Responsiveness to Lipopolysaccharide Changes According to the Aging of Periodontal Ligament Fibroblasts

  • Jun, Ji-Hae (Department of Pharmacology and Dental Therapeutics, College of Dentistry, Dental Research Institute, Seoul National University) ;
  • Kim, Gwan-Shik (Department of Pharmacology and Dental Therapeutics, College of Dentistry, Dental Research Institute, Seoul National University) ;
  • Woo, Kyung-Mi (Department of Pharmacology and Dental Therapeutics, College of Dentistry, Dental Research Institute, Seoul National University) ;
  • Min, Byung-Moo (Department of Oral Biochemistry, College of Dentistry, Dental Research Institute, Seoul National University) ;
  • Baek, Jeong-Hwa (Department of Pharmacology and Dental Therapeutics, College of Dentistry, Dental Research Institute, Seoul National University)
  • Published : 2005.03.31

Abstract

The elderly suffer from an impaired immune function being obvious in a higher susceptibility to infections. Although the inflammatory cells are the major immunomodulatory cells, fibroblasts also secrete a variety of inflammatory cytokines and chemokines. Therefore periodontal tissue aging might playa role in development and progress of periodontitis. In this study, we investigated the effect of in vitro periodontal ligament cellular aging on the inflammatory cytokines, chemokines, and matrix metalloprotease(MMP)-2 expression induced by lipopolysaccharide(LPS) treatment. Three different cell populations were used; passages 4-5, 14-15, and 24-25 (at passage 27, more than 90% cells were replicative senescent). LPS increased the expression of interleukin(IL)-1${\beta}$, IL-6, and tumor necrosis factor-${\alpha}$, IL-8, RANTES, and MMP-2. However, the order of induction folds were passages 14-15 > 4-5 > 24-25. While the expression level of Toll-like receptor(TLR) 4 decreased according to the increase in passage number, the level of TLR2 was highest at passages 14-15 and then decreased at passages 24-25. While the spontaneous expression of IL-8 decreased according to the increase in passage number, that of RANTES and proMMP-2 increased according to the increase in passage number. These results suggest that the aging of periodontal ligament fibroblasts differentially affect the role as immunomodulatory cells in response to periodontopathic bacteria and therefore might be another risk factor of periodontitis progression.

Keywords

References

  1. Agarwal, S., Piesco, N.P., Johns, L.P. and Riccelli, A.E.: Differential expression of IL-1 beta, TNF-alpha, IL-6, and IL-8 in human monocytes in response to lipopolysaccharides from different microbes. J. Dent. Res. 74:1057-1065, 1995 https://doi.org/10.1177/00220345950740040501
  2. Assuma, R., Oates, T., Cochran, D., Amar, S. and Graves, D.T.: IL-1 and TNF antagonists inhibit the inflammatory response and bone loss in experimental periodontitis. J. Immunol. 160:403-409, 1998
  3. Butcher, S.K., Chahal, H., Nayak, L., Sinclair, A., Henriquez, N.V., Sapey, E., O'Mahony, D. and Lord, J.M.: Senescence in innate immune responses: reduced neutrophil phagocytic capacity and CD16 expression in elderly humans. J Leukoc. Biol. 70:881-886, 2001
  4. D'Ambrosio, D., Panina-Bordignon, P. and Sinigaglia, F.: Chemokine receptors in inflammation: an overview. J. Immunol. Methods 273:3-13, 2003 https://doi.org/10.1016/S0022-1759(02)00414-3
  5. Dimri, G.P., Lee, X., Basile, G., Acosta, M., Scott, G., Roskelley, C., Medrano, E.E., Linskens, M., Rubelj, I. and Pereira-Smith, O. et al.: A biomarker that identifies senescent human cells in culture and in aging skin in vivo. Proc. Natl. Acad. Sci. U.S.A. 92:9363-9367, 1995 https://doi.org/10.1073/pnas.92.20.9363
  6. Doria, G. and Frasca, D.: Genes, immunity, and senescence: looking for a link. Immunol. Rev. 160: 159-170, 1997 https://doi.org/10.1111/j.1600-065X.1997.tb01036.x
  7. Fransson, C., Mooney, J., Kinane, D.F. and Berglundh, T.: Differences in the inflammatory response in young and old human subjects during the course of experimental gingivitis. J. Clin. Periodontol. 26:453-460, 1999 https://doi.org/10.1034/j.1600-051X.1999.260707.x
  8. Gabriel, P., Cakrnan, I. and Rink, L.: Overproduction of monokines by leukocytes after stimulation with lipopolysaccharide in the elderly. Exp. Gerontol. 37:235-247, 2002 https://doi.org/10.1016/S0531-5565(01)00189-9
  9. Gamonal, J., Acevedo, A., Bascones, A., Jorge, O. and Silva, A.: Levels of interleukin-1 beta, -8, and -10 and RANTES in gingival crevicular fluid and cell populations in adult periodontitis patients and the effect of periodontal treatment. J. Periodontol. 71:1535-1545, 2000 https://doi.org/10.1902/jop.2000.71.10.1535
  10. Gamonal, J., Acevedo, A., Bascones, A., Jorge, O. and Silva, A.: Characterization of cellular infiltrate, detection of chemokine receptor CCR5 and interleukin-8 and RANTES chemokines in adult periodontitis. J. Periodontal Res. 36: 194-203, 2001 https://doi.org/10.1034/j.1600-0765.2001.360309.x
  11. Goseki, T., Shimizu, N., Iwasawa, T., Takiguchi, H. and Abiko, Y: Effects of in vitro cellular aging on alkaline phosphatase, cathepsin activities and collagen secretion of human periodontal ligament derived cells. Mech. Ageing Dev. 91:171-183, 1996 https://doi.org/10.1016/S0047-6374(96)01785-X
  12. Graves, D.T.: The potential role of chemokines and inflammatory cytokines in periodontal disease progression. Clin. Infect. Dis. 28:482-490, 1999 https://doi.org/10.1086/515178
  13. Haffajee, A.D., Socransky, S.S., Lindhe, J., Kent, R.L., Okamoto, H. and Yoneyama, T.: Clinical risk indicators for periodontal attachment loss. J. Clin. Periodontol. 18: 117-125, 1991 https://doi.org/10.1111/j.1600-051X.1991.tb01700.x
  14. Hayflick, L.: The limited in vitro lifetime of human diploid cell strains. Exp. Cell. Res. 37:614-636, 1965 https://doi.org/10.1016/0014-4827(65)90211-9
  15. Heumann, D. and Roger, T.: Initial responses to endotoxins and Gram-negative bacteria. Clin. Chim. Acta. 323:59-72. 2002 https://doi.org/10.1016/S0009-8981(02)00180-8
  16. Hsu, H.C., Shi, J., Yang, P., Xu, X., Dodd, C., Matsuki, Y., Zhang, H.G. and Mountz, J.D.: Activated CD8(+) T cells from aged mice exhibit decreased activation-induced cell death. Mech. Ageing Dev. 122: 1663-1684, 2001 https://doi.org/10.1016/S0047-6374(01)00279-2
  17. Jiang, Y., Russell, T.R., Graves, D.T., Cheng, H., Nong, S.H. and Levitz, S.M.: Monocyte chemoattractant protein 1 and interleukin-8 production in mononuclear cells stimulated by oral microorganisms. Infect. Immun. 64:4450-4455, 1996
  18. Kabashima, H., Yoneda, M., Nagata, K., Hirofuji, T. and Maeda, K.: The presence of chemokine (MCP-1, MIP-1alpha, MIP-1beta, IP-10, RANTES)-positive cells and chemokine receptor (CCR5, CXCR3)-positive cells in inflamed human gingival tissues. Cytokine 20:70-77, 2002 https://doi.org/10.1006/cyto.2002.1985
  19. Kim, H., You, S., Farris, J., Kong, B.W., Christman, S.A., Foster, L.K. and Foster, D.N.: Expression profiles of p53-, p16(INK4a)-, and telomere- regulating genes in replicative senescent primary human, mouse, and chicken fibroblast cells. Exp. Cell. Res. 272:199-208, 2002 https://doi.org/10.1006/excr.2001.5420
  20. Mariani, E., Pulsatelli, L., Meneghetti, A., Dolzani, P., Mazzetti, I., Neri, S., Ravaglia, G., Forti, P. and Facchini, A.: Different IL-8 production by T and NK lymphocytes in elderly subjects. Mech. Ageing Dev.122:1383-1395, 2001 https://doi.org/10.1016/S0047-6374(01)00270-6
  21. Mariani, E., Pulsatelli, L., Neri, S., Dolzani, P., Meneghetti, A., Silvestri, T., Ravaglia, G., Forti, P., Cattini, L. and Facchini, A.: RANTES and MIP-1alpha production by T lymphocytes, monocytes and NK cells from nonagenarian subjects. Exp. Gerontol. 37:219-226, 2002 https://doi.org/10.1016/S0531-5565(01)00187-5
  22. Masada, M.P., Persson, R., Kenney, J.,S, Lee, S.W., Page, R.C. and Allison, A.C.: Measurement of interleukin-1 alpha and -1 beta in gingival crevicular fluid: implications for the pathogenesis of periodontal disease. J. Periodontal Res. 25:156-163, 1990 https://doi.org/10.1111/j.1600-0765.1990.tb01038.x
  23. McCulloch, C.A. and Bordin, S.: Role of fibroblast subpopulations in periodontal physiology and pathology. J. Periodontal Res. 26: 144-154, 1991 https://doi.org/10.1111/j.1600-0765.1991.tb01638.x
  24. Miller, R.A.: The aging immune system: primer and prospectus. Science 73:70-74, 1996
  25. Mochizuki, K., Yamaguchi, M. and Abiko, Y.: Enhancement of LPS-stimulated plasminogen activator production in aged gingival fibroblasts. J. Periodontal. Res. 34:251-260, 1999 https://doi.org/10.1111/j.1600-0765.1999.tb02251.x
  26. Nishimura, F., Terranova, V.P., Braithwaite, M., Orman, R., Ohyama, H., Mineshiba, J., Chou, H.H., Takashiba, S. and Murayama, Y.: Comparison of in vitro proliferative capacity of human periodontal ligament cells in juvenile and aged donors. Oral Dis. 3:162-166, 1997 https://doi.org/10.1111/j.1601-0825.1997.tb00029.x
  27. Offenbacher, S., Collins, J.G. and Heasman, P.A.: Diagnostic potential of host response mediators. Adv. Dent. Res. 7: 175-181, 1993 https://doi.org/10.1177/08959374930070020801
  28. Ogura, N., Matsuda, U., Tanaka, F., Shibata, Y., Takiguchi, H. and Abiko, Y.: In vitro senescence enhances IL-6 production in human gingival fibroblasts induced by lipopolysaccharide from Campylobacter rectus. Mech. Ageing Dev. 87:47-59, 1996 https://doi.org/10.1016/0047-6374(96)01701-0
  29. Okada, H. and Murakami, S.: Cytokine expression in periodontal health and disease. Crit. Rev. Oral Biol. Med. 9:248-266, 1998 https://doi.org/10.1177/10454411980090030101
  30. Okamura, H., Yamaguchi, M. and Abiko, Y.: Enhancement of lipopolysaccharide -stimulated PGE2 and IL-1 beta production in gingival fibroblast cells from old rats. Exp. Gerontol. 34:379-392, 1999 https://doi.org/10.1016/S0531-5565(99)00006-6
  31. Papapanou, P.N., Lindhe, J., Sterrett, J.D. and Eneroth, L.: Considerations on the contribution of ageing to loss of periodontal tissue support. J. Clin. Periodontol. 18:611-615, 1991 https://doi.org/10.1111/j.1600-051X.1991.tb00098.x
  32. Park, S.Y., Kim, Y.H., Jun, J.H., Kim, G.S., Min, B.M. and Baek, J.H.: P.gingivalis and A. actinomycetemcomitans LPS differentially regulate the expression of inflammatory cytokines and chemokines in gingival fibroblast and periodontal ligament fibroblast. Int. J. Oral. Biol. 29:27-36, 2004
  33. Pulsatelli, L., Meliconi, R., Mazzetti, I., Dolzani, P., Meneghetti, A., Neri, S., Silvestri, T., Ravaglia, G., Forti, P., Facchini, A. and Mariani, E.: Chemokine production by peripheral blood mononuclear cells in elderly subjects. Mech. Ageing Dev.121:89-100, 2000 https://doi.org/10.1016/S0047-6374(00)00200-1
  34. Renshaw, M., Rockwell, J., Engleman, C., Gewirtz, A., Katz, J. and Sambhara, S.: Cutting edge: impaired Toll-like receptor expression and function in aging. J. Immunol. 169:4697-4701, 2002 https://doi.org/10.4049/jimmunol.169.9.4697
  35. Saito, H., Sherwood, E.R., Varma, T.K. and Evers, B.M.: Effects of aging on mortality, hypothermia, and cytokine induction in mice with endotoxemia or sepsis. Mech. Ageing Dev. 124:1047-1058, 2003 https://doi.org/10.1016/j.mad.2003.08.002
  36. Sandhir, R., Puri, V., Klein, R.M. and Berman, N.E.: Differential expression of cytokines and chemokines during secondary neuron death following brain injury in old and young mice. Neurosci. Lett. 369(1):28-32, 2004 https://doi.org/10.1016/j.neulet.2004.07.032
  37. Sawa, Y., Phillips, A., Hollard, .J, Yoshida, S. and Braithwaite, M.W.: Impairment of osteoca1cin production in senescent periodontal ligament fibroblasts. Tissue Cell 32: 198-204, 2000 https://doi.org/10.1054/tice.2000.0104
  38. Socransky, S.S. and Haffajee, A.D.: Effect of therapy on periodontal infections. J. Periodontol. 64:754-759, 1993 https://doi.org/10.1902/jop.1993.64.8s.754
  39. Takeda, K., Kaisho, T. and Akira, S.: Toll-like receptors. Ann. Rev. Immunol. 21:335-376, 2003 https://doi.org/10.1146/annurev.immunol.21.120601.141126
  40. Takiguchi, H., Yamaguchi, M., Okamura, H. and Abiko, Y.: Contribution of IL-1 beta to the enhancement of Campylobacter rectus lipopolysaccharide -stimulated PGE2 production in old gingival fibroblasts in vitro. Mech. Ageing Dev. 98:75-90, 1997 https://doi.org/10.1016/S0047-6374(97)00068-7
  41. Tamura, M., Tokuda, M., Nagaoka, S. and Takada, H.: Lipopolysaccharides of Bacteroides intermedius (Prevotella intermedia) and Bacteroides (Porphyromonas) gingivalis induce interleukin-8 gene expression in human gingival fibroblast cultures. Infect. Immun. 60:4932-4937, 1992
  42. Wang, P.L., Ohura, K., Fujii, T., Oido-Mori, M., Kowashi, Y., Kikuchi, M., Suetsugu, Y. and Tanaka, J.: DNA microarray analysis of human gingival fibroblasts from healthy and inflammatory gingival tissues. Biochem. Biophys. Res. Commun. 305:970-973, 2003 https://doi.org/10.1016/S0006-291X(03)00821-0
  43. Yamaji, Y., Kubota, T., Sasaguri, K., Sato, S., Suzuki, Y., Kumada, H. and Umemoto, T.: Inflammatory cytokine gene expression in human periodontal ligament fibroblasts stimulated with bacterial lipopolysaccharides. Infect. Immun. 63:3576-3581, 1995
  44. Zheng, F., Cheng, Q.L., Plati, A.R., Ye, S.Q., Berho, M., Banerjee, A, Potier, M., Jaimes, E.A., Yu, H., Guan, Y.F., Hao, C.M., Striker, L.J. and Striker, G.E.: The glomerulosclerosis of aging in females: contribution of the pro inflammatory mesangial cell phenotype to macrophage infiltration. Am. J. Pathol. 165: 1789-1798, 2004 https://doi.org/10.1016/S0002-9440(10)63434-7
  45. Zhong, W.W., Burke, P.A., Hand, A.T., Walsh, M.J., Hughes, L.A. and Forse, R.A: Regulation of cytokine mRNA expression in lipopolysaccharide- stimulated human macrophages. Arch. Surg. 128:158-163, 1993 https://doi.org/10.1001/archsurg.1993.01420140035006