Gingival tissue transcriptomes in experimental gingivitis.

AIMS We investigated the sequential gene expression in the gingiva during the induction and resolution of experimental gingivitis. MATERIAL AND METHODS Twenty periodontally and systemically healthy non-smoking volunteers participated in a 3-week experimental gingivitis protocol, followed by debridement and 2-week regular plaque control. We recorded clinical indices and harvested gingival tissue samples from four interproximal palatal sites in half of the participants at baseline, Day 7, Day 14 and Day 21 (the "induction phase"), and at Day 21, Day 25, Day 30 and Day 35 in the other half (the "resolution phase"). RNA was extracted, amplified, reversed transcribed, amplified, labelled and hybridized using Affymetrix Human Genome U133Plus2.0 microarrays. Paired t-tests compared gene expression changes between consecutive time points. Gene ontology analyses summarized the expression patterns into biologically relevant categories. RESULTS The median gingival index was 0 at baseline, 2 at Day 21 and 1 at Day 35. Differential gene regulation peaked during the third week of induction and the first 4 days of resolution. Leucocyte transmigration, cell adhesion and antigen processing/presentation were the top differentially regulated pathways. CONCLUSIONS Transcriptomic studies enhance our understanding of the pathobiology of the reversible inflammatory gingival lesion and provide a detailed account of the dynamic tissue responses during the induction and resolution of experimental gingivitis.

[1]  S. Socransky,et al.  Effect of various chlorhexidine regimens on salivary bacteria and de novo plaque formation. , 2003, Journal of clinical periodontology.

[2]  R. Page,et al.  Histopathologic features of the initial and early stages of experimental gingivitis in man. , 1975, Journal of periodontal research.

[3]  H Löe,et al.  The Gingival Index, the Plaque Index and the Retention Index Systems. , 1967, Journal of periodontology.

[4]  P. Pavlidis,et al.  Granulocyte chemotactic protein 2 (gcp-2/cxcl6) complements interleukin-8 in periodontal disease. , 2009, Journal of periodontal research.

[5]  Matthew R. Thompson,et al.  ATF3 transcription factor and its emerging roles in immunity and cancer , 2009, Journal of Molecular Medicine.

[6]  D. Irvine,et al.  Homeostatic Lymphoid Chemokines Synergize with Adhesion Ligands to Trigger T and B Lymphocyte Chemokinesis1 , 2006, The Journal of Immunology.

[7]  J. Inoue,et al.  TNF Receptor Family Member BCMA (B Cell Maturation) Associates with TNF Receptor-Associated Factor (TRAF) 1, TRAF2, and TRAF3 and Activates NF-κB, Elk-1, c-Jun N-Terminal Kinase, and p38 Mitogen-Activated Protein Kinase1 , 2000, The Journal of Immunology.

[8]  Ryan T Demmer,et al.  Bioinformatics techniques in microarray research: applied microarray data analysis using R and SAS software. , 2010, Methods in molecular biology.

[9]  R. Newcombe,et al.  Evaluation of a mouthrinse containing chlorhexidine and fluoride as an adjunct to oral hygiene. , 1993, Journal of clinical periodontology.

[10]  N. Lang,et al.  Variability of histologic criteria in clinically healthy human gingiva. , 1987, Journal of periodontal research.

[11]  Purvesh Khatri,et al.  Onto-Tools: new additions and improvements in 2006 , 2007, Nucleic Acids Res..

[12]  C. Daly,et al.  Effect of localized experimental gingivitis on early supragingival plaque accumulation. , 1996, Journal of clinical periodontology.

[13]  Hua Lu,et al.  Interferon-Inducible Protein IFIXα1 Functions as a Negative Regulator of HDM2 , 2006, Molecular and Cellular Biology.

[14]  R. Deinzer,et al.  Comparison of experimental gingivitis with persistent gingivitis: differences in clinical parameters and cytokine concentrations. , 2007, Journal of periodontal research.

[15]  P. Pavlidis,et al.  Subgingival bacterial colonization profiles correlate with gingival tissue gene expression , 2009, BMC Microbiology.

[16]  H. Löe,et al.  EXPERIMENTAL GINGIVITIS IN MAN. , 1965, The Journal of periodontology.

[17]  Hendrik Veelken,et al.  CCL19 is a specific ligand of the constitutively recycling atypical human chemokine receptor CRAM‐B , 2010, Immunology.

[18]  P. Hull,et al.  The influence of experimental gingivitis on plaque formation. , 1977, Journal of clinical periodontology.

[19]  J. Lindhe,et al.  The effect of triclosan on developing gingivitis. , 1995, Journal of Clinical Periodontology.

[20]  K. Preissner,et al.  The Neutrophil-specific Antigen CD177 Is a Counter-receptor for Platelet Endothelial Cell Adhesion Molecule-1 (CD31)* , 2007, Journal of Biological Chemistry.

[21]  M. Quirynen,et al.  Effect of different chlorhexidine formulations in mouthrinses on de novo plaque formation. , 2008, Journal of clinical periodontology.

[22]  B. Harfe,et al.  TMEM16 Proteins Produce Volume-regulated Chloride Currents That Are Reduced in Mice Lacking TMEM16A* , 2009, The Journal of Biological Chemistry.

[23]  R. Petrie,et al.  Colocalization of the B Cell Receptor and CD20 Followed by Activation-Dependent Dissociation in Distinct Lipid Rafts1 , 2002, The Journal of Immunology.

[24]  W. Moore,et al.  Bacteriology of experimental gingivitis in young adult humans , 1982, Infection and immunity.

[25]  J. Levy The Unexpected Pleiotropic Activities of RANTES , 2009, The Journal of Immunology.

[26]  Martin Braddock,et al.  The transcription factor Egr-1: a potential drug in wound healing and tissue repair , 2001, Annals of medicine.

[27]  L. Trombelli,et al.  Modulation of clinical expression of plaque-induced gingivitis. I. Background review and rationale. , 2004, Journal of clinical periodontology.

[28]  G. Krohne,et al.  LEM-Domain proteins: new insights into lamin-interacting proteins. , 2007, International review of cytology.

[29]  J. Aitken,et al.  Experimental gingivitis in humans. A histochemical and immunological characterization of the lymphoid cell subpopulations. , 1983, Journal of periodontal research.

[30]  G. Seymour,et al.  In situ demonstration of natural killer (NK) cells in human gingival tissue. , 1986, Journal of periodontology.

[31]  C. Scapoli,et al.  Experimental gingivitis: reproducibility of plaque accumulation and gingival inflammation parameters in selected populations during a repeat trial. , 2008, Journal of clinical periodontology.

[32]  K. Tabeta,et al.  Interleukin-10 gene promoter polymorphism in Japanese patients with adult and early-onset periodontitis. , 2001, Journal of clinical periodontology.

[33]  F. Abbas,et al.  Experimental gingivitis in relation to susceptibility to periodontal disease. II. Phase-contrast microbiological features and some host-response observations. , 1986, Journal of clinical periodontology.

[34]  S. Socransky,et al.  "Checkerboard" DNA-DNA hybridization. , 1994, BioTechniques.

[35]  B. Lorber Role of NG2 in Development and Regeneration , 2006, The Journal of Neuroscience.

[36]  P. Khatri,et al.  A systems biology approach for pathway level analysis. , 2007, Genome research.

[37]  M. Thornhill,et al.  Endothelial cell leukocyte adhesion molecule-1 (ELAM-1) and intercellular adhesion molecule-1 (ICAM-1) expression in gingival tissue during health and experimentally-induced gingivitis. , 1992, Journal of periodontal research.

[38]  Rafael A Irizarry,et al.  Exploration, normalization, and summaries of high density oligonucleotide array probe level data. , 2003, Biostatistics.

[39]  M. Colonna,et al.  Cutting Edge: CD96 (Tactile) Promotes NK Cell-Target Cell Adhesion by Interacting with the Poliovirus Receptor (CD155) , 2004, The Journal of Immunology.

[40]  S. Syed,et al.  Bacteriology of Human Experimental Gingivitis: Effect of Plaque and Gingivitis Score , 1978, Infection and immunity.

[41]  I. Lamster,et al.  Development of a biochemical profile for gingival crevicular fluid. Methodological considerations and evaluation of collagen-degrading and ground substance-degrading enzyme activity during experimental gingivitis. , 1985, Journal of periodontology.

[42]  S. Offenbacher,et al.  Changes in crevicular fluid levels of interleukin‐1β, leukotriene B4, prostaglandin E2, thromboxane B2 and tumour necrosis factor α in experimental gingivitis in humans , 1993 .

[43]  Hua Lu,et al.  Interferon-inducible protein IFIXalpha1 functions as a negative regulator of HDM2. , 2006, Molecular and cellular biology.

[44]  L. Kjeldsen,et al.  Identification of human cysteine‐rich secretory protein 3 (CRISP‐3) as a matrix protein in a subset of peroxidase‐negative granules of neutrophils and in the granules of eosinophils , 2002, Journal of leukocyte biology.

[45]  Checkerboard assessments of serum antibodies to oral microbiota as surrogate markers of clinical periodontal status. , 2008, Journal of clinical periodontology.

[46]  P. Papapanou,et al.  The use of gene arrays in deciphering the pathobiology of periodontal diseases. , 2010, Methods in molecular biology.

[47]  W. Moore,et al.  Bacteriology of experimental gingivitis in children , 1984, Infection and immunity.

[48]  M. Timmerman,et al.  Plaque inhibition of two commercially available chlorhexidine mouthrinses. , 2005, Journal of clinical periodontology.

[49]  J. Preisser,et al.  Changes in gingival crevicular fluid inflammatory mediator levels during the induction and resolution of experimental gingivitis in humans. , 2010, Journal of clinical periodontology.

[50]  Gordon Barr,et al.  Proteomic Analysis of a Noninvasive Human Model of Acute Inflammation and Its Resolution: The Twenty-one Day Gingivitis Model , 2010, Journal of proteome research.

[51]  V. Bennett,et al.  Adducin: Ca++-dependent association with sites of cell-cell contact , 1989, The Journal of cell biology.

[52]  C. C. King,et al.  Induction and antimicrobial activity of platelet basic protein derivatives in human monocytes , 2004, Journal of leukocyte biology.

[53]  M. Timmerman,et al.  Oral microbiota in subjects with a weak or strong response in experimental gingivitis. , 1995, Journal of clinical periodontology.

[54]  Paul Pavlidis,et al.  Transcriptomes in Healthy and Diseased Gingival Tissues , 2008 .

[55]  A. Biesbrock,et al.  Gingival transcriptome patterns during induction and resolution of experimental gingivitis in humans. , 2009, Journal of periodontology.

[56]  A. Krieg,et al.  Toll‐like receptors 7, 8, and 9: linking innate immunity to autoimmunity , 2007, Immunological reviews.

[57]  D. Kinane,et al.  Differences in the inflammatory response in young and old human subjects during the course of experimental gingivitis. , 1999, Journal of clinical periodontology.

[58]  J. Wennström Mouthrinses in "experimental gingivitis" studies. , 1988, Journal of clinical periodontology.

[59]  Paul Pavlidis,et al.  Gene expression signatures in chronic and aggressive periodontitis: a pilot study. , 2004, European journal of oral sciences.

[60]  T. Ley,et al.  Differential expression of granzymes A and B in human cytotoxic lymphocyte subsets and T regulatory cells. , 2004, Blood.

[61]  M. Thornhill,et al.  Immunocytochemical characterization of cellular infiltrate, related endothelial changes and determination of GCF acute-phase proteins during human experimental gingivitis. , 1991, Journal of periodontal research.

[62]  P. Angel,et al.  The transcription factor Fos: a Janus-type regulator in health and disease. , 2009, Histology and histopathology.

[63]  S. Offenbacher,et al.  Changes in crevicular fluid levels of interleukin-1 beta, leukotriene B4, prostaglandin E2, thromboxane B2 and tumour necrosis factor alpha in experimental gingivitis in humans. , 1993, Journal of periodontal research.

[64]  L. Bonewald,et al.  Osteoinductive factor inhibits formation of human osteoclast-like cells. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[65]  Schroeder He,et al.  Pathogenesis of inflammatory periodontal disease. A summary of current work. , 1976 .