Casein and mucin have been shown to improve the erosion-protective properties of the pellicle when applied in combination. The aim of this in vitro study was to optimize the concentrations of these 2 proteins to achieve a maximum protective effect. For the 2 parts of this study, we prepared a total of 195 human enamel specimens and randomly assigned them to 13 groups, corresponding to 11 different casein-mucin concentration-combinations tested and 2 negative control groups (humid chamber). They underwent 5 cycles, consisting of pellicle formation from human whole saliva (2 h, 30°C), modification of the pellicle with casein and mucin in different concentrations (immersion in protein solutions for 2 h, 30°C), and erosion for 1 min in citric acid (0.65%, pH 3.5, 30°C). Surface microhardness (SMH), surface reflection intensity (SRI), and in the first part also calcium release were monitored during the cycling process, and analyzed with Kruskal-Wallis and post hoc Dunn’s tests. The results suggest that the best concentrations to achieve the highest erosion-protective effect are 3.0% casein and 0.81% mucin, which lead to a significant protection as measured by SMH as well as SRI compared to the unmodified pellicle. For the calcium release, no significant differences were found. This concentration combination corresponds to a general raise of the protein concentrations and a change in the molar ratio of the proteins as compared to earlier studies. Casein and mucin could now be incorporated at the determined concentration as natural ingredients in oral care products designed to protect against erosion.

1.
Barbour
ME
,
Shellis
RP
,
Parker
DM
,
Allen
GC
,
Addy
M
.
Inhibition of hydroxyapatite dissolution by whole casein: the effects of pH, protein concentration, calcium, and ionic strength
.
Eur J Oral Sci
.
2008
Oct
;
116
(
5
):
473
8
.
[PubMed]
0909-8836
2.
Batista
GR
,
Rocha Gomes Torres
C
,
Sener
B
,
Attin
T
,
Wiegand
A
.
Artificial saliva formulations versus human saliva pretreatment in dental erosion experiments
.
Caries Res
.
2016
;
50
(
1
):
78
86
.
[PubMed]
0008-6568
3.
Baumann
T
,
Kozik
J
,
Lussi
A
,
Carvalho
TS
.
Erosion protection conferred by whole human saliva, dialysed saliva, and artificial saliva
.
Sci Rep
.
2016
Oct
;
6
(
1
):
34760
.
[PubMed]
2045-2322
4.
Carvalho
TS
,
Baumann
T
,
Lussi
A
.
A new hand-held optical reflectometer to measure enamel erosion: correlation with surface hardness and calcium release
.
Sci Rep
.
2016
Apr
;
6
(
1
):
25259
.
[PubMed]
2045-2322
5.
Carvalho
TS
,
Colon
P
,
Ganss
C
,
Huysmans
MC
,
Lussi
A
,
Schlueter
N
, et al
Consensus report of the European Federation of Conservative Dentistry: erosive tooth wear—diagnosis and management
.
Clin Oral Investig
.
2015
Sep
;
19
(
7
):
1557
61
.
[PubMed]
1432-6981
6.
Cheaib
Z
,
Lussi
A
.
Impact of acquired enamel pellicle modification on initial dental erosion
.
Caries Res
.
2011
;
45
(
2
):
107
12
.
[PubMed]
0008-6568
7.
Cheaib
Z
,
Rakmathulina
E
,
Lussi
A
,
Eick
S
.
Impact of acquired pellicle modification on adhesion of early colonizers
.
Caries Res
.
2015
;
49
(
6
):
626
32
.
[PubMed]
0008-6568
8.
de Kruif
CG
.
Supra-aggregates of casein micelles as a prelude to coagulation
.
J Dairy Sci
.
1998
;
81
(
11
):
3019
28
. 0022-0302
9.
Derrien
M
,
van Passel
MW
,
van de Bovenkamp
JH
,
Schipper
RG
,
de Vos
WM
,
Dekker
J
.
Mucin-bacterial interactions in the human oral cavity and digestive tract
.
Gut Microbes
.
2010
Jul
;
1
(
4
):
254
68
.
[PubMed]
1949-0976
10.
Esser
D
,
Alvarez-Llamas
G
,
de Vries
MP
,
Weening
D
,
Vonk
RJ
,
Roelofsen
H
.
Sample stability and protein composition of saliva: implications for its use as a diagnostic fluid
.
Biomark Insights
.
2008
Feb
;
3
:
25
7
.
[PubMed]
1177-2719
11.
Fujikawa
H
,
Matsuyama
K
,
Uchiyama
A
,
Nakashima
S
,
Ujiie
T
.
Influence of salivary macromolecules and fluoride on enamel lesion remineralization in vitro
.
Caries Res
.
2008
;
42
(
1
):
37
45
.
[PubMed]
0008-6568
12.
Guggenheim
B
,
Schmid
R
,
Aeschlimann
JM
,
Berrocal
R
,
Neeser
JR
.
Powdered milk micellar casein prevents oral colonization by Streptococcus sobrinus and dental caries in rats: a basis for the caries-protective effect of dairy products
.
Caries Res
.
1999
Nov-Dec
;
33
(
6
):
446
54
.
[PubMed]
0008-6568
13.
Hall
AF
,
Buchanan
CA
,
Millett
DT
,
Creanor
SL
,
Strang
R
,
Foye
RH
.
The effect of saliva on enamel and dentine erosion
.
J Dent
.
1999
Jul
;
27
(
5
):
333
9
.
[PubMed]
0300-5712
14.
Hannig
M
,
Hannig
C
.
The pellicle and erosion
.
Monogr Oral Sci
.
2014
;
25
:
206
14
.
[PubMed]
0077-0892
15.
Hannig
M
,
Joiner
A
.
The structure, function and properties of the acquired pellicle
.
Monogr Oral Sci
.
2006
;
19
:
29
64
.
[PubMed]
0077-0892
16.
Hannig
M
.
Ultrastructural investigation of pellicle morphogenesis at two different intraoral sites during a 24-h period
.
Clin Oral Investig
.
1999
Jun
;
3
(
2
):
88
95
.
[PubMed]
1432-6981
17.
Hara
AT
,
Zero
DT
.
The potential of saliva in protecting against dental erosion
.
Monogr Oral Sci
.
2014
;
25
:
197
205
.
[PubMed]
0077-0892
18.
Jager
DH
,
Vieira
AM
,
Ligtenberg
AJ
,
Bronkhorst
E
,
Huysmans
MC
,
Vissink
A
.
Effect of salivary factors on the susceptibility of hydroxyapatite to early erosion
.
Caries Res
.
2011
;
45
(
6
):
532
7
.
[PubMed]
0008-6568
19.
Kapsimalis
P
,
Rosenthal
SL
,
Updegrave
W
,
Evans
R
,
Hurley
B
,
Cobe
HM
.
The relationship between caries activity, flow rate, total nitrogen and the mucin content of saliva
.
J Oral Med
.
1966
Jul
;
21
(
3
):
107
10
.
[PubMed]
0022-3247
20.
Larsen
MJ
.
Chemical events during tooth dissolution
.
J Dent Res
.
1990
Feb
;
69
(
Spec No
):
575
80
.
[PubMed]
0022-0345
21.
Marchin
S
,
Putaux
JL
,
Pignon
F
,
Léonil
J
.
Effects of the environmental factors on the casein micelle structure studied by cryo transmission electron microscopy and small-angle x-ray scattering/ultrasmall-angle x-ray scattering
.
J Chem Phys
.
2007
Jan
;
126
(
4
):
045101
.
[PubMed]
0021-9606
22.
Murray
PA
,
Prakobphol
A
,
Lee
T
,
Hoover
CI
,
Fisher
SJ
.
Adherence of oral streptococci to salivary glycoproteins
.
Infect Immun
.
1992
Jan
;
60
(
1
):
31
8
.
[PubMed]
0019-9567
23.
Rakhmatullina
E
,
Bossen
A
,
Höschele
C
,
Wang
X
,
Beyeler
B
,
Meier
C
, et al
Application of the specular and diffuse reflection analysis for in vitro diagnostics of dental erosion: correlation with enamel softening, roughness, and calcium release
.
J Biomed Opt
.
2011
Oct
;
16
(
10
):
107002
.
[PubMed]
1083-3668
24.
Rayment
SA
,
Liu
B
,
Offner
GD
,
Oppenheim
FG
,
Troxler
RF
.
Immunoquantification of human salivary mucins MG1 and MG2 in stimulated whole saliva: factors influencing mucin levels
.
J Dent Res
.
2000
Oct
;
79
(
10
):
1765
72
.
[PubMed]
0022-0345
25.
Schlueter
N
,
Hara
A
,
Shellis
RP
,
Ganss
C
.
Methods for the measurement and characterization of erosion in enamel and dentine
.
Caries Res
.
2011
;
45
Suppl 1
:
13
23
.
[PubMed]
0008-6568
26.
Senapati
S
,
Das
S
,
Batra
SK
.
Mucin-interacting proteins: from function to therapeutics
.
Trends Biochem Sci
.
2010
Apr
;
35
(
4
):
236
45
.
[PubMed]
0968-0004
27.
Siqueira
WL
,
Custodio
W
,
McDonald
EE
.
New insights into the composition and functions of the acquired enamel pellicle
.
J Dent Res
.
2012
Dec
;
91
(
12
):
1110
8
.
[PubMed]
0022-0345
28.
Svendsen
IE
,
Lindh
L
,
Elofsson
U
,
Arnebrant
T
.
Studies on the exchange of early pellicle proteins by mucin and whole saliva
.
J Colloid Interface Sci
.
2008
May
;
321
(
1
):
52
9
.
[PubMed]
0021-9797
29.
Trejo
R
,
Dokland
T
,
Jurat-Fuentes
J
,
Harte
F
.
Cryo-transmission electron tomography of native casein micelles from bovine milk
.
J Dairy Sci
.
2011
Dec
;
94
(
12
):
5770
5
.
[PubMed]
0022-0302
30.
Vacca-Smith
AM
,
Van Wuyckhuyse
BC
,
Tabak
LA
,
Bowen
WH
.
The effect of milk and casein proteins on the adherence of Streptococcus mutans to saliva-coated hydroxyapatite
.
Arch Oral Biol
.
1994
Dec
;
39
(
12
):
1063
9
.
[PubMed]
0003-9969
31.
Ventura
TM
,
Cassiano
LP
,
Souza E Silva
CM
,
Taira
EA
,
Leite
AL
,
Rios
D
, et al
The proteomic profile of the acquired enamel pellicle according to its location in the dental arches
.
Arch Oral Biol
.
2017
Jul
;
79
:
20
9
.
[PubMed]
0003-9969
32.
Vitkov
L
,
Hannig
M
,
Nekrashevych
Y
,
Krautgartner
WD
.
Supramolecular pellicle precursors
.
Eur J Oral Sci
.
2004
Aug
;
112
(
4
):
320
5
.
[PubMed]
0909-8836
33.
Vitorino
R
,
Calheiros-Lobo
MJ
,
Williams
J
,
Ferrer-Correia
AJ
,
Tomer
KB
,
Duarte
JA
, et al
Peptidomic analysis of human acquired enamel pellicle
.
Biomed Chromatogr
.
2007
Nov
;
21
(
11
):
1107
17
.
[PubMed]
0269-3879
34.
West
NX
,
Davies
M
,
Amaechi
BT
.
In vitro and in situ erosion models for evaluating tooth substance loss
.
Caries Res
.
2011
;
45
(
s1
Suppl 1
):
43
52
.
[PubMed]
0008-6568
35.
Yao
Y
,
Berg
EA
,
Costello
CE
,
Troxler
RF
,
Oppenheim
FG
.
Identification of protein components in human acquired enamel pellicle and whole saliva using novel proteomics approaches
.
J Biol Chem
.
2003
Feb
;
278
(
7
):
5300
8
.
[PubMed]
0021-9258
36.
Yao
Y
,
Grogan
J
,
Zehnder
M
,
Lendenmann
U
,
Nam
B
,
Wu
Z
, et al
Compositional analysis of human acquired enamel pellicle by mass spectrometry
.
Arch Oral Biol
.
2001
Apr
;
46
(
4
):
293
303
.
[PubMed]
0003-9969
37.
Zahradnik
RT
,
Moreno
EC
,
Burke
EJ
.
Effect of salivary pellicle on enamel subsurface demineralization in vitro
.
J Dent Res
.
1976
Jul-Aug
;
55
(
4
):
664
70
.
[PubMed]
0022-0345
38.
Zero
DT
,
Rahbek
I
,
Fu
J
,
Proskin
HM
,
Featherstone
JD
.
Comparison of the iodide permeability test, the surface microhardness test, and mineral dissolution of bovine enamel following acid challenge
.
Caries Res
.
1990
;
24
(
3
):
181
8
.
[PubMed]
0008-6568
39.
Zhang
YF
,
Zheng
J
,
Zheng
L
,
Shi
XY
,
Qian
LM
,
Zhou
ZR
.
Effect of adsorption time on the lubricating properties of the salivary pellicle on human tooth enamel
.
Wear
.
2013
;
301
(
1-2
):
300
7
. 0043-1648
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