Porphyromonas gingivalis infection increases osteoclastic bone resorption and osteoblastic bone formation in a periodontitis mouse model

BMC Oral Health

Gena Tribble¹  Todd Rigney¹  Jun Ju²  Wenjian Zhang² 

[1] Department of Periodontics and Dental Hygiene, University of Texas School of Dentistry at Houston, 7500 Cambridge Street, Houston 77054, TX, USA;Department of Diagnostic and Biomedical Sciences, 7500 Cambridge Street, Suite 5366, Houston 77054, TX, USA

关键词: Bone histomorphometry;    Micro-CT;    Invasion;    P. gingivalis;    Osteoblasts;   

Others  :  1109608
DOI  :  10.1186/1472-6831-14-89

 received in 2014-04-24, accepted in 2014-07-10,  发布年份 2014

【 摘 要 】

Background

Porphyromonas gingivalis has been shown to invade osteoblasts and inhibit their differentiation and mineralization in vitro. However, it is unclear if P. gingivalis can invade osteoblasts in vivo and how this would affect alveolar osteoblast/osteoclast dynamics. This study aims to answer these questions using a periodontitis mouse model under repetitive P. gingivalis inoculations.

Methods

For 3-month-old BALB/cByJ female mice, 10⁹ CFU of P. gingivalis were inoculated onto the gingival margin of maxillary molars 4 times at 2-day intervals. After 2 weeks, another 4 inoculations at 2-day intervals were applied. Calcein was injected 7 and 2 days before sacrificing animals to label the newly formed bone. Four weeks after final inoculation, mice were sacrificed and maxilla collected. Immunohistochemistry, micro-CT, and bone histomorphometry were performed on the specimens. Sham infection with only vehicle was the control.

Results

P. gingivalis was found to invade gingival epithelia, periodontal ligament fibroblasts, and alveolar osteoblasts. Micro-CT showed alveolar bone resorption and significant reduction of bone mineral density and content in the infected mice compared to the controls. Bone histomorphometry showed a decrease in osteoblasts, an increase in osteoclasts and bone resorption, and a surprisingly increased osteoblastic bone formation in the infected mice compared to the controls.

Conclusions

P. gingivalis invades alveolar osteoblasts in the periodontitis mouse model and cause alveolar bone loss. Although P. gingivalis appears to suppress osteoblast pool and enhance osteoclastic bone resorption, the bone formation capacity is temporarily elevated in the infected mice, possibly via some anti-microbial compensational mechanisms.

【 授权许可】

   
2014 Zhang et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Burt B: Position paper: epidemiology of periodontal diseases. J Periodontol 2005, 76(8):1406-1419.
• [2]Locker D, Matear D, Stephens M, Jokovic A: Oral health-related quality of life of a population of medically compromised elderly people. Community Dent Hlth 2002, 19(2):90-97.
• [3]Socransky SS, Haffajee AD: The bacterial etiology of destructive periodontal disease: current concepts. J Periodontol 1992, 63(4 Suppl):322-331.
• [4]Holt SC, Kesavalu L, Walker S, Genco CA: Virulence factors of Porphyromonas gingivalis. Periodontol 2000 1999, 20:168-238.
• [5]Azuma H, Kido J, Ikedo D, Kataoka M, Nagata T: Substance P enhances the inhibition of osteoblastic cell differentiation induced by lipopolysaccharide from Porphyromonas gingivalis. J Periodontol 2004, 75(7):974-981.
• [6]Kadono H, Kido J, Kataoka M, Yamauchi N, Nagata T: Inhibition of osteoblastic cell differentiation by lipopolysaccharide extract from Porphyromonas gingivalis. Infect Immun 1999, 67(6):2841-2846.
• [7]Kato T, Tsuda T, Inaba H, Kawai S, Okahashi N, Shibata Y, Abiko Y, Amano A: Porphyromonas gingivalis gingipains cause G(1) arrest in osteoblastic/stromal cells. Oral Microbiol Immun 2008, 23(2):158-164.
• [8]Kim CS, Choi SH, Choi BK, Chai JK, Park JB, Kim CK, Cho KS: The effect of recombinant human bone morphogenetic protein-4 on the osteoblastic differentiation of mouse calvarial cells affected by Porphyromonas gingivalis. J Periodontol 2002, 73(10):1126-1132.
• [9]Loomer PM, Ellen RP, Tenenbaum HC: Characterization of inhibitory effects of suspected periodontopathogens on osteogenesis in vitro. Infect Immun 1995, 63(9):3287-3296.
• [10]Loomer PM, Sigusch B, Sukhu B, Ellen RP, Tenenbaum HC: Direct effects of metabolic products and sonicated extracts of Porphyromonas gingivalis 2561 on osteogenesis in vitro. Infect Immun 1994, 62(4):1289-1297.
• [11]Choi BK, Moon SY, Cha JH, Kim KW, Yoo YJ: Prostaglandin E(2) is a main mediator in receptor activator of nuclear factor-kappaB ligand-dependent osteoclastogenesis induced by Porphyromonas gingivalis, Treponema denticola, and Treponema socranskii. J Periodontol 2005, 76(5):813-820.
• [12]Okahashi N, Inaba H, Nakagawa I, Yamamura T, Kuboniwa M, Nakayama K, Hamada S, Amano A: Porphyromonas gingivalis induces receptor activator of NF-kappaB ligand expression in osteoblasts through the activator protein 1 pathway. Infect Immun 2004, 72(3):1706-1714.
• [13]Wang YH, Jiang J, Zhu Q, AlAnezi AZ, Clark RB, Jiang X, Rowe DW, Nichols FC: Porphyromonas gingivalis lipids inhibit osteoblastic differentiation and function. Infect Immun 2010, 78(9):3726-3735.
• [14]Yasuhara R, Miyamoto Y, Takami M, Imamura T, Potempa J, Yoshimura K, Kamijo R: Lysine-specific gingipain promotes lipopolysaccharide- and active-vitamin D3-induced osteoclast differentiation by degrading osteoprotegerin. Biochem J 2009, 419(1):159-166.
• [15]Zhang W, Swearingen EB, Ju J, Rigney T, Tribble GD: Porphyromonas gingivalis invades osteoblasts and inhibits bone formation. Microbes Infect 2010, 12(11):838-845.
• [16]Zhang W, Ju J, Rigney T, Tribble GD: Fimbriae of Porphyromonas gingivalis are important for initial invasion of osteoblasts, but not for inhibition of their differentiation and mineralization. J Periodontol 2011, 82(6):909-916.
• [17]Frank RM, Voegel JC: Bacterial bone resorption in advanced cases of human periodontitis. J Periodontal Res 1978, 13(3):251-261.
• [18]Baker PJ, Evans RT, Roopenian DC: Oral infection with Porphyromonas gingivalis and induced alveolar bone loss in immunocompetent and severe combined immunodeficient mice. Arch Oral Biol 1994, 39(12):1035-1040.
• [19]Baker PJ, Dixon M, Roopenian DC: Genetic control of susceptibility to Porphyromonas gingivalis-induced alveolar bone loss in mice. Infect Immun 2000, 68(10):5864-5868.
• [20]Hart GT, Shaffer DJ, Akilesh S, Brown AC, Moran L, Roopenian DC, Baker PJ: Quantitative gene expression profiling implicates genes for susceptibility and resistance to alveolar bone loss. Infect Immun 2004, 72(8):4471-4479.
• [21]Pathirana RD, O’Brien-Simpson NM, Brammar GC, Slakeski N, Reynolds EC: Kgp and RgpB, but not RgpA, are important for Porphyromonas gingivalis virulence in the murine periodontitis model. Infect Immun 2007, 75(3):1436-1442.
• [22]O’Brien-Simpson NM, Pathirana RD, Paolini RA, Chen YY, Veith PD, Tam V, Ally N, Pike RN, Reynolds EC: An immune response directed to proteinase and adhesin functional epitopes protects against Porphyromonas gingivalis-induced periodontal bone loss. J Immunol 2005, 175(6):3980-3989.
• [23]Feldkamp LA, Davis LC, Kress JW: Practical cone-beam algorithm. J Opt Soc Am 1984, A1:612-619.
• [24]Park CH, Abramson ZR, Taba M Jr, Jin Q, Chang J, Kreider JM, Goldstein SA, Giannobile WV: Three-dimensional micro-computed tomographic imaging of alveolar bone in experimental bone loss or repair. J Periodontol 2007, 78(2):273-281.
• [25]Parfitt AM, Drezner MK, Glorieux FH, Kanis JA, Malluche H, Meunier PJ, Ott SM, Recker RR: Bone histomorphometry: standardization of nomenclature, symbols, and units. Report of the ASBMR Histomorphometry Nomenclature Committee. J Bone Miner Res 1987, 2(6):595-610.
• [26]Klausen B: Microbiological and immunological aspects of experimental periodontal disease in rats: a review article. J Periodontol 1991, 62(1):59-73.
• [27]Baker PJ: Genetic control of the immune response in pathogenesis. J Periodontol 2005, 76(11 Suppl):2042-2046.
• [28]Genco CA, Van Dyke T, Amar S: Animal models for Porphyromonas gingivalis-mediated periodontal disease. Trends Microbiol 1998, 6(11):444-449.
• [29]Kimura S, Nagai A, Onitsuka T, Koga T, Fujiwara T, Kaya H, Hamada S: Induction of experimental periodontitis in mice with Porphyromonas gingivalis-adhered ligatures. J Periodontol 2000, 71(7):1167-1173.
• [30]Polak D, Wilensky A, Shapira L, Halabi A, Goldstein D, Weiss EI, Houri-Haddad Y: Mouse model of experimental periodontitis induced by Porphyromonas gingivalis/Fusobacterium nucleatum infection: bone loss and host response. J Clin Periodontol 2009, 36(5):406-410.
• [31]Socransky SS, Haffajee AD: Periodontal microbial ecology. J Periodontol 2000 2005, 38:135-187.
• [32]Duncan MJ, Nakao S, Skobe Z, Xie H: Interactions of Porphyromonas gingivalis with epithelial cells. Infect Immun 1993, 61(5):2260-2265.
• [33]Lamont RJ, Chan A, Belton CM, Izutsu KT, Vasel D, Weinberg A: Porphyromonas gingivalis invasion of gingival epithelial cells. Infect Immun 1995, 63(10):3878-3885.
• [34]Lamont RJ, Oda D, Persson RE, Persson GR: Interaction of Porphyromonas gingivalis with gingival epithelial cells maintained in culture. Oral Microbiol Immun 1992, 7(6):364-367.
• [35]Sandros J, Papapanou PN, Nannmark U, Dahlen G: Porphyromonas gingivalis invades human pocket epithelium in vitro. J Periodontal Res 1994, 29(1):62-69.
• [36]Andrian E, Grenier D, Rouabhia M: In vitro models of tissue penetration and destruction by Porphyromonas gingivalis. Infect Immun 2004, 72(8):4689-4698.
• [37]Colombo AV, da Silva CM, Haffajee A, Colombo AP: Identification of intracellular oral species within human crevicular epithelial cells from subjects with chronic periodontitis by fluorescence in situ hybridization. J Periodontal Res 2007, 42(3):236-243.
• [38]Rudney JD, Chen R, Sedgewick GJ: Intracellular Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis in buccal epithelial cells collected from human subjects. Infect Immun 2001, 69(4):2700-2707.
• [39]Rudney JD, Chen R, Sedgewick GJ: Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Tannerella forsythensis are components of a polymicrobial intracellular flora within human buccal cells. J Dent Res 2005, 84(1):59-63.
• [40]Christersson LA: Actinobacillus actinomycetemcomitans and localized juvenile periodontitis. Clinical, microbiologic and histologic studies. Swed Dent J Suppl 1993, 90:1-46.
• [41]Saglie FR: Scanning electron microscope and intragingival microorganisms in periodontal diseases. Scanning Microsc 1988, 2(3):1535-1540.
• [42]Manor A, Lebendiger M, Shiffer A, Tovel H: Bacterial invasion of periodontal tissues in advanced periodontitis in humans. J Periodontol 1984, 55(10):567-573.
• [43]Potempa J, Banbula A, Travis J: Role of bacterial proteinases in matrix destruction and modulation of host responses. Periodontol 2000 2000, 24:153-192.
• [44]Berezow AB, Ernst RK, Coats SR, Braham PH, Karimi-Naser LM, Darveau RP: The structurally similar, penta-acylated lipopolysaccharides of Porphyromonas gingivalis and Bacteroides elicit strikingly different innate immune responses. Micro Pathogenesis 2009, 47(2):68-77.
• [45]Okugawa T, Kaneko T, Yoshimura A, Silverman N, Hara Y: NOD1 and NOD2 mediate sensing of periodontal pathogens. J Dent Res 2010, 89(2):186-191.
• [46]Peyret-Lacombe A, Brunel G, Watts M, Charveron M, Duplan H: TLR2 sensing of F. nucleatum and S. sanguinis distinctly triggered gingival innate response. Cytokine 2009, 46(2):201-210.
• [47]Yilmaz O, Sater AA, Yao L, Koutouzis T, Pettengill M, Ojcius DM: ATP-dependent activation of an inflammasome in primary gingival epithelial cells infected by Porphyromonas gingivalis. Cell Microbiol 2010, 12(2):188-198.
• [48]Darveau RP, Belton CM, Reife RA, Lamont RJ: Local chemokine paralysis, a novel pathogenic mechanism for Porphyromonas gingivalis. Infect Immun 1998, 66(4):1660-1665.
• [49]Taxman DJ, Swanson KV, Broglie PM, Wen H, Holley-Guthrie E, Huang MT, Callaway JB, Eitas TK, Duncan JA, Ting JP: Porphyromonas gingivalis mediates inflammasome repression in polymicrobial cultures through a novel mechanism involving reduced endocytosis. J Biol Chem 2012, 287(39):32791-32799.
• [50]Li CH, Amar S: Morphometric, histomorphometric, and microcomputed tomographic analysis of periodontal inflammatory lesions in a murine model. J Periodontol 2007, 78(6):1120-1128.
• [51]Zhang W, Ju J, Rigney T, Tribble G: Integrin alpha5beta1-fimbriae binding and actin rearrangement are essential for Porphyromonas gingivalis invasion of osteoblasts and subsequent activation of the JNK pathway. BMC Microbiol 2013, 13:5. doi:10.1186/1471-2180-1113-1185