Dental caries is a multifactorial biofilm disease and frequent sucrose consumption increases its’ risk. Xylitol, a five-carbon polyol sweetener, has advantageous effects on dental health. First by replacing sucrose and increasing salivary flow, but also by specifically antagonizing the growth of Streptococcus mutans and decreasing the amount of plaque. Here, an in vitro caries simulator model was used to evaluate the effect of sucrose and xylitol on adherence to hydroxyapatite (HA) by three S. mutans strains and one Streptococcus sobrinus strain, and their quantities in artificial saliva (AS).

1. Krista Salli1
, Sofia D. Forssten1
, Sampo Lahtinen1
, Arthur C. Ouwehand1
INTRODUCTION
METHODS
RESULTS
DISCUSSION
Xylitol alters sucrose supported S. mutans
biofilm in a caries simulator model
1
DuPont Nutrition & Health
Active Nutrition
Sokeritehtaantie 20
02460 KANTVIK, FINLAND
E-mail: krista.salli@dupont.com
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Dental caries is a multifactorial biofilm disease and frequent sucrose consumption increases its’ risk. Xylitol, a five-carbon polyol sweetener,
has advantageous effects on dental health. First by replacing sucrose and increasing salivary flow, but also by specifically antagonizing the
growth of Streptococcus mutans and decreasing the amount of plaque. Here, an in vitro caries simulator model was used to evaluate the
effect of sucrose and xylitol on adherence to hydroxyapatite (HA) by three S. mutans strains and one Streptococcus sobrinus strain, and their
quantities in artificial saliva (AS).
The simulator model mimics the oral cavity (Fig. 1). It consists of a continuous flow of AS1
, which supports the growth of bacteria similarly to
human whole saliva, constant temperature, mixing, and HA discs as a model teeth and adhesive support for the bacteria2,3
. DNA was extract-
ed from bacteria attached on the HA discs and from AS samples and bacteria quantified using real-time quantitative polymerase chain reac-
tion (qPCR). The statistical differences between groups were determined using one-way ANOVA and Tukey’s multiple comparison test.
This continuous culture biofilm model showed that sucrose promotes and xylitol diminishes bacterial colonization and proliferation in a young
biofilm. These results indicate that xylitol affects the adhesion of S. mutans on the HA. The model is suitable for studying the impact of test
agents on the adhesion of plaque, and is therefore useful as a potential screening tool prior to clinical trials of caries preventive agents. For
caries prevention it is important to find ways to counteract biofilm formation without disturbing commensal oral microbiota.
References:
1. Forssten SD, Björklund M, Ouwehand AC. Streptococcus mutans, caries and simulation models. Nutrients 2010 Mar;2(3):290-8
2. Björklund M, Ouwehand AC, Forssten SD. Improved artificial saliva for studing the cariogenic effects of carbohydrates Curr Microbiol. 2011 Jul;63(1):46-9.
3. Salli KM and Ouwehand AC. The use of in vitro model systems to study dental biofilms associated with caries: a short review. J Oral Microbiol. 2015 Mar 3;7:26149.
Figure 2.
Bacterial quantities from HA discs
A) Streptococcus mutans DSM 20523,
B) Streptococcus sobrinus DSM 20381 and clinical S.
mutans isolates C) 2366 and D) 117. *p < 0.05 in comparison to AS,
#p < 0.05 in comparison to 1% sucrose.
Figure 1.
Schematic diagram of the dental simulator (reproduced
from3
) 1. AS 2. Pump 3. Simulation vessel 4. Sample
collection during the simulation 5. Outlet pump 6. Waste
+4 °C +37 °C
2
1
5
6
3 4
A.
B)
D)C)
artificialsaliva
1
%
sucrose
2
%
xylitol
2
%
xylitol+
1
%
sucrose
0
2
4
6
8
Meanlog10S.mutans/HAdisc
artificialsaliva
1
%
sucrose
2
%
xylitol
2
%
xylitol+
1
%
sucrose
0
2
4
6
8
Meanlog10Streptococcus/HAdisc
artificialsaliva
1
%
sucrose
2
%
xylitol
2
%
xylitol+
1
%
sucrose
0
2
4
6
8
Meanlog10S.mutans/HAdisc
artificialsaliva
1
%
sucrose
2
%
xylitol
2
%
xylitol+
1
%
sucrose
0
2
4
6
8
Meanlog10S.mutans/HAdisc
*
*
*
#
* *
*
* *
**
A)
artificialsaliva
1
%
sucrose
2
%
xylitol
2
%
xylitol+
1
%
sucrose
0
2
4
6
8
Meanlog10S.mutans/mlAS
artificialsaliva
1
%
sucrose
2
%
xylitol
2
%
xylitol+
1
%
sucrose
0
2
4
6
8
Meanlog10Streptococcus/mlAS
artificialsaliva
1
%
sucrose
2
%
xylitol
2
%
xylitol+
1
%
sucrose
0
2
4
6
8
Meanlog10S.mutans/mlAS
artificialsaliva
1
%
sucrose
2
%
xylitol
2
%
xylitol+
1
%
sucrose
0
2
4
6
8
Meanlog10S.mutans/mlAS
B)
C) D)
* #
*
*
*
**
*
A)
Figure 2.
Bacterial quantities from AS
A) Streptococcus mutans DSM 20523, B) Streptococcus
sobrinus DSM 20381 and clinical S. mutans isolates C)
2366 and D) 117. *p < 0.05 in comparison to AS, #p < 0.01 in comparison to
1% sucrose.
In comparison to unsupplemented AS, sucrose (1%) increased colonization of the tested bacteria to the HA (p<0.0001) with all studied bacte-
rial strains, while 2% xylitol diminished it in them all (p < 0.05), with the exception of S. mutans clinical isolate 117 (Fig. 2). Xylitol also de-
creased the planktonic bacterial quantities (p<0.01) with the exception of clinical isolate 117 (Fig. 3). The combination of xylitol (2%) and su-
crose (1%) reduced the bacterial quantity in AS (p<0.001) and less colonization of HA was observed (p<0.05) with the S. mutans clinical iso-
late 2366 (Fig. 2C and 3C).