Skip to main content

Advertisement

SpringerLink
Log in

Osteoclast formation, inflammation, and matrix metalloproteinase-9 are downregulated in bone repair following root canal treatment in dogs teeth

  • Original Article
  • Published:
Clinical Oral Investigations Aims and scope Submit manuscript

Abstract

Objectives

The aim of this study was to investigate the inflammatory infiltrate, osteoclast formation, and expression of MMP-9 during the healing phase following root canal treatment in teeth with apical periodontitis.

Materials and methods

Apical periodontitis was induced in dogs teeth, and root canal treatment was performed in a single visit or using calcium hydroxide as intracanal medication. One hundred and eighty days following treatment the presence of inflammation was examined, and the tissues were stained to detect osteoclasts by means of a tartrate resistant alkaline phosphatase (TRAP) assay. Synthesis of MMP-9 was detected using Western blotting and immunohistochemistry.

Results

Teeth with apical periodontitis that had root canal therapy performed in a single visit presented a higher synthesis of MMP-9 compared with root canal treatment using calcium hydroxide. Treatment with calcium hydroxide resulted in a reduced amount of inflammatory cells and MMP-9 positive cells. Osteoclast formation, the number of MMP-9 positive osteoclasts and cementocytes, was reduced following root canal treatment, regardless of the root canal treatment protocol used.

Conclusion

Root canal treatment reduced the amount of inflammatory cells and osteoclasts in periapical area. The use of calcium hydroxide as intracanal medication resulted in a lower synthesis of MMP-9, though the number of osteoclasts and MMP-9 positive osteoclasts were similar between the groups.

Clinical relevance

Periapical bone repair following root canal treatment is impacted by therapy performed either in single visit or using calcium hydroxide dressing measured by inflammatory cell recruitment, osteoclast formation, and MMP-9 synthesis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  1. Martón IJ, Kiss C (2000) Protective and destructive immune reactions in apical periodontitis. Oral Microb Immunol 15:139–150

    Article  Google Scholar 

  2. Paula-Silva FWG, Ribeiro-Santos FR, Petean IBF, Manfrin Arnez MF, Almeida-Junior LA, Carvalho FK, Silva LABD, Faccioli LH (2020) Root canal contamination or exposure to lipopolysaccharide differentially modulate prostaglandin E 2 and leukotriene B 4 signaling in apical periodontitis. J Appl Oral Sci 28:e20190699

    Article  PubMed  PubMed Central  Google Scholar 

  3. Eberhard J, Plagmann HC (1999) Changes in the periodontal membrane due to apical periodontitis. J Endod 25:486–489

    Article  PubMed  Google Scholar 

  4. Paula-Silva FWG, da Silva LA, Kapila YL (2010) Matrix metalloproteinase expression in teeth with apical periodontitis is differentially modulated by the modality of root canal treatment. J Endod 36:231–237

    Article  PubMed  PubMed Central  Google Scholar 

  5. Cohenca N, Silva LAB, Silva RAB, Nelson-Filho P, Heilborn C, Watanabe E, Saraiva MCP (2013) Microbiological evaluation of different irrigation protocols on root canal disinfection in teeth with apical periodontitis: an in vivo study. Braz Dent J 24:467–473

    Article  PubMed  Google Scholar 

  6. Cohenca N, Romualdo PC, da Silva LA, da Silva RA, de Queiroz AM, De Rossi A, Nelson-Filho P (2015) Tissue response to root canal irrigation systems in dogs’ teeth with apical periodontitis. Clin Oral Investig 19:1147–1156

    Article  PubMed  Google Scholar 

  7. Jesus SF, Cohenca N, Romualdo PC, Nelson-Filho P, Queiroz AMD, Sousa-Neto MD, Silva LAB (2019) Radiographic and immunohistochemical evaluation of root canal treatment using different irrigation systems. Braz Dent J 30:123–132

    Article  PubMed  Google Scholar 

  8. Paula-Silva FWG, D’Silva NJ, Silva LAB, Kapila YL (2009) High matrix metalloproteinase activity is a hallmark of periapical granulomas. J Endod 35:1234–1242

    Article  PubMed  Google Scholar 

  9. Paula-Silva FWG, Arnez MFM, Petean IBF, Almeida-Junior LA, da Silva RAB, da Silva LAB, Faccioli LH (2020) Effects of 5-lipoxygenase gene disruption on inflammation, osteoclastogenesis and bone resorption in polymicrobial apical periodontitis. Arch Oral Biol 112:104670

    Article  PubMed  Google Scholar 

  10. Teronen O, Salo T, Konttinen YT, Rifkin B, Vernillo A, Ramamurthy NS, Kjeldsen L, Borregaard N, Hietanen J, Sorsa T (1995) Identification and characterization of gelatinase / type IV collagenase in jaw cysts. J Oral Pathol Med 24:78–84

    Article  PubMed  Google Scholar 

  11. Shin SJ, Lee J, Baek SH, Lin SS (2002) Tissue levels of matrix metalloproteinases in pulps and periapical lesions. J Endod 28:313–315

    Article  PubMed  Google Scholar 

  12. Wahlgren J, Salo T, Teronen O, Luoto H, Sorsa T, Tjäderhane L (2002) Matrix metalloproteinase-8 (MMP-8) in pulpal and periapical inflammation and periapical root- canal exudates. Int Endod J 35:897–904

    Article  PubMed  Google Scholar 

  13. Álvares PR, Arruda JAA, Silva LPD, Nascimento GJFD, Silveira MFD, Sobral APV (2017) Immunohistochemical expression of TGF-β1 and MMP-9 in periapical lesions. Braz Oral Res 31:e51

    Article  PubMed  Google Scholar 

  14. Andrade ALDL, Santos EM, Carmo AF, Freitas RA, Galvão HC (2017) Analysis of tryptase-positive mast cells and immunoexpression of MMP-9 and MMP-13 in periapical lesions. Int Endod J 50:446–454

    Article  PubMed  Google Scholar 

  15. Tay JY, Bay BH, Yeo JF, Harris M, Meghji S, Dheen ST (2004) Identification of RANKL in osteolytic lesions of the facial skeleton. J Dent Res 83:349–353

    Article  PubMed  Google Scholar 

  16. Trouvin AP, Goeb V (2010) Receptor activator of nuclear factor-kappa B ligand and osteoprotegerin: maintaining the balance to prevent bone loss. Clin Interv Aging 5:345–354

    PubMed  PubMed Central  Google Scholar 

  17. Carneiro E, Menezes R, Garlet GP, Garcia RB, Bramante CM, Figueira R, Sogayar M, Granjeiro JM (2009) Expression analysis of matrix metalloproteinase-9 in epithelialized and nonepithelialized apical periodontitis lesions. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 107:127–132

    Article  PubMed  Google Scholar 

  18. Campos K, Gomes CC, Farias LC, Silva RM, Letra A, Gomez RS (2016) DNA methylation of MMP9 is associated with high levels of MMP-9 messenger RNA in periapical inflammatory lesions. J Endod 42:127–130

    Article  PubMed  Google Scholar 

  19. Martinho FC, Teixeira FF, Cardoso FG, Ferreira NS, Nascimento GG, Carvalho CA, Valera MC (2016) Clinical investigation of matrix metalloproteinases, tissue inhibitors of matrix metalloproteinases, and matrix metalloproteinase/tissue inhibitors of matrix metalloproteinase complexes and their networks in apical periodontitis. J Endod 42:1082–1088

    Article  PubMed  Google Scholar 

  20. Guo J, Zeng X, Miao J, Liu C, Wei F, Liu D, Liu Y (2019) MiRNA-218 regulates osteoclast differentiation and inflammation response in periodontitis rats through Mmp9. Cell Microb 21:e12979

    Article  Google Scholar 

  21. Leonardo MR, Almeida WA, Ito IY, Silva LAB (1994) Radiographic and microbiologic evaluation of posttreatment apical and periapical repair of root canals of dogs’ teeth with experimentally induced chronic lesion. Oral Surg Oral Med Oral Pathol 78:232–238

    Article  PubMed  Google Scholar 

  22. Fabricius L, Dahlén G, Sundqvist G, Happonen RP, Möller AJ (2006) Influence of residual bacteria on periapical tissue healing after chemomechanical treatment and root filling of experimentally infected monkey teeth. Eur J Oral Sci 114:278–285

    Article  PubMed  Google Scholar 

  23. Takahama A Jr, Rôças IN, Faustino ISP, Alves FRF, Azevedo RS, Gomes CC, Araújo-Filho WR, Siqueira JF Jr (2018) Association between bacteria occurring in the apical canal system and expression of bone resorbing mediators and matrix metalloproteinases in apical periodontitis. Int Endod J 51:738–746

    Article  PubMed  Google Scholar 

  24. Paula-Silva FWG, Santamaria-Junior M, Leonardo MR, Consolaro A, da Silva LA (2009) Cone-beam computerized tomographic, radiographic, and histologic evaluation of periapical repair in dogs’ post-endodontic treatment. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 108:796–805

    Article  PubMed  Google Scholar 

  25. International Organization for Standardization (1997). Dentistry – Preclinical evaluation of biocompatibility of medical devices used in dentistry – Test methods for dental materials. (ISO Standard No. 7405)

  26. Birkedal-Hansen H (1993) Role of matrix metalloproteinases in human periodontal diseases. J Periodontal 64:474–484

    Google Scholar 

  27. Hong CY, Lin SK, Kok SH, Cheng SJ, Lee MS, Wang TM, Chen CS, Lin LD, Wang JS (2004) The role of lipopolysaccharide in infectious bone resorption of periapical lesion. J Oral Pathol Med 33:162–169

    Article  PubMed  Google Scholar 

  28. 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 42:547–551

    Article  PubMed  Google Scholar 

  29. D'addazio G, Artese L, Piccirilli M, Perfetti G (2014) Role of matrix metalloproteinases in radicular cysts and periapical granulomas. Minerva Stomatol 63:411–420

    PubMed  Google Scholar 

  30. Torres AFC, Antunes LS, Oliveira NF, Küchler EC, Gomes CC, Antunes LAA (2020). Genetic polymorphism and expression of matrix metalloproteinases and tissue inhibitors of metalloproteinases in periapical lesions: systematic review. J Endod 46:3-11.e1

  31. Corotti MV, Zambuzzi WF, Paiva KB, Menezes R, Pinto LC, Lara VS, Granjeiro JM (2009) Immunolocalization of matrix metalloproteinases-2 and -9 during apical periodontitis development. Arch Oral Biol 54:764–771

    Article  PubMed  Google Scholar 

  32. Carvalho CAT, Hasna AA, Carvalho AS, Vilela PDGF, Ramos LP, Valera MC, Oliveira LD (2020) Clinical study of sodium hypochlorite, Polymyxin B and limewater effect on MMP-3,-8,-9 in apical periodontitis. Braz Dent J 31:116–121

    Article  PubMed  Google Scholar 

  33. Leonardo MR, Salgado AA, da Silva LA, Tanomaru Filho M (2003) Apical and periapical repair of dogs’ teeth with periapical lesions after endodontic treatment with different root canal sealers. Pesqui Odontol Bras 17:69–74

    Article  PubMed  Google Scholar 

  34. Holland R, Otoboni Filho JA, de Souza V, Nery MJ, Bernabé PF, Dezan E Jr (2003) A comparison of one versus two appointment endodontic therapy in dogs' teeth with apical periodontitis. J Endod 29:121–124

    Article  PubMed  Google Scholar 

  35. Holland R, Sant'Anna Júnior A, Souza V, Dezan Junior E, Otoboni Filho JA, Bernabé PF, Nery MJ, Murata SS (2005) Influence of apical patency and filling material on healing process of dogs' teeth with vital pulp after root canal therapy. Braz Dent J 16:9–16

    Article  PubMed  Google Scholar 

  36. Arnold M, Ricucci D, Siqueira JF Jr (2013) Infection in a complex network of apical ramifications as the cause of persistent apical periodontitis: a case report. J Endod 39:1179–1184

    Article  PubMed  Google Scholar 

  37. Duque TM, Prado M, Herrera DR, Gomes BPFA (2019) Periodontal and endodontic infectious/inflammatory profile in primary periodontal lesions with secondary endodontic involvement after a calcium hydroxide-based intracanal medication. Clin Oral Invest 23:53–63

    Article  Google Scholar 

  38. Safavi KE, Nichols FC (1993) Effect of calcium hydroxide on bacterial lipopolysaccharide. J Endod 19:76–78

    Article  PubMed  Google Scholar 

  39. Nelson-Filho P, Leonardo MR, Silva LAB, Assed S (2002) Radiographic evaluation of the effect of endotoxin (LPS) plus calcium hydroxide on apical and periapical tissues of dogs. J Endod 28:694–696

    Article  PubMed  Google Scholar 

  40. Silva LABD, Silva RABD, Branco LGS, Navarro VP, Nelson-Filho P (2008) Quantitative radiographic evaluation of periapical bone resorption in dog's teeth contaminated with bacterial endotoxin (LPS) associated or not with calcium hydroxide. Braz Dent J 19:296–300

    Article  PubMed  Google Scholar 

  41. Özdemir MB, Karataş E, Albayrak M, Bayır Y (2019) Effect of intracanal medicaments on matrix metalloproteinase-9 and vasoactive intestinal peptide secretion in periapical lesions of re-treated canals: a randomized controlled clinical study. Clin Oral Investig 23:921–928

    Article  PubMed  Google Scholar 

  42. Wan C, Yuan G, Yang J, Sun Q, Zhang L, Zhang J, Chen Z (2014) MMP9 deficiency increased the size of experimentally induced apical periodontitis. J Endod 40:658–664

    Article  PubMed  Google Scholar 

  43. Engsig MT, Chen QJ, Vu TH, Pedersen AC, Therkidsen B, Lund LR, Henriksen K, Lenhard T, Foged NT, Werb Z, Delaissé JM (2000) Matrix metalloproteinase 9 and vascular endothelial growth factor are essential for osteoclast recruitment into developing long bones. J Cell Biol 51:879–889

    Article  Google Scholar 

  44. Zhang H, Liu L, Jiang C, Pan K, Deng J, Wan C (2020) MMP9 protects against LPS-induced inflammation in osteoblasts. Innate Immun 26:259–269

    Article  PubMed  Google Scholar 

  45. Barreiros D, Filho NP, Paula-Silva FWG, Oliveira KMH, Lucisano MP, Rossi A, Silva LAB, Küchler EC, Silva RAB (2018) MMP2 and MMP9 are associated with apical periodontitis progression and might be modulated by TLR2 and MyD88. Braz Dent J 29:43–47

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

The authors thank São Paulo Research Foundation for financial support (Fellowship to RP #2009/16882–7 and Grant to FWGPS #2019/02060–7).

Funding

The work was supported by the Department of Pediatric Clinics, School of Dentistry of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brazil, and São Paulo Research Foundation by a fellowship to author Renato Petille and a travel grant to author Francisco Wanderley Garcia Paula-Silva.

Author information

Authors and Affiliations

  1. Department of Pediatric Clinics, School of Dentistry of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brazil

    Francisco Wanderley Garcia Paula-Silva, Maya Fernanda Manfrin Arnez, Giuliana de Campos Chaves Lamarque, Renato Petille, Fernanda Regina Ribeiro-Santos, Marina Fernandes de Sena, Paulo Nelson-Filho & Léa Assed Bezerra da Silva

  2. Universidade de Pernambuco, Arco Verde, PE, Brazil

    Fernanda Regina Ribeiro-Santos

Authors
  1. Francisco Wanderley Garcia Paula-Silva
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Maya Fernanda Manfrin Arnez
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Giuliana de Campos Chaves Lamarque
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Renato Petille
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Fernanda Regina Ribeiro-Santos
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Marina Fernandes de Sena
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Paulo Nelson-Filho
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Léa Assed Bezerra da Silva
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Francisco Wanderley Garcia Paula-Silva.

Ethics declarations

Conflict of interest

Author Francisco Wanderley Garcia Paula-Silva declares that he has no conflict of interest. Author Maya Fernanda Manfrin Arnez declares that she has no conflict of interest. Author Giuliana de Campos Chaves Lamarque declares that she has no conflict of interest. Author Renato Petille declares that he has no conflict of interest. Author Fernanda Regina Ribeiro-Santos declares that she has no conflict of interest. Author Marina Fernandes de Sena declares that she has no conflict of interest. Author Paulo Nelson-Filho declares that he has no conflict of interest. Author Léa Assed Bezerra da Silva declares that she has no conflict of interest.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. The study was approved by the Institutional Animal Research Ethics Committee (process #2007.1.192.53.6).

Informed consent

For this type of study, formal consent is not required.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Paula-Silva, F.W.G., Arnez, M.F.M., de Campos Chaves Lamarque, G. et al. Osteoclast formation, inflammation, and matrix metalloproteinase-9 are downregulated in bone repair following root canal treatment in dogs teeth. Clin Oral Invest 25, 4699–4707 (2021). https://doi.org/10.1007/s00784-021-03784-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00784-021-03784-0

Keywords

Search

Navigation