• Users Online: 125
  • Home
  • Print this page
  • Email this page
Home About us Editorial board Ahead of print Current issue Search Archives Submit article Instructions Subscribe Contacts Login 

 Table of Contents  
Year : 2019  |  Volume : 14  |  Issue : 3  |  Page : 237-240

Studies on the Association of DEFB1 Gene Polymorphism (rs11362 and rs1799946) and Dental Caries in South Indian Population

1 Faculty of Allied Health Sciences, Chettinad Hospital and Research Institute (CHRI), Chettinad Academy of Research and Education (CARE), Kelambakkam, Tamil Nadu, India
2 Department of Periodontics, Chettinad Dental College and Research Institute (CDCRI), Kelambakkam, Tamil Nadu, India

Date of Submission04-Jun-2019
Date of Decision25-Jul-2019
Date of Acceptance10-Aug-2019
Date of Web Publication2-May-2020

Correspondence Address:
Dr. Gowtham Kumar Subbaraj
Faculty of Allied Health Sciences, Chettinad Academy of Research and Education, OMR, Kelambakkam - 603 103, Tamil Nadu
Login to access the Email id

Source of Support: None, Conflict of Interest: None

DOI: 10.4103/jdmimsu.jdmimsu_102_17

Rights and Permissions

Background: Caries development in the teeth is multifactorial diseases where the environment, and the genetic factors play an important role in the onset and recurrence. Defensins are small, cationic, cysteine-rich peptides found to be active against microorganisms including bacteria, fungi and many non-enveloped and enveloped viruses. It play a significant role in defense against numerous pathogens, and they are deliberated part of the innate immunity. They have usually been considered to contribute to mucosal health. Defensin (DEFB1) gene polymorphisms may modulate human beta defensin (HBD-1) levels might be related to pathogenesis of numerous ailments. Aim: The present investigation was aimed to understand the association of two single-nucleotide polymorphisms (rs1799946 and rs11362) of DEFB1 gene with dental caries. Results: we could found that the single-nucleotide polymorphism (SNP) rs11362 is positively associated with the progression of dental caries in the tested population. Conclusion: our analysis possibly identified the accentuate relationship between dental caries and the SNP rs11362 polymorphism of the defensin gene (DEFB1), innate immune players in the oral area.

Keywords: β-defensins, dental caries, mucosal immunity, polymorphism

How to cite this article:
Akilpprienka B, Malarkodi M, Vivedharani R, Varghese S, Anitha V, Rajamanickam K, Subbaraj GK. Studies on the Association of DEFB1 Gene Polymorphism (rs11362 and rs1799946) and Dental Caries in South Indian Population. J Datta Meghe Inst Med Sci Univ 2019;14:237-40

How to cite this URL:
Akilpprienka B, Malarkodi M, Vivedharani R, Varghese S, Anitha V, Rajamanickam K, Subbaraj GK. Studies on the Association of DEFB1 Gene Polymorphism (rs11362 and rs1799946) and Dental Caries in South Indian Population. J Datta Meghe Inst Med Sci Univ [serial online] 2019 [cited 2020 Sep 25];14:237-40. Available from: http://www.journaldmims.com/text.asp?2019/14/3/237/283572

  Introduction Top

Dental caries is a chronic noncommunicable multifactorial disease that results in the demineralization of the inorganic part (enamel, dentin, and cementum) and destruction of the organic (pulp) substance of the teeth. It is a lifestyle-related disease where the initiation and progression is a complex and dynamic process involving the interaction between the bacterial microflora (cariogenic bacteria), the substrate and the host factors such as tooth structure, salivary composition, and genetic factors along with time inducing the formation of carious lesions on the susceptible tooth.[1] One of the major factors for the onset of dental caries includes the host-derived microbial flora. The oral cavity is home to more than 250 types of microorganisms.[2] Some of the microbial pathogens that are associated with caries are Streptococcus vestibularis, Streptococcus salivarius, Streptococcus sobrinus, Streptococcus parasanguinis and Veillonella atipica, Veillonella dispar, Veillonella parvula, whose elevated levels is noted in a significant number of caries patients. However, in these patients, low levels of Streptococcus mutans were found. S. sobrinus is closely associated with S. mutans and is found to be associated with childhood caries. S. vestibularis and S. salivarius are salivarius groups with similar structures and studies reveals that Streptococcus salivariusis highly cariogenic than S. vestibularis.[3]

Genetic factors play a significant role in the onset of dental caries and numerous studies shown that there are several genetic factors underlying to cause dental caries. Defensins are a family of microbicidal and cytotoxic peptides that are made by host neutrophils. It is a 2–6 kDa cationic microbicidal peptides which are active against many Gram-positive and Gram-negative bacteria, fungi, and even on enveloped viruses.[4] Large concentrations of these defensins are found to be present in the whole body in the healthy as well as inflamed tissues which also contributes the first line of defence mechanism against the pathogens in the oral region. In 2006, Dale et al.[5] reported that decreased levels of α-defensins and development of dental caries in the children. The genes which encode for β-defensins are found to have single-nucleotide polymorphisms (SNPs).[6] The presence of SNPs in the coding region is likely to affect the resistance against infections which occur because of the alterations in the protein sequence and therefore give rise to different biological functions.[6] Hence, the present investigation was aimed to reveal the genetic factors in the development of caries and its recurrence and to validate the association between single-nucleotide polymorphisms (SNP) (rs11362 and rs1799946) with dental caries in the tested population.

  Materials And Methods Top

Collection of samples

A total number of 48 blood samples were collected from the Department of Periodontics, Chettinad Dental College and Research Institute, Kelambakkam, Tamil Nadu, India. It includes 33 patient and 15 control samples (participants without dental caries). The study was conducted as per the ethical norms followed by the institution (157/IHEC/12-16 dated January 21, 2017).

Decayed missing filled teeth index assessment

The decayed missing filled teeth (DMFT) index screening was developed by Henry T Klein, Carrole E Palmer, and Knutson J W, 1938. Occlusal and smooth surface caries detection is mostly done by measuring the number of teeth decayed, a number of teeth missing due to caries and number of filled teeth present in the oral cavity.[7]

Isolation of genomic DNA by Miller's method

The DNA was isolated from human blood by Miller's method.[8]

Polymerase chain reaction

The SNP detection was done on two SNPs (rs11362 and rs1799946) by polymerase chain reaction. The study was done on both control and case DNA samples. Polymerase chain reaction (PCR) amplification reaction (20 μl) was performed using Taq polymerase, template DNA, forward primer, reverse primer, Taq buffer, template DNA, and Milli-Q water. PCR Primers used for -52 G > A (rs1799946) Forward primer 5'-CAGCTCAGCCTCCAAACG-3'; Reverse primer 5'-GCAGGTACCAGAGCTTACC-3’ and the product Size: 148 bp and PCR Primers for -20G > A (rs11362) Forward primer 5'-CCAGTTCCTGAAATCCTCG-3'; reverse primer 5’ TGCAGGTACCAGAGCTTAC-3'and the product size: 119 bp.

  Results Top

The various mean ± standard deviation data of DMFT index, DT index, MT index, and FT index among the gender group is presented in [Figure 1]. The average DMFT score of the whole population was found to be 4.10 ± 3.4. There is a difference in the mean score of DMFT among males and females. Our study shows that males have high DMFT scores when compared to females [Figure 1]. However, literature has already reported that females are more exposed to dental caries than males.[9]
Figure 1 : Frequency of DMFT Index in control and patients with dental caries

Click here to view

PCR was done to the isolated DNA for both the SNPs (rs1799946 and rs11362). After the run, PCR products were analyzed by running into 2% agarose gel electrophoresis along with control ladder (100 bp) in one lane. [Figure 2] shows the data obtained after PCR reaction of the DNA sample obtained from the study participants. After the agarose gel electrophoresis, bands were observed from the corresponding positions from patients with dental caries as well as from the normal healthy controls for SNP rs1799946 (-52 region) and SNP rs11362 (-20 region) [Figure 2]. [Table 1] shows two DEFB1 gene polymorphism (rs11362 and rs1799946) and its association with dental caries in different populations.
Figure 2: Frequencies of DEFB1 rs179946 (-52 regions) and rs11362 (-20 regions) polymorphism in control and participants with dental caries

Click here to view
Table 1: SNP (rs11362 and rs1799946) associated with caries among different ethnic groups

Click here to view

The given [Figure 3] shows association of various single nucleotide polymorphisms with dental caries. The forest plot illustrates the different DEFBI gene polymorphism with different odds ratio. The DEFB1 gene polymorphism rs11362 have shown high odds ratio revealing a positive association in the progression of dental caries. Similar to the previous studies the present study have also shown a high risk of developing dental caries. Whereas another DEFBI gene polymorphism rs 1799946 didn't show any association with dental caries in tested population.
Figure 3: Forest plot of various single-nucleotide polymorphisms associated with dental caries showing different odds ratio with 95% confidence interval

Click here to view

Statistical analysis

Of the 15 positive samples 9 were found in -52 regions while the other 6 were found in -20 boundaries. We used Chi-square to test that each of the SNPs and the values were 0.897 with the positive predictive value of 18.18% for -52 regions. The Chi-values were 0.678 with the positive predictive value of 9.09% for -20 regions.[10],[11],[12],[14]

  Discussion Top

The DEFB1 gene is known to be expressed in almost a wide variety of mouse tissues which includes the airways similar to the human gene.[15] Loo et al.[16] observed that the DEFB1 SNP at -1654 position leads to a change of G to A substitution and a valine to isoleucine substitution at the polypeptide position. This was most frequently found in the patients suffering from moderate to severe chronic periodontitis when compared to the healthy group. The A/A genotype is mostly found in the patients with chronic periodontitis whereas the G/G genotype was present in the control group. The genotypic makeup should be content to an individual and in the case of any difference in the polymorphic status of the genes arise only due to the circulation of the body. A significant association of the rare an allele of DEFB1 variant was found with an increased risk of periodontal disease.[13]

The β-defensins are known to follow several ways to maintain the homeostatic levels of the commensal bacteria as well as to inhibit the pathogenic microbial colonization present in the oral cavity.[17] The pathogens upregulate the expression of β-defensins encoding genes. Several studies identified that the hBD-2, hBD-3, and hBD-4 is able to activate the human keratinocytes to increase its own gene expression and protein production of interleukin (IL)-6, monocyte chemoattractant protein-1, RANTES (regulated on activation normal T-cell expressed and secreted), macrophage inflammatory protein-3 alpha, and the IL-10 interferon gamma-induced protein 10.[18]

The genetic associations in which these particular SNPs were found to be haplotypes or single.[19] It has been proved by many investigators that the polymorphism in the DEFB1 boosted the DMFT and DMFS score by a 5-fold, and also revealed that the differences in the major histocompatibility complex can influence the cause of dental caries in children.[20] Liu et al.,[21] in his study have stated that the salivary antimicrobial peptides can be used as a biomarker for the early diagnosis of periodontitis. The human β-defensins are known to be involved in several processes. The β-defensins is known to promote wound healing,[22] and also it plays a vital role in the metastasis and also in the tumor growth. In one of the studies, it has been shown that the hBD-1, hBD-2, and hBD-3 were found to be more expressed in the bisphosphonate-associated osteomyelitis of the jaw.

  Conclusion Top

In this present investigation, the association of two-single nucleotide polymorphisms (rs1799946 and rs11362) of DEFB1 gene with dental caries was investigated. From the results, we could conclude that the SNP rs11362 is positively associated with the progression of dental caries in the tested population. Literature shows that other candidate SNP, rs1799946 is also associated with caries in various population, but we were unable to replicate the same results from the tested samples. The study population should definitely be increased, and more extensive study should be done to accentuate the relationship between dental caries and the β-Defensin (DEFB1) gene polymorphism.

Financial support and sponsorship

The authors record their sincere gratitude to Prof. Dr. R. Murugesan, Director -Research. This work was carried out by receiving the financial support provided by the management of Chettinad Academy of Research and Education (CARE).

Conflicts of interest

There are no conflicts of interest.

  References Top

White SH, Wimley WC, Selsted ME. Structure, function, and membrane integration of defensins. Curr Opin Struct Biol 1995;5:521-7.  Back to cited text no. 1
Miller SA, Dykes DD, Polesky HF. A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Res 1988;16:1215.  Back to cited text no. 2
Morrison GM, Davidson DJ, Kilanowski FM, Borthwick DW, Crook K, Maxwell AI, et al. Mouse beta defensin-1 is a functional homolog of human beta defensin-1. Mamm Genome 1998;9:453-7.  Back to cited text no. 3
Cypriano S, de Sousa Mda L, Wada RS. Evaluation of simplified DMFT indices in epidemiological surveys of dental caries. Rev Saude Publica 2005;39:285-92.  Back to cited text no. 4
Dale BA, Tao R, Kimball JR, Jurevic RJ. Oral antimicrobial peptides and biological control of caries. BMC Oral Health 2006;6 Suppl 1:S13.  Back to cited text no. 5
Schaefer R. Foundations of Global Genetic Optimization. Springer-Verlag Berlin Heidelberg. Springer Publishing Company, Incorporated; 2007.  Back to cited text no. 6
Niyonsaba F, Ushio H, Nakano N, Ng W, Sayama K, Hashimoto K, et al. Antimicrobial peptides human beta-defensins stimulate epidermal keratinocyte migration, proliferation and production of proinflammatory cytokines and chemokines. J Invest Dermatol 2007;127:594-604.  Back to cited text no. 7
van Dijk A, Veldhuizen EJ, Haagsman HP. Avian defensins. Vet Immunol Immunopathol 2008;124:1-8.  Back to cited text no. 8
Usha C, Sathyanarayanan R. Dental caries – A complete changeover (Part I). J Conserv Dent 2009;12:46-54.  Back to cited text no. 9
[PUBMED]  [Full text]  
Loo WT, Bai LJ, Fan CB, Yue Y, Dou YD, Wang M, et al. Clinical application of human β-defensin and CD14 gene polymorphism in evaluating the status of chronic inflammation. J Transl Med 2012;10 Suppl 1:S9.  Back to cited text no. 10
Krasone K, Lāce B, Akota I, Care R, Deeley K, Küchler EC, et al. Genetic variation in the promoter region of beta-defensin 1 (DEFB 1) is associated with high caries experience in children born with cleft lip and palate. Acta Odontol Scand 2014;72:235-40.  Back to cited text no. 11
Liu Z, Liu Y, Song Y, Zhang X, Wang S, Wang Z. Systemic oxidative stress biomarkers in chronic periodontitis: A meta-analysis. Dis Markers 2014;2014:931083.  Back to cited text no. 12
Naslavsky MS, Crovella S, Lima Filho JL, Rocha CR. The sound of silence: Human beta-defensin-1 gene untranslated SNPs change the predicted mRNA secondary structure in a length-dependent manner. Immunol Lett 2010;129:53-5.  Back to cited text no. 13
Caufield PW, Schön CN, Saraithong P, Li Y, Argimón S. Oral Lactobacilli and dental caries: A model for niche adaptation in humans. J Dent Res 2015;94:110S-8S.  Back to cited text no. 14
Schaefer AS, Richter GM, Nothnagel M, Laine ML, Rühling A, Schäfer C, et al. A 3’ UTR transition within DEFB1 is associated with chronic and aggressive periodontitis. Genes Immun 2010;11:45-54.  Back to cited text no. 15
Ozturk A, Famili P, Vieira AR. The antimicrobial peptide DEFB1 is associated with caries. J Dent Res 2010;89:631-6.  Back to cited text no. 16
Diamond G, Ryan L. Beta-defensins: What are they really doing in the oral cavity? Oral Dis 2011;17:628-35.  Back to cited text no. 17
Stockmann P, Wehrhan F, Schwarz-Furlan S, Stelzle F, Trabert S, Neukam FW, et al. Increased human defensine levels hint at an inflammatory etiology of bisphosphonate-associated osteonecrosis of the jaw: An immunohistological study. J Transl Med 2011;9:135.  Back to cited text no. 18
Ikuta T, Inagaki Y, Tanaka K, Saito T, Nakajima Y, Bando M, et al. Gene polymorphism of β-defensin-1 is associated with susceptibility to periodontitis in Japanese. Odontology 2015;103:66-74.  Back to cited text no. 19
Navarra CO, Robino A, Pirastu N, Bevilacqua L, Gasparini P, Di Lenarda R, et al. Caries and innate immunity: DEFB1 gene polymorphisms and caries susceptibility in genetic isolates from North-Eastern Italy. Caries Res 2016;50:589-94.  Back to cited text no. 20
Li X, Duan D, Yang J, Wang P, Han B, Zhao L, et al. The expression of human β-defensins (hBD-1, hBD-2, hBD-3, hBD-4) in gingival epithelia. Arch Oral Biol 2016;66:15-21.  Back to cited text no. 21
Zupin L, Robino A, Navarra CO, Pirastu N, Di Lenarda R, Gasparini P, et al. LTF and DEFB1 polymorphisms are associated with susceptibility toward chronic periodontitis development. Oral Dis 2017;23:1001-8.  Back to cited text no. 22


  [Figure 1], [Figure 2], [Figure 3]

  [Table 1]


Similar in PUBMED
   Search Pubmed for
   Search in Google Scholar for
 Related articles
Access Statistics
Email Alert *
Add to My List *
* Registration required (free)

  In this article
Materials And Me...
Article Figures
Article Tables

 Article Access Statistics
    PDF Downloaded27    
    Comments [Add]    

Recommend this journal