maximal shape variation. Succeeding components are
standard deviation: 3.66 years). Subjects were roughly evenly orthogonal to preceding components but with decreasing variance,
distributed within this age range as can be seen in the age thus representing less important variations in shape. From
histogram in Fig. 2. Prior to scanning, height and weight of the corresponding landmarks we created the PCA-based shape model
subjects were additionally collected by the expert on forensic age Mshp by rst performing full Procrustes analysis [44] with respect
estimation responsible for the study. to translation, rotation and scale to achieve a model where aligned
shapes are centred around the same origin, have the same 3D
2.3. MR image acquisition orientation and are normalized in size. This model solely contains
variations in shape due to anatomical differences in a population,
Subjects were examined on clinical 3 T MRI scanners (Tim Trio/ i.e. a manubrium shape space according to the denition of Kendall
Skyra, Siemens Healthineers, Erlangen, Germany). Images of the [45]. After this Generalized Procrustes analysis step, PCA was
manubrium were acquired in supine position with a neck coil performed using Matlab R2014a, thus achieving a parametric
(Siemens Healthineers, Erlangen, Germany). T1-weighted images model of shape where each 3D mesh can be represented exactly
of the upper thorax region were obtained using a 3D VIBE sequence using a projection onto the principal components.
with water excitation (FOV = 240 mm, TR/TE = 9.77/3.72 ms, voxel
size 0.9  0.9  0.9 mm3). The sequence was oriented on a coronal 2.5. Linking manubrium development with age
oblique plane, parallel to the sternal bone. To avoid pulsation
artifacts from the heart and vessels, saturation bands were applied We rst investigated the correlation of each manubrium feature
in caudal and posterior position of the 3D region of interest. Fig. 1 (conventional linear measurements Muw, Mlw, Mh, Mt in cm,
shows two representative MR images of 13.02 and 23.71 year old surface area Msur in cm2, volume Mvol in cm3 and the most
subjects, respectively. MR images of manubria included in this prominent shape change described by the rst principal compo-
study did not show a visible fusion at the manubriosternal joint nent of the PCA model M1shp ) and each subject feature (height Sh in
and those images which presented lateral ossications were cm and weight Sw in kg) with known chronological age. Unity-
incorporated, however, these ossications were not included into based normalization was used to scale these variables into the
the analysis (Fig. 9, right). range between 0 (minimum value) and 1 (maximum value). For
those two features which showed highest correlation with age
2.4. Manubrium characterization (Mvol, Msur), we additionally performed linear regression analysis
to describe how changes in these variables are related with age in
From the acquired T1-weighted images, we characterized our target age range of subjects.
manubria by indices of width, height, thickness, volume, surface We further explored the relationship of individual PCA shape
area and shape variations. Therefore, a volumetric manubrium components together with the highest correlated individual
segmentation was semi-automatically performed for each image manubrium and subject features and chronological age using
by using the active learning based segmentation method from multiple linear regression (MLR) analysis. Therefore, in a rst step
TurtleSeg software1 [37,38]. Due to inhomogeneities in the MR we identied an appropriate number of principal shape compo-
images, some borders of the manubrium were difcult to get nents to restrict further age regression analysis to the most
detected automatically by the software, therefore some segmen- relevant shape variables. Besides the determined shape compo-
tations were manually corrected using ITKSnap software2 [39]. nents Mshp, additional candidate regressors for MLR were
From the semi-automatically segmented manubria (red mask in manubrium features Mvol, Msur as well as subject-specic features
top row of Fig. 1), 3D meshes were created with ITKSnap using the Sh and Sw . To rule out collinearities between multiple explanatory

1, last accessed 11.04.2017.
2 3, last accessed 11.04.2017., last accessed 11.04.2017.
24 N.P. Martnez Vera et al. / Forensic Science International 277 (2017) 2129

variables, Besley collinearity diagnostics [46] was performed.

Potential correlations were tested by singular values (s), condition
index (condIdx) and variance decomposition proportions
(VarDecomp). Finally, MLR models were chosen to be constituted
by variables which presented signicant correlation with age in the
linear correlation analysis and which showed independence of
each other in the collinearity tests. From these variables, different
combinations were used to t a number of multivariate models
that allow regression of chronological age.
In our nal experiments, 13-fold cross-validation analysis was
performed for all candidate MLR models to evaluate the ability to
predict age on images not contained in the dataset from which the
respective model is computed, thus indicating practical generali-
zation capabilities of the models. For this purpose, the data was
divided into ve age groups (1316 years, 1619 years, 1922 years,
2225 years) and 13 tests were conducted. Each test partitioned
the manubria set into two subsets, the test group with two
manubria per age group and the training group with the remaining
120 manubria. The test group was created by randomly drawing
from the whole dataset, such that each image was used once as a
test image during the 13 folds, but taking the distribution among
age groups into account. During each cross-validation fold, the
shape model Mshp was created from the training group and used to
predict the age of the subjects in the test group. This procedure was
repeated for all investigated MLR models. The absolute differences
between estimated age and chronological age for all subjects in the
test group were computed in each cross-validation round, and
summarized by calculating the median and the maximal absolute
difference. These values were then used in the statistical analysis.

2.6. Statistical analysis

Statistical analysis was performed using Matlab R2014a. To

measure the linear correlation between each manubrium and
subject variable and age, the Pearson's r coefcient was used.
Goodness of t of regression models was analysed using R2 values,
together with their statistical signicance. In MLR analysis, R2 is
inuenced by the number of variables. Adding any regressor
variable to an MLR model generates a bias towards a smaller error
and therefore a higher R2. To take this into account when
Fig. 3. Strength of pairwise linear relationships indicated by Pearson's linear
comparing models, in the MLR experiments goodness of t was correlation coefcients. Manubrium shape is represented only by the rst principal
determined by the adjusted coefcient of determination R2 . component (M1shp ) of the PCA model. Data were normalized between 0
Additionally, the standard error of estimate (SEE) was used to (minimum value) and 1 (maximum value). All correlations (highlighted in
assess different MLR models. In our cross-validation experiment, red) were statistically signicant with p < 0.01. (For interpretation of the
references to colour in this gure legend, the reader is referred to the web
the prediction measure R2p was used to evaluate the ability of a version of this article.)
tted regression model derived from a training group to predict the
age of the associated test group. In this part of the study, all models
to be compared had the same number of variables, such that conventional linear manubrium measurements as dened in [32]
unadjusted R2 could be used for comparison. Details of all used showed correlations with a Pearson's r coefcient larger than 0.5,
statistical measures can be found in Appendix A. A paired t-test i.e. upper and lower manubrium width Muw (r = 0.66), Mlw
was used for comparison between age prediction candidate (r = 0.66). Highest correlation with age was found for surface area
models. A p-value less than 0.05 was considered statistically Msur (r = 0.73) and volume Mvol (r = 0.71) of the manubria, while
signicant, both in correlation/regression as well as t-test analysis the rst principal component of the manubrium shape model
for comparing models. (M1shp ) showed only low correlation with age (r = 0.25).
Individual linear regression models of upper and lower
3. Results manubrium width, surface area and volume with age were
statistically signicant with respect to the linear tting
3.1. Analysis of individual manubrium variables
(Muw : R2 0:43, p < 0.01; Mlw : R2 0:43, p < 0.01;
2 2
The scatter plots in Fig. 3 show paired variables formed from the Mvol : R 0:50, p < 0.01; Msur : R 0:53, p < 0.01). The two most
individual manubrium and subject features and CA. All correla- promising linear regression models (Mvol, Msur) are plotted in
tions (highlighted in red) are statistically signicant, with a p-value detail in Fig. 4, together with their 95% prediction intervals. Both
<0.01. The histograms of each single variable appear next to each models indicated similar variation with age (Msur(x) = 0.04  x
scatter plot, showing the range of values for the normalized  0.25, Mvol(x) = 0.04  x  0.25). Values outside the prediction
variable to reveal the underlying distribution. Two of the four intervals came from two subjects whose BMI (as derived from Sh
N.P. Martnez Vera et al. / Forensic Science International 277 (2017) 2129 25

Fig. 6. Eigenvalues of the rst 60 eigenvectors of the shape PCA-model. In red, the
rst 13 principal components which explain 96% of the shape variation, providing
Fig. 4. Regression of manubrium surface area (Msur) and volume (Mvol) over age. a suitable trade-off between model exibility and compactness. (For interpretation
Dotted and dash-dotted lines indicate the 95% prediction intervals for surface area of the references to colour in this gure legend, the reader is referred to the web
and volume, respectively. version of this article.)

and Sw ) fell into obese (BMI = 34.32 kg/m2) and underweight Besley collinearity diagnostics assessed the strength and
(BMI = 18.19 kg/m2) categories. sources of collinearity among the manubrium (shape, volume
and surface area) and subject (height and weight) features in the
3.2. Shape variation and MLR analysis MLR model. Strong multicollinearity existed between the manu-
brium variables Msur and Mvol (see Fig. 7). From the test of
To investigate the hypothesis that shape variations of the collinearity and the MLR analysis, the best candidate models to
manubrium are related to age, we analysed the PCA based shape build a predictive age model are summarized in Table 1 together
model with respect to the used number of principal components in with their goodness of t and SEE when used on all available
an MLR setup. Starting from a very low coefcient of determination manubria. Models which included shape with surface area or with
of the manubrium shape model when using just the rst principal
surface area divided by height yielded high R2 values (p < 0.01,
component (M1shp : p < 0:01, R2 0:05), the inclusion of more
R2 0:71). Similar results were obtained using models constituted
principal components increased R until saturation was reached at by shape and volume (p < 0.01, R2 0:70), however an improve-
13 included components (Mshp : p < 0.01, R2 0:60). This behav- ment was found when the volume was divided by the height of the
iour is illustrated in Fig. 5. The magnitudes of the eigenvalues of subject (p < 0.01, R2 0:72). Lowest R2 values (R2  0:60) were
Mshp are shown in Fig. 6. With 13 principal components, 96% of found in models where the subject's weight was included. The
the shape variation could be explained, while the rest can be
assumed to come from noise as seen by its low eigenvalue
magnitudes, thus providing a suitable trade-off between model
exibility and compactness.

Fig. 7. Strength and sources of collinearity among the manubrium and subject
variables in a multiple linear regression model. Manubrium shape is represented by
using only the rst principal component (M1shp ). Variance-decomposition
proportions with a condition index (condIdx) larger than the tolerance 30
are shown. The variance-decomposition proportions exceeding the default
Fig. 5. Adjusted coefcients of multiple determination R2 obtained when the tolerance proportion 0.5 (tolProp) are indicated by red markers. Results
shape models were correlated with age. These models were constituted from indicated that surface area (Msur) and volume (Mvol) features presented
one to 60 principal components. Dashed line points out the subsets with 13 linear dependency (condIdx = 36.9). (For interpretation of the references to
principal components, where a large improvement in R2 was observed before colour in this gure legend, the reader is referred to the web version of this
going into saturation. article.)
26 N.P. Martnez Vera et al. / Forensic Science International 277 (2017) 2129

Table 1
Multiple linear regression (MLR) models for age estimation. Results of best
performing MLR models in bold.

MLR model R2 SEE/years

Y  Mshp + Mvol 0.70** 2.12

Y  Mshp + Msur 0.71** 2.12
Y  Mshp + Mvol/Sh 0.72** 2.08
Y  Mshp + Msur/Sh 0.71** 2.07
Y  Mshp Mvol =Sw 0.63** 2.39
Y  Mshp Msur =Sw 0.60** 2.50

R2 = adjusted coefcient of determination; SEE = standard error of estimate;

Y = estimated age; Mshp = manubrium shape; Mvol = manubrium volume;
Msur = manubrium surface area; Sh = subject height; Sw = subject weight.
p-value <0.01.

standard error of estimate for these models was larger than two
years, with a small improvement when the height of the subject
was taken into account (Y  MshpMsur/Sh: SEE = 2.08 years; Fig. 8. Each box-whisker plot represents the difference between estimated SA and
Y  MshpMvol/Sh: SEE = 2.07 years). CA for subjects from the specied age group. Between 16 and 22 years of
chronological age, the median absolute difference of 1619 and 1922 years age
groups are 0.97 and 1.12 years, respectively.
3.3. Predictive models
years and 1.12 years, respectively, however with very different
Table 2 sums up the results of the cross-validation experi- distributions (75% percentile was 2.63 and 1.63 for the two groups,
ments with different predictive models. Models with the respectively). For the youngest age group, absolute differences of
inuence of subject's height produced higher values of the 75% of the age predictions were less than 3 years (median 1.75
predictive coefcient of determination (p < 0.01; R2p 0:67) years), while in the oldest group the median was highest with 2.31
compared to the models where only manubrium features were years. Outliers in the age group between 19 and 22 years came from
included (p < 0.01; R2p 0:65) and the models normalized by the overweight subjects (BMI = 34.32 kg/m2), subjects possessing a
manubrium with a rounder shape as expected at the same age
weight of the subject (p < 0.01; R2p  0:55). Regarding median
(Fig. 9, left), and subjects which presented lateral ossications
absolute difference of predicted and known chronological age, the
(Fig. 9, right).
model including both manubrium shape and surface area features
together with subject height predicted CA more accurately
4. Discussion
(1.18  0.41 years) than the same model but with the manubrium
volume feature (1.29  0.32 years). Nonetheless, the maximal
This retrospective study presented the manubrium bone as a
absolute age difference was smaller using the model which
candidate for age estimation in males during adolescence and early
included volume (4.76 years) than for the model where the surface
adulthood, especially focusing on the forensically important age
area of the manubrium was included (4.92 years). A statistically
range between 13 and 25 years, i.e. around majority age. To the best
signicant difference with p < 0.01 was found in median and
of our knowledge, we were the rst to propose the study of
maximal age difference when comparing those two models. Using
manubrium morphometrics using MRI data, which has the benet
either of the two methods with highest R2p , 17.57% of subjects of not involving ionizing radiation during acquisition. Age
younger than 18 years were estimated above the 18 years threshold prediction was studied in relation with shape variations of the
and 8.93% of the subjects older than 18 years were predicted as manubrium as derived from principal component analysis. This
younger than 18 years. Results of age estimation specic to the age extends upon previous morphometric studies [32,35] by using the
groups 13 years x< 16 years, 16 years x< 19 years, 19 years entire shape information instead of a limited user selected set of
x< 22 years and 22 years x< 25 years using the predictive model landmarks.
Y  MshpMsur/Sh can be observed in the box-whisker plots of The investigated 130 subjects cover an age range from 13.01 to
Fig. 8. Two age groups, between 16 and 19 years and between 19 24.98 years (approximately 10 subjects per year), where strong
and 22 years, showed a similar median absolute difference of 0.97 changes due to manubrium development may be expected. In
agreement with the literature, statistics of individual manubrium
features showed that volume and surface area of the manubria in
Table 2
our dataset correlated with age. According to Sandoz et al. [33], the
Predictive models for age estimation from cross-validation experiments. Results of
best performing predictive models in bold. volume of the sternum showed an increase between 0 and 15 year
old subjects, which they modelled using an exponential function.
MLR model R2p Std Median/year Std Max./year
After the age of 13 years, it is reasonable to consider that this
Y  Mshp + Mvol 0.65 **
0.16 1.39 0.28 5.25 exponential increase ceases, and the consequences of growth
Y  Mshp + Msur 0.65** 0.13 1.38 0.33 5.13 decline after puberty. So our hypothesis was to select a linear
Y  Mshp + Mvol/Sh 0.67** 0.14 1.29 0.32 4.76 regression model to describe the change of the different variables
Y  Mshp + Msur/Sh 0.67** 0.12 1.18 0.41 4.92
Y  Mshp Mvol =Sw 0.56** 0.12 1.66 0.87 6.35
along this age range (Fig. 4). Results afrmed this hypothesis with
Y  Mshp Msur =Sw 0.53** 0.15 1.79 0.93 6.83 statistically signicant changes over age both in volume and
surface area.
Y = estimated age; Mshp = manubrium shape; Mvol = manubrium volume; Msur =
The manubrium is a attened bone in an antero-posterior
manubrium surface area; Sh = subject height; Sw = subject weight; R2p = predictive
direction, with the posterior surface being smoother than the
coefcient of determination; Std = standard deviation; median and maximal
(Max.) absolute difference of predicted and known CA for the methods anterior one. The superior region is broader than the inferior,
selected with larger R2 values in the MLR analysis.
and the superior border is rounder and smoother than the
p < 0.01. roughened inferior border [47]. We found that the use of such
N.P. Martnez Vera et al. / Forensic Science International 277 (2017) 2129 27

Fig. 9. Two examples of T1-weighted coronal MRI slices of individuals which presented outlier cases. At the left side, the manubrium of a subject which presented more
rounded shape compared to others with a similar age (20.57 years). At the right side, a 19.35 years old subject which showed lateral ossications (yellow arrows). Our
predictive model estimated these subjects 17.42 and 23.76 years old, respectively. Anatomical structures: M = manubrium, C = clavicle. (For interpretation of the references to
colour in this gure legend, the reader is referred to the web version of this article.)

conventional linear measurements (i.e. distances between differences compared to other subjects with similar age. This
landmarks) describing width, height or thickness of the indicates that the two models with highest R2p are highly inuenced
manubrium are not enough to quantify the entire complex by such outlier cases. Although results obtained from all models
morphology of this bone. While Weaver at al. already are similar, we have selected the model which includes shape,
characterized the sternal shape variation independent from surface area and subject height because of its lowest median age
sternal size [35], in this work we extended their study by not estimation error (1.18 years). However, in cases in which human
only evaluating distances between landmarks, but taking remains have been mutilated and the stature of the subject is not
advantage of the whole shape variation of the manubria in available, the two models constituted only by manubrium features,
our dataset. After including 13 principal components into our i.e. without subject features, may also be practical methods for age
shape model, shape variations of the manubrium from MR estimation.
images showed statistically signicant correlation with age When dividing the data into four groups according to age and
(see Fig. 5). Using PCA, the full dimensionality of the shape- using the predictive model which showed the smallest median age
space is reduced to the more compact shape-space spanned by estimation error (see Fig. 8), manubria coming from subjects in the
13 principal components, these ones retaining the most youngest and oldest age groups showed the largest median
important shape variations of the manubrium within the entire difference to chronological age. This indicates that only small
age range under study. Generally, the shape tended mainly to morphological changes occur in these age ranges and our model
change from an oval shape in early childhood to an angular and lacks the specicity to detect them. However, manubria from
trapezoidal shape (Fig. 1) in young adults driven by the individuals between 16 and 22 years, resembling a very important
ossication process that takes place during manubrium age range in practical forensic age estimation, were predicted with
development [29,35]. a median error of 0.97 and 1.12 years, respectively, suggesting that
Different models composed of manubrium and subject features strong morphological changes occur and could be detected by our
were investigated using MLR analysis. Results from these models model. There was a small but considerable percentage (17.57%) of
indicated that changes in the manubrium shape described with 13 subjects younger than 18 years that were estimated older than 18,
principal components together with changes in volume or surface indicating that these models currently have to be considered with
area of the manubrium normalized by the body height of the care for establishing age of majority, which is 18 years in the legal
subject were highly age-dependent (Table 1). However, models systems of many countries.
including subject weight did not give any new information about
manubrium changes. The accuracy of predictions measured by SEE 4.1. Limitations
showed that these models presented in the best case 2.07 years of
prediction error. Due to its retrospective design and its connection to a forensic
To test prediction accuracy and generalization capabilities of age estimation study of adolescents and young adults, for this
the regression models in a more practically oriented setup, a 13- study only volunteers between 13 and 25 years were investigated.
fold cross-validation was carried out (Table 2). Models predicted For this age range, strong shape changes due to manubrium
age with median absolute differences to the known ground-truth development can be expected. Extension to other age ranges would
age smaller than 1.80 years. These prediction results conrm that have to be explored in follow-up studies, however, for examining
strong changes in manubrium morphology occur in this age range, younger children the need for sedation or anesthesia to enable the
which can be used for age estimation. However, unfortunately the currently time-consuming acquisition of MRI data clearly is a
maximal age differences were also large. When examining those major limitation.
manubria more closely, for which large differences occur, we see Despite our MR protocol being adapted for MCE imaging, the
that most outliers result from overweight subjects, from manubria manubrium bone was well depicted in our whole dataset of MR
presenting lateral ossications or from signicant shape images. However, some edges of the manubria in a number of MR
28 N.P. Martnez Vera et al. / Forensic Science International 277 (2017) 2129

images, mostly at the bottom of this structure, could not be Appendix A. Details of statistical analysis
detected reliably by the segmentation software. This issue is a
result of the MRI scans not being optimized for the manubrium but The adjusted coefcient of determination (R2 ) that assesses
for the clavicle. So, in these cases manual image correction was goodness of t in the MLR experiments was dened as:
performed which suffers from observer variability. Consequently,  
future work will focus on showing the reproducibility of the n1  
R2 1  1  R2 ;
manual image segmentation by letting another observer repeat the np
ground truth segmentations and studying inter-observer variabili-
where the number of model coefcients (p) is used in computing
ty. This resembles a large effort and was not yet considered, since
our aim was rst to verify our hypothesis that shape variations are R2 and n is the number of subjects.
in principle related with age. Following formula was used to calculate the standard error of
Currently, clinical applicability of our method is limited by the estimate (SEE) for all two degrees of freedom MLR models:
tedious and time-consuming semi-automatic manubrium seg- v
uX yi;CA  yi;SA 2
mentation step. While all subsequent steps like 3D mesh SEE t
generation, non-rigid point registration and projection into PCA i
shape space are already fully automatic, developing a software
pipeline for routine clinical use will require further research on where yi,CA is the CA, yi,SA is the estimated skeletal age of subject i,
automatic segmentation of the manubrium from MRI data. Here, and n the number of subjects in our sample.
statistical shape and appearance models [48] as well as recent In our cross-validation experiment involving predictive models,
machine learning methods for anatomical landmark localization R2p for a model t was calculated by
[49,50] are promising approaches to follow. We see the work-ow Pn
yi;CA  yi;SA 2
described in this manuscript as a rst step towards establishing R2p 1  Pi1 ;
n 2
such an automated pipeline. i1 yi;CA  yCA
Subjects with a manubrium whose shape differed considerably
where n, yi,CA and yCA were the number of subjects, the CA of
from the rest of our population at a comparable age, resulted in a
subject i and the mean age in the random training subset,
large age estimation error. Selthofer et al. suggested a standard
respectively. Using the model derived from a training group, the
sternum shape (trapezoid shape) which was present in more than
estimated SA of subject i in the test group was computed to give yi,
two thirds of his analysed samples, with the remaining third being
divided into two groups, triangular and quadrangular shapes [32].
So future work will need to pre-classify these three different types
of shapes and dene them separately in our PCA model.
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