ABSTRACT
BACKGROUND: Periodontitis is a chronic inflammatory disease caused by the microbiota of the periodontal pocket. We investigated the association between subgingival bacterial profiles and gene expression patterns in gingival tissues of patients with periodontitis. A total of 120 patients undergoing periodontal surgery contributed with a minimum of two interproximal gingival papillae (range 2-4) from a maxillary posterior region. Prior to tissue harvesting, subgingival plaque samples were collected from the mesial and distal aspects of each tissue sample. Gingival tissue RNA was extracted, reverse-transcribed, labeled, and hybridized with whole-genome microarrays (310 in total). Plaque samples were analyzed using checkerboard DNA-DNA hybridizations with respect to 11 bacterial species. Random effects linear regression models considered bacterial levels as exposure and expression profiles as outcome variables. Gene Ontology analyses summarized the expression patterns into biologically relevant categories. RESULTS: Wide inter-species variation was noted in the number of differentially expressed gingival tissue genes according to subgingival bacterial levels: Using a Bonferroni correction (p < 9.15 x 10(-7)), 9,392 probe sets were differentially associated with levels of Tannerella forsythia, 8,537 with Porphyromonas gingivalis, 6,460 with Aggregatibacter actinomycetemcomitans, 506 with Eikenella corrodens and only 8 with Actinomyces naeslundii. Cluster analysis identified commonalities and differences among tissue gene expression patterns differentially regulated according to bacterial levels. CONCLUSION: Our findings suggest that the microbial content of the periodontal pocket is a determinant of gene expression in the gingival tissues and provide new insights into the differential ability of periodontal species to elicit a local host response.
Subject(s)
Dental Plaque/microbiology , Gene Expression Profiling , Gingiva/metabolism , Periodontal Pocket/genetics , Adolescent , Adult , Aged , Bacteria/growth & development , Bacteria/isolation & purification , Cluster Analysis , Dental Plaque/genetics , Female , Gene Expression Regulation , Gingiva/microbiology , Humans , Linear Models , Male , Middle Aged , Oligonucleotide Array Sequence Analysis , Periodontal Pocket/microbiology , Species Specificity , Young AdultABSTRACT
AIMS: We investigated the effect of comprehensive periodontal therapy on the levels of multiple systemic inflammatory biomarkers. MATERIAL AND METHODS: Thirty patients with severe periodontitis received comprehensive periodontal therapy within a 6-week period. Blood samples were obtained at: 1-week pre-therapy (T1), therapy initiation (T2), treatment completion (T3), and 4 weeks thereafter (T4). We assessed the plasma concentrations of 19 biomarkers using multiplex assays, and serum IgG antibodies to periodontal bacteria using checkerboard immunoblotting. At T2 and T4, dental plaque samples were analysed using checkerboard hybridizations. RESULTS: At T3, PAI-1, sE-selectin, sVCAM-1, MMP-9, myeloperoxidase, and a composite summary inflammatory score (SIS) were significantly reduced. However, only sE-selectin, sICAM, and serum amyloid P sustained a reduction at T4. Responses were highly variable: analyses of SIS slopes between baseline and T4 showed that approximately 1/3 and 1/4 of the patients experienced a marked reduction and a pronounced increase in systemic inflammation, respectively, while the remainder were seemingly unchanged. Changes in inflammatory markers correlated poorly with clinical, microbiological and serological markers of periodontitis. CONCLUSIONS: Periodontal therapy resulted in an overall reduction of systemic inflammation, but the responses were inconsistent across subjects and largely not sustainable. The determinants of this substantial heterogeneity need to be explored further.
Subject(s)
Cardiovascular Diseases/blood , Inflammation Mediators/blood , Inflammation/blood , Periodontitis/blood , Periodontitis/therapy , Adiponectin/blood , Adolescent , Adult , Aged , Alveolar Bone Loss/blood , Alveolar Bone Loss/surgery , Alveolar Bone Loss/therapy , Antibodies, Bacterial/blood , C-Reactive Protein/analysis , Dental Plaque/microbiology , Dental Scaling , E-Selectin/blood , Female , Humans , Intercellular Adhesion Molecule-1/blood , Interleukins/blood , Linear Models , Male , Matrix Metalloproteinase 9/blood , Middle Aged , Oral Hygiene/education , Periodontitis/surgery , Peroxidase/blood , Plasminogen Activator Inhibitor 1/blood , Prospective Studies , Serum Amyloid P-Component/analysis , Tumor Necrosis Factor-alpha/blood , Vascular Cell Adhesion Molecule-1/blood , Young AdultABSTRACT
BACKGROUND: Clinical and radiographic measures are gold standards for diagnosing periodontitis but offer little information regarding the pathogenesis of the disease. We hypothesized that a comparison of gene expression signatures between healthy and diseased gingival tissues would provide novel insights in the pathobiology of periodontitis and would inform the design of future studies. METHODS: Ninety systemically healthy non-smokers with moderate to advanced periodontitis (63 with chronic periodontitis and 27 with aggressive periodontitis) each contributed at least two diseased interproximal papillae (with bleeding on probing [BOP], probing depth [PD] > or =4 mm, and attachment loss [AL] > or =3 mm) and a healthy papilla, if available (no BOP, PD < or =4 mm, and AL < or =2 mm). RNA was extracted, amplified, reverse-transcribed, labeled, and hybridized with whole genome microarrays. Differential expression was assayed in 247 individual tissue samples (183 from diseased sites and 64 from healthy sites) using a standard mixed-effects linear model approach, with patient effects considered random with a normal distribution and gingival tissue status considered a two-level fixed effect. Gene ontology analysis classified the expression patterns into biologically relevant categories. RESULTS: Transcriptome analysis revealed that 12,744 probe sets were differentially expressed after adjusting for multiple comparisons (P <9.15 x 10(7)). Of those, 5,295 were upregulated and 7,449 were downregulated in disease compared to health. Gene ontology analysis identified 61 differentially expressed groups (adjusted P <0.05), including apoptosis, antimicrobial humoral response, antigen presentation, regulation of metabolic processes, signal transduction, and angiogenesis. CONCLUSION: Gingival tissue transcriptomes provide a valuable scientific tool for further hypothesis-driven studies of the pathobiology of periodontitis.
Subject(s)
Gene Expression Profiling , Gene Expression Regulation/physiology , Gingiva/metabolism , Periodontitis/metabolism , Periodontium/metabolism , Adolescent , Adult , Aged , Case-Control Studies , Chronic Disease , Female , Humans , Male , Middle Aged , Periodontitis/physiopathology , RNA/analysis , Reference Values , Severity of Illness Index , Young AdultSubject(s)
Dentists , Mass Screening , Primary Health Care , Dental Offices , Diagnosis , Humans , Primary Prevention , Secondary PreventionABSTRACT
Periodontitis is an inflammatory disease of bacterial origin that results in the progressive destruction of the tissues that support the teeth, specifically the gingiva, periodontal ligament, and alveolar bone. The diagnosis of periodontal disease currently relies almost exclusively on clinical parameters and traditional dental radiography. In this article, the authors review current diagnostic techniques and present new approaches and technologies that are being developed to improve assessment of this common condition.
Subject(s)
Periodontal Diseases/diagnosis , Periodontal Index , Periodontics/methods , Diagnosis, Oral/instrumentation , Diagnosis, Oral/methods , Diagnosis, Oral/trends , Humans , Periodontics/instrumentation , Periodontics/trends , Severity of Illness IndexABSTRACT
AIMS: We investigated the effects of periodontal therapy on gene expression of peripheral blood monocytes. METHODS: Fifteen patients with periodontitis gave blood samples at four time points: 1 week before periodontal treatment (#1), at treatment initiation (baseline, #2), 6-week (#3) and 10-week post-baseline (#4). At baseline and 10 weeks, periodontal status was recorded and subgingival plaque samples were obtained. Periodontal therapy (periodontal surgery and extractions without adjunctive antibiotics) was completed within 6 weeks. At each time point, serum concentrations of 19 biomarkers were determined. Peripheral blood monocytes were purified, RNA was extracted, reverse-transcribed, labelled and hybridized with AffymetrixU133Plus2.0 chips. Expression profiles were analysed using linear random-effects models. Further analysis of gene ontology terms summarized the expression patterns into biologically relevant categories. Differential expression of selected genes was confirmed by real-time reverse transcriptase-polymerase chain reaction in a subset of patients. RESULTS: Treatment resulted in a substantial improvement in clinical periodontal status and reduction in the levels of several periodontal pathogens. Expression profiling over time revealed more than 11,000 probe sets differentially expressed at a false discovery rate of <0.05. Approximately 1/3 of the patients showed substantial changes in expression in genes relevant to innate immunity, apoptosis and cell signalling. CONCLUSIONS: The data suggest that periodontal therapy may alter monocytic gene expression in a manner consistent with a systemic anti-inflammatory effect.