International Journal of Dental Health Concerns (2015), 1, 1-4
Doi: 10.15713/ins.ijdhc.5
ORIGINAL ARTICLE
Effect of Acid and Fluoride Release from Four Glass Ionomer Cements on
Streptococcus mutans: An In Vitro Study
Simpy Amit Mahuli1, Amit Vasant Mahuli1, R. Subramaniam2, Mahesh Hiregoudar3,
G. M. Prashant4, G. N. Chandu4
Department of Public Health Dentistry, NIMS
Dental College & Hospital, Jaipur, Rajasthan,
India.
2
Department of Public Health Dentistry,
Indira Gandhi Institute of Dental Sciences,
Kothamangalam, Kerala, India.
3
Department of Public Health Dentistry, Coorg
Institute of Dental Sciences, Virajpet, Karnataka,
India.
4
Department of Public Health Dentistry, College
of Dental Sciences, Davangere, Karnataka, India.
Correspondence: Dr. Simpy Amit Mahuli,
Department of Public Health Dentistry, NIMS
Dental College & Hospital, Jaipur - 303121,
Rajasthan, India. Phone: +91-8107965195,
Email: [email protected]
How to Cite:
Mahuli SA, Mahuli AV, Subramaniam R,
Hiregoudar M, Prashant GM, Chandu GN.
Effect of acid and fluoride release from four
glass ionomer cements on Streptococcus mutans:
An in vitro study. Int J Dent Health Concern
2015;1:1-4.
Received: 27.12.2014
Accepted: 17.02.2015
1
ABSTRACT
Objectives: The objective was to evaluate fluoride and acid release from four glass
ionomer cements (GICs) over time and to evaluate and compare their effect on
Streptococcus mutans over time. Materials and Methods: The fluoride and acid release
from the prepared samples (GICs, Miracle Mix, Fuji IX, d-tech®, Ketac Molar) were
measured daily using digital fluoride meter and pH meter, respectively, after placing them
in deionized distilled water for 15 days. The water was replaced daily. The samples were
aged by placing them in distilled water for 1, 3, 5, and 7 days. The antimicrobial activity
was assessed by measuring the zone of inhibition around the samples (fresh and aged)
placed on the bacterial cultures after 48 h of incubation. Mean and standard deviation
were calculated. Pearson’s correlation coefficient was used. P < 0.05 was considered
statistically significant. Results: The antimicrobial activity of the four GICs was greatest
immediately after the cements were mixed. The greatest fluoride and acid release was from
d-tech. Conclusion: Both acid and fluoride release that contributed to the antibacterial
activity of the Glass ionomer cements reduced significantly as the cement aged.
Key words: Acid release, antibacterial activity, fluoride release, glass ionomer cement.
INTRODUCTION
Glass Ionomer Restorative Cement (d-tech® - dtechasia, Pune)
and Ketac Molar were used as the study materials. Pure culture of
S. mutans was obtained from Microbial Type Culture Collection
(MTCC), Chandigarh, India. Ethical clearance was obtained for
Institutional Review Board.
Atraumatic restorative treatment (ART) has been developed for
the treatment of caries in parts of the world with limited resources
through the use of dental hand instruments and glass ionomer
cement (GIC). However, hand instruments do not remove
carious dentin as effectively as rotary burs. Thus, cavities treated
by ART may have residual bacteria.[1] Hence, it is highly desirable
for GIC to possess antibacterial activity. The antibacterial activity
of GICs has been attributed to the longevity of the fluoride
release. Recent research has also shown that the antibacterial
property of GICs also depends on the acid release from the GIC.
However, both fluoride and acid release decrease progressively
as the cement ages.[2] Hence, this study was conducted with the
following objectives: (i) To determine the fluoride and acid
release from GICs over time and (ii) to evaluate and compare the
relationship of the fluoride and acid release from the four GICs
with their activity against Streptococcus mutans over time.
Evaluation of Fluoride and Acid Release
The cements were mixed according to manufacturer’s instructions
and placed in molds to create samples measuring 5 mm diameter
and 3 mm height. The samples were placed in 50 ml of deionized
distilled water and left undisturbed for 24 h. After 24 h, the
fluoride content and pH in each water sample were determined.
The fluoride content was measured using ion electrode method
with total ionic strength adjusted buffer (Orion 9409) and the
pH was measured using the digital pH meter. The procedure was
repeated daily for the next 15 days.
Evaluation of Activity against S. mutans
The microorganism S. mutans was used. Strains were grown in
the brain heart infusion broth and incubated anaerobically for 18 h
at 37°C. Strains were then grown and sub-cultured on blood agar.
Prepared samples were placed in 50 ml of deionized distilled
water and were left undisturbed for 1, 3, 5, 7, 9, and 11 days. The
MATERIALS AND METHODS
An In vitro study was carried out. The study was conducted over a
period of 3 months. Four GICs, GIC (Fuji IX GP), Miracle Mix,
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Effect of Acid and Fluoride Release from Four GICs on Streptococcus mutans … Mahuli, et al.
the toxicity of the material against the bacteria tested and the
diffusibility of the material across the culture medium used. This
method allows measurement of the inhibitory activity against
bacteria colonizing a surface, which imparts clinical relevance
if in vitro data are being extrapolated to oral bacteria colonizing
around or on restorative materials.[3]
The results of this study show that the antibacterial property of
GICs decreased as the acid release from the material decreased.
This finding is consistent with a similar study conducted by
Fischman and Tinanoff in USA in 1994.[2] Fraga et al. conducted
a study in Brazil in 1996 to test the antibacterial effects of
photocured GICs during setting and found a similar relation
between the antibacterial activity and acid release from GICs.[3]
Studies conducted by Marczuk-Kolada et al. in Poland in 2006
and Berg et al. in USA in 1988 to evaluate the fluoride release
from GICs and its effect on the growth of S. mutans around the
material, show results similar to this study.[4,5]
Studies conducted by Tobias et al. in the UK in 1985 and
Seppä et al. in Norway in 1993 showed that the maximum
antibacterial activity was exhibited by freshly mixed samples
of restorative materials and there was drastic reduction in the
antibacterial activity after day 1. These results confirm the
findings of the present study.[6,7]
Yap et al. in Singapore in 1999 showed that the amount of
fluoride release from GIC decreased drastically after 1-week,
which is consistent with the results of the present study.[8] In
another study conducted in Japan by Nakajo et al., indicate that
GIC fillings in the oral cavity, reduce bacterial acid production,
and the subsequent bacterial growth may decrease, creating the
cario-static environment.[9]
In a study conducted by Chau et al., results suggest that the
anti-cariogenic biofilm activity of GICs is closely correlated with
their fluoride release rate during biofilm formation.[10,11]
The antimicrobial activity of the GICs could be related to
the synergistic action of acid and fluoride release of the cement
water was replaced every 24 h. Samples of each of the four GICs,
either fresh or aged, were placed on the inoculated agar plates.
Inhibition zones were measured after 48 h of incubation at 37°C.
Statistical Analysis
Mean and standard deviation of the fluoride release, the pH and
the zone of inhibition of the different samples were calculated.
Pearson’s correlation coefficient was used for correlating the
fluoride and acid release from the four GICs with the change in
their activity against S. mutans over time. P < 0.05 was considered
statistically significant. The analysis of data was done by SPSS
version 16.0 (Statistical Package Software).
RESULTS
The antimicrobial activity of the four GICs was greatest
immediately after the cements were mixed. The fluoride release
from all the four cements was maximum in the first 24 h after
which it reduced drastically to similar levels [Figure 1]. The
greatest fluoride release was from d-tech, followed by Miracle
Mix, Ketac Molar, and Fuji IX. Similar results were shown by all
the samples with respect to the acid release. The greatest acid
release was from d-tech followed by Miracle Mix and Fuji IX,
and the least was from Ketac Molar [Figure 2]. The antibacterial
property of all the test materials decreased as the fluoride release
from the materials decreased [Figure 3]. The antibacterial
property of the test materials reduced as the acid release from the
test materials reduced [Figure 4].
DISCUSSION
7.4
7.2
Fuji IX
7
Miracle Mix
Ketac Molar
d-Tech
pH
FLUORIDE RELEASE ( in ppm)
The agar diffusion test has been widely used to evaluate the
antibacterial property of dental materials. In this method,
the zones of inhibition provided by the materials depend on
20
19
18
17
16
15
14
13
12
11
10
9
8
7
6
5
4
3
2
1
0
IJDHC
FUJI IX
6.8
MIRACLE MIX
KETAC MOLAR
6.6
d-TECH
6.4
6.2
6
1
2
3
4
5
6
7
8
9
DAYS
10 11 12 13 14 15
Figure 1: Fluoride release over time from the different materials
1
2
3
4
5
6
7
8 9
DAYS
10 11 12 13 14 15
Figure 2: Acid release from the different materials over time
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Effect of Acid and Fluoride Release from Four GICs on Streptococcus mutans … Mahuli, et al.
IJDHC
Figure 3: Correlation between the fluoride release and the antibacterial activity of the different materials
Figure 4: Correlation between the acid release and the antibacterial activity of the different materials
samples, since the bacterial inhibitory effect of fluoride increases
as the pH decreases.
antibacterial activity of GICs reduce significantly as the cement
ages, thereby reducing its antibacterial activity.
CONCLUSIONS
ACKNOWLEDGMENT
The acid release, fluoride release, and the antibacterial activity
were at their peak immediately after mixing the cements. Both
the factors, acid and fluoride release that contribute to the
The authors report no conflicts of interest related to this
study. The authors would like to thank Dr. V.V. Subba Reddy,
Director, College of Dental Sciences, Staff of the Department of
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Effect of Acid and Fluoride Release from Four GICs on Streptococcus mutans … Mahuli, et al.
Oral Pathology and Microbiology, College of Dental Sciences,
Davangere, India, and Staff of the Department of Environmental
Engineering, Bapuji Institute of Engineering and Technology,
Davangere, India, for their help and support in carrying out this
research.
IJDHC
Glass ionomer/silver cermet restorations on interproximal
bacterial growth. J Dent Res 1988;67:1280-336.
6. Tobias RS, Browne RM, Wilson CA. Antibacterial activity of
dental restorative materials. Int Endod J 1985;18:161-71.
7. Seppä L, Forss H, Ogaard B. The effect of fluoride application
on fluoride release and the antibacterial action of glass
ionomers. J Dent Res 1993;72:1310-4.
8. Yap AU, Khor E, Foo SH. Fluoride release and antibacterial
properties of new-generation tooth-colored restoratives.
Oper Dent 1999;24:297-305.
9. Nakajo K, Imazato S, Takahashi Y, Kiba W, Ebisu S,
Takahashi N. Fluoride released from glass-ionomer cement
is responsible to inhibit the acid production of caries-related
oral streptococci. Dent Mater 2009;25:703-8.
10. Chau NP, Pandit S, Cai JN, Lee MH, Jeon JG. Relationship
between fluoride release rate and anti-cariogenic
biofilm activity of glass ionomer cements. Dent Mater
2015;31:e100‑8.
11. Mayanagi G, Igarashi K, Washio J, Domon-Tawaraya H,
Takahashi N. Effect of fluoride-releasing restorative
materials on bacteria-induced pH fall at the bacteria-material
interface: An in vitro model study. J Dent 2014;42:15-20.
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