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Mode of action studies of a new desensitizing mouthwashcontaining 0.8% arginine, PVM/MA copolymer,pyrophosphates, and 0.05% sodium fluoride
Sarita V. Mello a,*, Evangelia Arvanitidou a, Michael A. Stranick a, Ramon Santana a,Yasemin Kutes b, Bryan Huey b
aColgate-Palmolive Technology Center, 909 River Road, Piscataway, NJ, USAbUniversity of Connecticut, Chemical, Materials & Biomolecular Engineering Department, 97 North Eagleville Road,
Unit 3136, Storrs, CT 06269-3136, USA
j o u r n a l o f d e n t i s t r y 4 1 s ( 2 0 1 3 ) s 1 2 – s 1 9
a r t i c l e i n f o
Article history:
Received 18 July 2012
Received in revised form
30 October 2012
Accepted 1 November 2012
Keywords:
Mouthwash
Arginine
Tooth sensitivity
Dentine occlusion
a b s t r a c t
Objective: The mode of action of an arginine mouthwash using the Pro-ArginTM Mouthwash
Technology, containing 0.8% arginine, PVM/MA copolymer, pyrophosphates and 0.05%
sodium fluoride, has been proposed and confirmed as occlusion using a variety of
in vitro techniques.
Methods: Quantitative and qualitative laboratory techniques were employed to investigate the
mode of action of the new arginine mouthwash. Confocal laser scanning microscopy (CSLM)
and atomic force microscopy (AFM) investigated a hydrated layer on dentine surface. Electron
spectroscopy for chemical analysis (ESCA), secondary ion mass spectroscopy (SIMS) and near-
infrared spectroscopy (NIR) provided information about its chemical nature.
Results: CLSM was used to observe the formation of a hydrated layer on exposed dentine
tubules upon application of the arginine mouthwash. Fluorescence studies confirmed pene-
tration of the hydrated layer in the inner walls of the dentinal tubules. The AFM investigation
confirmed the affinity of the arginine mouthwash for the dentine surface, supporting its
adhesive nature. NIR showed the deposition of arginine after several mouthwash applications,
and ESCA/SIMS detected the presence of phosphate groups and organic acid groups, indicating
the deposition of copolymer and pyrophosphates along with arginine.
Conclusion: The studies presented in this paper support occlusion of the dentine surface
upon the deposition of an arginine-rich layer together with copolymer and phosphate ions
from an alcohol-free mouthwash containing 0.8% arginine, PVM/MA copolymer, pyropho-
sphates and 0.05% sodium fluoride.
# 2012 Elsevier Ltd.
Available online at www.sciencedirect.com
journal homepage: www.intl.elsevierhealth.com/journals/jden
Open access under CC BY-NC-ND license.
1. Introduction
Tooth sensitivity has been recognized as one of the earliest
maladies of the mouth reported in history. Tooth sensitivity is
* Corresponding author at: Colgate-Palmolive Technology Center, 909
fax: +1 7328787365.E-mail address: sarita_mello@colpal.com (S.V. Mello).
0300-5712 # 2012 Elsevier Ltd.
http://dx.doi.org/10.1016/j.jdent.2012.11.001
Open access under CC BY-NC-ND license.
a common name for dentine hypersensitivity or root sensitiv-
ity. It is estimated that up to 57% of the population in
the United States suffers from dentine hypersensitivity.1
Dentine hypersensitivity is characterized by short, sharp pain
arising from exposed dentine in response to stimuli, typically
River Road, Piscataway, NJ 08855-1343, USA. Tel.: +1 7328787883;
j o u r n a l o f d e n t i s t r y 4 1 s ( 2 0 1 3 ) s 1 2 – s 1 9 S13
thermal, evaporative, tactile, osmotic or chemical and which
cannot be ascribed to any other dental defect or pathology.2
Diverse in-office treatments have been reported to address
dentine hypersensitivity, such as desensitizing pastes, fluo-
ride varnishes, and restorative resins.3–13 Over-the-counter
products that treat dentine hypersensitivity are normally
toothpastes formulated with potassium ions or dentine
occlusion agents.14–21 The mode of action of desensitizing
products can be divided in two main groups: (1) nerve
desensitization by potassium ions and (2) decrease in dentine
permeability by occlusion agents. Historically, marketed anti-
hypersensitivity toothpastes were recommended for a mini-
mum of two weeks use, in order to allow the accumulation of
an effective concentration of potassium ions to desensitize the
nerve endings. The over-the-counter occlusion treatment
approach has been traditionally reported as the deposition of
solid particles or mineralized crystals, such as stannous,
strontium and oxalates.4,5 More recently, a breakthrough
occlusion technology containing arginine and calcium car-
bonate has been formulated into toothpaste and in-office
products.7,20–24
In vitro investigations of dentine occlusion systems are
commonly carried out via Pashley’s hydraulic conductance
method25–29 and surface analysis techniques.3,23 Also, Kolker
et al.30 studied the in vitro performance of different synthetic
resins, showing effective reduction of hydraulic conductance
in the range of 40–60%, for different coatings with diverse
chemical compositions. In contrast to mineral solids occlu-
sion, the mechanism of action of adhesive coatings depends
on the physical adsorption of organic materials, their diffusion
through porous dentine, the chemical bonding or curing and
electrostatic interactions. The viscosity and contact time of
the polymeric layer are also important to allow the coating
effect (film-forming) and ensure enough cohesive forces to
increase the life-time of the coating. Most recently, hydraulic
conductance studies have demonstrated a significant reduc-
tion in fluid flow for a toothpaste23 and for a mouthwash.31
Although a variety of dental products have been employed
successfully to address dentine hypersensitivity, there is still
an absence of effective mouthwash products and only a few
scientific papers have been published.32–35 Herein we present
the in vitro evaluation of a new mouthwash designed to
effectively reduce dentine hypersensitivity through the
occlusion of the dentine surface via a clinically proven
formulation of arginine, PVM/MA copolymer and pyropho-
sphates.36,37 Polymethylvinyl ether/maleic acid (PVM/MA)
copolymer and pyrophosphates have been associated with
dentine occlusion in toothpaste formulations by Miller et al.38
and Mason et al.39 based on in vitro measurements of
hydraulic conductance and surface analysis techniques. The
mechanism of occlusion presented here differs from Miller
et al.38 due to several factors, especially the presence of
arginine, other formula ingredients, pH, and delivery in the
absence of brushing (toothpaste vs mouthwash). The mode of
action of the new mouthwash containing the Pro-ArginTM
Mouthwash Technology is based on occlusion of the dentine
tubules from the deposition of arginine, PVM/MA copolymer
and polyphosphates onto the dentine, which after repeated
application, decreases dentine permeability and reduces pain-
causing stimuli. The mechanism of action was investigated via
several laboratory techniques, namely confocal laser scanning
microscopy (CSLM), atomic force microscopy (AFM), electron
spectroscopy for chemical analysis (ESCA), secondary ion
mass spectroscopy (SIMS) and near-infrared spectroscopy
(NIR).
2. Materials and methods
2.1. Test products
The test mouthwash (‘‘arginine’’) consisted of 0.8% arginine,
polyvinylmethyl ether/maleic acid (PVM/MA) copolymer,
pyrophosphates and 0.05% sodium fluoride (Colgate-Palmol-
ive Company, New York, NY) in an alcohol-free base. The
control mouthwash (negative control) (Colgate-Palmolive
Company, New York, NY) did not contain any of the
ingredients mentioned above. In addition, a commercial
fluoride toothpaste (Colgate-Palmolive Company, New York,
NY) was employed during one of the evaluations.
2.2. Dentine disc preparation
Dentine discs used in all tests were prepared from extracted
human molars. An average of 3 discs per molar were cut and
prepared accordingly to each to be used in different experi-
ments. At least 6 discs were randomly assigned per sample in
all tests. Specimens, 0.8 mm thick, were obtained by using a
slow-speed saw turning a diamond wafering blade (Isomet,
Lake Bluf, USA). Afterwards, discs were sanded 600 grit paper
discs (Carbimet, Buehler, Lake Bluf, USA) and polished with
soft plane cloth (P4000, Silicon Carbide using a Variable Speed
Grinder-Polisher EcoMet1 3000, Buehler, Lake Bluf, USA).
Each dentine disc was sonicated in deionized water and
dipped into 6% (w/v) citric acid for 1 min to remove the smear
layer.28
2.3. Surface and chemical characterization
2.3.1. Confocal laser scanning microscopy (CLSM)A Leica TCS SP confocal laser scanning microscope equipped
with a spectral detection system was used to visualize the
dentine surfaces. The 488 nm line from an argon laser along
with a PLO APO 50� objective was used in all experiments.
Dentine discs were prepared according to the procedure
described above. Images were taken with a 50� objective and
optical zoom of 2�. Distinct treatment procedures were
developed for each detection mode in order to assess the
relevant aspect of the coating mechanism, i.e. a thicker
coating to allow the tubule blocking visualization (10 applica-
tions) and a thinner coating (2 applications) for the fluor-
ophore detection inside the tubules.
Reflectance mode: on dentine discs pre-conditioned in
phosphate buffer saline (PBS), 15 mL of mouthwash was
applied 10 times for 1 min, with intervals of 10 min
between each application.
Fluorescent mode: fluorescein–cadaverine (molecular probes)
was added to both mouthwash samples prior to treatment,
to a final concentration of 0.4 mM. A mouthwash aliquot of
j o u r n a l o f d e n t i s t r y 4 1 s ( 2 0 1 3 ) s 1 2 – s 1 9S14
15 mL was added to the dentine surface and left to dry for
10 min. The same procedure was repeated one more time
followed by a gentle rinse with PBS.
2.3.2. Atomic force microscopy (AFM)Atomic force microscopy (AFM) is a high-resolution, non-
destructive laboratory technique that allows imaging of nano
structures by mapping topography and/or other properties
using a tip with a nanoscale apex.40 When working with soft
samples such as polymer, biological and dental materials,41–
43 the AFM is typically operated in ‘AC’ mode, where the probe
is vibrated near a specimen such that they measurably
interact �10,000–300,000 times per second. A constant
interaction amplitude (and thus force gradient) is then
maintained by raising or lowering the tip as needed, which
when scanning the sample thus provides images of the
surface height with sub-nanometer resolution in the x, y, and
z directions.44
For the specimens considered here, the arginine mouth-
wash was coated on tooth substrates with a spin coating
technique to create uniform layers.45 Each layer was spun with
3000 rpm for 30 s. The samples were then mounted on a glass
slide and kept at 37 8C until the images were taken.
2.3.3. Near infrared (NIR)Baseline spectra were taken prior to arginine mouthwash.
Samples were evaluated in triplicate. Treatment was per-
formed as follows:
1. 10 mL of mouthwash was applied to disc surface and left for
10 min on a hot plate at 37 8C.
2. Disc was rinsed with PBS buffer (3� dipping for 1 s).
3. Buffer excess was removed with air stream parallel to the
disc surface and taken immediately to the NIR instru-
ment.
4. Following spectra acquisition, a new mouthwash treatment
was applied to the disc.
2.3.4. Electron spectroscopy for chemical analysis (ESCA) andsecondary ion mass spectroscopy (SIMS)All dentine discs were analysed prior to treatment to obtain
baseline compositions. For ESCA, three separate areas per disc
were analysed, while for SIMS one area encompassing most of
the disc was analysed. The same discs were analysed by both
ESCA and SIMS. For the untreated dentine, the ESCA results
represent the average compositions for the four discs studied,
while the SIMS values are the averages for two of the discs
studied. The ESCA and SIMS results shown for the treated
discs represent the average values for two discs of each
treatment regimen.
The Test treatment consisted of the commercial fluoride
toothpaste and the arginine mouthwash, and the control
treatment consisted of the commercial fluoride toothpaste
and the negative control mouthwash.
Treatments were performed as follows:
1. Dentine surface was wetted with PBS.
2. Surface was gently brushed with toothpaste for 30 s.
3. Disc was air dried for 2 min, and then moved to a beaker
containing PBS with mild agitation for 1 min.
4. Disc was removed from PBS, tapped dry on the back
(untreated surface) to remove excess fluid and then placed
in beaker containing mouthwash for 2 min under mild
agitation.
5. Disc was removed from the mouthwash and air dried for
2 min. Excess mouthwash was gently removed with a small
brush.
6. Two drops of PBS was placed on the surface and the excess
of liquid was gently removed from the surface with a small
soft brush. Film was air dried for 30–40 min. Steps 4–6 were
repeated once.
3. Results
3.1. CLSM
Images shown in Figs. 1 and 2 represent dentine discs after ten
1-min treatments using the arginine mouthwash and the
negative control mouthwash, respectively. There was a 10-
min interval between each treatment. Reflectance mode
(Fig. 1a and b) was used to assess the surface effect while
fluorescent mode (Fig. 2a and b) investigated the penetration
of the Arginine mouthwash into open tubules. Fig. 1a shows a
continuous image after treatment with the arginine mouth-
wash, indicating complete coverage of the open dentine
tubules. The solid line of the xz view indicates the surface
effect, i.e. the light source cannot penetrate the tubules. Fig. 1b
shows the open tubules before and after treatment with
negative control mouthwash. A non-continuous line on the xz
view confirms the openness of the tubules.
Fig. 2a shows the fluorescence images taken after treat-
ment with the arginine mouthwash. The bright points indicate
the presence of the arginine mouthwash throughout the
surface. The side view shows the penetration of the arginine
mouthwash inside the tubule walls, indicating the affinity of
the mouthwash to the dentine surface. The same experiment
was performed with the negative control mouthwash shown
in Fig. 2b. In contrast to the arginine mouthwash, very little
evidence of mouthwash on the surface (xy image) in the
tubules (xz image) could be visualized for the negative control
mouthwash. These images prove that the arginine mouth-
wash penetrates inside the tubules as well as deposits on the
surface.
3.2. NIR
Arginine has a unique fingerprint in the NIR region
(�1530 nm), allowing its identification on the dentine surface.
Fig. 3 shows the spectra obtained on the dentine surface before
treatment (baseline) and after 3, 7 and 13 treatments with the
arginine mouthwash. The presence of arginine molecules was
identified by the 3rd treatment, and their concentration
increased upon deposition with subsequent treatments.
3.3. AFM
The ability of the Arginine mouthwash to deposit onto
dentine surfaces has also been explored by AFM. The high
resolution of AFM even allowed for imaging the fine structure
in the vicinity of a single dentine tubule (Fig. 4). The
Fig. 1 – (a) CLSM images (reflectance mode) of dentine surface treated with arginine mouthwash. Image scale
100 mm T 100 mm. (b) CLSM images (reflectance) of dentine surface treated with negative control mouthwash. Image scale
100 mm T 100 mm.
j o u r n a l o f d e n t i s t r y 4 1 s ( 2 0 1 3 ) s 1 2 – s 1 9 S15
5 mm � 5 mm images of single tubules, before and after
coating, indicate that the arginine mouthwash is deposited
onto the dentine surface. The occlusion occurs by a buildup
mechanism of the arginine mouthwash in the tubules after
several applications.
3.4. ESCA/SIMS
ESCA/SIMS assessed the chemical composition on dentine
after treatment with a mouthwash in conjunction with a
commercial fluoride toothpaste, in order to mimic human use.
As shown in Table 1, the compositions of the discs before
treatment were consistent with those for demineralized,
Table 1 – ESCA results.
Sample Atomic
C O N Ca
ESCA – surface composition
Untreated dentine
2, 3, 10, 11 Avg. 61.30 21.16 16.30 0.74
Fluoride toothpaste/Negative Control MW – 20 treatments
2, 10 Avg. 36.88 39.85 5.77 9.43
Fluoride toothpaste/Arginine MW – 20 treatments
3, 11 Avg. 58.03 30.04 2.28 2.24
untreated dentine: high surface protein with low calcium (Ca)
and phosphorus (P) levels. After treatment, the sodium (Na)
levels for the Test discs were greater than those for Negative
Control discs. Potassium (K) was detected for Test discs only.
The P/Ca ratio was also greater for Test discs compared to
negative control discs. The Na, K and P/Ca results indicate the
deposition onto the dentine surface of the tetrapotassium
pyrophosphate and tetrasodium pyrophosphate (pyropho-
sphates) present only in the arginine mouthwash. Elevated
carbon (C) levels with reduced N, Ca, and P were observed for
the Test discs compared to the negative control discs. This is
indicative of the presence of an organic material on the Test
discs. Also, the nitrogen (N) spectra for the Test discs differed
percent Ratio
P Na F Si K P/Ca N+/N
0.50 – – 0.00 – 0.67 0.030
6.92 0.25 0.44 0.93 – 0.71 0.019
2.97 1.16 0.11 1.86 1.30 1.49 0.095
Fig. 2 – (a) CLSM images (fluorescence mode) of dentine surface treated with arginine mouthwash. Image scale
100 mm T 100 mm. (b) CLSM images (fluorescence mode) of dentine surface treated with the negative control mouthwash.
Image scale 100 mm T 100 mm.
j o u r n a l o f d e n t i s t r y 4 1 s ( 2 0 1 3 ) s 1 2 – s 1 9S16
in line shape from those for the negative control discs, because
of the greater amount of charged nitrogen, N+. This difference
is reflected in the N+/N ratios shown in Table 1. Higher N+ is
consistent with that observed in data for arginine reference
Fig. 3 – NIR spectra of dentine surface t
films, and indicates the presence of arginine on the surfaces of
the Test discs. SIMS analysis (Table 2) of the treated dentine
revealed the presence of a mass peak at 175 amu after
treatment with the Test treatment. This peak was not
reated with Arginine mouthwash.
Fig. 4 – AFM topography images of a dentinal tubule before and after treatment with the arginine mouthwash.
j o u r n a l o f d e n t i s t r y 4 1 s ( 2 0 1 3 ) s 1 2 – s 1 9 S17
observed on untreated dentine or on dentine treated with the
negative control mouthwash. The mass peak at 175 amu is
consistent with that for the arginine ion and confirms
deposition of arginine on the surfaces of the Test treated
discs. SIMS analysis of PVM/MA copolymer reference films
revealed mass peaks from fragments of the copolymer at 45
and 59 amu. SIMS analysis of the Test treated discs revealed an
increase in the 45/43 and 59/43 peak intensity ratios, compared
to the negative control treated discs. The increases in these
ratios above the background values for the baseline suggests
PVM/MA copolymer deposition.
4. Discussion
A new alcohol-free mouthwash, with 0.8% arginine, PVM/MA
copolymer, pyrophosphates, and 0.05% sodium fluoride has
been designed to alleviate dentine hypersensitivity by occlu-
sion. To the best of our knowledge, this is the first time
a mouthwash has been effective on addressing dentine
hypersensitivity via occlusion mechanism with a unique
combination of a complex formed by copolymer/aminoacid/
pyrophosphate salts. Laboratory studies indicate the forma-
tion a micro-adhesive layer on the dentine surface comprised
arginine, PVM/MA copolymer and pyrophosphates. Arginine
and the PVM/MA copolymer form an adhesive complex, which
Table 2 – SIMS results.
Sample Gantrez pe
45/43
Untreated dentine
10, 11 Avg. 0.098
Fluoride toothpaste/Negative Control MW – 20 treatments
2, 10 Avg. 0.069
Fluoride toothpaste/Arginine MW – 20 treatments
3,11 avg. 1.823
PVM/MA copolymer – fragment positive ions at 45 and 59 amu; hydroca
175 amu.
deposits onto the dentine surface. The pyrophosphates
promote the complex’s stability in solution and are deposited
together onto dentine surface, leading to a coherent layer after
multiple mouthwash applications to dentine specimens. The
overall result is an arginine/copolymer/phosphate deposit on
the demineralized dentine surface. Experimental challenges
of characterizing a hydrated polymeric layer have been
addressed by using a range of qualitative and quantitative
techniques already established to investigate dentine occlu-
sion as shown in the work of Petrou et al.24.
In addition, hydraulic conductance studies have shown
that this new mouthwash provides a significant reduction in
fluid flow, consistent with an occlusion mechanism.31 The
occlusion of dentine tubules with this arginine mouthwash is
the basis of the dentine hypersensitivity relief demonstrated
in two clinical studies.36,37 In this paper, CLSM confirmed the
ability of the Arginine mouthwash to form a layer on the
dentine surface (reflectance) as well as within the internal
walls of the dentine tubules (fluorescence). Based on the
fluorophore chemistry, which is designed to interact with
carboxylic groups, the fluorescence detected is due to arginine
and copolymer deposition inside the dentine tubules as well as
on the dentine surface. One important aspect of the CLSM
study was the ability to study the deposition of the arginine
mouthwash onto dentine in open air and after a relatively
short time after treatment (30 min). Arginine is drawn to the
ak intensity ratios Arginine peak (amu)
59/43 175
0.164 �
0.041 �
0.514 +
rbon reference peak at 43 amu. Arginine – molecular positive ion at
j o u r n a l o f d e n t i s t r y 4 1 s ( 2 0 1 3 ) s 1 2 – s 1 9S18
dentine surface, serving as the anchor for subsequent
copolymer deposition. Studies using NIR spectroscopy confirm
arginine deposition on the dentine surface upon application of
the arginine mouthwash. As seen using AFM, the images of a
single tubule indicate that the arginine mouthwash is present
on the dentine surface, and occlusion happens by a buildup
mechanism of the arginine mouthwash in the tubules after
several applications. This may explain the lack of a hyper-
sensitivity effect within the first 30 min of using the arginine
mouthwash for the first time.37
Finally, the ESCA and SIMS results indicate the presence of
an organic material on the discs with the Test treatment. Also,
the nitrogen spectra for these discs differed in line shape from
those for the negative control treated discs. The discs treated
with the Test treatment containing the arginine mouthwash
exhibited a greater amount of charged nitrogen than the
Negative Control treated discs.
5. Conclusion
A new hypersensitivity mouthwash based on the Pro-ArginTM
Mouthwash Technology containing 0.8% arginine, PVM/MA
copolymer, pyrophosphates and 0.05% sodium fluoride has
been developed. In vitro studies confirm the mechanism of
action as occlusion. CLSM captured images of the hydrated
layer on the dentine surface. Fluorescence studies confirmed
penetration of the hydrated layer in the inner walls of the
dentine tubules. ESCA confirmed deposition of arginine and
phosphates onto the dentine surface. AFM confirmed this
observation, even identifying tubules that are spanned by the
mouthwash. SIMS analysis confirmed the presence of arginine
and organic groups which are indicative of copolymer
deposition. NIR confirmed the presence of arginine on dentine
surface upon treatment with the arginine mouthwash. The
occlusion from the arginine mouthwash has been achieved
due to a formula design that allows for the deposition of
arginine and phosphates embedded in a thin polymeric
matrix. This unique formulation is an effective system to
address dentine hypersensitivity.
Conflicts of interest
None declared.
r e f e r e n c e s
1. Irwin CR, McCusker P. Prevalence of dentin hypersensitivityin a general dental population. Journal of the Irish DentalAssociation 1997;43:7–9.
2. Dababneh RH, Khouri AT, Addy M. Dentinhypersensitivity—an enigma? A review of terminology,epidemiology, mechanisms, aetiology and management.British Dental Journal 1999;187:606–11.
3. Addy M. Dentin hypersensitivity: definition prevalencedistribution and aetiology. In: Addy M, Embery G, Edgar WM,Orchardson R, editors. Tooth wear and sensitivity: clinicaladvances in restorative dentistry. London: Martin Dunitz; 2000.p. 239–48.
4. Orchardson R, Gillam DG. Managing dentin hypersensitivity.Journal of American Dental Association 2006;137:990–8.
5. Cummins D. Recent advances in dentin hypersensitivity:clinically proven treatments for instant and lastingsensitivity relief. American Journal of Dentistry 2010;23:3A–13A.
6. MacCarthy D. Dentin hypersensitivity: a review of theliterature. Journal of the Irish Dental Association 2004;50:36–41.
7. Cummins D. The efficacy of a new dentifrice containing8.0% arginine, calcium carbonate, and 1450 ppm fluoride indelivering instant and lasting relief of dentinhypersensitivity. Journal of Clinical Dentistry 2009;20:109–14.
8. Trowbridge HO, Silver DR. A review of current approaches toin-office management of tooth hypersensitivity. DentalClinics of North America 1990;34:561–81.
9. Gaffar A. Treating hypersensitivity with fluoride varnish.Compendium of Continuing Education in Dentistry 1999;20(Suppl.1):27–33.
10. Corona SA, Nascimento TN, Catirse AB, Lizarelli RF, DinelliW, Palma-Dibb RG. Clinical evaluation of low-level lasertherapy and fluoride varnish for treating cervical dentinalhypersensitivity. Journal of Oral Rehabilitation 2003;30:1183–9.
11. Prati C, Cervellati F, Sanasi V, Montebugnoli L. Treatment ofcervical dentin hypersensitivity with resin adhesives: 4-week evaluation. American Journal of Dentistry 2001;14:378–82.
12. Baysan A, Lynch E. Treatment of cervical sensitivity with aroot sealant. American Journal of Dentistry 2003;16:135–8.
13. Kielbasa AM, Attin T, Hellwig E, Schade-Brittinger C. In vivostudy on the effectiveness of a lacquer containing CaF2/NaFin treating dentine hypersensitivity. Clinical OralInvestigations 1997;1:95–9.
14. Kanapka JA. Over-the-counter dentifrices in the treatmentof tooth hypersensitivity. Review of clinical studies. DentalClinics of North America 1990;34:545–60.
15. Schiff T, Zhang YP, DeVizio W, Stewart B, Chaknis P, PetroneME, et al. A randomized clinical trial of the desensitizingefficacy of three dentifrices. Compendium of ContinuingEducation in Dentistry Supplement 2000;27:4–10.
16. Sowinski JA, Battista GW, Petrone ME, Chaknis P, Zhang YP,DeVizio W, et al. A new desensitizing dentifrice: an 8-weekclinical investigation. Compendium of Continuing Education inDentistry 2000:11–6.
17. Schiff T, Bonta Y, Proskin HM, DeVizio W, Petrone ME, VolpeAR. Desensitizing efficacy of a new dentifrice containing5.0% potassium nitrate and 0.454% stannous fluoride.American Journal of Dentistry 2000;13:111–5.
18. Sowinski JA, Bonta Y, Battista GW, Petrone D, DeVizio W,Petrone ME, et al. Desensitizing efficacy of Colgate sensitivemaximum strength and fresh mint Sensodyne dentifrices.American Journal of Dentistry 2000;13:116–20.
19. Sowinski JA, Ayad F, Petrone ME, DeVizio W, Volpe AR,Ellwood R, et al. Comparative investigations of thedesensitising efficacy of a new dentifrice. Journal of ClinicalPeriodontology 2001;28:1032–6.
20. Schiff T, Delgado E, Zhang YP, Cummins D, DeVizio W, MateoLR. Clinical evaluation of the efficacy of an in-officedesensitizing paste containing 8.0% arginine and calciumcarbonate in providing instant and lasting relief of dentinhypersensitivity. American Journal of Dentistry 2009;22:8A–15A.
21. Hamlin D, Williams KP, Delgado E, Zhang YP, DeVizio W,Mateo LR. Clinical evaluation of the efficacy of adesensitizing paste containing 8.0% arginine and calciumcarbonate for the in-office relief of dentin hypersensitivityassociated with dental prophylaxis. American Journal ofDentistry 2009;22:16A–20A.
22. Ayad F, Ayad N, Zhang YP, Cummins D, DeVizio W,Cummins D, et al. Comparing the efficacy in reducing dentinhypersensitivity of a new toothpaste containing 8.0%
j o u r n a l o f d e n t i s t r y 4 1 s ( 2 0 1 3 ) s 1 2 – s 1 9 S19
arginine, calcium carbonate, and 1450 ppm fluoride to acommercial sensitive toothpaste containing 2% potassiumion: an eight-week clinical study on Canadian adults. TheJournal of Clinical Dentistry 2009;20:10–6.
23. Lavender S, Petrou I, Heu R, Stranick M, Cummins D,Kilpatrick L, et al. Mode of action studies of a newdesensitizing dentifrice, based upon the Pro-ArginTM
technology, with a gentle whitening benefit. American Journalof Dentistry 2010;23:14A–9A.
24. Petrou I, Heu R, Stranick M, Lavender S, Zaidel L, CumminsD, et al. A breakthrough therapy for dentin hypersensitivity:how dental products containing 8% arginine and calciumcarbonate work to deliver effective relief of sensitive teeth.Journal of Clinical Dentistry 2009;20:23–31.
25. Greenhill J, Pashley D. The effects of desensitizing agents onthe hydraulic conductance of human dentin in vitro. Journalof Dental Research 1981;60:686–98.
26. Pashley D, O’Meara JA, Kepler EE, Galloway SE,Thompson SM, Stewart FP. Dentin permeability. Effects ofdesensitizing dentifrices in vitro. Journal of Periodontology1984;55:522–5.
27. Pashley D. Dentin permeability, sensitivity and treatmentthrough tubule occlusion. Journal of Endodontics 1986;12:465–74.
28. Pashley DH, Agee K, Zhang Y, Smith A, Tavss E, Gambogi RJ.The effects of outward forced convective flow on inwarddiffusion of potassium across human dentin. AmericanJournal of Dentistry 2002;5:256–61.
29. Sauro S, Pashley DH, Montanari M, Chersoni S, Carvalho R,Toledano M, et al. Effect of simulated pulpal pressure ondentin permeability and adhesion of self-etch adhesives.Dental Materials 2007;23(6):705–13.
30. Kolker JL, Vargas MA, Armstrong SR, Dawson DV.Effect of desensitizing agents on dentin permeability anddentin tubule occlusion. Journal of Adhesive Dentistry2002;4:211–21.
31. Mello SV, Arvanitidou E, Vandeven M. The development of anew desensitising mouthwash containing arginine, PVM/MA copolymer, pyrophosphates, and sodium fluoride—Ahydraulic conductance study. Journal of Dentistry2013;41S:S20–5.
32. Gillam DG, Bulman JS, Jackson RJ, Newman HN. Efficacy of apotassium nitrate mouthwash in alleviating cervicaldentine sensitivity (CDS). Journal of Clinical Periodontology1996;23:993–7.
33. Pereira R, Chava VK. Efficacy of a 3% potassium nitratedesensitizing mouthwash in the treatment of dentinalhypersensitivity. Journal of Periodontology 2001;72:1720–5.
34. Yates R, West N, Addy M, Marlow I. The effects of apotassiumcitrate, cetylpyridinium chloride, sodium fluoridemouthrinse on den-tine hypersensitivity, plaque andgingivitis: a placebo controlled study. Journal of ClinicalPeriodontology 1998;25:813–20.
35. Yates RJ, Newcombe RG, Addy M. Dentine hypersensitivity:a randomized, double-blind placebo-controlled study of theefficacy of a fluoride-sensitive teeth mouthrinse. Journal ofClinical Periodontology 2004;31:885–9.
36. Hu D, Mateo LR, Stewart B, Zhang YP, Panagakos F. Efficacyof a mouthwash containing 0.8% arginine, PVM/MAcopolymer, pyrophosphates, and 0.05% sodium fluoridecompared to a negative control mouthwash on dentinhypersensitivity reduction. A randomized clinical trial..Journal of Dentistry 2013;41S:S26–33.
37. Boneta AE, Galan RM, Mateo LR, Stewart B, Mello S,Arvanitidou E, et al. Efficacy in reducing dentinehypersensitivity of a regimen using a toothpaste containing8% arginine and calcium carbonate, a mouthwashcontaining 0.8% arginine, pyrophosphate and PVM/MAcopolymer. Journal of Dentistry 2013;41S:S42–9.
38. Miller S, GaffarA. Sullivan R, Heu R, Truong T, Stranick M.Evaluation of a new dentifrice for the treatment of sensitiveteeth. Journal of Clinical Dentistry 1994;5:71–9.
39. Mason S, Levan A, Crawford F, Fisher S, Gaffar A. Evaluationof tartar control dentifrices in in vitro models of dentinsensitivity. Clinical Preventive Dentistry 1991;13:6–12.
40. Binning G, Quate CF, Gerber Ch. Atomic force microscope.Physical Review Letters 1986;56:930.
41. Verma S, Huey BD, Burgess DJ. Scanning probe microscopymethod for nanosuspension stabilizer selection. Langmuir2009;25:12481–7.
42. Penna R, Mante F, Huey BD, Ghafari J. Comparison of surfacetreated orthodontic bands: evaluation of shear force andsurface roughness. American Journal of Orthodontics andDentofacial Orthopedics 1998;114:162–5.
43. Advincula MC, Patel PA, Mather PT, Underhill D, Huey BD,Goldberg AJ. Directed mineralization on polyelectrolytemultilayer films. MRS proceedings-975E-mechanics of biologicaland bio-inspired materials. 2006.
44. Bonnell DA, Huey BD. In: Bonnell DA, editor. Scanning probemicroscopy & spectroscopy: theory, techniques and applications.New York: Wiley-VCH; 2001.
45. Schubert DW, Dunkel T. Spin coating from a molecularpoint of view: its concentration regimes, influence of molarmass and distribution. Materials Research Innovations2003;7:312–21.
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