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NovaMin

Tubule occlusion of a Research


NovaMin-containing Report
dentifrice compared to Research Institution:
Recaldent-containing NovaMin Technology Inc.
(February 2006)

dentifrice – a Remin/Demin Principal Investigator:


Anora Burwell, MS
study in vitro.
Abstract: An in vitro study was conducted to determine the ability of two different materials to physically occlude
dentinal tubules (DenShield™ and GC Tooth Mousse dentifrices) when repeatedly challenged with an acidic (demineralizing)
environment. The active ingredients in DenShield™ and GC Tooth Mousse (NovaMin® and RECALDENT™, respectively)
are both presented to consumers as sources of minerals that are naturally found in teeth. Brushing with both products should, in
theory, aid in remineralizing tooth surfaces and occluding exposed tubules during the daily remineralization/demineralization
(remin/demin) cycle that takes place in the mouth.
Root portions of bovine incisors were progressively ground, polished and acid-etched to expose tubules in the dentin
layer. The root dentin samples were randomly assigned to one of three treatment groups: Control (brushed with DI water), GC
Tooth Mousse, or DenShield™ and were subjected to the following protocol every day for a total of 10 days: 1) 30-minute soak
in demineralizing solution, 2) 2-minute brush with the appropriate dentifrice, 3) 6-hour soak in artificial saliva, 4) 30-minute
soak in demineralizing solution, 5) 2-minute brush with the appropriate dentifrice, 6) 15-hour (overnight) soak in artificial
saliva. At the end of the 10-day remin/demin cycle the root dentin samples were prepared for analysis with scanning electron
microscopy (SEM). Quantitative tubule occlusion data were independently obtained by three people who examined each SEM
image and recorded the number of open, partially occluded, and total visible (open + partially occluded) tubules.
Treatment with both dentifrices significantly reduced the number of open tubules compared to the control group. The
average number of open tubules visible in an SEM image (60 x 44µm) dropped from 48 in the control group to 28 in GC Tooth
Mousse-treated samples and less than 1 in samples treated with DenShield™. Samples treated with GC Tooth Mousse had a
large proportion of partially occluded tubules and very few fully occluded tubules present on the dentin surface. The number of
open, partially occluded, and total visible tubules seen in treated with DenShield™ were significantly less than in the GC Tooth
Mousse-treated samples.

The current study has demonstrated that the NovaMin® particles in the DenShield™ dentifrice are superior to the
RECALDENT™ in GC Tooth Mousse in depositing a mineral layer on dentin that will occlude dentinal tubules and resist
Introduction designed to test this theory using an in flowing on the grinding wheel during
vitro model of this daily remin- all grinding and polishing steps. The
Recently, a 10-day remineralization/ eralization/demineralization cycle. samples were ultrasonicated three
demineralization (‘remin/demin’) times in fresh DI water for 15 minutes
study was conducted with the goal of Materials and Methods each. The smear layer was removed
comparing the longer-term remin- by soaking the samples in a 40% o-
eralization benefits of DenShield™ Preparation of Bovine Root Dentin phosphoric acid solution for 15
toothpaste with those of GC Tooth Samples minutes. The cleaning procedure was
Mousse. The active ingredients in The root portion of 15 then repeated by ultrasonicating the
DenShield™ and GC Tooth Mousse bovine incisors were separated from samples an additional three times in
(NovaMin® and RECALDENT™, the crown portion at the cementum- DI water for 15 minutes each.
respectively) are both presented to enamel junction (CEJ) using a Prepared samples were refrigerated
consumers as sources of minerals that diamond-embedded blade and a low- overnight in TRIS buffer before
are naturally found in teeth. Brushing speed saw. The root portions of the testing was performed.
with both products should, in theory, teeth were progressively ground at
aid in remineralizing tooth surfaces low speeds using 320, 600, and 1200 Preparation of Remineralizing
during the daily remineralization/ grit silicon carbide abrasive papers to Solution
demineralization cycle that takes expose and polish the dentin layer.
place in the mouth. This study was Fresh deionized (DI) water was kept
Artificial saliva (remineralizing sol- 15:30-16:00: All teeth soaked in parametric ANOVA on Ranks test
ution) was prepared to have the demineralizing solution at 37°C was used to detect significance.
following composition: When significance was found, pair-
16:00-17:00: All teeth removed from wise comparisons were made using
2.200g/L Gastric Mucin demineralizing solution, brushed the nonparametric Student-Newman-
0.381g/L NaCl using a soft-bristled toothbrush with Keuls test (significant if p<0.05).
0.213g/L CaCl2•2H2O the appropriate dentifrice, and gently Data are reported as mean ± standard
0.738g/L K2HPO4•3H2O rinsed with deionized (DI) water. error of the mean.
1.114g/L KCl
17:00-08:00: All teeth soaked in Results
Each ingredient was added separately artificial saliva at 37°C Figures 1 – 3 (following page) contain
to stirring DI water and was allowed representative SEM images of teeth
to dissolve completely before the next from each of the three treatment
ingredient was added. The solution groups. It is clear that the polished,
was heated to 37°C and the pH was SEM Analysis acid-etched bovine dentin samples
adjusted to 7.00 using 85% lactic At the end of the 10-day study, the contain open tubules (Figs. 1a and
acid. teeth were dried at 37°C and mounted 1b). All five control samples were
for analysis using scanning electron very uniform in appearance and in the
Preparation of Demineralizing microscopy (SEM). Three teeth from number of tubules present.
Solution each treatment group were mounted
Demineralizing solution was prepared on carbon mounts and coated with Figures 2a and 2b show the samples
to have the following composition: carbon coating and the remaining two from the GC Tooth Mousse treatment
teeth from each treatment group were group after 10 days of remin/demin
2.2mM CaCl2•2H2O mounted on aluminum mounts and cycling. A film can be seen on the
2.2mM NaH2PO4•7H2O coated with a gold/palladium coating. dentin and there is significantly more
0.05M Lactic Acid SEM images were obtained at 500X fibrous-appearing material on the
0.5ppm Fluoride and 2000X from three randomly surface compared to the control
chosen spots on each tooth surface. samples. There is also evidence of
Each ingredient was added separately some tubule occlusion (top and lower
to stirring DI water and was allowed Quantitative Evaluation of Tubule right in Fig. 2a and lower portion of
to dissolve completely before the next Occlusion Fig. 2b).
ingredient was added. The solution Tubule counting was performed on all
was heated to 37°C and the pH was SEM images that were obtained at Figures 3a and 3b show the
adjusted to 4.52 using 50% NaOH 2000X (15 images per treatment DenShield™-treated dentin samples
solution. group). Three people separately after 10 days of remin/demin cycling.
examined each image and counted the Most of the exposed dentin tubules
Remineralization/Demineralization number of open tubules and partially have been occluded and there is
Study Protocol occluded tubules using previously significant build-up of a reaction layer
The 15 root dentin samples were agreed-upon criteria. Tubules that on the surface of the dentin.
randomly assigned to one of three could not be fully visualized on the
treatment groups: 1) Control (brushed SEM images were not counted.
with DI water), 2) GC Tooth Mousse,
or 3) DenShield™ and were subjected For each SEM image the tubule
to the following protocol every day counts from the three people were
for a total of 10 days: averaged together to obtain the
number of open, partially occluded,
08:00-08:30: All teeth soaked in and total visible (open + partially
demineralizing solution at 37°C occluded) tubules (n=3). For each
treatment group, the data from the 15
08:30-09:30: All teeth removed from SEM images were averaged together
demineralizing solution, brushed to obtain the average number of open,
using a soft-bristled toothbrush with partially occluded, and total visible
the appropriate dentifrice, and gently tubules (n=15).
rinsed with deionized (DI) water
Data were statistically analyzed using
09:30-15:30: All teeth soaked in SigmaPlot software (v9.01, Systat
artificial saliva at 37°C Software, Inc.). Data did not have a
normal distribution so the non-
Results

Figures 1a and 1b: SEM Images of Control Teeth Brushed with DI Water (2000X)

Figures 2a and 2b: SEM Images of Teeth Brushed with GC Tooth Mousse (2000X)

Figures 3a and 3b: SEM Images of Teeth Brushed with DenShield™ (2000X)
GC Tooth Mousse DenShield

* Tubule count differs significantly from Control (P<0.001)


# Tubule count differs significantly from GC Tooth Mousse (p<0.001)

Figure 4: Quantitative Tubule Occlusion Data


Figure 4 contains the results of the clearly demonstrate that the reactivity
quantitative evaluation of tubule of the NovaMin® particles in the The SEM and tubule occlusion data
occlusion after 10 days of DenShield™ dentifrice occluded the presented here clearly demonstrate the
remin/demin cycling. The samples vast majority of dentin tubules and difference in performance of the
from the GC Tooth Mousse and was much more effective than the surface layers deposited by the two
DenShield™ treatment groups had competitive GC Tooth Mousse. dentifrices. The GC Tooth Mousse
significantly less total visible and leaves a thin film on the structural
open tubules than the control samples. While a definitive explanation for the dentin after 10 days, but there is very
There were more partially occluded observed differences must await little occlusion of the dentin tubules.
tubules visible on the GC Tooth further detailed surface analysis, it is In comparison, the NovaMin®
Mousse-treated samples than the likely that the performance particles rapidly form a tenacious
control samples while the differences of the two dentifrices are HCA layer on the dentin that resists
DenShield™-treated samples due to very different mechanisms of the repeated acid challenges.
contained the same number of action. GC Tooth Mousse contains
partially occluded tubules as the casein phosphopeptide Conclusions
control group. The number of open, (RECALDENT™) that carries The current study has demonstrated
partially occluded, and total visible amorphous calcium phosphate. When that the NovaMin® particles are
tubules seen in samples from the the peptide complex binds to plaque superior to the casein phosphopeptide
DenShield™ treatment group were or the tooth surface it is said to deliver (RECALDENT™) in depositing a
significantly less than seen in the GC bio-available calcium and phosphate mineral layer on dentin that will
Tooth Mousse-treated samples. for remineralization, resulting in occlude dentinal tubules and resist
occlusion of dentin tubules. demineralization by repeated acid
Discussion challenges. This study supports the
The use of in vitro remineralization/ In comparison, the NovaMin®- previous clinical studies that have
demineralization methods has been containing DenShield™ deposits fine demonstrated superior reductions in
well accepted by the dental particles onto the dentin or enamel dentinal hypersensitivity in patients
community in assessing the efficacy surface and a series of reactions treated with NovaMin®-containing
of materials to physically occlude occurs over time that results in the dentifrices compared with other
tubules and to remineralize the formation of a crystalline, commercially available products.
surface of both dentin and enamel. In hydroxycarbonate apatite (HCA)
this study, two products that claim to layer. This HCA layer is chemically
treat dentine hypersensitivity were and structurally similar to natural
compared side-by-side in a 10 day enamel and dentin and is more
remineralization/demineralization resistant to acid challenges than the
protocol. The qualitative and amorphous calcium phosphate
quantitative results presented here deposited by the GC Tooth Mousse.