, Volume 42, Issue 3, pp 1117–1128 | Cite as

Periodontal Injection of Lipopolysaccharide Promotes Arthritis Development in Mice

  • Anna ScanuEmail author
  • Chiara Giraudo
  • Francesca Galuppini
  • Vanni Lazzarin
  • Gianmaria Pennelli
  • Stefano Sivolella
  • Edoardo Stellini
  • Francesca Oliviero
  • Paola Galozzi
  • Massimo Rugge
  • Roberto Stramare
  • Roberto Luisetto
  • Leonardo Punzi


This study evaluated the arthritogenic effect of lipopolysaccharide (LPS) in a mouse model of periodontal disease. Periodontitis was induced in wild-type CD1 mice by nine LPS injections (10 or 50 ng) into the maxillary mucosa. Untreated mice or injected with LPS at the tail were used as controls. Two weeks after final inoculation, mice were sacrificed to collect blood, maxilla, and paw samples. Development and progression of periodontitis and arthritis were monitored using clinical assessment, micro-computed tomography (micro-CT), ultrasound (US), and histological analysis. CXCL1, IL-1β, IL-6, TNF-α, and anti-citrullinated peptide antibodies (ACPA) serum levels were determined by enzyme immunoassay. Ankle swelling and inflammation manifested after the 5th periodontal injection of 50 ng of LPS and progressed until the end of experiments. Periodontal injection of 10 ng of LPS and LPS tail injection did not induce paw changes. Therefore, the subsequent assessments were conducted only in mice periodontally injected with 50 ng of LPS. Maxillary micro-CT and histological analysis showed that LPS-induced alveolar bone resorption and vascular proliferation in periodontal tissue, but not inflammation. US and histology revealed increased joint space, leukocyte infiltration, synovial proliferation, and mild cartilage and bone destruction in the paws of mice orally injected. Cytokines and ACPA showed a trend towards an increase in LPS mice. This study shows that arthritis and periodontal disease can co-occur in wild-type mice after periodontal injection of LPS at optimal dose. Our model may be useful to improve the understanding of the mechanisms linking periodontitis and arthritis.


periodontitis arthritis inflammation mouse model periodontal disease animal model 



This work was supported by Institutional Research Funds “DOR1615309/16” and “DOR1784408/17” from the University of Padova.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.


  1. 1.
    Kassebaum, N.J., E. Bernabé, M. Dahiya, B. Bhandari, C.J. Murray, and W. Marcenes. 2014. Global burden of severe periodontitis in 1990-2010: a systematic review and meta-regression. Journal of Dental Research 93: 1045–1053.PubMedPubMedCentralGoogle Scholar
  2. 2.
    Pihlstrom, B.L., B.S. Michalowicz, and N.W. Johnson. 2005. Periodontal diseases. Lancet 366: 1809–1820.PubMedGoogle Scholar
  3. 3.
    Holt, S.C., J. Ebersole, J. Felton, M. Brunsvold, and K.S. Kornman. 1988. Implantation of Bacteroides gingivalis in nonhuman primates initiates progression of periodontitis. Science 239: 55–57.PubMedGoogle Scholar
  4. 4.
    Kadono, H., J. Kido, M. Kataoka, N. Yamauchi, and T. Nagata. 1999. Inhibition of osteoblastic cell differentiation by lipopolysaccharide extract from Porphyromonas gingivalis. Infection and Immunity 67: 2841–2846.PubMedPubMedCentralGoogle Scholar
  5. 5.
    Kadowaki, T., K. Nakayama, K. Okamoto, N. Abe, A. Baba, Y. Shi, D.B. Ratnayake, and K. Yamamoto. 2000. Porphyromonas gingivalis proteinases as virulence determinants in progression of periodontal diseases. The Journal of Biochemistry 128: 153–159.PubMedGoogle Scholar
  6. 6.
    McGraw, W.T., J. Potempa, D. Farley, and J. Travis. 1999. Purification, characterization, and sequence analysis of a potential virulence factor from Porphyromonas gingivalis, peptidylarginine deiminase. Infection and Immunity 67: 3248–3256.PubMedPubMedCentralGoogle Scholar
  7. 7.
    Kassem, A., P. Henning, P. Lundberg, P.P. Souza, C. Lindholm, and U.H. Lerner. 2015. Porphyromonas gingivalis stimulates bone resorption by enhancing RANKL (receptor activator of NF-κB ligand) through activation of Toll-like receptor 2 in osteoblasts. Journal of Biological Chemistry 290: 20147–20158.PubMedGoogle Scholar
  8. 8.
    Jiang, Y., C.K. Mehta, T.Y. Hsu, and F.F. Alsulaimani. 2002. Bacteria induce osteoclastogenesis via an osteoblast-independent pathway. Infection and Immunity 70: 3143–3148.PubMedPubMedCentralGoogle Scholar
  9. 9.
    Rogers, J.E., F. Li, D.D. Coatney, C. Rossa, P. Bronson, J.M. Krieder, W.V. Giannobile, and K.L. Kirkwood. 2007. Actinobacillus actinomycetemcomitans lipopolysaccharide-mediated experimental bone loss model for aggressive periodontitis. Journal of Periodontology 78: 550–558.PubMedPubMedCentralGoogle Scholar
  10. 10.
    Garcia de Aquino, S., F.R. Manzolli Leite, D.R. Stach-Machado, J.A. Francisco da Silva, L.C. Spolidorio, and C. Rossa Jr. 2009. Signaling pathways associated with the expression of inflammatory mediators activated during the course of two models of experimental periodontitis. Life Sciences 84: 745–754.PubMedGoogle Scholar
  11. 11.
    Pizzo, G., R. Guiglia, L. Lo Russo, and G. Campisi. 2010. Dentistry and internal medicine: from the focal infection theory to the periodontal medicine concept. European Journal of Internal Medicine 21: 496–502.PubMedGoogle Scholar
  12. 12.
    de Pablo, P., I.L. Chapple, C.D. Buckley, and T. Dietrich. 2009. Periodontitis in systemic rheumatic diseases. Nature Reviews Rheumatology 5: 218–224.PubMedGoogle Scholar
  13. 13.
    Mikuls, T.R., J.B. Payne, F. Yu, G.M. Thiele, R.J. Reynolds, G.W. Cannon, J. Markt, D. McGowan, G.S. Kerr, R.S. Redman, A. Reimold, G. Griffiths, M. Beatty, S.M. Gonzalez, D.A. Bergman, B.C. Hamilton III, A.R. Erickson, J. Sokolove, W.H. Robinson, C. Walker, F. Chandad, and J.R. O'Dell. 2014. Periodontitis and Porphyromonas gingivalis in patients with rheumatoid arthritis. Arthritis & Rheumatology 66: 1090–1100.Google Scholar
  14. 14.
    Kässer, U.R., C. Gleissner, F. Dehne, A. Michel, B. Willershausen-Zönnchen, and W.W. Bolten. 1997. Risk for periodontal disease in patients with longstanding rheumatoid arthritis. Arthritis and Rheumatism 40: 2248–2251.PubMedGoogle Scholar
  15. 15.
    Scher, J.U., C. Ubeda, M. Equinda, R. Khanin, Y. Buischi, A. Viale, L. Lipuma, M. Attur, M.H. Pillinger, G. Weissmann, D.R. Littman, E.G. Pamer, W.A. Bretz, and S.B. Abramson. 2012. Periodontal disease and the oral microbiota in new-onset rheumatoid arthritis. Arthritis and Rheumatism 64: 3083–3094.PubMedPubMedCentralGoogle Scholar
  16. 16.
    de Smit, M., J. Westra, A. Vissink, B. Doornbos-van der Meer, E. Brouwer, and A.J. van Winkelhoff. 2012. Periodontitis in established rheumatoid arthritis patients: a cross-sectional clinical, microbiological and serological study. Arthritis Research & Therapy 14: R222.Google Scholar
  17. 17.
    Potempa, J., P. Mydel, and J. Koziel. 2017. The case for periodontitis in the pathogenesis of rheumatoid arthritis. Nature Reviews Rheumatology 13: 606–620.PubMedGoogle Scholar
  18. 18.
    Lundberg, K., N. Wegner, T. Yucel-Lindberg, and P.J. Venables. 2010. Periodontitis in RA—the citrullinated enolase connection. Nature Reviews Rheumatology 6: 727–730.PubMedGoogle Scholar
  19. 19.
    Stabholz, A., W.A. Soskolne, and L. Shapira. 2010. Genetic and environmental risk factors for chronic periodontitis and aggressive periodontitis. Periodontology 2000 (53): 138–153.Google Scholar
  20. 20.
    Di Giuseppe, D., A. Discacciati, N. Orsini, and A. Wolk. 2014. Cigarette smoking and risk of rheumatoid arthritis: a dose-response meta-analysis. Arthritis Research & Therapy 16: R61.Google Scholar
  21. 21.
    Fisher, B.A., A.J. Cartwright, A.M. Quirke, P. de Pablo, D. Romaguera, S. Panico, A. Mattiello, D. Gavrila, C. Navarro, C. Sacerdote, P. Vineis, R. Tumino, D.F. Lappin, D. Apatzidou, S. Culshaw, J. Potempa, D.S. Michaud, E. Riboli, and P.J. Venables. 2015. Smoking, Porphyromonas gingivalis and the immune response to citrullinated autoantigens before the clinical onset of rheumatoid arthritis in a southern European nested case-control study. BMC Musculoskeletal Disorders 16: 331.PubMedPubMedCentralGoogle Scholar
  22. 22.
    Maciel, S.S., M. Feres, T.E. Gonçalves, G.S. Zimmermann, H.D. da Silva, L.C. Figueiredo, and P.M. Duarte. 2016. Does obesity influence the subgingival microbiota composition in periodontal health and disease? Journal of Clinical Periodontology 43: 1003–1012.PubMedGoogle Scholar
  23. 23.
    Chaffee, B.W., and S.J. Weston. 2010. Association between chronic periodontal disease and obesity: a systematic review and meta-analysis. Journal of Periodontology 81: 1708–1724.PubMedPubMedCentralGoogle Scholar
  24. 24.
    Qin, B., M. Yang, H. Fu, N. Ma, T. Wei, Q. Tang, Z. Hu, Y. Liang, Z. Yang, and R. Zhong. 2015. Body mass index and the risk of rheumatoid arthritis: a systematic review and dose-response meta-analysis. Arthritis Research & Therapy 17: 86.Google Scholar
  25. 25.
    Cheng, Z., J. Meade, K. Mankia, P. Emery, and D.A. Devine. 2017. Periodontal disease and periodontal bacteria as triggers for rheumatoid arthritis. Best Practice & Research in Clinical Rheumatology 31: 19–30.Google Scholar
  26. 26.
    Maresz, K.J., A. Hellvard, A. Sroka, K. Adamowicz, E. Bielecka, J. Koziel, K. Gawron, D. Mizgalska, K.A. Marcinska, M. Benedyk, K. Pyrc, A.M. Quirke, R. Jonsson, S. Alzabin, P.J. Venables, K.A. Nguyen, P. Mydel, and J. Potempa. 2013. Porphyromonas gingivalis facilitates the development and progression of destructive arthritis through its unique bacterial peptidylarginine deiminase (PAD). PLoS Pathogens 9: e1003627.PubMedPubMedCentralGoogle Scholar
  27. 27.
    Marchesan, J.T., E.A. Gerow, R. Schaff, A.D. Taut, S.Y. Shin, J. Sugai, D. Brand, A. Burberry, J. Jorns, S.K. Lundy, G. Nuñez, D.A. Fox, and W.V. Giannobile. 2013. Porphyromonas gingivalis oral infection exacerbates the development and severity of collagen-induced arthritis. Arthritis Research & Therapy 15: R186.Google Scholar
  28. 28.
    de Aquino, S.G., S. Abdollahi-Roodsaz, M.I. Koenders, F.A. van de Loo, G.J. Pruijn, R.J. Marijnissen, B. Walgreen, M.M. Helsen, L.A. van den Bersselaar, R.S. de Molon, M.J. Avila Campos, F.Q. Cunha, J.A. Cirelli, and W.B. van den Berg. 2014. Periodontal pathogens directly promote autoimmune experimental arthritis by inducing a TLR2- and IL-1-driven Th17 response. The Journal of Immunology 192: 4103–4111.PubMedGoogle Scholar
  29. 29.
    Chukkapalli, S., M. Rivera-Kweh, P. Gehlot, I. Velsko, I. Bhattacharyya, S.J. Calise, M. Satoh, E.K. Chan, J. Holoshitz, and L. Kesavalu. 2016. Periodontal bacterial colonization in synovial tissues exacerbates collagen-induced arthritis in B10.RIII mice. Arthritis Research & Therapy 18: 161.Google Scholar
  30. 30.
    Jung, H., S.M. Jung, Y.A. Rim, N. Park, Y. Nam, J. Lee, S.H. Park, and J.H. Ju. 2017. Arthritic role of Porphyromonas gingivalis in collagen-induced arthritis mice. PLoS One 12: e0188698.PubMedPubMedCentralGoogle Scholar
  31. 31.
    Corrêa, M.G., S.B. Sacchetti, F.V. Ribeiro, S.P. Pimentel, R.C. Casarin, F.R. Cirano, and M.Z. Casati. 2017. Periodontitis increases rheumatic factor serum levels and citrullinated proteins in gingival tissues and alter cytokine balance in arthritic rats. PLoS One 12: e0174442.PubMedPubMedCentralGoogle Scholar
  32. 32.
    Sandal, I., A. Karydis, J. Luo, A. Prislovsky, K.B. Whittington, E.F. Rosloniec, C. Dong, D.V. Novack, P. Mydel, S.G. Zheng, M.Z. Radic, and D.D. Brand. 2016. Bone loss and aggravated autoimmune arthritis in HLA-DRβ1-bearing humanized mice following oral challenge with Porphyromonas gingivalis. Arthritis Research & Therapy 18: 249.Google Scholar
  33. 33.
    Gully, N., R. Bright, V. Marino, C. Marchant, M. Cantley, D. Haynes, C. Butler, S. Dashper, E. Reynolds, and M. Bartold. 2014. Porphyromonas gingivalis peptidylarginine deiminase, a key contributor in the pathogenesis of experimental periodontal disease and experimental arthritis. PLoS One 9: e100838.PubMedPubMedCentralGoogle Scholar
  34. 34.
    Yamakawa, M., K. Ouhara, M. Kajiya, S. Munenaga, M. Kittaka, S. Yamasaki, K. Takeda, K. Takeshita, N. Mizuno, T. Fujita, E. Sugiyama, and H. Kurihara. 2016. Porphyromonas gingivalis infection exacerbates the onset of rheumatoid arthritis in SKG mice. Clinical & Experimental Immunology 186: 177–189.Google Scholar
  35. 35.
    de Aquino, S.G., J. Talbot, F. Sônego, W.M. Turato, R. Grespan, M.J. Avila-Campos, F.Q. Cunha, and J.A. Cirelli. 2017. The aggravation of arthritis by periodontitis is dependent of IL-17 receptor A activation. Journal of Clinical Periodontology 44: 881–891.PubMedGoogle Scholar
  36. 36.
    Bevaart, L., M.J. Vervoordeldonk, and P.P. Tak. 2010. Collagen-induced arthritis in mice. Methods in Molecular Biology 602: 181–192. Humana Press.Google Scholar
  37. 37.
    de Molon, R.S., E.D. de Avila, and J.A. Cirelli. 2013. Host responses induced by different animal models of periodontal disease: a literature review. Journal of Investigative and Clinical Dentistry 4: 211–218.PubMedGoogle Scholar
  38. 38.
    Oz, H.S., and D.A. Puleo. 2011. Animal models for periodontal disease. Journal of Biomedicine and Biotechnology 2011: 754857.PubMedPubMedCentralGoogle Scholar
  39. 39.
    Yamaguchi, M., T. Ukai, T. Kaneko, M. Yoshinaga, M. Yokoyama, Y. Ozaki, and Y. Hara. 2008. T cells are able to promote lipopolysaccharide-induced bone resorption in mice in the absence of B cells. Journal of Periodontal Research 43: 549–555.PubMedGoogle Scholar
  40. 40.
    Park, J., J.D. Cha, K.M. Choi, K.Y. Lee, K.M. Han, and Y.S. Jang. 2017. Fucoidan inhibits LPS-induced inflammation in vitro and during the acute response in vivo. International Immunopharmacology 43: 91–98.PubMedGoogle Scholar
  41. 41.
    Elburki, M.S., C. Rossa Jr., M.R. Guimarães-Stabili, H.M. Lee, F.A. Curylofo-Zotti, F. Johnson, and L.M. Golub. 2017. A chemically modified curcumin (CMC 2.24) inhibits nuclear factor κB activation and inflammatory bone loss in murine models of LPS-induced experimental periodontitis and diabetes-associated natural periodontitis. Inflammation 40: 1436–1449.PubMedGoogle Scholar
  42. 42.
    Saito, T., Y. Yamamoto, G.G. Feng, Y. Kazaoka, Y. Fujiwara, and H. Kinoshita. 2017. Lidocaine prevents oxidative stress-induced endothelial dysfunction of the systemic artery in rats with intermittent periodontal inflammation. Anesthesia & Analgesia 124: 2054–2062.Google Scholar
  43. 43.
    Eriksson, K., E. Lönnblom, G. Tour, A. Kats, P. Mydel, P. Georgsson, C. Hultgren, N. Kharlamova, U. Norin, J. Jönsson, A. Lundmark, A. Hellvard, K. Lundberg, L. Jansson, R. Holmdahl, and T. Yucel-Lindberg. 2016. Effects by periodontitis on pristane-induced arthritis in rats. Journal of Translational Medicine 14: 311.PubMedPubMedCentralGoogle Scholar
  44. 44.
    Bas, S., S. Genevay, O. Meyer, and C. Gabay. 2003. Anti-cyclic citrullinated peptide antibodies, IgM and IgA rheumatoid factors in the diagnosis and prognosis of rheumatoid arthritis. Rheumatology (Oxford) 42: 677–680.Google Scholar
  45. 45.
    Aletaha, D., T. Neogi, A.J. Silman, J. Funovits, D.T. Felson, C.O. Bingham 3rd, N.S. Birnbaum, G.R. Burmester, V.P. Bykerk, M.D. Cohen, B. Combe, K.H. Costenbader, M. Dougados, P. Emery, G. Ferraccioli, J.M. Hazes, K. Hobbs, T.W. Huizinga, A. Kavanaugh, J. Kay, T.K. Kvien, T. Laing, P. Mease, H.A. Ménard, L.W. Moreland, R.L. Naden, T. Pincus, J.S. Smolen, E. Stanislawska-Biernat, D. Symmons, P.P. Tak, K.S. Upchurch, J. Vencovsky, F. Wolfe, and G. Hawker. 2010. 2010 rheumatoid arthritis classification criteria: an American College of Rheumatology/European League Against Rheumatism collaborative initiative. Annals of the Rheumatic Diseases 69: 1580–1588.PubMedGoogle Scholar
  46. 46.
    Egeberg, A., L. Mallbris, G. Gislason, P.R. Hansen, and U. Mrowietz. 2017. Risk of periodontitis in patients with psoriasis and psoriatic arthritis. Journal of the European Academy of Dermatology and Venereology 31: 288–293.PubMedGoogle Scholar
  47. 47.
    Üstün, K., U. Sezer, B. Kısacık, S.Z. Şenyurt, E.Ç. Özdemir, G. Kimyon, Y. Pehlivan, K. Erciyas, and A.M. Onat. 2013. Periodontal disease in patients with psoriatic arthritis. Inflammation 36: 665–669.PubMedGoogle Scholar
  48. 48.
    Fuggle, N.R., T.O. Smith, A. Kaul, and N. Sofat. 2016. Hand to mouth: a systematic review and meta-analysis of the association between rheumatoid arthritis and periodontitis. Frontiers in Immunology 7: 80.PubMedPubMedCentralGoogle Scholar
  49. 49.
    Damjanovska, L., M.M. Thabet, E.W. Levarth, G. Stoeken-Rijsbergen, E.I. van der Voort, R.E. Toes, T.W. Huizinga, and A.H. van der Helm-van Mil. 2010. Diagnostic value of anti-MCV antibodies in differentiating early inflammatory arthritis. Annals of the Rheumatic Diseases 69: 730–732.PubMedGoogle Scholar
  50. 50.
    Kobayashi, T., and H. Yoshie. 2015. Host responses in the link between periodontitis and rheumatoid arthritis. Current Oral Health Reports 2: 1–8.PubMedGoogle Scholar
  51. 51.
    van den Berg, W.B. 2009. Lessons from animal models of arthritis over the past decade. Arthritis Research & Therapy 11: 250.Google Scholar
  52. 52.
    Bessis, N., P. Decker, E. Assier, L. Semerano, and M.C. Boissier. 2017. Arthritis models: usefulness and interpretation. Seminars in Immunopathology 39: 469–486.PubMedGoogle Scholar
  53. 53.
    Scanu, A., R. Luisetto, F. Oliviero, L. Gruaz, P. Sfriso, D. Burger, and L. Punzi. 2015. High-density lipoproteins inhibit urate crystal-induced inflammation in mice. Annals of the Rheumatic Diseases 74: 587–594.PubMedGoogle Scholar
  54. 54.
    Payne, J.B., L.M. Golub, G.M. Thiele, and T.R. Mikuls. 2015. The link between periodontitis and rheumatoid arthritis: a periodontist’s perspective. Current Oral Health Reports 2: 20–29.PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Anna Scanu
    • 1
    Email author
  • Chiara Giraudo
    • 2
  • Francesca Galuppini
    • 3
    • 4
  • Vanni Lazzarin
    • 3
  • Gianmaria Pennelli
    • 3
  • Stefano Sivolella
    • 5
  • Edoardo Stellini
    • 5
  • Francesca Oliviero
    • 1
  • Paola Galozzi
    • 1
  • Massimo Rugge
    • 3
  • Roberto Stramare
    • 2
  • Roberto Luisetto
    • 6
  • Leonardo Punzi
    • 7
  1. 1.Rheumatology Unit, Department of Medicine – DIMEDUniversity of PadovaPadovaItaly
  2. 2.Radiology Unit, Department of Medicine – DIMEDUniversity of PadovaPadovaItaly
  3. 3.Surgical Pathology Unit, Department of Medicine – DIMEDUniversity of PadovaPadovaItaly
  4. 4.Department of Women’s and Children’s HealthPadovaItaly
  5. 5.Dentistry Section, Department of NeurosciencesUniversity of PadovaPadovaItaly
  6. 6.Department of Surgical Oncological and Gastroenterological SciencesUniversity of PadovaPadovaItaly
  7. 7.Centre for Gout and Metabolic Bone and Joint Diseases, RheumatologySS Giovanni and Paolo HospitalVeniceItaly

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