Skip to main content
Top
Published in: BMC Oral Health 1/2018

Open Access 01-12-2018 | Research article

Molecular and clinical analyses of Helicobacter pylori colonization in inflamed dental pulp

Authors: Ryota Nomura, Yuko Ogaya, Saaya Matayoshi, Yumiko Morita, Kazuhiko Nakano

Published in: BMC Oral Health | Issue 1/2018

Login to get access

Abstract

Background

Recently, dental pulp has been considered a possible source of infection of Helicobacter pylori (H. pylori) in children. We previously developed a novel PCR system for H. pylori detection with high specificity and sensitivity using primer sets constructed based on the complete genome information for 48 H. pylori strains. This PCR system showed high sensitivity with a detection limit of 1–10 cells when serial dilutions of H. pylori genomic DNA were used as templates. However, the detection limit was lower (102–103 cells) when H. pylori bacterial DNA was detected from inflamed pulp specimens. Thus, we further refined the system using a nested PCR method, which was much more sensitive than the previous single PCR method. In addition, we examined the distribution and virulence of H. pylori in inflamed pulp tissue.

Methods

Nested PCR system was constructed using primer sets designed from the complete genome information of 48 H. pylori strains. The detection limit of the nested PCR system was 1–10 cells using both H. pylori genomic DNA and bacterial DNA isolated from inflamed pulp specimens. Next, distribution of H. pylori was examined using 131 inflamed pulp specimens with the nested PCR system. In addition, association between the detection of H. pylori and clinical information regarding endodontic-infected teeth were investigated. Furthermore, adhesion property of H. pylori strains to human dental fibroblast cells was examined.

Results

H. pylori was present in 38.9% of inflamed pulp specimens using the nested PCR system. H. pylori was shown to be predominantly detected in primary teeth rather than permanent teeth. In addition, samplings of the inflamed pulp were performed twice from the same teeth at 1- or 2-week intervals, which revealed that H. pylori was detected in most specimens in both samplings. Furthermore, H. pylori strains showed adhesion property to human dental fibroblast cells.

Conclusion

Our results suggest that H. pylori colonizes inflamed pulp in approximately 40% of all cases through adhesion to human dental fibroblast cells.
Literature
2.
go back to reference Prasanthi CH, Prasanthi NL, Manikiran SS, Rama-Rao NN. Focus on current trends in the treatment of Helicobacter pylori infection: an update. Inter J Pharm Sci Rev Res. 2011;1:42–51. Prasanthi CH, Prasanthi NL, Manikiran SS, Rama-Rao NN. Focus on current trends in the treatment of Helicobacter pylori infection: an update. Inter J Pharm Sci Rev Res. 2011;1:42–51.
3.
go back to reference Westblom TU, Bhatt BD. Diagnosis of Helicobacter pylori infection. Curr Topics Microbiol Immunol. 1999;241:215–35. Westblom TU, Bhatt BD. Diagnosis of Helicobacter pylori infection. Curr Topics Microbiol Immunol. 1999;241:215–35.
4.
go back to reference Silva DG, Tinoco EM, Rocha GA, Rocha AM, Guerra JB, Saraiva IE, Queiroz DM. Helicobacter pylori transiently in the mouth may participate in the transmission of infection. Mem Inst Oswaldo Cruz. 2010;105:657–60.CrossRefPubMed Silva DG, Tinoco EM, Rocha GA, Rocha AM, Guerra JB, Saraiva IE, Queiroz DM. Helicobacter pylori transiently in the mouth may participate in the transmission of infection. Mem Inst Oswaldo Cruz. 2010;105:657–60.CrossRefPubMed
5.
go back to reference Ogaya Y, Nomura R, Watanabe Y, Nakano K. Detection of Helicobacter pylori DNA in inflamed dental pulp specimens from Japanese children and adolescents. J Med Microbiol. 2015;64:117–23.CrossRefPubMed Ogaya Y, Nomura R, Watanabe Y, Nakano K. Detection of Helicobacter pylori DNA in inflamed dental pulp specimens from Japanese children and adolescents. J Med Microbiol. 2015;64:117–23.CrossRefPubMed
6.
go back to reference Mapstone NP, Lynch DA, Lewis FA, Axon AT, Tompkins DS, Dixon MF, Quirke P. Identification of Helicobacter pylori DNA in the mouths and stomachs of patients with gastritis using PCR. J Clin Pathol. 1993;46:540–3.CrossRefPubMedPubMedCentral Mapstone NP, Lynch DA, Lewis FA, Axon AT, Tompkins DS, Dixon MF, Quirke P. Identification of Helicobacter pylori DNA in the mouths and stomachs of patients with gastritis using PCR. J Clin Pathol. 1993;46:540–3.CrossRefPubMedPubMedCentral
7.
go back to reference Miyabayashi H, Furihata K, Shimizu T, Ueno I, Akamatsu T. Influence of oral Helicobacter pylori on the success of eradication therapy against gastric Helicobacter pylori. Helicobacter. 2000;5:30–7.CrossRefPubMed Miyabayashi H, Furihata K, Shimizu T, Ueno I, Akamatsu T. Influence of oral Helicobacter pylori on the success of eradication therapy against gastric Helicobacter pylori. Helicobacter. 2000;5:30–7.CrossRefPubMed
8.
go back to reference Wang J, Chi DS, Laffan JJ, Li C, Ferguson DA Jr, Litchfield P, Thomas E. Comparison of cytotoxin genotypes of Helicobacter pylori in stomach and saliva. Dig Dis Sci. 2002;47:1850–6.CrossRefPubMed Wang J, Chi DS, Laffan JJ, Li C, Ferguson DA Jr, Litchfield P, Thomas E. Comparison of cytotoxin genotypes of Helicobacter pylori in stomach and saliva. Dig Dis Sci. 2002;47:1850–6.CrossRefPubMed
9.
go back to reference Park CY, Kwak M, Gutierrez O, Graham DY, Yamaoka Y. Comparison of genotyping Helicobacter pylori directly from biopsy specimens and genotyping from bacterial cultures. J Clin Microbiol. 2003;41:3336–8.CrossRefPubMedPubMedCentral Park CY, Kwak M, Gutierrez O, Graham DY, Yamaoka Y. Comparison of genotyping Helicobacter pylori directly from biopsy specimens and genotyping from bacterial cultures. J Clin Microbiol. 2003;41:3336–8.CrossRefPubMedPubMedCentral
10.
go back to reference Smith SI, Oyedeji KS, Arigbabu AO, Cantet F, Megraud F, Ojo OO, Uwaifo AO, Otegbayo JA, Ola SO, Coker AO. Comparison of three PCR methods for detection of Helicobacter pylori DNA and detection of cagA gene in gastric biopsy specimens. World J Gastroenterol. 2004;10:1958–60.CrossRefPubMedPubMedCentral Smith SI, Oyedeji KS, Arigbabu AO, Cantet F, Megraud F, Ojo OO, Uwaifo AO, Otegbayo JA, Ola SO, Coker AO. Comparison of three PCR methods for detection of Helicobacter pylori DNA and detection of cagA gene in gastric biopsy specimens. World J Gastroenterol. 2004;10:1958–60.CrossRefPubMedPubMedCentral
11.
go back to reference Sugimoto M, Wu JY, Abudayyeh S, Hoffman J, Brahem H, Al-Khatib K, Yamaoka Y, Graham DY. Unreliability of results of PCR detection of Helicobacter pylori in clinical or environmental samples. J Clin Microbiol. 2009;47:738–42.CrossRefPubMedPubMedCentral Sugimoto M, Wu JY, Abudayyeh S, Hoffman J, Brahem H, Al-Khatib K, Yamaoka Y, Graham DY. Unreliability of results of PCR detection of Helicobacter pylori in clinical or environmental samples. J Clin Microbiol. 2009;47:738–42.CrossRefPubMedPubMedCentral
12.
go back to reference Aksit Bıcak D, Akyuz S, Kıratlı B, Usta M, Urganci N, Alev B, Yarat A, Sahin F. The investigation of Helicobacter pylori in the dental biofilm and saliva samples of children with dyspeptic complaints. BMC Oral Health. 2017;17:67.CrossRefPubMedPubMedCentral Aksit Bıcak D, Akyuz S, Kıratlı B, Usta M, Urganci N, Alev B, Yarat A, Sahin F. The investigation of Helicobacter pylori in the dental biofilm and saliva samples of children with dyspeptic complaints. BMC Oral Health. 2017;17:67.CrossRefPubMedPubMedCentral
13.
go back to reference Hirsch C, Tegtmeyer N, Rohde M, Rowland M, Oyarzabal OA, Backert S. Live Helicobacter pylori in the root canal of endodontic-infected deciduous teeth. J Gastroenterol. 2012;47:936–40.CrossRefPubMed Hirsch C, Tegtmeyer N, Rohde M, Rowland M, Oyarzabal OA, Backert S. Live Helicobacter pylori in the root canal of endodontic-infected deciduous teeth. J Gastroenterol. 2012;47:936–40.CrossRefPubMed
14.
go back to reference Amano A, Nakagawa I, Kataoka K, Morisaki I, Hamada S. Distribution of Porphyromonas gingivalis strains with fimA genotypes in periodontitis patients. J Clin Microbiol. 1999;37:1426–30.PubMedPubMedCentral Amano A, Nakagawa I, Kataoka K, Morisaki I, Hamada S. Distribution of Porphyromonas gingivalis strains with fimA genotypes in periodontitis patients. J Clin Microbiol. 1999;37:1426–30.PubMedPubMedCentral
15.
go back to reference Nakano K, Inaba H, Nomura R, Nemoto H, Takeda M, Yoshioka H, Matsue H, Takahashi T, Taniguchi K, et al. Detection of cariogenic Streptococcus mutans in extirpated heart valve and atheromatous plaque specimens. J Clin Microbiol. 2006;44:3313–7.CrossRefPubMedPubMedCentral Nakano K, Inaba H, Nomura R, Nemoto H, Takeda M, Yoshioka H, Matsue H, Takahashi T, Taniguchi K, et al. Detection of cariogenic Streptococcus mutans in extirpated heart valve and atheromatous plaque specimens. J Clin Microbiol. 2006;44:3313–7.CrossRefPubMedPubMedCentral
16.
go back to reference Nomura R, Ogaya Y, Nakano K. Contribution of the collagen-binding proteins of Streptococcus mutans to bacterial colonization of inflamed dental pulp. PLoS One. 2016;11:e0159613.CrossRefPubMedPubMedCentral Nomura R, Ogaya Y, Nakano K. Contribution of the collagen-binding proteins of Streptococcus mutans to bacterial colonization of inflamed dental pulp. PLoS One. 2016;11:e0159613.CrossRefPubMedPubMedCentral
17.
go back to reference Robert J, Palmer JR. Composition and development of oral bacterial communities. Periodontol. 2013;64:20–39. Robert J, Palmer JR. Composition and development of oral bacterial communities. Periodontol. 2013;64:20–39.
18.
go back to reference Peek RM Jr, Miller GG, Tham KT, Pérez-Pérez GI, Cover TL, Atherton JC, Dunn GD, Blaser MJ. Detection of Helicobacter pylori gene expression in human gastric mucosa. J Clin Microbiol. 1995;33:28–32.PubMedPubMedCentral Peek RM Jr, Miller GG, Tham KT, Pérez-Pérez GI, Cover TL, Atherton JC, Dunn GD, Blaser MJ. Detection of Helicobacter pylori gene expression in human gastric mucosa. J Clin Microbiol. 1995;33:28–32.PubMedPubMedCentral
19.
go back to reference Yamamoto Y. PCR in diagnosis of infection: detection of bacteria in cerebrospinal fluids. Clin Diagn Lab Immunol. 2002;9:508–14.PubMedPubMedCentral Yamamoto Y. PCR in diagnosis of infection: detection of bacteria in cerebrospinal fluids. Clin Diagn Lab Immunol. 2002;9:508–14.PubMedPubMedCentral
20.
go back to reference Takahashi T, Nakayama T, Tamura M, Ogawa K, Tsuda H, Morita A, Hara M, Togo M, Shiota H, et al. Nested polymerase chain reaction for assessing the clinical course of tuberculous meningitis. Neurology. 2005;64:1789–93.CrossRefPubMed Takahashi T, Nakayama T, Tamura M, Ogawa K, Tsuda H, Morita A, Hara M, Togo M, Shiota H, et al. Nested polymerase chain reaction for assessing the clinical course of tuberculous meningitis. Neurology. 2005;64:1789–93.CrossRefPubMed
21.
go back to reference Umeda M, Kobayashi H, Takeuchi Y, Hayashi J, Morotome-Hayashi Y, Yano K, Aoki A, Ohkusa T, Ishikawa I. High prevalence of Helicobacter pylori detected by PCR in the oral cavities of periodontitis patients. J Periodontol. 2003;74:129–34.CrossRefPubMed Umeda M, Kobayashi H, Takeuchi Y, Hayashi J, Morotome-Hayashi Y, Yano K, Aoki A, Ohkusa T, Ishikawa I. High prevalence of Helicobacter pylori detected by PCR in the oral cavities of periodontitis patients. J Periodontol. 2003;74:129–34.CrossRefPubMed
22.
go back to reference Oh JD, Kling-Bäckhed H, Giannakis M, Xu J, Fulton RS, Fulton LA, Cordum HS, Wang C, Elliott G, et al. The complete genome sequence of a chronic atrophic gastritis Helicobacter pylori strain: evolution during disease progression. Proc Natl Acad Sci U S A. 2006;103:9999–10004.CrossRefPubMedPubMedCentral Oh JD, Kling-Bäckhed H, Giannakis M, Xu J, Fulton RS, Fulton LA, Cordum HS, Wang C, Elliott G, et al. The complete genome sequence of a chronic atrophic gastritis Helicobacter pylori strain: evolution during disease progression. Proc Natl Acad Sci U S A. 2006;103:9999–10004.CrossRefPubMedPubMedCentral
23.
go back to reference Ribet D, Cossart P. How bacterial pathogens colonize their hosts and invade deeper tissues. Microbes Infect. 2015;17:173–83.CrossRefPubMed Ribet D, Cossart P. How bacterial pathogens colonize their hosts and invade deeper tissues. Microbes Infect. 2015;17:173–83.CrossRefPubMed
24.
go back to reference Sheu BS, Yang HB, Yeh YC, Wu JJ. Helicobacter pylori colonization of the human gastric epithelium: a bug’s first step is a novel target for us. J Gastroenterol Hepatol. 2010;25:26–32.CrossRefPubMed Sheu BS, Yang HB, Yeh YC, Wu JJ. Helicobacter pylori colonization of the human gastric epithelium: a bug’s first step is a novel target for us. J Gastroenterol Hepatol. 2010;25:26–32.CrossRefPubMed
25.
go back to reference Jones KR, Whitmire JM, Merrell DS. A tale of two toxins: Helicobacter Pylori CagA and VacA modulate host pathways that impact disease. Front Microbiol. 2010;1:115.CrossRefPubMedPubMedCentral Jones KR, Whitmire JM, Merrell DS. A tale of two toxins: Helicobacter Pylori CagA and VacA modulate host pathways that impact disease. Front Microbiol. 2010;1:115.CrossRefPubMedPubMedCentral
26.
go back to reference Rieder G, Hatz RA, Moran AP, Walz A, Stolte M, Enders G. Role of adherence in interleukin-8 induction in Helicobacter pylori-associated gastritis. Infect Immun. 1997;65:3622–30.PubMedPubMedCentral Rieder G, Hatz RA, Moran AP, Walz A, Stolte M, Enders G. Role of adherence in interleukin-8 induction in Helicobacter pylori-associated gastritis. Infect Immun. 1997;65:3622–30.PubMedPubMedCentral
27.
go back to reference Segal ED, Lange C, Covacci A, Tompkins LS, Falkow S. Induction of host signal transduction pathways by Helicobacter pylori. Proc Natl Acad Sci U S A. 1997;94:7595–9.CrossRefPubMedPubMedCentral Segal ED, Lange C, Covacci A, Tompkins LS, Falkow S. Induction of host signal transduction pathways by Helicobacter pylori. Proc Natl Acad Sci U S A. 1997;94:7595–9.CrossRefPubMedPubMedCentral
28.
go back to reference Ghosh P, Sarkar A, Ganguly M, Raghwan Alam J, De R, Mukhopadhyay AK. Helicobacter pylori strains harboring babA2 from Indian sub population are associated with increased virulence in ex vivo study. Gut Pathog. 2016;8:1.CrossRefPubMedPubMedCentral Ghosh P, Sarkar A, Ganguly M, Raghwan Alam J, De R, Mukhopadhyay AK. Helicobacter pylori strains harboring babA2 from Indian sub population are associated with increased virulence in ex vivo study. Gut Pathog. 2016;8:1.CrossRefPubMedPubMedCentral
Metadata
Title
Molecular and clinical analyses of Helicobacter pylori colonization in inflamed dental pulp
Authors
Ryota Nomura
Yuko Ogaya
Saaya Matayoshi
Yumiko Morita
Kazuhiko Nakano
Publication date
01-12-2018
Publisher
BioMed Central
Published in
BMC Oral Health / Issue 1/2018
Electronic ISSN: 1472-6831
DOI
https://doi.org/10.1186/s12903-018-0526-2

Other articles of this Issue 1/2018

BMC Oral Health 1/2018 Go to the issue