Expression of Toll-like receptors 2 and 4 and its association with matrix metalloproteinases in symptomatic and asymptomatic apical periodontitis
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To determine Toll-like receptors (TLR)2 and TLR4 expression levels and associate them with matrix metalloproteinases (MMPs) in asymptomatic apical periodontitis (AAP), symptomatic apical periodontitis (SAP), and healthy controls. Apical tissue/lesion samples were obtained from chronic AAP (n = 35) and SAP (n = 29), and healthy periodontal ligament (HPL, n = 10) with indication of tooth extraction, respectively. mRNA expression levels of TLR2, TLR4, MMP-1, MMP-2, MMP-8, and MMP-13 were determined by real-time reverse-transcription polymerase chain reaction. The data were analyzed with Kruskal-Wallis and Dunn’s pot hoc test (p < 0.05). The correlation coefficient was obtained using the Spearman correlation (p < 0.05). TLR2, MMP-1, MMP-2, and MMP-13 mRNA levels were the highest in SAP followed by AAP and controls (p < 0.05). TLR4 and MMP-8 were over expressed in AAP and SAP compared to HPL (p < 0.05). TLR2 positively correlated with TLR4, MMP-1, MMP-8, and MMP-13 in SAP (p < 0.05). TLR2 and TLR4 are overexpressed in apical lesions versus healthy periodontal ligament and correlate with collagenolytic MMPs. Particularly, TLR2 is overexpressed in SAP in association with MMP-1, MMP-8, and MMP-13. Our results suggest that the activation of TLR2 along with MMP overexpression might contribute to SAP clinical presentation and progression. TLRs, MMPs, and their interaction can explain the clinical presentations and evolution of apical periodontitis and might represent key targets for new diagnostic and treatment approaches.
KeywordsMatrix metalloproteinases Periapical periodontitis Toll-like receptor 2 Toll-like receptor 4
We thank Peter Gebicke-Haerter and Andrés Tittarelli for their valuable support with bioanalyzer equipment. Alejandra Fernández thanks the fellowship CONICYT 21181377, from the Chilean Government.
The work was supported by FONDECYT grant no. 1160741.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.
Informed consent was obtained from all individual participants included in the study.
- 2.Croitoru IC, CraiToiu S, Petcu CM, Mihailescu OA, Pascu RM, Bobic AG, Agop Forna D, CraiToiu MM (2016) Clinical, imagistic and histopathological study of chronic apical periodontitis. Romanian J Morphol Embryol 57(2 Suppl):719–728Google Scholar
- 3.Salinas-Munoz M, Garrido-Flores M, Baeza M, Huaman-Chipana P, Garcia-Sesnich J, Bologna R, Vernal R, Hernandez M (2017) Bone resorptive activity in symptomatic and asymptomatic apical lesions of endodontic origin. Clin Oral Investig 21(8):2613–2618. https://doi.org/10.1007/s00784-017-2062-x CrossRefPubMedPubMedCentralGoogle Scholar
- 6.de Oliveira Rde C, Beghini M, Borges CR, Alves PM, de Araujo MS, Pereira SA, Rodrigues V Jr, Rodrigues DB (2014) Higher expression of galectin-3 and galectin-9 in periapical granulomas than in radicular cysts and an increased toll-like receptor-2 and toll-like receptor-4 expression are associated with reactivation of periapical inflammation. J Endod 40(2):199–203. https://doi.org/10.1016/j.joen.2013.10.031 CrossRefPubMedGoogle Scholar
- 7.Kassem A, Henning P, Lundberg P, Souza PP, Lindholm C, Lerner UH (2015) Porphyromonas gingivalis stimulates bone resorption by enhancing RANKL (receptor activator of NF-kappaB ligand) through activation of toll-like receptor 2 in osteoblasts. J Biol Chem 290(33):20147–20158. https://doi.org/10.1074/jbc.M115.655787 CrossRefPubMedPubMedCentralGoogle Scholar
- 8.Itoh K, Udagawa N, Kobayashi K, Suda K, Li X, Takami M, Okahashi N, Nishihara T, Takahashi N (2003) Lipopolysaccharide promotes the survival of osteoclasts via Toll-like receptor 4, but cytokine production of osteoclasts in response to lipopolysaccharide is different from that of macrophages. J Immunol 170(7):3688–3695CrossRefGoogle Scholar
- 9.Tang L, Zhou XD, Wang Q, Zhang L, Wang Y, Li XY, Huang DM (2011) Expression of TRAF6 and pro-inflammatory cytokines through activation of TLR2, TLR4, NOD1, and NOD2 in human periodontal ligament fibroblasts. Arch Oral Biol 56(10):1064–1072. https://doi.org/10.1016/j.archoralbio.2011.02.020 CrossRefPubMedGoogle Scholar
- 10.Barreiros D, Nelson PF, Paula-Silva FWG, Oliveira KMH, Lucisano MP, Rossi A, Silva LAB, Kuchler EC, Silva RAB (2018) MMP2 and MMP9 are associated with apical periodontitis progression and might be modulated by TLR2 and MyD88. Braz Dent J 29(1):43–47. https://doi.org/10.1590/0103-6440201801731 CrossRefPubMedGoogle Scholar
- 14.Mundi Burgos V, Dezerega Piwonka A, Osorio Alfaro C, Dutzan Muñoz N, Franco Martínez ME, Ortega Pinto AV, Hernandez Rios M (2011) Inmunodetección de metaloproteinasas de matriz extracelular (MMPs) -2, -9, -13, -14 en lesiones apicales asociadas con periotontitis apical asintomática. Rev Clin Periodoncia Implantol Rehabil Oral 4(1):17–21CrossRefGoogle Scholar
- 15.Baeza M, Garrido M, Hernandez-Rios P, Dezerega A, Garcia-Sesnich J, Strauss F, Aitken JP, Lesaffre E, Vanbelle S, Gamonal J, Brignardello-Petersen R, Tervahartiala T, Sorsa T, Hernandez M (2016) Diagnostic accuracy for apical and chronic periodontitis biomarkers in gingival crevicular fluid: an exploratory study. J Clin Periodontol 43(1):34–45. https://doi.org/10.1111/jcpe.12479 CrossRefPubMedGoogle Scholar
- 16.Pereira Faustino IS, Azevedo RS, Takahama A Jr (2016) Metalloproteinases 2 and 9 immunoexpression in periapical lesions from primary endodontic infection: possible relationship with the histopathological diagnosis and the presence of pain. J Endod 42(4):547–551. https://doi.org/10.1016/j.joen.2015.12.020 CrossRefPubMedGoogle Scholar
- 18.Lisboa RA, Andrade MV, Cunha-Melo JR (2013) Toll-like receptor activation and mechanical force stimulation promote the secretion of matrix metalloproteinases 1, 3 and 10 of human periodontal fibroblasts via p38, JNK and NF-kB. Arch Oral Biol 58(6):731–739. https://doi.org/10.1016/j.archoralbio.2012.12.009 CrossRefPubMedGoogle Scholar
- 23.Rider D, Furusho H, Xu S, Trachtenberg AJ, Kuo WP, Hirai K, Susa M, Bahammam L, Stashenko P, Fujimura A, Sasaki H (2016) Elevated CD14 (cluster of differentiation 14) and toll-like receptor (TLR) 4 signaling deteriorate periapical inflammation in TLR2 deficient mice. Anat Rec (Hoboken) 299(9):1281–1292. https://doi.org/10.1002/ar.23383 CrossRefGoogle Scholar
- 24.Cavalla F, Reyes M, Vernal R, Alvarez C, Paredes R, Garcia-Sesnich J, Infante M, Farina V, Barron I, Hernandez M (2013) High levels of CXC ligand 12/stromal cell-derived factor 1 in apical lesions of endodontic origin associated with mast cell infiltration. J Endod 39(10):1234–1239. https://doi.org/10.1016/j.joen.2013.06.020 CrossRefPubMedGoogle Scholar
- 25.Garrido M, Dezerega A, Bordagaray MJ, Reyes M, Vernal R, Melgar-Rodriguez S, Ciuchi P, Paredes R, Garcia-Sesnich J, Ahumada-Montalva P, Hernandez M (2015) C-reactive protein expression is up-regulated in apical lesions of endodontic origin in association with interleukin-6. J Endod 41(4):464–469. https://doi.org/10.1016/j.joen.2014.12.021 CrossRefPubMedGoogle Scholar
- 28.Buzoglu HD, Unal H, Ulger C, Mert S, Kucukyildirim S, Er N (2009) The zymographic evaluation of gelatinase (MMP-2 and -9) levels in acute and chronic periapical abscesses. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 108(5):e121–e126. https://doi.org/10.1016/j.tripleo.2009.07.014 CrossRefPubMedGoogle Scholar
- 34.Hadziabdic N, Kurtovic-Kozaric A, Pojskic N, Sulejmanagic N, Todorovic L (2016) Gene-expression analysis of matrix metalloproteinases 1 and 2 and their tissue inhibitors in chronic periapical inflammatory lesions. J Oral Pathol Med 45(3):224–230. https://doi.org/10.1111/jop.12347 CrossRefPubMedGoogle Scholar
- 35.Trombone AP, Cavalla F, Silveira EM, Andreo CB, Francisconi CF, Fonseca AC, Letra A, Silva RM, Garlet GP (2016) MMP1-1607 polymorphism increases the risk for periapical lesion development through the upregulation MMP-1 expression in association with pro-inflammatory milieu elements. J Appl Oral Sci 24(4):366–375. https://doi.org/10.1590/1678-775720160112 CrossRefPubMedPubMedCentralGoogle Scholar
- 36.Shin SJ, Lee W, Lee JI, Baek SH, Kum KY, Shon WJ, Bae KS (2011) Matrix metalloproteinase-8 and substance P levels in gingival crevicular fluid during endodontic treatment of painful, nonvital teeth. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 112(4):548–554. https://doi.org/10.1016/j.tripleo.2011.04.026 CrossRefPubMedGoogle Scholar
- 37.Kuula H, Salo T, Pirila E, Tuomainen AM, Jauhiainen M, Uitto VJ, Tjaderhane L, Pussinen PJ, Sorsa T (2009) Local and systemic responses in matrix metalloproteinase 8-deficient mice during porphyromonas gingivalis-induced periodontitis. Infect Immun 77(2):850–859. https://doi.org/10.1128/IAI.00873-08 CrossRefPubMedGoogle Scholar
- 44.Osorio C, Cavalla F, Paula-Lima A, Diaz-Araya G, Vernal R, Ahumada P, Gamonal J, Hernandez M (2015) H2 O2 activates matrix metalloproteinases through the nuclear factor kappa B pathway and Ca(2+) signals in human periodontal fibroblasts. J Periodontal Res 50(6):798–806. https://doi.org/10.1111/jre.12267 CrossRefPubMedGoogle Scholar
- 49.Senturk RA, Sezgin Y, Bulut S, Ozdemir BH (2018) The effects of smoking on the expression of gelatinases in chronic periodontitis: a cross-sectional study. Braz Oral Res 32:e114. https://doi.org/10.1590/1807-3107bor-2018.vol32.0114 CrossRefPubMedGoogle Scholar