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Archives of Environmental & Occupational Health, Vol. 64, No.

4, 2009
Copyright C 2009 Heldref Publications
Mercury, Copper, and Zinc
Concentrations in Extracted Human
Teeth
Rita de C assia Villar Brigato; Liovando Marciano Costa, PhD; Marcelo Rocha da
Costa, MD; Neuza Maria Souza Picorelli Assis, DDS; Claudio Hideki Kubo, DDS
ABSTRACT. Amalgam has been used as a lling material for over 150 years. Mercury, copper, and
zinc are present in restoration. The aim of this study was to compare mercury, copper, and zinc
concentrations in extracted human teeth with amalgam restorations and teeth without restorations.
Thirty-two teeth, 15 restored with dental amalgam and 17 without restorations, were chemically
analyzed in an Optima 3300 DV (Perkin Elmer) plasma emission spectrometer. Mercury, copper,
and zinc were found in human teeth regardless of the presence of amalgam restorations. The highest
mercury concentrations were found in the coronary portions of the teeth with amalgam restorations.
Copper concentrations were very high. Zinc concentrations in the teeth without restoration were
lower than those seen in the coronary portion of the teeth with restorations.
KEYWORDS: contamination, heavy metals, human teeth
S
ome heavy metals, such as cadmium (Cd), mercury
(Hg), and lead (Pb), have been linked to human in-
fertility.
1
Other metals, such as copper (Cu) and zinc
(Zn), necessary for human homeostasis, are supplied by the
daily diet.
Although amalgam has been used as a lling material
for over 150 years, one of its components, mercury, a heavy
metal, has long been known for its human toxicity. According
to the World Health Organization,
2
mercury vapor released
from dental amalgams is the greatest source of human ex-
posure to mercury, followed by air, sh, ordinary food, and
drinking water.
Mercury vapors may cause acute intoxication character-
ized by weakness, metallic taste, nausea, vomiting, diar-
rhea, dyspnea, and coughing. Chronic exposure produces
a neurasthenic syndrome along with goiter, gingivitis, and
excessive salivation.
3
Small amounts of mercury found in
Rita de C assia Villar Brigato and Neuza Maria Souza Picorelli Assis are with the Federal University of Juiz de Fora, Restorative
Dentistry, Juiz de Fora, Brazil. Liovando Marciano Costa is with the Universidade Federal de Vicosa, Solos, Vicosa, Brazil. Marcelo Rocha
da Costa is a Family Doctor of Vicosa Municipality, Minas Gerais, Brazil. Claudio Hideki Kubo is with the S ao Paulo State University,
Restorative Dentistry, S ao Jos e dos Campos, Brazil.
several areas of the dental pulp of rats exposed to mercury
vapor.
4
Copper is found in all human tissues and it is an impor-
tant catalyst of hemoglobin synthesis. Because it favors the
connection of collagen bers and bone formation, copper is
important in tissue sustentation.
5
High-copper-content amal-
gam alloys have become the favorite restoration material be-
cause of their improved mechanical properties and corrosion
behavior, and better marginal sealing, as found in clinical
trials comparing them with traditional low-copper-content
alloys.
6
Microscopically moderate to severe pulp reactions
were nd in most hematoxylin-eosinstained monkey teeth
exposed to copper amalgam.
7
In an in vitro study of decidu-
ous teeth, obliteration of the dentine tubules with reduction
of dentin permeability due to copper irritation of the dental
pulp was observed. In vitro studies blame copper release from
restorations for the cytotoxic effect of the copper amalgam.
8
266 Archives of Environmental & Occupational Health
Zinc in amalgam alloys has a main role as an antioxidant,
binding oxygen and reducing oxide formation. Nevertheless,
zinc may lead to abnormal amalgam expansion if the latter
is condensed in the presence of humidity.
9
Nowadays, amalgam alloy is still often used and heavy
metals are present in the restorations. Therefore, we aimed
to compare mercury, copper, and zinc concentrations in ex-
tracted human teeth with amalgam restorations and teeth
without restorations.
MATERIAL AND METHODS
Thirty-two human teeth (26 molars, 3 premolars, 2 in-
cisors, and 1 cuspid) extracted for several reasons were ob-
tained from 24 patients of the Surgical Clinic of the Dental
School of the Federal University of Juiz de Fora, Mi-
nas Gerais, Brazil. Fifteen teeth were lled with amalgam
restorations and 17 teeth without restorations. The research
project was approved by the Ethics Committee of the Uni-
versity Hospital (protocol 184023/2002, group III). The pa-
tients gave their written informed consent. Each patient was
interviewed specically about occupational history (areas of
metal prospection, light bulb industry, pharmaceutical in-
dustry, hospital or dental clinic sterilization services, dentist
assistant), bruxism, and sh intake (including quantity and
frequency). The patients underwent an oral examination and
had their odontograms obtained, including the number of
teeth with amalgam restorations and the number of missing
teeth.
The teeth were kept for 48 hours in metal-free individual
containers (lled with previously analyzed 10% formalin),
and then rinsed with deionized water, dried with paper towel,
and nally kept in individualized plastic bags sealed with
adhesive tape. The samples were individually described as to
the presence or absence of caries, presence and type (classes
I, II, V) of amalgam restoration, and presence of incisal or
occlusal wear. Restorations were removed with a number
06 spherical tungsten-carbide bur under high-speed rotation
with refrigeration. Any material protecting the dentine-pulp
complex was noted and then removed under refrigeration.
The teeth were cut with a double face diamond exible disk
(KG Sorensen reference 7020), mounted in microengine and
straight handpiece, without refrigeration. The coronary por-
tion of each tooth was longitudinally cut in 3 parts, a central
portion with 2 lateral ones being obtained (the latter corre-
sponding to the mesial and distal portions of the tooth). The
root was separated from the crown for further fragmentation
in a green-quartz pestle and mortar device.
Chemical analyses were done by the Soil Department,
Soil Chemistry Division, Federal University of Vicosa,
Minas Gerais, Brazil. Sample readings were obtained with an
Optima 3300 DV (inductively coupled plasma [ICP]; Perkin
Elmer) plasma emission espectrometer. All the glassware
used were initially kept in 2%hydrochloric acid for 24 hours,
then rinsed in distilled-deionized water, and dry-heated to
avoid contamination.
The material for analysis of each tooth consisted of 1
central and 1 lateral portions of the crown, and 2 random
portions of the root. The samples were divided into 4 groups:
group 1 (G1), coronal portion of the teeth with restorations;
group 2 (G2), radicular portion of the teeth with restora-
tion; group 3 (G3), coronal portion of the teeth without
restorations; and group 4 (G4), radicular portion of the teeth
without restoration.
The fragments were weighed (on aluminized paper) on a
Marte AL200Cscale, and put in labeled 75-mLtest tubes, to-
taling 128 samples, with 4 control tubes (to allow for sample
contamination). The fragments ranged from 0.200 to 0.600
g. Concentrations were calculated according to the mass of
each fragment.
Twenty milliliters of a 1:1 nitric acid solution (50% nitric
acid and 50% deionized water) were used to open each sam-
ple. The test tubes were put in a digestor block (40 tubes at
a time) at 150

C (Tecnal TE 007A temperature controller)


for 45 minutes. The samples were then ltered (glass lter
for 25-mL volumetric ask) through Framex quantitative l-
ter paper (11 cmash: 0.00007 g; folded and cut to t the
ltration funnel). The ltered volume was completed with
deionized water to 25 mL. The volumetric asks were man-
ually agitated and the samples transferred to closed asks.
Standard curves for mercury, copper, and zinc were drawn.
Sample readings were used to calculate the concentrations
according to the dilutions, results being expressed in ppm.
RESULTS
Mercury data
Table 1 shows that the coronary portion of the human teeth
with restoration (G1) had statistically signicant higher mer-
cury concentrations. No statistically signicant differences
were observed in G2, G3, and G4 (Tukeys test).
Copper data
The highest copper concentration was detected in the coro-
nary portion of the human teeth with restorations (G1), as
shown in Table 1. There were no differences in copper con-
centration in G1, G3, and G4. There was signicant reduction
in the copper concentration of the samples from the radicular
portion of the teeth with restoration, in comparison with the
coronary portion.
Zinc data
The highest zinc concentration was seen in samples from
the coronary portion of human teeth with restoration (G1),
as shown in Table 1. However, there was no difference in the
zinc concentration in samples from G2, G3, and G4 (Tukeys
test).
COMMENT
The metals studied (Hg, Cu, and Zn) were found in the
dental structure, regardless of the presence of amalgam
Winter 2009, Vol. 64, No. 4 267
Table 1.-Mean and Standard Deviation of the Concentrations of Each Metal
Concentration (ppm)
Mercury Copper Zinc
Restauration Portion M SD n M SD n M SD n
Present Coronary
(G1)
78.88
a
70.19 30 481.63
a,b
178.23 28 452.93
a
395.77 26
Radicular
(G2)
28.07
b
13.25 30 280.41
b
94.94 30 37.41
b
36.18 28
Absent Coronary
(G3)
9.05
b
4.11 34 402.47
a,b
219.8 34 115.27
b
50.32 34
Radicular
(G4)
13.21
b
7.4 34 417.54
a
206.51 30 121.89
b
14.73 34
Note. n =number of samples used for the statistcal analysis. Means followed by similar letters in the same column do not differ statistically (Tukeys
test).
restorations. Age and sex were not considered in this study.
The 19% of the teeth without amalgam restoration were ob-
tained from patients who did not have any amalgam restora-
tion in their mouths, the mercury found must come from
other sources. There are several alternative sources of mer-
cury contamination
6
: metallic mercury (Hg

) in the air and


sh; inorganic mercury (Hg
2+
) in the air, water, and food; and
methylmercury in sh. Eighty-one percent of the analyzed
teeth came from patients who rarely ate sh.
The highest mercury concentrations were seen in the coro-
nary portion of the teeth with amalgam restorations (78.88
70.19 ppm), a nding suggesting that amalgam restorations
may contaminate the tooth with mercury. Mercury from the
restoration is known to penetrate the dental structure.
6
Dentin
underlying amalgam restorations has shown mercury, which
may be responsible for tooth discoloration. Yet the possi-
bility of toxic reactions from these trace amounts found or
sensitivity to the dissolution of mercury salts on the amalgam
surface is but remote.
6
Considering the mercury concentrations in the teeth with-
out restoration (9.05 4.11 ppm), our results were similar
to those found by Haller et al,
10
who reported mercury con-
centrations above 5.9 ppm in human dentin of teeth without
amalgam restorations. Those authors stated that long-term
mercury exposure can be measured by dentin levels.
10
Chewing and bruxism contribute to the wear of amalgam
restorations, thus releasing mercury that can be ingested.
6
Yet, because 75% of the teeth analyzed were from patients
without bruxism and the amalgam restorations were previ-
ously removed, we could not relate bruxism with higher or
lower mercury impregnation in the teeth or restorations.
Mercury and its compounds are deemed responsible for
occupational exposure.
11
However, we could not relate mer-
cury contamination to occupational exposure, as none of our
patients had worked in areas of metal prospection, light bulb
industry, pharmaceutical industry, hospital or dental clinic
sterilization services, or as dentist assistants.
Of the 15 teeth with restorations, only 4 (26.7%) did not
have material protecting the dentine-pulp complex, which
means that 11 (73.3%) had some kind of material protect-
ing the dentine-pulp complex. According to an autometallo-
graphic study, calcium hydroxide cement does not prevent
mercury penetration in decalcied and parafn-embedded
monkey teeth, as mercury was also present in the pulp of
amalgam-restored teeth 1 year after cavity restoration.
12
An electronic probe was used by other authors to study
amalgam-restored human teeth that had been extracted, and
no mercury was found in the dentin.
13,14
Conversely, Nixon
et al,
15
using neutron activation analysis, found mercury in
both healthy erupted teeth (2.61 ppm) and nonerupted teeth
(1.11 ppm). Healthy erupted teeth in close contact with amal-
gam restorations had much higher mercury concentrations,
ranging from 279 to 1600 ppm in the external enamel. The
high mercury concentration was suggested to be due to vapor
contamination during manipulation or mercury ion migration
from amalgam restorations in close contact with the teeth.
15
The different methods used for metal determination may ac-
count for the divergent results reported.
Although no signicant difference in copper concentration
was found, regardless of the portion (coronary/radicular) or
the presence of amalgamrestoration, copper concentration in
the teeth without restoration was very high (402.47 219.80
ppm) in comparison with another study, which found a mean
10 g/g for copper content in healthy teeth of people from
different geological areas of Finland.
16
Copper may have an
important role in bone formation and tooth calcication. The
literature states that copper is commonly found in teeth, but
in a concentration that varies from tooth to tooth.
17
Derise
et al, using atomic absorption spectrophotometry, also found
low copper concentrations in healthy human teeth (8.1
0.03 ppm in the enamel, and 7.3 0.1 ppm in the dentin).
18
In another study, Derise and Ritchey found a copper concen-
tration of 11.7 1.9 ppm in the enamel of healthy teeth from
people aged 25 years or above.
19
Copper was a remarkable
268 Archives of Environmental & Occupational Health
component of tooth.
17
Soremark and Samsahl used gamma-
ray spectrophotometry to study healthy human teeth that had
been extracted, and found low copper concentrations (0.21
0.10 ppm).
20
Although Tvinnereimet al found zinc in deciduous teeth,
21
they could not correlate zinc concentrations with drinking
water contamination or industrial waste. Zinc concentrations
varied according to caries level, tooth type, and radicular
length. Some high zinc levels could not be readily accounted
for, although the authors suggested that such may have been
due to zinc-containing restorations. In this study, zinc con-
centrations in the teeth without restorations was signicantly
lower than that seen in the coronary portion of teeth with
amalgam restorations, pointing to the restorations as a pos-
sible zinc source. As for the zinc concentrations found in the
teeth without restorations (115.27 50.32 g), the values
were similar to those found by Lappalainen and Knuuttila
16
in healthy human teeth: 187 48.0. Derise and Ritchey found
a zinc concentration of 180.3 3.8 ppm in the enamel, and
170.4 83 ppm in the dentin of healthy teeth from patients
aged 25 years or above, with age-dependent statistically sig-
nicant differences.
19
Soremark and Samsahl found a zinc
concentration of 199 78.1 ppm in healthy teeth.
20
Conclusion
We found mercury, copper, and zinc in human teeth that
had been extracted, regardless of the presence of amalgam
restorations.
The highest mercury concentrations were found in the
coronary portions of the teeth with amalgam restorations,
suggesting that the latter may be a source of tooth contami-
nation.
Copper concentrations in the teeth without restorations
were very high when compared with those found by other
authors.
Zinc concentrations in the teeth without restoration were
lower than those seen in the coronary portion of the teeth with
restorations. It is suggested that zinc might come from the
amalgam restorations.
**********
For comments and further information, address correspondence to L. M.
Costa, Universidade Federal de Vicosa, Solos, Campus da UFV s/n, Vicosa,
36571000 Brazil.
E-mail: liovandomc@yahoo.com.br
**********
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Winter 2009, Vol. 64, No. 4 269
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