Journal of Dentistry (2004) 32, 41–46

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Effect of Er:YAG laser and organic matrix

on porosity changes in human enamel
Deng Yinga, G.K. Chuahb, Chin-Ying S. Hsua,*

a
Department of Preventive Dentistry, Faculty of Dentistry, National University of Singapore,
5 Lower Kent Ridge Road, Singapore 119074, Singapore
b
Department of Chemistry, Faculty of Science, National University of Singapore, Singapore, Singapore

Received 29 October 2002; revised 25 June 2003; accepted 5 August 2003

KEYWORDS Summary Objectives. Cariostatic effects of lasers have been well documented in the
Er:YAG laser; Porosity; past few decades; however, the mechanisms remain unclear. Our previous study
Organic materials; revealed the statistically significant effect of organic matrix (OM) in the laser-induced
Human enamel caries prevention. The aim of the present study is to further investigate the role of
organic matters in the laser-induced porosity changes.
Methods. Enamel powder from five sound molars was randomly divided into two
samples. One sample was treated with , 10% NaClO solution to remove OM and the
other was left untreated. Both groups were then subjected to irradiation by an Er:YAG
laser. The samples were characterized by thermogravimetric analysis (TGA) and N2
physico-adsorption.
Results. TGA results confirmed that the NaClO treatment had at least partially
removed the OM in enamel powder. The surface area and pore volume of normal
enamel powder decreased significantly after laser treatment. In contrast, in the
NaClO-treated enamel powder, laser treatment did not significantly decrease the
surface area and the pore volume remained almost unchanged. The substantial
difference between the normal enamel powder and the NaClO-treated enamel powder
after laser treatment confirmed the laser-induced blocking of OM in the micro-
diffusion pathway in enamel.
Conclusion. The findings in this study seemed to substantiate the ‘organic blocking
theory’ as one of the mechanisms in the laser-induced caries prevention.
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Introduction out of the enamel microstructures.1 – 3 The diffusion

Caries formation has been shown to involve two
major processes: (1) the dissolution of enamel
apatite crystals, and (2) diffusion of ions, such as
calcium, phosphate, and hydrogen ions, into and
*Corresponding author. Tel.: þ 65-779-5555x1658; fax: þ 65-
773-2602.
E-mail address: pndhsus@nus.edu.sg
process was regarded as more important in the
control of caries formation.4
The physical and chemical composition of
enamel may dictate the efficiency of diffusion.
Enamel is a complex porous solid, composed of long
thin crystals of hydroxyapatite-like mineral sur-
rounded by a matrix of water and organic material.
The crystal arrangement of enamel gives rise to
two main categories of pores corresponding to

0300-5712/$ - see front matter q 2003 Elsevier Ltd. All rights reserved.
doi:10.1016/S0300-5712(03)00138-6
42
the spaces between single crystals in the prism
cores and those between prisms. The larger pores
are presumably related to spaces in the interpris-
matic regions and the smaller pores are probably
associated with the spaces within the prisms
themselves.2,5 Before acid can react and dissolve
the apatite crystal within the enamel, the acid has
to diffuse to the crystal surfaces through the
interprismatic and intercrystalline spaces.4
The mechanism of laser-induced retardation of
enamel demineralization has been studied for
several decades. While the high energy laser-
induced melting and fusion of enamel hydroxyapa-
tite (HA) crystals may be effective in inhibition of
enamel demineralization; however, the high tem-
perature (above 1000 8C) resulting from high-energy
laser irradiation may cause a temperature rise
potentially harmful to the underlying dentin
and/or pulp. A recent study has shown that low
energy laser is effective in inhibition of enamel
demineralization, where the temperature induced
by laser was estimated to be lower than 400 8C.6 It
was proposed as the ‘organic blocking’ theory that
the partial denaturation of organic matrix (OM) may
block the diffusion pathway in enamel, resulting in a
retardation of enamel demineralization. The block-
ing of diffusion pathway may affect the porosity and
micro-surface area of enamel.
The purpose of this study is to further test the
‘organic blocking theory’ by comparing the laser-
induced porosity change in enamel powder with and
without prior removal of organic matrix.
Materials and methods
Sample preparation
Five sound teeth (two premolars and three molars)
were selected, cleaned, and dried in air. The
enamel parts of the five teeth were then cut by an
Alloy grinder (DEMCO, Dental Maintenance Co., Inc.
Bonsall, CA, USA), micro-tomed, mixed together,
and then ground to powder at 0 8C. The enamel
powder was filtered through a 140-mesh sieve,
collected, and stored in a desiccator at room
temperature under vacuum for 2 days. Thereafter,
the powder was randomly divided into two samples.
One sample was left untreated and labeled as
‘normal’ enamel. Another sample, 207.6 mg, was
treated with a 150 ml commercial , 10% NaClO
solution (Fluka 71696) for 4 h at room temperature
with mild stirring (300 rpm). The powder was then
filtered off, washed intensively with distilled water,
dehydrated five times with absolute alcohol for
D. Ying et al.
1 min each, dried in desiccator under vacuum for 3
days and thus labeled as ‘NaClO-treated’ enamel.
Laser treatment
Both the normal enamel powder and the NaClO-
treated enamel powder were irradiated by an
Er:YAG laser (KEYe, KAVO, Germany) with an
energy density of 6.25 J/cm2, 4 Hz, and a 2-mm
beam spot and 5 pulses per spot. The laser was
consecutively applied over an area of 1.57 cm2 of
the evenly spread powder. The irradiated samples
were labeled as ‘laser-treated’ enamel and ‘NaClO
and laser-treated’ enamel, respectively.
Characterization
The amount of organic materials in the enamel was
determined from the weight loss by thermogravi-
metric analysis (TGA). The sample was heated at a
ramp of 10 8C/min to 700 8C with an airflow of
70 ml/min in Dupont SDT 2920 thermal analyzer
(Dupont, USA). Multipoint BET surface area of
sample was obtained from N2 adsorption measure-
ment at 2 196 8C with a NOVA 2000 porosimeter
(Quantachrome, USA), and the pore size distri-
bution was calculated using the Barrett – Joyner–
Halenda (BJH) method. About 0.15 g of sample was
used in the measurement.
Establishment of systemic error of porosity
measurement and statistic analysis
For evaluation of repeatability of porosity measure-
ment, another 10 sound molars were selected,
cleaned, and dried in air. No carious nor defective
areas were found using the stereomicroscope. The
enamel parts of the 10 teeth were then cut, ground,
sieved and dried as mentioned in ‘Sample prep-
aration’. Two samples, sample A and B with 200 mg
each, were randomly selected from the dried
enamel powder and the porosity was measured.
The measurement was repeated five times for each
sample. Between measurements, the two samples
were stored in a desiccator at room temperature
under vacuum for 2 days.
The standard deviation of the porosity measure-
ment was calculated from the five-time repeated
data set. One-sample t-test was then used to
evaluate the statistical significance of the changes
of porosity and surface area after laser treatments.
A usual alpha level of 0.05 was chosen to determine
the statistical significance of the results.
Effect of Er:YAG laser and organic matrix on porosity changes in human enamel
Results
Thermogravimetric analysis
In TGA, the samples showed a small initial weight
loss peak below 50 8C, another peak between 50 and
200 8C, and a broad peak between 200 and 600 8C
(Fig. 1). The percentage weight loss at the different
temperature ranges is shown in Table 1. The weight
loss between room temperature to 200 8C was 1.05%
for the normal enamel and 1.15% for the NaClO-
treated enamel powder. From 200 to 600 8C, it was
2.05% for the normal enamel powder and 1.95% for
the NaClO-treated enamel powder.
Porosity
From the statistical analysis of the five repeated
measurements of the enamel porosity, the standard
deviation of the surface area in group A and B is 0.42
and 0.33 m2/g, respectively. The standard devi-
ation for pore volume in group A and B is 0.0021,
0.0023 cm3/g, respectively.
The nitrogen adsorption/desorption isotherms
revealed that the enamel powders are essentially of
Figure 1 TGA – DTA curves of enamel powder: (a)
normal (b) NaClO-treated.
43
Table 1 The percentage weight losses of enamel powder
measured from TGA curves.
Temperature R.T. 2 200 8C 200–600 8C
Weight loss of 1.05 2.05
normal enamel
powder (%)
Weight loss of 1.15 1.95
NaClO-treated enamel powder (%)
low porosity. The hysteresis loops were character-
istic of slit-like pores (Fig. 2). The pore size
distribution of all samples showed two peaks with
a narrow one around 40 Å in diameter and a broad
one from 50 to 140 Å (Fig. 3). Treatment with NaClO
did not change the pore volume; however, the
surface area increased 15% ðP , 0:001Þ (Table 2).
After laser treatment, the total pore volume of
the normal enamel sample decreased significantly,
from 0.024 to 0.020 cm3/g ðP ¼ 0:023Þ; and the
surface area decreased significantly, from 6.83 to
6.02 m2/g ðP ¼ 0:027Þ: Pores with diameters around
40 Å, in particular, were significantly reduced after
laser treatment (Fig. 3(a)). In contrast to the
normal enamel sample, laser irradiation did not
result in a significant decrease of the pore volume in
the NaClO-treated sample. Although the surface
area was reduced by about 5%, this laser-induced
reduction is only about half of that in the normal
enamel sample and did not reach the statistically
significant level ðP ¼ 0:11Þ:
Discussions
TGA confirmed the changes of water sorption and
organic matter after NaClO treatment. The weight
loss of enamel between 0 and 200 8C can be
assigned to the removal of adsorbed or ‘loosely
bound’ water.7 The NaClO-treated sample had a
slightly higher weight loss in this temperature
range. This may be due to the increase in the
total surface area after NaClO treatment so that the
adsorption of water molecules is increased. A
similar result has been shown in a previous study
on bovine dentine.8 In the temperature range of
200 –600 8C, the weight loss is mainly due to the
decomposition of organic matter, crystalline water,
and part of carbonate compounds.7,9 – 11 Enamel
proteins were reported to decompose from 280 8C
onwards, with the maximum rate at 340 8C.12 The
NaClO-treated sample suffered a smaller weight
loss of 1.95% as compared to the untreated sample,
2.05%. As NaClO treatment does not change the
content of inorganic components, this difference of
44
Figure 2 Adsorption/desorption isotherms of enamel powder: (a) normal (b) NaClO-treated (c) laser-treated (d)
NaClO and laser-treated.
0.1% is most probably due to the loss of organic
matters. The total organic matters in enamel is
reported to be around 0.3 – 1 wt%.13 – 16 The smaller
value measured in this study indicates that the
organic matter, especially those located in the
deeper layers of sound enamel powder, was not
totally removed by the NaClO treatment.
The enamel sample was found to possess pores
with diameters from 30 to 140 Å. This is in
agreement with previous reports. Orams et al.17
showed that pore diameter of enamel was within
the range 10 – 100 Å using the electron microscope.
Zahradnik and Moreno18 obtained a value of 32 Å for
molars using water vapour sorption isotherm at
25 8C. The specific surface area of 6.83 m2/g,
measured in this study, for the normal enamel
powder was considerably higher than the value of
0.3 – 0.4 m2/g for ground enamel, measured using
krypton adsorption.19 However, the surface area of
3.4 – 4.5 m2/g for enamel section, enumerated by
the isothermal water vapour sorption, was closer to
the result in this study.18 The difference in the
reported surface area may be due to various
accuracy levels of different equipments and
D. Ying et al.
methods, and the different hydroxyapatite crystal-
line organization and water content of the samples.
The increase in adsorbed water may lead to lower
surface area.
After laser treatment, the normal enamel
showed a decrease in both the surface area and
the pore volume. This may be due to fusion and
sealing of the enamel micropores as a result of
laser-induced blocking of the organic matrix and/or
melting of the hydroxyapatite crystals.6,20 – 21 Since
no new phases were detected by X-ray diffraction in
our pilot study, the fusion and melting of hydro-
xyapatite crystals can be ruled out and the effect of
laser irradiation seems to be limited to a partial
blocking of the pores with concomitant decrease in
the surface area. This result is important as surface
area is a key factor for many chemical reactions,
including enamel demineralization. This finding
would also help explaining why organic matter
may impart a protective effect on enamel deminer-
alization. 22,23 In contrast, the NaClO-treated
enamel, where there was partial removal of organic
matters, did not show any significant reduction in
its surface area nor pore volume after
Effect of Er:YAG laser and organic matrix on porosity changes in human enamel 45

Conclusions

The organic matters, causing a statistically signifi-

cant decrease in the pore volume and surface area
in enamel after laser irradiation, may be one of the
key players in the laser-induced blocking of diffu-
sion pathway and the subsequent prevention of
enamel demineralization.

Acknowledgements

This project is financially supported by the research

grant ‘R-222-000-004-112’, of the National Univer-
sity of Singapore.

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