Objective: Saliva is the most important biological factor to protect against erosive tooth wear (ETW). Gastroesophageal reflux disease (GERD) patients have an increased risk of ETW due to the frequent presence of intrinsic acids in the oral cavity. Remarkably, not all GERD patients suffer from ETW, which might be due to differences in the composition of the saliva. Methods: This study compared the proteomic profile of saliva in patients (1) with GERD and ETW (basic erosive wear examination, BEWE, score ≥9; GE group) and (2) with GERD without ETW (BEWE = 0; GNE group) using shotgun label-free quantitative proteomic analysis nLC-ESI-MS/MS. The ability of hemoglobin (Hb) to protect against initial enamel erosion caused by a daily 10-s immersion of enamel specimens in 0.01 M HCl (pH 2.3) for 3 days was evaluated in vitro for proof of concept. Surface hardness change was used as response variable. Results: The differential expression of Hb subunits was significantly increased in the GNE group versus the GE group, in particular the Hb α-subunit that showed a >22-fold increase. Expressions of serum albumin (4.5-fold) and isoforms of cytoskeletal keratin type II (>3-fold) were also increased in the GNE group. Proteinase inhibitors, such as α1-antitrypsin and α2-macroglobulin, were only identified in the GNE group. In vitro, Hb (1.0 and 4.0 mg/mL) significantly reduced initial enamel erosion compared to a negative control after 3 days. Conclusions: Our results indicate that many proteins, with special emphasis on Hb, may be involved in the resistance of GERD patients to the occurrence of ETW. These proteins may be candidates for inclusion in dental products to protect against ETW.

1.
Bartlett
D
,
Ganss
C
,
Lussi
A
.
Basic Erosive Wear Examination (BEWE): a new scoring system for scientific and clinical needs
.
Clin Oral Investig
.
2008
Mar
;
12
(
S1
Suppl 1
):
S65
8
.
[PubMed]
1432-6981
2.
Bateman
A
,
Martin
MJ
,
Orchard
S
,
Magrane
M
,
Alpi
E
,
Bely
B
, et al;
UniProt Consortium
.
UniProt: a worldwide hub of protein knowledge
.
Nucleic Acids Res
.
2019
Jan
;
47
D1
:
D506
15
.
[PubMed]
0305-1048
3.
Bragulla
HH
,
Homberger
DG
.
Structure and functions of keratin proteins in simple, stratified, keratinized and cornified epithelia
.
J Anat
.
2009
Apr
;
214
(
4
):
516
59
.
[PubMed]
0021-8782
4.
Buzalaf
MA
,
Charone
S
,
Tjäderhane
L
.
Role of host-derived proteinases in dentine caries and erosion
.
Caries Res
.
2015
;
49
Suppl 1
:
30
7
.
[PubMed]
0008-6568
5.
Buzalaf
MA
,
Hannas
AR
,
Kato
MT
.
Saliva and dental erosion
.
J Appl Oral Sci
.
2012
Sep-Oct
;
20
(
5
):
493
502
.
[PubMed]
1678-7757
6.
Buzalaf
MA
,
Magalhães
AC
,
Rios
D
.
Prevention of erosive tooth wear: targeting nutritional and patient-related risks factors
.
Br Dent J
.
2018
Mar
;
224
(
5
):
371
8
.
[PubMed]
0007-0610
7.
Cheaib
Z
,
Lussi
A
.
Impact of acquired enamel pellicle modification on initial dental erosion
.
Caries Res
.
2011
;
45
(
2
):
107
12
.
[PubMed]
0008-6568
8.
Delecrode
TR
,
Siqueira
WL
,
Zaidan
FC
,
Bellini
MR
,
Moffa
EB
,
Mussi
MC
, et al
Identification of acid-resistant proteins in acquired enamel pellicle
.
J Dent
.
2015
Dec
;
43
(
12
):
1470
5
.
[PubMed]
0300-5712
9.
Dent
J
,
El-Serag
HB
,
Wallander
MA
,
Johansson
S
.
Epidemiology of gastro-oesophageal reflux disease: a systematic review
.
Gut
.
2005
May
;
54
(
5
):
710
7
.
[PubMed]
0017-5749
10.
Gell
DA
.
Structure and function of haemoglobins
.
Blood Cells Mol Dis
.
2018
May
;
70
:
13
42
.
[PubMed]
1079-9796
11.
Hannig
C
,
Hannig
M
,
Attin
T
.
Enzymes in the acquired enamel pellicle
.
Eur J Oral Sci
.
2005
Feb
;
113
(
1
):
2
13
.
[PubMed]
0909-8836
12.
Hannig
C
,
Spitzmüller
B
,
Miller
M
,
Hellwig
E
,
Hannig
M
.
Intrinsic enzymatic crosslinking and maturation of the in situ pellicle
.
Arch Oral Biol
.
2008
May
;
53
(
5
):
416
22
.
[PubMed]
0003-9969
13.
Hemingway
CA
,
Shellis
RP
,
Parker
DM
,
Addy
M
,
Barbour
ME
.
Inhibition of hydroxyapatite dissolution by ovalbumin as a function of pH, calcium concentration, protein concentration and acid type
.
Caries Res
.
2008
;
42
(
5
):
348
53
.
[PubMed]
0008-6568
14.
Kawasaki
T
,
Ikeda
K
,
Takahashi
S
,
Kuboki
Y
.
Further study of hydroxyapatite high-performance liquid chromatography using both proteins and nucleic acids, and a new technique to increase chromatographic efficiency
.
Eur J Biochem
.
1986
Mar
;
155
(
2
):
249
57
.
[PubMed]
0014-2956
15.
Kawasaki
T
,
Niikura
M
,
Takahashi
S
,
Kobayashi
W
.
Strontium-phosphate hydroxyapatite high-performance liquid chromatography
.
Biochem Int
.
1987
Dec
;
15
(
6
):
1137
49
.
[PubMed]
0158-5231
16.
Kawasaki
T
,
Takahashi
S
,
Ikeda
K
.
Hydroxyapatite high-performance liquid chromatography: column performance for proteins
.
Eur J Biochem
.
1985
Oct
;
152
(
2
):
361
71
.
[PubMed]
0014-2956
17.
Kim
TH
,
Lee
KJ
,
Yeo
M
,
Kim
DK
,
Cho
SW
.
Pepsin detection in the sputum/saliva for the diagnosis of gastroesophageal reflux disease in patients with clinically suspected atypical gastroesophageal reflux disease symptoms
.
Digestion
.
2008
;
77
(
3-4
):
201
6
.
[PubMed]
0012-2823
18.
Kosoric
J
,
Hector
MP
,
Anderson
P
.
The influence of proteins on demineralization kinetics of hydroxyapatite aggregates
.
J Biomed Mater Res A
.
2010
Sep
;
94
(
3
):
972
7
.
[PubMed]
1549-3296
19.
Martini
T
,
Rios
D
,
Cassiano
LP
,
Silva
CM
,
Taira
EA
,
Ventura
TM
, et al
Proteomics of acquired pellicle in gastroesophageal reflux disease patients with or without erosive tooth wear
.
J Dent
.
2019
Feb
;
81
:
64
9
.
[PubMed]
0300-5712
20.
Meurman
JH
,
Toskala
J
,
Nuutinen
P
,
Klemetti
E
.
Oral and dental manifestations in gastroesophageal reflux disease
.
Oral Surg Oral Med Oral Pathol
.
1994
Nov
;
78
(
5
):
583
9
.
[PubMed]
0030-4220
21.
Milosevic
A
,
Young
PJ
,
Lennon
MA
.
The prevalence of tooth wear in 14-year-old school children in Liverpool
.
Community Dent Health
.
1994
Jun
;
11
(
2
):
83
6
.
[PubMed]
0265-539X
22.
Moazzez
R
,
Bartlett
D
,
Anggiansah
A
.
Dental erosion, gastro-oesophageal reflux disease and saliva: how are they related?
J Dent
.
2004
Aug
;
32
(
6
):
489
94
.
[PubMed]
0300-5712
23.
Muñoz
JV
,
Herreros
B
,
Sanchiz
V
,
Amoros
C
,
Hernandez
V
,
Pascual
I
, et al
Dental and periodontal lesions in patients with gastro-oesophageal reflux disease
.
Dig Liver Dis
.
2003
Jul
;
35
(
7
):
461
7
.
[PubMed]
1590-8658
24.
Preetha
A
,
Sujatha
D
,
Patil
BA
,
Hegde
S
.
Oral manifestations in gastroesophageal reflux disease
.
Gen Dent
.
2015
May-Jun
;
63
(
3
):
e27
31
.
[PubMed]
0363-6771
25.
Santiago
AC
,
Khan
ZN
,
Miguel
MC
,
Gironda
CC
,
Soares-Costa
A
,
Pelá
VT
, et al
A new sugarcane cystatin strongly binds to dental enamel and reduces erosion
.
J Dent Res
.
2017
Aug
;
96
(
9
):
1051
7
.
[PubMed]
0022-0345
26.
Schlueter
N
,
Ganss
C
,
Pötschke
S
,
Klimek
J
,
Hannig
C
.
Enzyme activities in the oral fluids of patients suffering from bulimia: a controlled clinical trial
.
Caries Res
.
2012
;
46
(
2
):
130
9
.
[PubMed]
0008-6568
27.
Schlueter
N
,
Hardt
M
,
Klimek
J
,
Ganss
C
.
Influence of the digestive enzymes trypsin and pepsin in vitro on the progression of erosion in dentine
.
Arch Oral Biol
.
2010
Apr
;
55
(
4
):
294
9
.
[PubMed]
0003-9969
28.
Schroeder
PL
,
Filler
SJ
,
Ramirez
B
,
Lazarchik
DA
,
Vaezi
MF
,
Richter
JE
.
Dental erosion and acid reflux disease
.
Ann Intern Med
.
1995
Jun
;
122
(
11
):
809
15
.
[PubMed]
0003-4819
29.
Schulz
BL
,
Cooper-White
J
,
Punyadeera
CK
.
Saliva proteome research: current status and future outlook
.
Crit Rev Biotechnol
.
2013
Sep
;
33
(
3
):
246
59
.
[PubMed]
0738-8551
30.
Schweigel
H
,
Wicht
M
,
Schwendicke
F
.
Salivary and pellicle proteome: A datamining analysis
.
Sci Rep
.
2016
Dec
;
6
(
1
):
38882
.
[PubMed]
2045-2322
31.
Siqueira
WL
,
Zhang
W
,
Helmerhorst
EJ
,
Gygi
SP
,
Oppenheim
FG
.
Identification of protein components in in vivo human acquired enamel pellicle using LC-ESI-MS/MS
.
J Proteome Res
.
2007
Jun
;
6
(
6
):
2152
60
.
[PubMed]
1535-3893
32.
Sujatha
S
,
Jalihal
U
,
Devi
Y
,
Rakesh
N
,
Chauhan
P
,
Sharma
S
.
Oral pH in gastroesophageal reflux disease
.
Indian J Gastroenterol
.
2016
May
;
35
(
3
):
186
9
.
[PubMed]
0254-8860
33.
Taira
EA
,
Ventura
TM
,
Cassiano
LP
,
Silva
CM
,
Martini
T
,
Leite
AL
, et al
Changes in the proteomic profile of acquired enamel pellicles as a function of their time of formation and hydrochloric acid exposure
.
Caries Res
.
2018
;
52
(
5
):
367
77
.
[PubMed]
0008-6568
34.
Tjäderhane
L
,
Buzalaf
MA
,
Carrilho
M
,
Chaussain
C
.
Matrix metalloproteinases and other matrix proteinases in relation to cariology: the era of ‘dentin degradomics’
.
Caries Res
.
2015
;
49
(
3
):
193
208
.
[PubMed]
0008-6568
35.
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
36.
Ventura
TM
,
Ribeiro
NR
,
Dionizio
AS
,
Sabino
IT
,
Buzalaf
MA
.
Standardization of a protocol for shotgun proteomic analysis of saliva
.
J Appl Oral Sci
.
2018
Jun
;
26
(
0
):
e20170561
.
[PubMed]
1678-7757
37.
Yu
YD
,
Zhu
YJ
,
Qi
C
,
Jiang
YY
,
Li
H
,
Wu
J
.
Hydroxyapatite nanorod-assembled porous hollow polyhedra as drug/protein carriers
.
J Colloid Interface Sci
.
2017
Jun
;
496
:
416
24
.
[PubMed]
0021-9797
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.
You do not currently have access to this content.