Abstract
The role of bacterial infections in the pathogenesis of rheumatoid arthritis (RA) has gained increasing interest. Patients with RA often exhibit periodontal disease, which is associated with pathogens like Porphyromonas gingivalis. The present study examines the direct effects of P. gingivalis on apoptosis of human chondrocytes (a feature of inflammatory joint diseases) as one can assume an interrelation of pathogenesis of RA and P. gingivalis infections. Primary chondrocytes were infected with P. gingivalis. Early apoptotic and dead cell analysis was performed using Annexin-V, 7AAD, and propidium iodide and examined by flow cytometry and fluorescence microscopy. Caspase activation and DNA fragmentation were determined by western blot analysis and TUNEL reaction. Flow cytometry and fluorescence microscopy demonstrated an increase of Annexin-V-positive early apoptotic chondrocytes after infection. Western blot showed upregulation of activated caspase-3 expression, and TUNEL reaction revealed considerable DNA fragmentation following infection. The data show that P. gingivalis promotes early and later stages of apoptosis of primary human chondrocytes, which might contribute to the joint damage seen in the pathogenesis of RA.
Similar content being viewed by others
References
Alamanos Y, Drosos A (2005) Epidemiology of adult rheumatoid arthritis. Autoimmun Rev 4:130–136
Smith RL, Merchant TC, Schurman DJ (1982) In vitro cartilage degradation by Escherichia coli and Staphylococcus aureus. Arthritis Rheum 25:441–446
Lee MS, Ueng SWN, Shih CH, Chao CCK (2001) Primary cultures of human chondrocytes are susceptible to low inocula of Staphylococcus aureus infection and undergo apoptosis. Scand J Infect Dis 33:47–50
Jasin HE (1983) Bacterial lipopolysaccharides induce in vitro degradation of cartilage matrix through chondrocyte activation. J Clin Invest 72:2014–2019
Rosenstein ED, Greenwald RA, Kushner LJ, Weissmann G (2004) Hypothesis: the humoral immune response to oral bacteria provides a stimulus for the development of rheumatoid arthritis. Inflammation 28:311–318
Moen K, Brun JG, Valen M et al (1996) Synovial inflammation in active rheumatoid arthritis and psoriatic arthritis facilitates trapping of a variety of oral bacterial DNAs. Clin Exp Rheumatol 24:656–663
Hara Y, Kaneko T, Yoshimura A, Kato I (1996) Serum rheumatoid factor induced by intraperitoneal administration of periodontopathic bacterial lipopolysaccharide in mice. J Periodont Res 31:502–507
de Pablo P, Dietrich T, McAlindon T (2007) Association of periodontal disease and tooth loss with rheumatoid arthritis in the US population. J Rheumatol 35:70–76
Pischon N, Pischon T, Kroger J et al (2008) Association among rheumatoid arthritis, oral hygiene, and periodontitis. J Periodontol 79:979–986
Pischon N, Pischon T, Gülmez E et al (2010) Periodontal disease in patients with ankylosing spondylitis. Ann Rheum Dis 69:34–38
Mercado F, Marshall RI, Klestov A, Barthold PM (2001) Relationship between rheumatoid arthritis and periodontitis. J Periodontol 72:779–787
de Pablo P, Chapple IL, Buckley CD, Dietrich T (2009) Periodontitis in systemic rheumatic diseases. Nat Rev Rheumatol 5:218–224
Haffajee AD, Socransky SS (1994) Microbial etiological agents of destructive periodontal diseases. Periodontology 2000 5:78–111
Deshpande RG, Khan MB, Genco CA (1998) Invasion of aortic and heart endothelial cells by Porphyromonas gingivalis. Infect Immun 66:5337–5343
Weinberg A, Belton CM, Yoonsuk P, Lamont R (1997) Role of fimbriae in Porphyromonas gingivalis invasion of gingival epithelial cells. Infect Immun 65:313–316
Walter C, Zahlten J, Schmeck B et al (2004) Porphyromonas gingivalis strain dependent activation of human endothelial cells. Infect Immun 72:5910–5918
Pischon N, Röhner E, Hocke A et al (2009) Effects of Porphyromonas gingivalis on cell cycle progression and apoptosis of primary human chondrocytes. Ann Rheum Dis 68:1902–1907
Dorn BR, Burks JN, Seifert KN, Progulske-Fox A (2000) Invasion of endothelial and epithelial cells by strains of Porphyromonas gingivalis. FEMS Microbiol Lett 187:139–144
Urnowey S, Ansai T, Bitko V, Nakayama K, Takehara T, Barik S (2006) Temporal activation of anti- and pro-apoptotic factors in human gingival fibroblasts infected with the periodontal pathogen, Porphyromonas gingivalis: potential role of bacterial proteases in host signalling. BMC Microbiol 6:26
McGraw WT, Potempa J, Farley D, Travis J (1999) Purification, characterization, and sequence analysis of a potential virulence factor from Porphyromonas gingivalis, peptidylarginine deiminase. Infect Immun 67:3248–3256
Klamfeldt A (1986) Degradation of bovine articular cartilage proteoglycans in vitro. The effect of Bacteroides gingivalis. Scand J Rheumatol 15:297–301
Ronnelid J, Lysholm J, Engstrom-Laurent A et al (1994) Local anti-type II collagen antibody production in rheumatoid arthritis synovial fluid. Arthritis Rheum 37:1023–1029
Kim HA, Song YW (1999) Apoptotic chondrocyte death in rheumatoid arthritis. Arthritis Rheum 42:1528–1537
Matsumoto S, Müller-Ladner U, Gay RE, Nishioka K, Gay S (1996) Ultrastructural demonstration of apoptosis, Fas and Bcl-2 expression of rheumatoid synovial fibroblasts. J Rheumatol 23:1345–1352
Zahlten J, Riep B, Nichols FC et al (2007) Porphyromonas gingivalis dihydroceramides induce apoptosis in endothelial cells. J Dent Res 86:635–640
Wilton JM, Hurst TJ, Scott EE (1993) Inhibition of polymorphonuclear leucocyte phagpcytosis by Porphyromonas gingivalis culture products in patients with adult periodontitis. Arch Oral Biol 38:285–289
Ogrendik M (2009) Rheumatoid arthritis is linked to oral bacteria: etiological association. Mod Rheumatol
Kokkonen H, Söderström I, Rocklöv J, Hallmans G, Lejon K, Rantapää Dahlqvist S (2010) Up-regulation of cytokines and chemokines predates the onset of rheumatoid arthritis. Arthritis Rheum 62:383–391
Forner L, Larsen T, Kilian M, Holmstrup P (2006) Incidence of bacteremia after chewing, tooth brushing and scaling in individuals with periodontal inflammation. J Clin Periodontol 33:401–407
Mikuls TR, Payne JB, Reinhardt RA et al (2009) Antibody responses to Porphyromonas gingivalis in subjects with rheumatoid arthritis and periodontitis. Int Immunopharmacol 9:38–42
Hengartner MO (2000) The biochemistry of apoptosis. Nature 407:770–776
Murray DA, Wilton JM (2003) Lipopolysaccharide from the periodontal pathogen Porphyromonas gingivalis prevents apoptosis of HL60-derived neutrophils in vitro. Infect Immun 71:7232–7235
Sheets SM, Potempa J, Travis J, Casiano CA, Fletcher HM (2005) Gingipains from Porphyromonas gingivalis W83 induce cell adhesion molecule cleavage and apoptosis in endothelial cells. Infect Immun 73:1543–1552
Geatch DR, Harris JI, Heasman PA, Taylor JJ (1999) In vitro studies of lymphocyte apoptosis induced by the periodontal pathogen Porphyromonas gingivalis. J Periodontal Res 34:70–78
Stathopoulou PG, Galicia JC, Benakanakere MR, Garcia CA, Potempa J, Kinane DF (2009) Porphyromonas gingivalis induce apoptosis in human gingival epithelial cells through a gingipain-dependent mechanism. BMC Microbiol 9:107
Porter AG, Jänicke RU (1999) Emerging roles of caspase-3 in apoptosis. Cell Death Differ 6:99–104
Brozovic S, Sahoo R, Barve S et al (2006) Porphyromonas gingivalis enhances FasL expression via up-regulation of NFkappaB-mediated gene transcription and induces apoptotic cell death in human gingival epithelial cells. Microbiology 152:797–806
Nakhjiri SF, Park Y, Yilmaz O et al (2001) Inhibition of epithelial cell apoptosis by Porphyromonas gingivalis. FEMS Microbiol Lett 200:145–149
Acknowledgements
This work was financially supported DFG GK 325 (Bonn, Germany), by DGP e.V. (Germany), GABA GmbH (Lörrach, Germany), and a Habilitation Fellowship from Charité-Universitätsmedizin (Berlin, Germany to N. P.).
Author information
Authors and Affiliations
Corresponding author
Additional information
E. Röhner and J. Detert contributed equally to this work.
The authors have stated that they have no conflict of interest.
Rights and permissions
About this article
Cite this article
Röhner, E., Detert, J., Kolar, P. et al. Induced Apoptosis of Chondrocytes by Porphyromonas gingivalis as a Possible Pathway for Cartilage Loss in Rheumatoid Arthritis. Calcif Tissue Int 87, 333–340 (2010). https://doi.org/10.1007/s00223-010-9389-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00223-010-9389-5