Archives of Oral Biology 108 (2019) 104522

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Archives of Oral Biology

journal homepage: www.elsevier.com/locate/archoralbio

Review

Dental caries: Genetic and protein interactions T

a,⁎ a b,c a
Tayla Cavallari , Letícia Yumi Arima , Adriano Ferrasa , Samuel Jorge Moysés ,
Simone Tetu Moysésa, Roberto Hirochi Heraib,d, , Renata Iani Wernecka,
⁎ ⁎

a
Graduate Program in Dentistry, Dentistry Department, School of Life Sciences, Pontifícia Universidade Católica do Paraná, Rua Imaculada Conceição 1155, Curitiba,
Paraná, Brazil, 80215-901
b
Graduate Program in Health Sciences (PPGCS), School of Medicine, Pontifícia Universidade Católica do Paraná (PUCPR), Rua Imaculada Conceição 1155, Curitiba,
Paraná, Brazil, 80215-901
c
Informatics Department, Universidade Estadual de Ponta Grossa (UEPG), Paraná, Brazil, 84030-900
d
Research Division, Lico Kaesemodel Institute, Paraná, Brazil, 80240-000

ARTICLE INFO ABSTRACT

Keywords: Objective: To present a genetic and protein interaction analysis associated with dental caries.
Dental diseases Material and Methods: The first step was to conduct a systematic literature review (SLR) through an electronic
Genetics database search. Case-controls that reported associations between genes and dental caries were the main type of
Epidemiology study design used as inclusion criteria, retrieved from the PubMed and the Virtual Health Library databases,
comprising the chronological range from 1982 to 2017. The SLR was guided by PRISMA protocol and the
methodological quality of the studies was established through Newcastle-Ottawa Scale (NOS). In the second step,
the String Protein Interaction (SPI) approach was used to analyze protein interaction (by esyN software) and also
the Ingenuity Pathway Analysis (IPA) to check biological pathways associated with dental caries genes.
Results: A total of 51 articles were included to perform this SLR, describing a number of 27 genes associated with
dental caries development. At the genetic level, 23 genes have at least one other gene with which they interact.
The genes TUFT1, VDR, TFIP11, LTF, HLA-DRB1, MMP2, MMP3 and MUC5B were shown to be connected in
interactive networks by at least 10 other genes.
Conclusion: It is essential to apprehend the multifactorial pattern of inheritance in human disease. This study
presents pathways which may be directly correlated with several dental caries phenotype and this contributes to
a better understanding of this disease, opening up a wider range of biotechnology options for its effective control
in the future.

1. Introduction 2010).
The genetic case-control study design has been frequently used by
Dental caries is a chronic disease with a multifactorial etiology. researchers worldwide to investigate different types of diseases and also
There is considerable evidence about the importance of environmental dental caries. These studies aim to compare affected and unaffected
and behavioral factors in dental decay development (Fejerskov, 2014). individuals as well as to test whether a particular allele occurs at a
However, the polarization of this disease is still occurring and im- significantly different frequency between paired groups (Izakovicova
portant variances among different population groups are evident Holla et al., 2017; Li, Hu, Zhou, Xie, & Zhang, 2015; Olszowski et al.,
(Tanner et al., 2013). 2017; Wendell et al., 2010; Valarini, Maciel, Moura, & Poli-Frederico,
Despite the available knowledge on several risk factors related to 2012; Choi et al., 2016; Cogulu et al., 2017; Ergöz et al., 2014).
the predisposition of dental caries, there may be some individual var- Several studies have already reported findings associating different
iations that might help to explain why some people exposed to the same genes with dental caries (Banderas-Tarabay, Zacarías-D’Oleire,
risk factors develop the disease and others do not. Therefore, studies Garduño-Estrada, Aceves-Luna, & González-Begné, 2002; Filho et al.,
have been conducted to better understand the nature of the genetic 2016; Haznedaroğlu et al., 2015; Kulkarni et al., 2013; Robino et al.,
influence on dental caries (Vieira, 2012; Werneck, Mira, & Trevilatto, 2015; Olszowski, Adler, Janiszewska-Olszowska, Safranow, & Chlubek,

Corresponding authors.
⁎

E-mail addresses: [email protected] (T. Cavallari), [email protected] (L.Y. Arima), [email protected] (A. Ferrasa),
[email protected] (S.J. Moysés), [email protected] (S. Tetu Moysés), [email protected] (R. Hirochi Herai), [email protected] (R. Iani Werneck).

https://doi.org/10.1016/j.archoralbio.2019.104522
Received 19 November 2018; Received in revised form 9 July 2019; Accepted 11 August 2019
0003-9969/ © 2019 Elsevier Ltd. All rights reserved.
T. Cavallari, et al. Archives of Oral Biology 108 (2019) 104522

Fig. 1. Flowchart of the systematic review for the screening of genetic variants associated with dental caries.

2015; Yu, Bixler, Goodman, Azen, & Karn, 1986; Hu et al., 2015), yet year_cluster:("2015″ OR "2014″ OR "2017″ OR "2013″ OR "2012″ OR
there is a shortage of knowledge regarding the metabolic pathways "2016″ OR "2011″ OR "2008″ OR "2010″ OR "2002″ OR "2005″ OR
involved and whether these pathways are directly correlated with "2004″ OR "2007″ OR "2009″ OR "2006″ OR "2000″ OR "2001″ OR
dental caries phenotype. It is important to understand the set of dif- "1993″ OR "1995″ OR "1998″ OR "1994″ OR "1999″ OR "2003″ OR
ferent pathways and associated genes driving similar dental pheno- "1990″ OR "1996″ OR "1991″ OR "1997″ OR "2018″ OR "1986″ OR
types, although different genes are affected. Since protein networks "1987″ OR "1989″ OR "1992″ OR "1983″ OR "1985″ OR "1982″ OR
consist of various types of interaction and regulation, networks re- "1988″ OR "1984″)).
flecting this complex scenario will provide a better understanding of the This research included genetic studies on humans, with no restric-
disease. Thus, the aim of this study was to present, for the first time, a tions regarding age or ethnicity. Case-control genetic studies were
genetic and protein interaction network analysis with the related dental primarily selected because this design has methodological strengths
caries genes assembled by means of a SLR. that often allow the most relevant candidate genes to be revealed.
Additionally, we also included well-conducted cross-sectional or cohort
studies, when affected and non-affected individuals for dental caries
2. Materials and methods
were separated and analyzed to compare the allelic and genotypic
frequencies between the groups. This SLR included articles published in
A SLR was performed to generate a catalog of genes already de-
Portuguese, English, French, or Spanish between January 1982 and
scribed in published works related to dental caries. The SLR followed
December 2017. The electronic publication databases were PubMed
the recommendations proposed in the Preferred Reporting Items for
and the Virtual Health Library (VHL), which included LILACS and
Systematic Reviews and Meta-Analyses (PRISMA) guidelines, and
Medline.
sought to answer the following question: “Which genes are associated
Two reviewers (T.C and L.Y.A) performed independent searches for
to the development of dental caries?” The approach used to structure
articles in the databases and selected the eligible articles by reading
the research question was based on the Population, Intervention,
their titles and abstracts. Following this first step selection, duplicate
Comparison and Outcome framework (PICO), following the parameters:
articles were excluded. The Newcastle-Ottawa Scale (NOS) was used to
P = genes
evaluate the methodological quality of the articles selected. The two
I = not applicable
reviewers read all articles in their entirety to determine whether they
C = individuals affected or unaffected by dental caries
met the criteria for inclusion in the review. In the event of disagree-
O = association with dental caries
ment, a third reviewer was consulted to help disentangle the issue and
The search strategy included the following keywords: dental caries
to reach consensus (R.I.W). Animal studies, narrative reviews, studies
OR dental decay OR dental caries susceptibility OR caries experience AND
that used a methodological design that did not adhere to the inclusion
genetic OR genetic epidemiology OR genes OR polymorphism. The fol-
criteria and studies with bacteria were excluded (Fig. 1). After com-
lowing search strategy was used in the PubMed database: (((dental
bining the findings of the SLR, the genes that showed association with
caries[Title/Abstract] OR dental decay[Title/Abstract]) OR dental
dental caries were then evaluated in relation to their metabolic
caries susceptibility[Title/Abstract]) OR caries experience[Title/
pathway. To perform the protein interaction analysis, it was used the
Abstract]) AND ((((genetic therapy[Title/Abstract] OR genes[Title/
String Protein Interaction (SPI) approach (von Mering et al., 2003). This
Abstract]) OR molecular epidemiology[Title/Abstract]) OR genetic
was used to detect interactions reported from curated databases, ex-
[Title/Abstract]) OR polymorphism[Title/Abstract]) AND ("1982/01/
perimentally determined and text-mining based interaction. Only
01″[PDAT] : "2017/12/31″[PDAT]). For the VHL, the following search
medium confidence interactions were reported (M = 0.4). For genetic
strategy was used: tw:((tw:(dental caries)) AND (tw:(genetic))) AND
interaction analysis, it was used the esyN software (Bean et al., 2014) to
(instance:"regional") AND (la:("en" OR "es" OR "pt" OR "fr") AND

2
 Table 1
Studies included in the SRL relating genetic and dental caries.
Study Country Genes studied Case/ Reported Average age Caries diagnosis criterion controlled risk Statistical analysis Summarized result NCO
control sample factors
T. Cavallari, et al.

calculation
Anderson & USA DB, PRP1, and PR 26/28 No Adults aged at least Control group with no caries history and No Chi-square No difference in the 6
Mandel, 1982 24 years old (Not case group with DMFT > 15 distribution of phenotypes
mention the
average)
Anderson et al., USA DB, PRP1, PR, PS, 47/46 No Not described Case group with at least 5 caries lesions No Chi-square No difference in the 6
1982 PMF, and PB distribution of phenotypes
*Yu et al., 1986 USA PR, PA PS, PMF, Total of No 9,7 DMFT No Regression coefficient The PR and PA genes may be 6
PIF, and DB 306 homogeneity test, associated with susceptibility
covariance analysis to dental caries
*Banderas- Mexico MG1, MG2, and Total of No 19 Control group: DMFT ≤4 and case group No ANOVA, t test, Pearson's DMFT ≥ 10 showed a 6
Tarabay et al., PRP1 120 DMFT: ≥10 correlation significant reduction of MG1,
2002 MG2 and PRP1 protein
Pehlivan et al., Turkey MBL 42/40 No 9.78 case group and Not reported No Chi-square No difference between groups 6
2005 10.7 control group
Slayton et al., USA AMELX, ENAM, 92/92 No Children from 3 to 5 Control group: DMFT ≤4 and control group No Chi-square There was no association 7
2005 AMBN, 4TUFT1, years old. (Not without evidence or history of caries
TFIP11, and KLK4 mention the (including white spot lesions)
average)
*Deeley et al., Guatemala AMELX, ENAM, 66/44 Yes 32.6 case group and Control group: DMFT ≤2 and case group No Chi-square and Student's AMELX may be related to the 8
2008 AMBN, TUFT1, 23.5 control group DMFT: ≥3. t susceptibility with dental
and TFIP11 caries.
*Patir et al., 2008 Turkey AMELX, ENAM, 91/82 No 4.8 case group and Case group: DMFT : ≥4 and control group No Chi-square and Student's AMELX, AMBN, and TUFT1 8
AMBN, TUFT1, 5.9 control group without evidence or history of caries t may be associated with dental
and TFIP11 (including white spot lesions) caries

3
*Azevedo et al., Brazil LTF 48/62 No 12 Case group DMFT:≥1 (white spots were No Chi-square Protective effect on dental 8
2010 considered), and control group with no caries experience
dental caries experience
*Ozturk et al., USA DEFB1 201/95 No 40.23 Younger than 30 years old (control group No Chi-square, Fisher's exact DEFB1 polymorphisms are 7
2010 with DMFT: < 14 and case group: DMFT test, multiple logistic potential markers for dental
≥14). Older than 30 years old (control regression, Z-test caries
group DMFT: < 9 and case group DMFT: ≥
9).
Brancher et al., Brazil LTF 25/25 Yes 12 Control group: DMFT = 0 and case group No Chi-square There was no association 7
2011 DMF: ≥4
* Kang et al., 2011 South Korea AMELX 87/33 No 21.3 case group and Control group with DMFT ≤2 and case Yes Fisher's exact test, SNPs of AMELX might be 7
23.2 control group group with DMFT ≥3 OH multiple logistic associated with dental caries
regression

Buczkowska-Radliska et al., Poland MUC7 31/22 No 20-21 years old (Not mention the average) Control group: DMFT≤7 and Yes Mann-Whitney U test, There was no association 7
2012 case group DMFT: ≥15 OH, B chi-square
*Olszowski et al., 2012 Poland AMELX, ENAM, MBL2, 380/376 No Children aged 5 years (deciduous dentition) and Control group: DMFT < 3 and No Chi-square and Fisher's MBL2 was associated with dental 7
and MASP2 13 years old (permanent dentition) case group DMFT: ≥3 exact test caries experience

*Shimizu et al., 2012 Philippines, Turkey, AMELX, ENAM, 742/ No 25,8 Children: control group DMFT = 0 to 2 and case No Chi-square, Fisher's AMELX, AMBN, TUFT1, and 8
Argentina, Brazil AMBN, TUFT1, and 1077 group: DMFT ≥3. Adolescents: control group DMFT exact test ENAM may be associated with
TFIP11 = 0 to 5 and case group: DMFT ≥ 6. Adults: control dental caries experience
group: DMFT = 0 to 8 and case group: DMFT ≥9
*Tannure, Küchler et al., Brazil MMP20 227/ No 8.7 case group Control group: DMFT = 0 and case group divided Yes Chi-square, t-test, MMP20 may be associated with 8
2012 161 and 9.4 control into low caries experience (DMFT =1, moderate OH, binary logistic dental caries
group (DMFT ≥ 2 and ≤ 3) and high (DMFT ≥ 4) DH regression
*Tannure, Küchler et al., Brazil MMP2, MMP9, 293/ No 8.8 case group Case group: DMFT ≥ 1 and control group: DMFT = 0 Yes Student's t, chi-square, MMP13 may be associated with 8
2012; Tannure, MMP13, and TIMP2 212 and 9.4 control OH, binary regression dental caries
Kuchler et al., 2012 group DH
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 Table 1 (continued)

Valarini et al., 2012 Brazil HLA-DR4, HLA-DQ4, 99/65 No 15.9 case group Case group: DMFT ≥1 and control group: DMFT = 0 Yes logistic regression, chi- There was no association 8
HLA-DQ5, HLA-DQ6 and 15.7 S square
and HLA-DQ2 control group
T. Cavallari, et al.

*Wang et al., 2012 USA ENAM, TUFT1, 82/251 No 5,2 Case group: DMFT ≥1 and control group: DMFT = 0 Yes Linear regression, DSPP, KLK4, and AQP5 may be 8
DSPP, KLK4, OH, S logistic regression associated with dental caries
MMP20, AQP5, and protection
SPP1
*Kulkarni et al., 2013 Canada GLUT2 and TAS1R2 Total of No 26,4 WHO, ICDAS, and interproximal radiographic Yes T test, chi-square, GLUT2 and TAS1R2 were 6
80 analyses OH, B, ANOVA associated with dental caries
G, ST susceptibility.
Gasse et al., 2013 France AMELX 212/ Yes 7.6 case group WHO, ICDAS, and interproximal radiographic Yes multivariate regression, There was no association in this 8
146 and 22 control OH, Fisher's exact test, odds population
group DH, S ratio
Yang et al., 2013 China MBL 62/68 No 3,5 Case group with ECC and control group with no No Student's t test, chi- There was no association of 8
history of caries square MBL with early childhood
caries
*Chaussain et al., 2014 France ENAM 212/ No 7.6 case group According to the WHO Yes Logistic regression, odds ENAN is a candidate gene for 8
146 and 22 control OH, ratio, multivariate susceptibility to dental caries
group DH, S analyses

Volckova et al., Czech LTF 482/155 No Children aged 11-13. Case group DMFT: ≥1 and Yes Fisher's exact test, There was no association in this 7
2014 Republic (Not mention the control group: DMFT = 0 G, B Pearson's test, odds population
average) ratio
Doetzer et al., Brazil LTF 346/331 Yes 12 Control group: DMFT = 0 Yes Chi-square, Fisher's LTF were associated with dental caries 8
2015 and case group: DMFT ≥1, G, B, ST exact test, Mantel protection
(low caries experience) or Haenszel's exact
DMFT ≤2 (high caries test
experience)

4
*Izakovicova Holla Czech GLUT2 and TAS1R2 482/155 No Children aged 11-13. Case group: DMFT ≥ 1 and Yes Fisher's exact test, GLUT2 and TAS1R2 may influence the 7
et al., 2015 Republic (Not mention the control group: DMFT = 0 G, B, OH, odds ratio, logistic risk for dental caries
average) B regression
*Li et al., 2015 China CA-VI 164/191 No 51.16 case group and Case group: DMFT ≥ 3 and Yes Chi-square, odds The ACA haplotype of the CA-VI gene 7
47.44 control group. control group: DMFT ≤ 2 S ratio, logistic may be associated with susceptibility
regression to dental caries
*Robino et al., Italy TAS1R2 and GLUT2 647 No 44.9 DMFT and panoramic Yes Association It was identified a direct association 8
2015 radiography analyses OH analysis, between variants in TAS1R2 and
Regression Analysis GLUT2 genes and caries prevalence
*Abbasolu et al., Turkey AMBN, AMELX, ENAM, KLK4, MMP20 136/123 No 4.6 Case group: DMFT≥1 and Yes Student's t, chi- The TT genotype (ALOX1) was 8
2015 TUFT1, DEFB1, LTF, and ALOX15 control group: DMFT = 0 OH, DH square, Fisher's associated as a risk factor for the
exact test, development of early childhood
multivariate caries. The GG (ENAM), AG and GG
analysis, logistic (KLK4), CT (LTF), and GG (TUFT1)
regression genotypes showed protection
*Antunes et al., Brazil MMP2, MMP3, MMP9, MMP20, TIMP1, 786 No 3.9 The presence of WSL (White Yes Chi-square, Fisher’s MMP9 and MMP20 are involved in 8
2015 and TIMP2 spot lesions and/or ECC) and OH, DH exact test WSL and ECC development
without disease (the absence
of WSL or ECC
*Olszowski et al., Poland ACE I/D 120/41 No Children aged 15 Case group DMF: ≥1 and No Chi-square, Fisher’s The DD genotype of ACE I/D 8
2015 years old. (Not control group DMFT = 0 exact test polymorphism might be protective
mention the average) against dental caries
*Hu et al., 2015 China VDR TaqI 264/219 No 50 Case group: DMFT ≥ 1 and No Odds ratio and chi- The T allele may be a genetic 7
control group: DMFT = 0 square determinant factor for the
development of caries disease
*Haznedaroglu Turkey TAS1R2 and TAS1R3 Total of 184 No Children aged According to the WHO Yes Fisher's exact test, Moderate caries disease experience 6
et al. 2015 between 7 and 12 OH, DH, chi-square, for the TAS1R3 rs307355 gene and
years (Not mention S Student's t, logistic high for the TAS1R2 rs35874116 gene
the average) regression test
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 Table 1 (continued)

*Gerreth et al., Poland ENAM 48/48 No Children aged 20-42 Diagnoses included No Chi-square, Fisher's Polymorphisms in the ENAM gene 8
2016 months. (Not cavitated lesions and non- exact test may be a predictor of caries disease in
mention the average) cavitated incipient initial children
T. Cavallari, et al.

lesions or white spots

*Saha et al., 2016 India AMELX 30/30 Yes Children aged 5-12 Case group: DMFT ≥4, No Chi-square, t-test High correlation with dental caries 7
years. (Not mention control group had no history
the average) of dental caries
*Filho et al., 2016 Brazil MMP20 184 No Not described the Low caries experience: No Chi-square, Fisher’s MMP20 rs1784418 C > T appears to 7
avarege. Children DMFT = 1. Moderate caries exact test protect against dental caries
aged 4-7 years. experience DMF= 2–3, and
high caries experience:
DMFT = 4 or higher
*Karayasheva Bulgaria MMP2 and MMP3 41/41 No Children aged 20-32 Control group: DMFT ≤ 5 Yes Chi-square, odds The genes are probably associated 7
et al., 2016 years. (Not mention and case group: DMFT ≥ 14 OH, DH, ratio with caries disease
the average) G, ST, B
*Linhartova Czech ACE I/D 561/182 No Children aged 13-15 Case group: DMFT ≥ 1 and Yes Chi-square, Fisher’s ACE I/D polymorphism may be 8
Borilova et al., Republic years. (Not mention control group: DMFT = 0. G, B exact test associated with caries in permanent
2016 the average) dentition
*Yildiz et al., 2016 Turkey AMELX, CA-VI, DEFB1, and TAS2R38 77/77 Yes 28.0 Control group: DMFT ≤ 5 Yes Chi-square, Polymorphisms in the CA6, DEFB1, 8
and case group: DMFT ≥ 14, OD, DH, multiple regression, and TAS2R38 genes associated with
B, ST Mann-Whitney U biofilm and lactobacilli can increase
test the risk of developing caries
*Cogulu et al., Turkey APAL, FOKL, CDX2 and TAQL 112/38 No 10.19 High caries risk group: Yes Chi-square, Fisher’s No statistically significant differences 8
2017 DMFT > 4, moderate caries OH, DH, exact test were detected
risk group: DMFT= 1–4) B
and caries-free group.
*Gerreth et al., Poland AMELX, AMBN, TUFT1, TFIP11, MMP20 and 48/48 No 30.26 mounths Dental evaluation concerned Yes Chi-square, Fisher’s Polymorphisms in AMELX, AMBN, 8
2017 KLK4 the occurrence of teeth with D,OH exact test TUFT1, KLK4 genes may be genetic

5
carious cavities as well as markers that contribute to dental
teeth with initial (incipient) caries
caries lesions (non
cavitated,white spot)
*Alyousef et al., Saudi MMP9, MBL2, MMP2, and TIMP2 102/100 No Children aged 5-13 According to the WHO Yes Chi-square, odds MBL2 gene was shown to be 8
2017 Arabia years. (Not mention OH,DH, S ratio, T-test associated with a high prevalence of
the average) caries
*Cavallari, Brazil MUC5B 100/100 Yes 18.43 case group and According to ICDAS. Control Yes Chi-square, Fisher’s Genetic polymorphisms of the MUC5B 8
Salomão et al., 18.52 control group group: zero score for all OH, DH, exact test gene can influence dental caries
2017 dental elements B, S development
*Cavallari, Brazil KLK4 100/100 Yes 18.43 case group and According to ICDAS. Control Yes Chi-square, Fisher’s Genetic variations in the KLK4 gene 7
Salomão et al., 18.52 control group group: zero score for all OH, DH, exact test may contribute to dental decay
2017 dental elements B, S
Lips et al., 2017 Brazil DEFB1 162/342 No 3.2 control group According to DMFT Yes Shapiro-Wilk test, There was no association 8
and 4.7 case group OH, DH, Chi-square, Fisher’s
B exact test
Olszowski et al., Poland FCN2 82/175 No 15 According to DMFT No Chi-square, Fisher’s There was no association 8
2017 exact test
Izakovicova Holla Czech VDR Taq I 235/153 No Children aged 13-15 Case group: DMFT ≥ 1 and Yes Chi-square, Fisher’s There was no association 8
et al., 2017 Republic years. (Not mention control group: DMFT = 0 B, G exact test
the average)
Kastovsky et al., Czech BMP2 and DLX3 Primary dentition No Children aged 2-6 According to DMFT No Chi-square, Fisher’s Variability in BMP2 and DLX3 was not 7
2017 Republic 113/83 years old (primary exact test associated with caries
Permanent dentition) and 13-15
dentition542/176 (permanent
dentition) (Not
mention the
average).
(continued on next page)
Archives of Oral Biology 108 (2019) 104522
T. Cavallari, et al. Archives of Oral Biology 108 (2019) 104522

8 show complex interaction, enhancing genetic interaction, suppressing

8
genetic interaction and physical interaction. It is based on several

be used as a risk factor of dental caries

VDR-Fok I gene polymorphisms may

ENAM variant (rs12640848) cannot

be associated with susceptibility to
permanent tooth caries in Chinese
biological databases available, such as KEGG (Kanehisa & Goto, 2000)
and Reactome (Fabregat et al., 2016). To check the biological pathways

in the Czech population

associated with a list of genes (regarding dental caries), it was used
Ingenuity Pathway Analysis (IPA, http://www.ingenuity.com) with
adolescent

default parameters.

3. Results

After applying the SLR protocol, it was found 549 articles in the
PubMed database and 737 in the VHL database. Next, by reading the
titles and abstracts, 61 articles were selected from PubMed and 39 from
Chi-square, Fisher’s

the VHL. Duplicated articles and those that did not meet the inclusion
Chi-square

exact test

criteria were excluded. A final number of 51 articles were included in

this SLR (Fig. 1). These articles where then scrutinized by full-text
analysis to collect the following information: authors and year of pub-
lication, country where the research was performed, studied genes,
number of participants in each group (case-control), average age of
individuals, dental caries diagnostic criteria used, controlled risk fac-
OH,
Yes

No
G

tors, statistical analyses performed, and the assigned score according to

the NOS, which are presented in Table 1.
Legends:Oral Hygiene Habits: OH; Dietary Habits: DH; Gingivitis: G; Biofilm: B; Socioeconomic/ demographic characteristic; Salivary Tests: ST.

The SLR counted in 54 different genes (ACE I/D, ALOX15, AMBN,

Case group:DMFT ≥1 and
control group DMFT = 0

AMELX, APAL, AQP5, BMP2, CA-VI, CDX2, DB, DEFB1, DLX3, DSPP,
According to DMFT

ENAM, FOKL, FCN2, GLUT2, HLA-DR4, HLA-DQ2, HLA-DQ3, HLA-

DQ4, HLA-DQ5, HLA-DQ6, KLK4, LTF, MASP2, MBL MBL2, MG1, MG2,
MMP2, MMP3, MMP9, MMP13, MMP20, MUC5B, MUC7, PA, PB, PIF,
PMF, PR, PRP1, OS, SPP1, TAS1R2, TAS1R3, TAS2R38, TIMP1, TIMP2,
TUFT1, TFIP11, VDR-TAQI, VDR-FOK). Furthermore, 27 of those genes
were found to be associated either with protection or risk of dental
caries (PRP1, PR, PA, MG1, MG2, AMELX, ENAM, TUFT1, KLK4, HLA-
12.24 case group and

mention the average)

dentition 13-15 (Not
12.20 control group

Primary dentition

DR4, TAS1R3, TAS2R38, MBL2, MMP20, MMP2, MMP9, MMP13,

3.8 Permanent

GLUT2, TAS1R2, CA-VI, DEFB1, ALOX15, VDR-TAQI, MMP3, CA6,

MUC5B,VDR-FOK) (Table 1).
Most studies analyzed genes related to enamel formation, develop-
ment and mineralization, including amelogenin (AMELX) (Kang, Yoon,
Lee, & Cho, 2011; Slayton, Cooper, & Marazita, 2005; Shimizu et al.,
No

No

2012; Gasse et al., 2013), tuftelin (TUFT1) (Partir et al., 2008), ame-
loblastin (AMBN) (Deeley et al., 2008; Shimizu et al., 2012), enamelin
Primary dentition

dentition541/177

(ENAN) (Gerreth, Zaorska, Zabel, Borysewicz-Lewicka, & Nowicki,

Permanent
200/200

109/78

2016; Linhartova Borilova et al., 2017), aquaporin (AQP5) (Wang et al.,

2012), kallikrein (KLK4) (Wang et al., 2012; Cavallari et al., 2017) and
metalloproteinase genes (Karayasheva, Glushkova, Boteva, Mitev, &
Kadiyska, 2016; Tannure, Kuchler et al., 2012; Tannure, Küchler et al.,
2012). Other studies analyzed genes related to the host immune re-
sponse, including mannose-binding lectin (MBL) (Alyousef et al., 2017;
Pehlivan et al., 2005) and defensin beta 1 (DEFB1) (Lips et al., 2017;
Ozturk, Famili, & Vieira, 2010). Genes related to the composition of
saliva were also examined, including salivary carbonic anhydrase VI
VDR-FOK

(CA6) (Li et al., 2015), lactotransferrin (LTF) (Azevedo et al., 2010;

ENAM

Brancher et al., 2011; Doetzer et al., 2015) and mucins (MUC7)

(Buczkowska-Radliska, Pol, Szmidt, & Bińczak-Kuleta, 2012) and
(MUC5B) (Cavallari, Moysés, Moysés, & Werneck, 2017; Cavallari,
*Articles with association for dental caries.

Salomão, Moysés, Moysés, & Werneck, 2017).

The major part of the included studies was conducted with adults.
However, studies with preschool children related to early childhood
caries were also explored (Bagherian, Nematollahi, Afshari, & Moheghi,
Republic

2008; Yang, Wang, & Qin, 2013; Abbasoğlu et al., 2015; Antunes et al.,
Czech
China

2016;Linhartova Borilova et al., 2016; Kastovsky et al., 2017; Gerreth,

Zaorska, Zabel, Borysewicz-Lewicka, & Nowicki, 2017).
Table 1 (continued)

The genes associated with dental caries also showed a correlation at

Borilova et al.,
Kong, & Chen,
*Yu, Jiang, Sun,

genetic and protein levels (Table 2). At the genetic level, it was found
Linhartova
2017

2017

that 23 genes have at least another interacting gene (Fig. 2). The genes
TUFT1, VDR, TFIP11, LTF, HLA-DRB1, MMP2, MMP3 and MUC5B are
all central elements (known as HUB), but they are connected to inter-
acting networks formed by at least 10 other genes. The genes TAS1R2,

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Table 2 Table 2 (continued)

Associated metabolic pathways of genes associated with dental caries.
Canonical Pathways Molecules
Canonical Pathways Molecules
Ovarian Cancer Signaling MMP2
Inhibition of Matrix Metalloproteases MMP20,MMP3,MMP13,MMP2 IL-12 Signaling and Production in ALOX15
HIF1α Signaling MMP20,MMP3,SLC2A2,MMP13,MMP2 Macrophages
Bladder Cancer Signaling MMP20,MMP3,MMP13,MMP2 Phagosome Maturation HLA-DRB1
Granulocyte Adhesion and Diapedesis MMP20,MMP3,MMP13,MMP2 Th2 Pathway HLA-DRB1
Agranulocyte Adhesion and Diapedesis MMP20,MMP3,MMP13,MMP2 Type II Diabetes Mellitus Signaling SLC2A2
Leukocyte Extravasation Signaling MMP20,MMP3,MMP13,MMP2 PKCθ Signaling in T Lymphocytes HLA-DRB1
Colorectal Cancer Metastasis Signaling MMP20,MMP3,MMP13,MMP2 GNRH Signaling MMP2
Atherosclerosis Signaling ALOX15,MMP3,MMP13 Cdc42 Signaling HLA-DRB1
Oncostatin M Signaling MMP3,MMP13 Acute Phase Response Signaling MBL2
Gustation Pathway TAS1R2,TAS2R38,TAS1R3 eNOS Signaling AQP5
IL-17 Signaling MMP3,MUC5B Th1 and Th2 Activation Pathway HLA-DRB1
Airway Pathology in Chronic MMP2 Role of NFAT in Regulation of the HLA-DRB1
Obstructive Pulmonary Disease Immune Response
Axonal Guidance Signaling MMP3,MMP13,MMP2 Regulation of the Epithelial- MMP2
Hepatic Fibrosis/Hepatic Stellate Cell MMP13,MMP2 Mesenchymal Transition Pathway
Activation Dendritic Cell Maturation HLA-DRB1
Maturity Onset Diabetes of Young SLC2A2 IL-8 Signaling MMP2
(MODY) Signaling
Osteoarthritis Pathway MMP3,MMP13
Differential Regulation of Cytokine DEFB1
Production in Intestinal Epithelial
TAS1R3, AMBN, MBL2, MUC2, DEFB1 and SLC2A2 form small net-
Cells by IL-17A and IL-17F works, having less than 10 connections and only some of them interact
Role of Osteoblasts, Osteoclasts and MMP3,MMP13 with the other genes. At the protein level, most proteins are shown to be
Chondrocytes in Rheumatoid interacting with each other (Fig. 3). These interactions are restricted to
Arthritis
only direct associations formed by four groups, with the biggest group
B Cell Development HLA-DRB1
Complement System MBL2 having six proteins, and then four, three and two proteins forming
Antigen Presentation Pathway HLA-DRB1 different groups (Fig. 3). The proteins MMP20, ENAM, AMELX, AMBN
Intrinsic Prothrombin Activation KLK4 have the highest number of interactions, a total of six, all identified by
Pathway text mining-based analysis. The protein interaction between TAS1R3
Role of IL-17 F in Allergic MMP13
Inflammatory Airway Diseases
and TAS1R2 was the pair having four different methods that confirmed
Neuroinflammation Signaling Pathway HLA-DRB1,MMP3 the interaction, the highest between all interactions found.
Role of Macrophages, Fibroblasts and MMP3,MMP13 This study also investigated the biological pathways associated with
Endothelial Cells in Rheumatoid dental caries and genes, and found several canonical pathways asso-
Arthritis
ciated with different cellular functions and processes, such as Antigen
Autoimmune Thyroid Disease Signaling HLA-DRB1
Graft-versus-Host Disease Signaling HLA-DRB1 Presentation Pathway, Inhibition of Matrix Metalloproteases, L-17
Docosahexaenoic Acid (DHA) Signaling ALOX15 Signaling and HIF1a Signaling, which are illustrated in Fig. 4 and de-
Nur77 Signaling in T Lymphocytes HLA-DRB1 scribed in Table 2.
Calcium-induced T Lymphocyte HLA-DRB1 Most of the dental caries-related genes have also been associated
Apoptosis
Eicosanoid Signaling ALOX15
with different pathologies (Table 3), such as amelogenesis (genes KLK4,
Role of IL-17A in Arthritis MMP13 MMP20, ENAM, AMBN, AMELX), abnormal color of incisor (gene
Glioma Invasiveness Signaling MMP2 AMBN), abnormal morphology of enamel and molar crown (AMBN and
MSP-RON Signaling Pathway KLK4 MMP20, and AMBN, respectively), lack of enamel (gene AMBN), den-
T Helper Cell Differentiation HLA-DRB1
tinogenesis imperfecta (gene DSPP) and congenital anomaly of tooth
VDR/RXR Activation VDR
IL-17A Signaling in Airway Cells MUC5B (genes AMBN, AMELX, DSPP, ENAM, KLK4, MMP20).
Allograft Rejection Signaling HLA-DRB1
TGF-β Signaling VDR 4. Discussion
HER-2 Signaling in Breast Cancer MMP2
Crosstalk between Dendritic Cells and HLA-DRB1
Natural Killer Cells Genetic association studies have aimed to determine whether ge-
IL-4 Signaling HLA-DRB1 netic factors may affect dental decay, along with other variables. For
Altered T Cell and B Cell Signaling in HLA-DRB1 this study, 51 articles published over a 35-year period were selected
Rheumatoid Arthritis and the genetic epidemiology of dental caries was investigated. These
OX40 Signaling Pathway HLA-DRB1
Communication between Innate and HLA-DRB1
studies provide scientific evidence that genetic factors are involved in
Adaptive Immune Cells the development of dental caries and that several genes are associated
Antioxidant Action of Vitamin C SLC2A2 with the disease.
Type I Diabetes Mellitus Signaling HLA-DRB1 The included studies were assessed by the NOS scale. This research
GPCR-Mediated Nutrient Sensing in TAS1R3
instrument helps to verify the quality of studies according to certain
Enteroendocrine Cells
Neuroprotective Role of THOP1 in KLK4 methodological properties (selection, comparability and exposure). All
Alzheimer's Disease studies reached the minimum quality score required by the scale.
iCOS-iCOSL Signaling in T Helper Cells HLA-DRB1 Some risk factors for dental caries were surveyed in most studies,
Role of Tissue Factor in Cancer MMP13 including visible plaque index, tooth brushing frequency, food intake
Phagosome Formation MBL2
CD28 Signaling in T Helper Cells HLA-DRB1
between meals and the buffering capacity of saliva. Various factors
Th1 Pathway HLA-DRB1 were significantly associated, however there were some studies that
Role of Pattern Recognition Receptors MBL2 only performed bivariate analyses, failing to test the effects of en-
in Recognition of Bacteria and vironmental variables along with genetic variables (Anderson &
Viruses
Mandel, 1982; Anderson, Lamberts, & Bruton, 1982).
Sample size estimation is an important method to minimize the

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T. Cavallari, et al. Archives of Oral Biology 108 (2019) 104522

Fig. 2. Genetic interaction of genes associated with dental caries.

Genes associated with caries disease are highlighted in blue. All genetic interactions are restricted to genes having associations previously described in literature by in
silico on experimental models.

Fig. 3. Protein-protein interaction network of associated genes with dental caries.

Interaction network was restricted to the caries-related genes and did not include indirect correlations.

probability of error. The size of a sample influences in the precision of Deeley et al., 2008; Kang et al., 2011; Olszowski, Adler, Janiszewska-
the estimates and in the power of the study to draw conclusions. There Olszowska, Safranow, & Kaczmarczyk, 2012; Shimizu et al., 2012;
was a variation in the number of participants among the included ar- Gasse et al., 2013; Abbasoğlu et al., 2015; Yildiz, Ermis, Calapoglu,
ticles. In this research, only a few studies reported sample size calcu- Celik, & Turel, 2016; Saha, Sood, Sandhu, Diwaker, & Upadhyaye,
lation (Table 1). Authors should generally be required to provide de- 2016) and ENAM (Slayton et al., 2005; Patir et al., 2008; Deeley et al.,
tailed information on the sample size calculation approach used when 2008; Olszowski et al., 2012; Chaussain et al., 2014) were the most
performing population studies. commonly studied genes. It is very important to replicate the findings of
After a careful analysis of the articles, it was found that 29 genes genetic studies in different populations to confirm the results. The
were replicated by different researches in a multiplicity of populations above replicated genes were studied in different populations, sug-
(ACE I/D, DB, PRP1, PR, PS, PMF, MBL, AMELX, ENAM, AMBN, TUFT1, gesting that the associations with dental caries were not at random. On
TFIP11, KLK4, HLA-DR4, HLA-DQ5, HLA-DQ6, HLA-DQ2, LTF, MBL2, the other hand, some genes were also searched for replication in other
MMP20, MMP2,MMP3, MMP9, TIMP2, GLUT2, TAS1R2, CA-6, DEFB1, research groups, but the associations were not confirmed, such as for
VDR-TAQI). The genes AMELX (Slayton et al., 2005; Patir et al., 2008; the gene LTF. Just to give an example, in a region of Brazil it was found

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T. Cavallari, et al. Archives of Oral Biology 108 (2019) 104522

Fig. 4. Canonical pathways of caries-disease associated genes.

an association of the gene LTF (Azevedo et al., 2010; Doetzer et al., composition. The protein encoded by the ENAM gene, for example, is
2015), whilst the same gene in the Czech Republic population had no involved in the mineralization and structural organization of enamel
association (Volckova et al., 2014). (Gerreth et al., 2015). The AMELX gene encodes a member of the
It is evident that the most cited candidates genes are more biolo- amelogenin family, which is involved in bio mineralization during
gically plausible to be tested as caries-related phenotype, since their tooth enamel development (Saha et al., 2016). Furthermore, this study
function is directly associated with enamel formation or saliva also found that the identified dental caries-related candidate genes are

Table 3
Systematically identified genes with dental caries associated with dental formation and pathologies.
Diseases or Functions Annotation Molecules

Abnormal morphology of ameloblasts AMBN,MMP20

Enlargement of dental pulp chamber DSPP
Tooth development AMELX,AQP5,ENAM,MMP20
Amelogenesis imperfecta AMBN,AMELX,ENAM,KLK4,MMP20
Amelogenesis imperfecta hypomaturation type AMELX,KLK4,MMP20
Amelogenesis imperfecta type IB ENAM
Amelogenesis imperfecta type IC ENAM
Amelogenesis imperfecta type IF AMBN
Amelogenesis imperfecta X-linked 1 AMELX
Autosomal dominant dentin dysplasia, type ii DSPP
Autosomal recessive amelogenesis imperfecta AMBN,ENAM,KLK4,MMP20
Hypomaturation type amelogenesis imperfecta type IIA1 KLK4
Hypomaturation type amelogenesis imperfecta type IIA2 MMP20
Hypoplastic amelogenesis imperfecta AMELX,ENAM
Hypoplastic/hypomaturation amelogenesis imperfecta X-linked 2 AMELX
Pigmented hypomaturation type amelogenesis imperfecta KLK4,MMP20
Abnormal color of incisor AMBN
Abnormal morphology of enamel AMBN,MMP20
Abnormal morphology of molar crown AMBN
Lack of enamel AMBN
Autosomal dominant dentinogenesis imperfecta Shields type II DSPP
Dentinogenesis imperfecta - Shields type III DSPP
Congenital anomaly of tooth AMBN,AMELX,DSPP,ENAM,KLK4,MMP20
Abnormal morphology of ameloblasts AMBN,MMP20

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T. Cavallari, et al. Archives of Oral Biology 108 (2019) 104522

possibly interacting in genetic and protein levels (Figs. 2 and 3). When Janus, & Krom, 2017; Moye, Zeng, & Burne, 2014). These micro-
the genetic interaction between them had been analyzed, it was dis- organisms within saliva microbiota are able to change metabolic
covered that several candidate genes have shown a complex interacting pathways associated with central carbon metabolism or carbohydrate
network, such as for the genes VDR, TF1P11, HLA-DRB1 and MMP2. metabolism.
In this review, the genes AMELX, KLK4, AMBN, TUFT1 have already Even though it is still challenging, Other approaches could be used
been associated in both dentitions. While this appears to be a trend, in combination to increase the current knowledge about the interaction
some studies have warned that the effects of genetic variation may of genes and proteins previously associated with dental caries, such as
impact primary and permanent dentitions differently, once human de- the ones based on UV crosslinking immunoprecipitation for gene and
ciduous and permanent teeth exhibit different developmental processes, protein interactions (Stork & Zheng, 2016) and two-hybrid for protein-
morphologies, histological characteristics and life cycles (Kim et al., protein interactions (Lin & Lai, 2017). Other methods to correlate
2014; Bayram et al., 2015). genotype-phenotype also involve the use of gain or loss-of-function
Some of the discussed genes, such as ENAM and AMELX, are also experiments, such as microRNAs (Williams, Cheng, Blenkiron, & Reid,
directly interacting at the protein level, including interactions with 2017), plasmid for gene overexpression (Moriya, 2015) and also the
several other important genes: DSPP, MMP20, KLK4, TUFT1 and AMBN. CRISPR/cas9 system for both of these previous investigations (Adli,
At this level, the gene ameloblastin (AMBN) is strongly connected to 2018).
seven other caries-associated protein coding genes, suggesting a key Knowledge of inherited genetic variation has a fundamental impact
role in dental caries phenotype. These data on genetic and protein in- on understanding human disease. New approaches to assess functional
teractions suggest that mutations in any of the detected genes could also significance of inherited genetic variation, to combine molecular ge-
be interfering in the network formed by these molecules, and then netics, epidemiology and bioinformatics, promise to enhance the re-
potentially leading to similar phenotypes of dental caries. producibility and plausibility of associations between genotypes and
Using the IPA approach, several genes have been previously asso- disease phenotype.
ciated with diverse cellular and metabolic functions (Table 3). For in- One important approach to better identify the genetic impact in
stance, a pathway was found the Th1, which is described to be activated dental caries is to look at the individuals with extreme DMFTs. In ad-
by IL-6 to promote local immune system response and might be in- dition, the ideal would be to select individuals from the same location,
volved in the formation of edema induced by the progressive in- having the same access level to fluoridated water, healthcare and with a
tradentinal penetration of bacteria (Farges et al., 2015). Another im- similar diet. In this SR, most of considered investigations have selected
portant pathway was foundthat is involved with dental microorganism the population study according to similar characteristics, however, only
activity (the complement system). It is known to play a significant role a few investigations have followed the gold standard when considering
in the activation of the immune system with the presence of gram-po- the criterion of diagnosis and crossed the genetic data with environ-
sitive oral bacteria (Tsai, Nilsson, McArthur, & Taichman, 1977). The mental variables. Considering the mentioned aspects, the molecules
complement system may also contribute to dentin-pulp regeneration by associated with dental caries were evaluated as controlled risk factors
recruiting pulp progenitors, via this pathway (Chmilewsky, Jeanneau, by distinct investigations, and only the following genes were included
Laurent, & About, 2014). The genes AMELX, AQP5, ENAM and MMP20 within studies considering extreme phenotypes: AMELX, MMP20,
were also associated with tooth development (Table 3) and it could be GLUT2, TAS1R2, LTF, MMP2, MMP3, CA6, DEFB1, TAS2R38 and CA-VI
hypothesized that mutations in some of these genes can also affect (Kang et al., 2011; Tannure, Kuchler et al., 2012; Tannure, Kchler et al.,
normal tooth formation and biogenesis, leading to a higher suscept- 2012; Kulkarni et al., 2013; Doetzer et al., 2015; Karayasheva et al.,
ibility to dental caries. 2016; Yildiz et al., 2016; Li et al., 2015). In our review, we also high-
To perform the correlations among genes, protein interactions, lighted that distinct genetic factors can play a considerable role in the
metabolic pathways and dental caries, it was initially investigated development of the disease. However, an important consideration is
through the literature the phenotypic characteristics associated with that a gene is no longer studied as an isolated entity but rather part of a
dental caries. Next, it was identified the pathways associated with the complex network, as supported by the interaction network analysis we
mapped characteristics, to finally verify whether the genes found by our performed, which showed that distinct genes are interacting and are
literature review could be implicating in the dental caries. Although involved in similar pathways. Together, our investigation suggests that
these data were based on in silico experiments using bioinformatics dental caries might not be inherited as a single gene defect, but instead
pipelines, it could also be useful to perform a functional analysis to as a result from distinct genetic modifications and by the gene-en-
provide insightful information between the genotype-phenotype asso- vironment interactions.”
ciations with dental caries. A novel and efficient approach is being used Both positive and negative results were presented in this systematic
to investigate progression patterns of dental caries, based on long- review. Negative results are important for the broader field where they
itudinal data that is contrasted to the use of cross sectional assessment are relevant, since observing unexpected results can always bring us
scores (Weber et al., 2018). In a recessive model, the approach was able some learning and save time and resources. The reporting of negative
to find a significant innovative association between mutations in the results can help other scientists adjust their research.
gene Kallikrein 4 (KLK4) with different caries sub-phenotypes. One limitation of this review is that, while there is clear evidence on
After conducting a multiple single-nucleotide polymorphism (SNP) the influence of individual and contextual risk factors contributing to
assay in the genes TAS2R38 and TAS1R2, another study found by the caries experience, some included articles (in compliance with the
transmission disequilibrium test (TDT) analysis, significant associations review protocol) are focused only on genetic polymorphisms associated
between these taste pathway genes with caries risk and/or protection with susceptibility to dental caries. Some of them are the precursors of
(Wendell et al., 2010). this research line and have a relative role in understanding the patho-
In another study, it was performed a genome-wide investigation to genesis of dental caries. Nonetheless, the scientific community should
calculate two quantitative measures that combines both the primary build upon an integrated and comprehensive approach to investigating
and permanent dentition, while adjusting for age effects (Govil et al., dental caries, since inadequate results from single-variable tests are
2018). Genes identified in peak linkage regions underline the im- well established.
portance of exploring potential relationships between caries and other Another limitation of this review was the interpretation of diag-
traits. nostic criteria for dental caries presented by the included articles. Most
Other studies also correlates the dental caries occurrence with dif- of the studies included in this SLR used WHO classification, which is
ferent microorganisms, including Streptococcus mutans, Granulicatella widely accepted in epidemiological surveys. Some investigations used
elegans, Veillonella spp., S. mutans and Bifidobacteriaceae spp. (Lof, the ICDAS criteria (International System for Detection and Evaluation

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Moysés and Moysés: designed the study and revised the manuscript Guatemalan-Mayan population. Caries Research, 42(1), 8–13. https://doi.org/10.
All the authors approved the final version. 1159/000111744.
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Declaration of Competing Interest
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