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Forensic or Archaeological Issue: Is Chemical Analysis of Dental Restorations

Helpful in Assessing Time Since Death and Identification of Skeletonized
Human Remains?

Article  in  Journal of Forensic Sciences · July 2013

DOI: 10.1111/1556-4029.12228 · Source: PubMed

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 J Forensic Sci, September 2013, Vol. 58, No. 5
doi: 10.1111/1556-4029.12228
TECHNICAL NOTE Available online at: onlinelibrary.wiley.com

ANTHROPOLOGY; ODONTOLOGY

Ksenija Zelic,1 D.M.D.; Danijela Djonic,1 Ph.D.; Olivera Neskovic,2 Ph.D.; Milovan Stoiljkovic,2 Ph.D.;
Slobodan Nikolic,3 Ph.D.; Vladimir Zivkovic,3 Ph.D.; and Marija Djuric,1 Ph.D.

Forensic or Archaeological Issue: Is Chemical

Analysis of Dental Restorations Helpful in
Assessing Time Since Death and Identification
of Skeletonized Human Remains?*

ABSTRACT: In 2011, small mass grave with completely skeletonized remains was discovered in Belgrade suburb. An eyewitness claimed
that skeletons belonged to German soldiers killed in WWII. Anthropologists were engaged to investigate whether the skeletal remains corre-
spond to the indicated German group or represent more recent case requiring court trial. Numerous dental restorations were noticed. Owing to
the fact that different dental materials were used in dental practice at certain times, the aim of this study was to explore whether analysis of
dental restorations could help in identification and estimation of time since death. Inductively coupled plasma optical emission spectrometry
revealed that dental fillings corresponded to copper amalgam, conventional silver amalgam, silicophosphate cement, and zinc phosphate cement.
Chemical results combined with anthropological and historical facts suggest that the individuals lived before the 1960s in country with well-
developed dental service at that time. Therefore, chemical analysis of dental fillings was useful to distinguish between skeletal remains that are
too old to be of forensic interest and the remains relevant to legal investigations.

KEYWORDS: forensic science, dental forensic, identification, dental fillings, time since death, mass grave

In routine work, forensic anthropologists are often faced with
the identification of skeletal remains found in mass graves from
the recent past. This is always a difficult task, especially when
many years have passed since the burial and when there is a lack
of archival information. First step in this procedure is to estimate
as accurately as possible the time since death (1). But determin-
ing the time of death with great precision is extremely difficult,
especially when body is completely skeletonized (2). Particu-
larly, it is difficult to distinguish whether skeletal remains belong
to archaeological context or represent more recent forensic case
relevant to legal investigations.
In the past century, the city of Belgrade has been a playground
for many battles (particularly during the Great War and World
War II), which left many victims. For many of them, the place of
burial has never been discovered. Periodically, during city recon-
structions or digging in the city areas, small mass graves have
been accidentally discovered, and forensic specialists were
invited to make analysis and possible identification of victims.
1 archaeology and forensics. This idea was based on the fact that
Laboratory for Anthropology, Institute of Anatomy, School of Medicine,
University of Belgrade, 4/2 Dr Subotica, 11000 Belgrade, Serbia.
2
Institute of Nuclear Sciences Vinca, University of Belgrade, P.O. Box
522, 11001 Belgrade, Serbia.
3
Institute of Forensic Medicine, School of Medicine, University of
Belgrade, 31a Deligradska, 11000 Belgrade, Serbia.
*Supported by The Ministry of Education and Science of The Republic of
Serbia, Grant No. 45005.
Received 31 May 2012; and in revised form 7 Sept. 2012; accepted 6 Oct.
2012.
The first question addressed to anthropologists is weather the
remains are of archaeological or forensic (medicolegal) signifi-
cance. A clear distinction between these two categories is not sta-
ted in the current legal documents in Serbia, so it is up to the
court to estimate potential forensic importance of each case (3).
In May 2011, a small mass grave with completely skeleton-
ized remains was accidentally discovered in a suburb of
Belgrade. An eyewitness who lived in the neighborhood at the
end of WWII indicated that skeletons most probably belonged to
a certain group of German soldiers killed by city defense forces.
Anthropologists were engaged by the local court to investigate
whether characteristics of the skeletal remains correspond to the
indicated German group or represent more recent forensic case.
After careful study of the classical anthropological markers of
identity and analysis of skeletal injuries, we have concentrated
on the dental status looking for additional evidence that could
be helpful in identification process. The aim of this study was to
explore whether chemical analysis of dental restorations could
be helpful in the identification process and estimation of the
time since death in such cases that are in the zone between
different dental materials of various compositions have been
used in the past century and that they were introduced in dental
practice and abandoned at certain times. Furthermore, the devel-
opment of dentistry in certain countries and areas was not
uniform, so we assumed that types of dental work found on
skeletal remains could help in distinction among the individuals
living in a region with well-developed dental service or those
from poor economic areas.

1284 © 2013 American Academy of Forensic Sciences

 ZELIC ET AL.
Materials and Methods
Location of the Grave and Testimonies of Witnesses
In May 2011, in the middle of digging in a yard of the family
house in a Belgrade suburb Visnjica, a small mass grave was
accidentally discovered. The police came to the scene, collected
skeletal remains, and transferred them to the Institute of Forensic
Medicine for anthropological analysis.
During the investigation of the scene, police found an old
person living in the neighborhood who stated that he was an
eyewitness of the incident connected to the mass grave.
He stated that on October 1944, after the liberation of
Belgrade, partisans executed six captured German soldiers near
the point where police found the skeletons 67 years later. The
witness was interrogated thoroughly, and it was determined that
the victims were middle-aged men and that they were shot in the
head.
Anthropological Analysis
The state of preservation of the skeletons was moderate. All
analyzed bones were bright yellow, dry, light, brittle, degreased,
and free of soft tissue. The minimum number of individuals was
five, determined by the number of skulls.
The sex was estimated by observation of the morphological
traits of pelvis and cranium (4,5). Age assessment was based on
morphological characteristics of pubic symphysis using the
Suchey–Brooks method (6). Stature was estimated by applying
the formula for the maximum length of long bones developed by
Trotter and Glesser (7). For each skeleton, morphological fea-
tures (cranial and postcranial) and evidence of signs of disease
and injury were recorded.
Dental Status
Dental analysis of the victims comprised morphological analy-
sis of present teeth, recording of the caries lesions and dental
fillings, and analysis of the orthopantomography of the present
jaws.
Six teeth with dental fillings were further chemically analyzed:
three amalgam fillings (A1—removed from the first left molar,
skull no. 1; A2—from the second right premolar skull no. 2, and
A3—from the first right molar skull no. 2), two cement fillings
(C1—taken from the second left premolar, skull no. 1, and C2—
from the second right premolar, skull no. 2), one dental base (L1
—taken from the first left molar, skull no. 1 after removing of
the amalgam A1 as it was underneath it).
The elemental composition of the dental fillings was deter-
mined using the technique of inductively coupled plasma optical
emission spectrometry (ICP-OES). The multichannel optical
emission spectrometer Spectroflame P was used, power was
2.5 kW at 27.12 MHz, and the wavelength ranged from 165 to
800 nm. Samples were gradually introduced in plasma by spray-
ing and analyzing calibration solutions. Calibration standards
were used in concentration ranges that match the content of the
studied elements. Standards have been tested using certified J.T.
Baker standards.
To determine the type of silver amalgam, one control amal-
gam was tested. We used nonconventional, high-copper amal-
gam, available in today’s dental practice. As the main distinction
between conventional and nonconventional amalgam is the
amount of copper, these data were inquired.
. DENTAL RESTORATIONS AND TIME SINCE DEATH 1285
Results
Anthropological analysis revealed that the remains belonged
to five persons, four males, and one individual of undetermined
sex. Epiphyseal union and teeth eruption indicated that all indi-
viduals were adult. A more detailed age determination was
possible in one skeleton, aged 45.6  10.4 (Suchey–Brooks
method). Estimated stature for three individuals was
181.63  3.37 cm, 180.37  3.37 cm, and 161.47  3.37 cm
(Trotter and Glesser method).
On four of five skulls, we found gunshot crater–shaped holes
(both entrance and exit on each skull). One of those skulls also
had additional keyhole-shaped defect, on the curved part of the
skull, probably due to tangential gunshot. The fifth skull had no
obvious gunshot injuries.
Dental Status
Four skulls were found with upper jaws, but one cranium had
all teeth lost ante mortem, so this one was not considered in our
study. No lower jaws were found.
Skull 1
In the upper jaws, the following teeth were present: on the
right side second premolar and first and second molar, and on
the left side second incisive, both premolars and all molars. All
other teeth have been lost postmortem except the third molar
(probably hypodontia). A small caries lesion was present on the
second molar on the right side. Amalgam fillings were present
on the first and second molar on the left side and on the first
molar on the right side, while cement fillings were found on the
second premolar on the left side. After removing amalgam filling
from the first molar (for further chemical analysis), we noticed
cement base underneath it. On orthopantomogram, it was notice-
able that the present incisive had endodontic treatment and that
the root has been filled partially (Fig 1). Furthermore, above the
root apex lunate, bone destructions were spotted (Fig 1). It was
probably caused by dentogenic infection. Similar destruction was
also found above the root of second left premolar probably with
the same cause. Also, on the second premolar on the right side,
a part of metal dowel core was detected (Fig 1).
Skull 2
This cranium contained both upper jaws with all teeth except
second incisor and canine (lost postmortem) and second premo-
lar (lost ante mortem), all on the left side. In the present teeth,
we noticed these fillings: amalgam (on third molar on the left
side, on both premolars, and on first molar on the right) and
cement filling (on the second premolar and on the first and third
molar on the right side). Caries destruction was present on the
third molar on the right side. On the remained premolar on the
left side, a part of artificial metal crown was found. On the same
tooth, on the orthopantomogram, root filling was noticed (Fig 1)
as well destruction of the bone above the root apex, probably
caused by dental root infection.
Skull 3
In both upper jaws, we found second premolar on the right
side with silver dental crown and third molar on the left side
with amalgam filling. First molar on the right side and first and
1286 JOURNAL OF FORENSIC SCIENCES

Silicophosphate cement

Silicophosphate cement
A1, amalgam removed from the first left molar, skull no. 1; A2, amalgam removed from the second right premolar, skull no. 2; A3, amalgam removed from the first right molar, skull no. 2; C1, cement filling
taken from the second left premolar, skull no. 1; C2, cement filling taken from the second right premolar, skull no. 2; L1, dental base taken from the first left molar, skull no. 1 after removing the amalgam A1
Chemical Composition

Zinc phosphate cement

Filling Material
According to

Copper amalgam

Silver amalgam
Silver amalgam
40.8  0.80

7.3  0.20
4.1  0.40
23.2  0.50
(Ca) mg/g
Calcium

0.00
Trace

133  10.00
2.1  0.10

8.6  0.20
(Sn) mg/g
Tin

Trace
0.00
0.00
69  1.00

80  1.00
(Si) mg/g
Silicon

Trace

0.00

0.00
0.00
FIG. 1––Orthopantomograms of skull 1 and skull 2: 1-amalgam, 2-cement, TABLE 1––Chemical analysis of dental fillings.
3-root filling, 4- bone destruction, 5- dowel core, 6-artificial metal crown,

112  4.00

69  5.00
Aluminum
(Al) mg/g
7-caries.

0.00

0.00

0.00
0.00
second molar on the left side had been extracted ante mortem.
All other teeth have been lost postmortem.
Phosphorus

52  4.00
70  5.00

41  2.00
(P) mg/g

Trace

Chemical Analysis
0.00
Results of chemical analysis of six dental fillings are shown 0.00
in Table 1.
Not quantified
Not quantified

Compounds from amalgam filling (A1) from the first skull

(Ag) mg/g

match the composition of copper amalgam because it contains

Silver

copper (Cu) and mercury (Hg) in sufficient amount and no silver

Trace
Trace
Trace

Trace

(Ag) or tin, which would be found in silver amalgam. Below

this amalgam, we found zinc phosphate cement (L1), and it was
used as dental base. This specimen contains zinc (Zn) and phos-
Not quantified
Not quantified

phorus (P) in weight ratio that match Harvard cement 860 mg

513  21.00
(Hg) mg/g
Mercury

of zinc oxide (ZnO) in 1 g of powder, 560 mg of phosphoric

acid (H3PO4) in 1 g of liquid, ratio 1.5:1(8). Also, it contained a
Trace
Trace

Trace

small amount of copper probably added to gain antiseptic effect

as it was used to be done (9).
Analyzed amalgam fillings from the skull 2 (A2 and A3) con-
1.8  0.10
44.4  0.40
413  5.00

0.56  0.09
0.18  0.03
153  5.00

tained all ingredients in notable amount, which would be found

(Zn) mg/g

in silver amalgams. In comparison with control high-copper

Zinc

amalgam containing 74  1 mg/g of copper, we found signifi-

cantly lower amount of copper (Table 1), which classifies it into
a group of conventional low-copper amalgams.
Cement fillings (C1 and C2) belong to a group of silicate
0.08  0.002
245  3.00
1.91  0.02
1.9  0.10

1.91  0.02
7.03  0.20
(Cu) mg/g

cements. In comparison with standard silicate cement (10), they

Copper

contained higher amount of zinc. This is most likely because they

as it was underneath it.

are a mixture of silicate and phosphate cement—silicophosphate

cement.
Case Filing

Discussion
Skull 1

Skull 2

The question addressed to anthropologists was whether the

A1

A2
A3
C1

C2
L1

human remains belonged to the victims from WWII, as it was

 ZELIC ET AL. . DENTAL RESTORATIONS AND TIME SINCE DEATH 1287

claimed in the testimony of the witness, or belonged to more

recent case requiring court trial. The place of burial, number of
individuals, estimated sex, type of injuries, and the state of pres-
ervation support the testimony that victims were soldiers exe-
cuted at the time of WWII. There were no additional data, such
as personal belongings, which would have helped in a more pre-
cise identification of the victims. Therefore, in this study, we
have concentrated on more detailed analysis of dental restora-
tions in an attempt to better estimate the time since death and
potentially to assume the place where these restorations could be
carried out.
During the historical development of dental materials, differ-
ent types of dental fillings were used. Differences in the compo-
sition of these materials are substantial, and this gives
investigators the opportunity to determine with great certainty
the type of material only by its chemical analysis.
The first filling we analyzed, found on the upper molar, was
made almost exclusively of mercury and copper, which suggests
that the used material was a copper amalgam. The finding of
green discoloration of bone surrounding tooth indicated, even at
first sight, that some abandoned type of restoration was in ques-
tion. Copper amalgam is a mixture of mercury and copper (11).
After its invention in 1859, it was used until the end of the 19th
and the beginning of the 20th centuries (11). It had good plastic-
ity, and opposed to silver amalgam that was most commonly
used, it could also be placed in a humid environment where sal-
iva could not be completely removed from the tooth preparation
cavity. This could explain why it was used in our case, because
dryness of tooth cavity of upper molar, prior to application of
filling material, was difficult to achieve. However, shortly after
the appearance of copper amalgam, scientists began to warn
about its side effects. In 1874, in one respectable journal of that FIG. 2––Cross section of first left molar extracted from the skull 1 after
time, Dental Cosmos recommended dentists to stop using it removing part of amalgam and dental base for chemical analysis.
because of its fast corrosion and loosing of metal ions (11). This
was also recommended in 1908 by J. V. Black, one of the most bone destruction above the root. There is no root canal treat-
distinguishing scientists in dentistry (11). Nevertheless, this ment, which would prevent the dentogenic infection and bone
material was still in use during the first three quarters of 20th destruction. This supports our suggestion that in this individual,
century for milk teeth only, because these teeth were not sup- dental work was done in haste.
posed to stay in the mouth for a long time. It was also used in Other two amalgams we analyzed were silver amalgams. The
war times by military dentists on the battlefields because the main components of the silver amalgam are silver, mercury, tin,
application of it was easier, faster, and cheaper than the use of zinc, and copper. However, by analyzing the amount of copper,
other dental filling materials available at the time (12). we could classify silver amalgam in high-copper or low-copper
On the same tooth, we found an unusually large amount of group. Namely, until the 1960s, low-copper (conventional) amal-
dental base (Fig 2). The base is used under the amalgam filling gams were in use, and in 1963, a new type of high-copper (non-
in the tooth preparation cavity and above the pulp cavity to pro- conventional) amalgam was invented (14). The main difference
tect intact pulp. In our case, the base was placed directly in the was in the quantity of copper. The conventional amalgam con-
pulp cavity, which means that the pulp cavity was open and that tained about 6% copper and the new type about 20% (14–16).
the pulp was damaged. In case of pulp damage, the strict proce- We quantified Cu in both the forensic and control sample, and
dure of its removal and root canal treatment must be performed. chemical analyses showed that the control sample contained sub-
Otherwise, an infection will develop and the tooth will be lost stantially more copper, while the forensic case corresponded to
(Fig 2). Restoration with copper amalgam and a lack of neces- the conventional amalgam.
sary root canal treatment suggest that the tooth was restored tem- The final two analyzed fillings belonged to a group of silicate
porarily or in haste. This could be expected in war time when it cements. The amount of all components found in these speci-
was only important to remove pain and to achieve more or less mens corresponded to the silicate cements (De Tray, Ash)
tooth function without taking into account the long-term progno- (10,17) except the amount of zinc. During the first half of 20th
sis. Chemical analysis of the base showed the ratio of Zn and P, century, silicates were widely used for front teeth, because of its
which corresponds to zinc phosphate cement. Also, we noticed tooth-like color, which meets the esthetic requirements. They
that the Zn/P ratio exactly corresponds to Harvard dental cement, were not convenient for back teeth because of their fragility
one of the oldest zinc phosphate cements (8). These zinc phos- (16). However, by adding the powder of zinc phosphate cement
phate cements were invented in 1860, in dentistry were intro- into silicate cement, it gains strength (losing the esthetic advanta-
duced in 1879, and are still in use (13). ges) and could be applied for back teeth as well (16). This was
One more tooth in this particular skull, second left premolar most likely performed in our case of two premolars, and it
(Fig 1), was incorrectly taken care of, which is revealed by large explains the higher concentration of Zn found by chemical
1288 JOURNAL OF FORENSIC SCIENCES
analysis. Silicate cements were invented in 1908 and were first
used in United States (13). Silicate and silicophosphate cements
were semipermanent fillings, so they had to be replaced after
4–5 years (13). Their use began to decrease with the invention
of acrylates in 1947, and when composites were introduced to
dentistry in 1962, the use of both silicate cements and acrylates
diminished dramatically and finally came to an end within few
years (13). Therefore, the use of silicophosphate cements in our
cases suggests that dental restorations were most likely carried
out before 1960s.
One of the leading countries in dentistry in the past century
was Germany. For example, in the 1930s, about 10.000 dentists
worked in Germany (18). The first academic dental clinic was
established in 1902, and by the 1930s, numerous similar clinics
were founded around the country (18). Historic data cited that in
1931, an impressive 6 million patients were treated by 528 den-
tal surgeons in 126 academic dental clinics in Germany (18). On
the other hand, in countries like Serbia in this period, dental ser-
vice was underdeveloped. Unqualified practitioners performed
dental service (predominately extractions), and the Faculty of
Dentistry was not founded until 1948 (12). One obvious example
of the difference between these two countries is the organization
of the dental service in WWII. While the German army had
6.000 dentists enlisted during the first months of conflict (18),
Serbian dentist service was not established until late in the 1944
(12). A report by the medical department of Supreme Headquar-
ter about dental service from December 1944 shows that there
were only three dental offices in Belgrade for the army with 22
dentists employed (12). In addition, the commandants would not
permit long-term absence to the soldiers, so the dentists mostly
performed extractions of teeth (12).
Among five skulls analyzed in the study, two demonstrated
considerable dental restorations, and this could probably be
assumed even for the third skull as the remaining teeth also had
dental restorations. Keeping in mind the historic period, this sug-
gests that individuals, although found in Serbia, most likely
came from a country with well-developed dental service. It also
contributes to the assumption that the remains belonged to Ger-
man soldiers befallen in Serbia during the war.
Based on chemical results and historical facts, we can assume
with high probability that the individuals we have studied, lived
after the 1910s and before the 1960s, had their dental work done
in a country with well-developed dental service and had some of
their dental fillings done in haste.
In conclusion, place of recovery, anthropological characteris-
tics (number of individuals, sex, age), pathological findings
(gunshot holes), and state of preservation of the remains (light,
dry, multicolored, degreased bones) resemble to the testimony of
eyewitness that skeletons belong to group of soldiers killed by
the end of World War II. The results of chemical analysis of
dental restorations were complementary source of information,
indicating more precisely time since death and probable region
View publication stats
where restorations were made. Therefore, chemical analyses of
dental fillings can be useful to distinguish between bones that
are too old to be of forensic interest and the remains that are rel-
evant to legal investigations.
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Additional information and reprint requests:
Marija Djuric, M.D., Ph.D.
Professor
Laboratory for Anthropology
Institute of Anatomy
School of Medicine
University of Belgrade
4/2 Dr Subotica
11000 Belgrade
Serbia
E-mail: marijadjuric5@gmail.com