Localized juvenile spongiotic gingival hyperplasia featuring unusual p16INK4A labeling and negative human papillomavirus status by polymerase chain reaction.

BACKGROUND: Localized juvenile spongiotic gingival hyperplasia (LJSGH) is a distinct type of gingival hyperplastic lesion with specific clinicopathologic features. Evaluation of the morphological characteristics of LJSGH indicates the potential role of human papillomavirus (HPV) infection as an underlying etiopathogenetic mechanism. METHODS: All cases of LJSGH from 2008 to present were retrieved. Clinical and demographic data were collected. HPV status was investigated by p16INK4A immunohistochemistry and HPV-Polymerase chain reaction (PCR). RESULTS: Twenty-one cases of LJSGH were identified, 14 (66.7%) affecting males and seven (33.3%) females (M:F = 2:1, age range: 8-36, mean: 13 years). All lesions were well-demarcated, exophytic, erythematous, and hemorrhagic with granular or slightly papillary surface. Preponderance for the maxillary gingiva (19, 90.5%) was observed. Two (9.5%) patients presented with recurrence 20 and 21 months after excision (mean follow-up: 18.7 months). Histopathologically, all LJSGH lesions featured epithelial hyperplasia with intense neutrophilic exocytosis and spongiosis. All cases demonstrated positivity for p16INK4A with the majority of specimens (47.6%) intensely decorated in >50% of the overlying epithelium with focal immunostaining observed in 47.6% and diffuse in 52.4%. Thirteen cases (61.9%) were negative for HPV DNA by PCR, while two (9.5%) were suspicious for the presence of low levels of HPV DNA but definitive genotyping was not possible. One case (4.8%) displayed positivity for HPV-31. The remaining five cases failed the PCR reaction. CONCLUSIONS: Human papillomavirus does not participate in the pathogenesis of LJSGH. P16INK4A expression in the absence of detectable HPV DNA can likely be attributed to the intense inflammation associated with LJSGH.

Irradiation by light-emitting diode light as an adjunct to facilitate healing of experimental periodontitis in vivo.

BACKGROUND AND OBJECTIVE: This study evaluated the biostimulatory effect of 660 nm light-emitting diode (LED) as an adjunct in the treatment of experimental periodontitis. MATERIAL AND METHODS: Ninety-six Sprague-Dawley rats underwent experimental periodontitis by placement of a silk ligature followed with or without additive Porphyromonas gingivalis lipopolysaccharide (Pg-LPS) injection. Irradiation with LED light was performed at varying energy densities of 5, 10 and 15 J/cm2, 1 d after debridement and detoxification. Rats were killed at 3, 7 and 14 d after irradiation with LED light, and the effect of irradiation was evaluated by descriptive histology and quantitative measurements of periodontal bone loss, inflammatory infiltration and cellular proliferation. RESULTS: Reduction of inflammation, accelerated collagen deposition and realignment was noted following irradiation with LED light at densities of 10 and 15 J/cm2, and temporary reduction of periodontal bone loss, as well as bundle bone apposition, was noted at day 3 in rats treated with 10 J/cm2 light. The biomodulatory effect was stronger in sites treated with Pg-LPS injection. In sites without Pg-LPS injection, temporary reduction of inflammation was noted in all LED light-irradiated specimens at day 3. No significant change in cellular proliferation was noted in any LED light-treated group. CONCLUSIONS: LED light (660 nm) with an energy density of 10 J/cm2 appeared suitable as an adjunct modality for periodontitis by temporarily reducing inflammation, facilitating collagen realignment and bundle bone deposition. Future studies will aim to amplify the biostimulatory effect of LED light by adding a supplementary medium or repeated irradiation.

Expression of peptidylarginine deiminase-2 and -4, citrullinated proteins and anti-citrullinated protein antibodies in human gingiva.

BACKGROUND AND OBJECTIVE: The presence of citrullinated proteins, and peptidylarginine deiminase types -2 (PAD-2) and -4 (PAD-4) in periodontal tissues, determine the presence of anti-cyclic citrullinated protein antibodies (anti-CCP) in gingival crevicular fluid (GCF) and compare the expression of these proteins between inflamed and non-inflamed sites. MATERIAL AND METHODS: Tissue sections were stained using antibodies against citrullinated proteins, PAD-2 and PAD-4. RT-PCR was performed to investigate PAD-2 and PAD-4 mRNA in inflamed and non-inflamed gingival tissues. Anti-CCP antibodies in gingival crevicular fluid were detected by ELISA. RESULTS: Citrullinated proteins, PAD-2 and PAD-4 were detected in gingiva. There was a correlation between inflammation and expression of these proteins. mRNAs for PAD-2 and PAD-4 were detected in both inflamed and non-inflamed gingival tissues. Antibodies to CCP were found mostly in the GCF of individuals with periodontitis. CONCLUSION: PAD-2 and PAD-4 (protein and mRNA) as well as citrullinated proteins are present in inflamed gingiva, and anti-CCP antibodies can be detected in the GCF of some patients. Tissue expression of citrullinated proteins and PAD increased with the severity of inflammation. The presence of anti-CCP antibodies in GCF was almost exclusive to a subset of patients with periodontitis. Increased expression of these proteins in inflamed gingiva lends support to the notion that periodontal inflammation contributes to the inflammatory burden in a similar way to rheumatoid arthritis.

Reliability of direct and indirect clinical attachment level measurements.

AIMS: To investigate the influence of subject-, tooth- and site-level characteristics on intra-examiner reproducibility of direct and indirect clinical attachment level (CALDIR and CALIND ) recordings, and to quantify the impact of less-than-perfect reliability on our ability to assess periodontitis progression. MATERIALS AND METHODS: Within a 1-week interval, a single examiner performed duplicate probing depth (PD), CALDIR and gingival recession (GR) recordings in six sites of all teeth present in 148 periodontitis patients. CALIND was calculated on the basis of the PD and GR. RESULTS: Agreement was observed in 65%, 62%, 69% and 84% of the duplicate CALDIR , CALIND , PD and GR recordings, respectively, and >95% of the differences were within ±1 mm. This study identified multiple predictors for CAL measurement errors ≥1 mm, including tooth and site location, presence of supra- and subgingival calculus, bleeding on probing and suppuration. Measurement errors were more likely in patients diagnosed with "extensive" rather than "less extensive" periodontitis. In over half of the patients, measurement error frequencies were too high to allow for detection of possible CAL changes ≥2 mm with a false-positive rate ≤5%. Detection of CAL changes with low false-positives was more likely using recordings obtained by the direct method. CONCLUSIONS: From a measurement error point of view, CALDIR recordings are preferable over CALIND .

Phagocytic cell activity and periodontitis in Down syndrome.

BACKGROUND: This study investigated the phagocytic function of peripheral granulocytes and monocytes from adult individuals with Down syndrome (DS) and assessed the relation between phagocytic function and periodontal status. METHODS: Fifty-five DS individuals (18-56 years old), 74 mentally retarded individuals, and 88 medically healthy controls (HC) participated in the study. Gingival inflammation index, plaque index, probing depth, periodontal attachment level (AL), and bleeding on probing were taken for each subject. Whole blood was collected for granulocyte/monocyte phagocytosis tests. Phagocytic function was determined by flow cytometry in terms of percentage of cells actively involved in phagocytosis, and phagocytic intensity (magnitude of the bacterial staining per cell). RESULTS: Phagocytic intensity of both granulocytes and monocytes was comparable in HC and DS subjects. While AL was directly related to phagocytic intensity of both granulocytes (r = 0.14, P = 0.03) and monocytes (r = 0.2, P = 0.003) in all subjects, this relationship was stronger in DS than in other subjects, even after controlling for known risk factors for periodontitis (P < 0.05). Monocyte phagocytic intensity was the only necessary predictor of AL (P = 0.003), indicating a similar relationship between AL and phagocytic activity in either cell type. CONCLUSIONS: While granulocyte and monocyte phagocytic intensities are similar in Down and non-DS individuals, phagocytic intensity was associated with more AL in DS than non-DS individuals.

Immunohistochemical localization of CD1a and S100 in gingival tissues of healthy and chronic periodontitis subjects.

OBJECTIVE: The aim of this study was to evaluate the presence and distribution of CD1a and S100 protein markers in states of gingival health and chronic periodontitis in human subjects. MATERIALS AND METHODS: Gingival tissue samples were derived from 10 healthy and 10 chronic periodontitis-affected human subjects. The presence and distribution of CD1a and S100 protein was assessed using immunohistochemistry, and the cell types involved in their expression was determined. RESULTS: The presence and distribution of CD1a was confined only to the gingival epithelium, whereas S100 was seen in the epithelium and connective tissue. However, increased expression of both CD1a and S100 protein was seen in periodontitis-affected gingival tissues compared with healthy gingiva. Immunohistochemistry demonstrated that CD1a- and S100-positive cells in the epithelium are Langerhans cells (LCs) and S100 positive cells in the connective tissue are dendritic cells (DCs). CONCLUSION: Our findings suggest the transition of CD1a-positive LCs to S100-positive DCs from epithelium to connective tissue in response to an antigenic challenge. Demonstration of increased number of S100-positive DCs in the gingival connective tissue in chronic periodontitis possibly suggests their involvement in bone resorption in addition to their antigen presentation property.

Comparison of two power interdental cleaning devices on the reduction of gingivitis.

OBJECTIVE: The objective of this study was to compare the reduction of gingivitis by two power interdental devices combined with a manual toothbrush. METHODS: Eighty-two subjects completed this randomized, four-week, single-blind, two-group parallel clinical study. Subjects were randomly assigned to one of two groups: Waterpik Water Flosser (WF) plus manual tooth brushing or Sonicare Air Floss (AF) plus manual tooth brushing. Subjects were provided written and verbal instructions for all products at the baseline visit and instructions were reviewed at the two-week (W2) visit. Data were evaluated for whole mouth, lingual, and facial areas for gingivitis and bleeding on probing. Plaque data were evaluated for whole mouth, lingual, facial, approximal, and marginal areas of the tooth. Gingivitis, bleeding on probing, and plaque were scored at baseline (BSL), two weeks, and four weeks (W4). RESULTS: Both groups showed significant reductions in gingivitis, bleeding on probing, and plaque from baseline for all regions and time points measured (p < 0.001). The WF group was significantly more effective than the AF group at reducing plaque and gingivitis at W2 and W4 for all areas measured (p <0.001). At W4, the WF group was 80% more effective than AF for whole mouth gingivitis reduction, and twice as effective for the lingual region. In terms of plaque removal at W4, the WF group was 70% more effective for whole mouth (50.9% vs. 30%), 60% for approximal area (76.7% vs. 48%), and 47% for facial (52.8% vs. 35.9%) surfaces. The WF was twice as effective for lingual areas and more than three times as effective for marginal areas vs. the AF group (p <0.001). Results for bleeding on probing showed the WF group was numerically better than the AF group for all areas and time points, with these improvements being statistically significance for whole mouth (p = 0.02) and facial area (p = 0.004) at W2, and for the facial area (p = 0.02) at W4. CONCLUSION: The Waterpik Water Flosser is significantly more effective than Sonicare Air Floss for reducing gingivitis and plaque.

Expression of transient receptor potential vanilloid receptor 1 and toll-like receptor 4 in aggressive periodontitis and in chronic periodontitis.

BACKGROUND AND OBJECTIVE: The objective of the present study was to evaluate the expression and the distribution of the transient receptor potential vanilloid receptor 1 (TRPV1) and of toll-like receptor 4 (TLR4) in tissue samples from patients with periodontal disease (aggressive periodontitis and chronic periodontitis) and from healthy controls. MATERIAL AND METHODS: Ten tissue samples from each disease group (aggressive periodontitis and chronic periodontitis) and from healthy subjects were obtained during routine oral surgical procedures. Subgingival specimens were collected from sites with advanced loss of support (probing depth>5mm) and specimens from the corresponding healthy controls were obtained during tooth extraction for orthodontic reasons or following surgical extraction of an impacted third molar. The distribution of TRPV1 and TLR4 receptors in human gingival tissue was studied by immunohistochemistry. RESULTS: Both TLR4 and TRPV1 were detected in gingival tissues from healthy subjects, and from patients with chronic periodontitis and aggressive periodontitis, particularly in gingival keratinocytes, fibroblasts, inflammatory cells and the endothelial lining of capillaries in connective tissues. Histologic examination of the samples from healthy controls disclosed that clinically healthy gingiva does not correspond to histologically healthy gingiva. Subsequently, these samples were redesignated as gingivitis samples. TRPV1 was down-regulated in all cell types in samples obtained from patients with chronic periodontitis compared to samples obtained from patients with gingivitis, whereas TLR4 was down-regulated only in the epithelium and in gingival fibroblasts. In contrast, the levels of these markers in patients with aggressive periodontitis were similar to those in healthy patients. CONCLUSION: Local expression of TRPV1 and TLR4 in gingival tissues may contribute to both physiological and pathological processes in the periodontium. Our data suggest that TRPV1 and TLR4 may play a role specifically in the pathophysiology of chronic periodontitis.

Dimensional changes of peri-implant soft tissue over 2 years with single-implant crowns in the anterior maxilla.

OBJECTIVE: To compare the peri-implant soft tissue dimensions after insertion of single-implant crowns in the anterior maxilla. MATERIALS AND METHODS: Twenty patients were accepted according to well-defined inclusion criteria and randomized to porcelain-fused-to-metal (PFM) or all-ceramic groups. Follow-up was at: Baseline (B), Crown Insertion (CI), 1-year (1Y), and 2-year (2Y). The following parameters were statistically analysed: distance implant shoulder to marginal peri-implant mucosa (DIM), papilla height (PH), width of keratinized mucosa (KM), crestal bone level (CBL), full mouth plaque score (FMPS), full mouth bleeding score (FMBS), and probing pocket depth. RESULTS: Between groups measurements for DIM, PH, KM, CBL, FMPS, and FMBS showed no statistically significant differences except the distal CBLs to adjacent tooth. DIM (mid-facial) decreased from B to CI remaining stable at 1Y and 2Y (p-value 0.0014). DIM mesial and distal aspects significantly increased from B to CI showing signs of stability at the 2Y. PH between B and CI increased at the mesial site and at the distal site, thereafter, peri-implant soft tissues were stable at the 2Y. CONCLUSION: The insertion of an implant crown affects the peri-implant mucosa morphology by an apical displacement at the mid-facial aspect and coronal at mesial and distal sites.

Effect of removing an impacted mandibular third molar on the periodontal status of the mandibular second molar.

PURPOSE: The aim of this study was to evaluate the change in the periodontal status of mandibular second molars after surgical extraction of adjacent impacted lower third molars. MATERIALS AND METHODS: The study was based on a 1-year follow-up of 48 patients (20 men and 28 women) recruited consecutively after the extraction of an impacted lower third molar. Panoramic radiographs were obtained and clinical examinations were carried out at baseline to determine the periodontal status (probing depth and dental plaque and gingival indices) both for the second molar and for the 4 posterior sextants. After surgical removal of the impacted mandibular third molars, all patients were assessed at 3, 6, 9, and 12 months for changes in periodontal status. RESULTS: The periodontal health of the second molar was found to improve gradually after third molar surgery in all clinical parameters. Probing depth was gradually reduced by about 0.6 mm quarterly, until a final depth of 2.6 ± 0.8 mm was attained. The relative risk of having a plaque index and gingival index coded as 0 (healthy) or 1 (minor problems) was about 10 times higher at the end of the follow-up than at baseline for both indices. The periodontal status of the 4 posterior sextants also improved gradually. Molar depth, according to the Pell and Gregory classes and types, seemed to be the main factor modulating both the baseline probing depth and the change in probing depth during follow-up. CONCLUSIONS: Our results suggest that the initial periodontal breakdown established on the distal surfaces of the second molars and in the periodontal health of the 4 posterior sextants can be significantly improved 1 year after surgical removal of the ipsilateral lower third molar.

C-reactive protein in gingival crevicular fluid may be indicative of systemic inflammation.

BACKGROUND AND AIM: Periodontitis is associated with elevated C-reactive protein (CRP) in both serum and gingival crevicular fluid (GCF). Although the liver is the primary source of CRP, extra-hepatic production of CRP has been reported. This study aimed to determine whether CRP in GCF is produced locally in the gingivae. MATERIALS AND METHODS: Gingivae and GCF were collected from non-periodontitis and periodontitis sites. Presence of CRP in gingivae was assessed by immunohistochemistry. CRP in GCF was measured using ELISA. Gene expression for CRP in gingivae was determined using real-time polymerase chain reaction. RESULTS: CRP was found in both the gingivae and GCF. No gingivae had detectable amounts of CRP mRNA. Not all patients with periodontitis had detectable levels of CRP in the GCF. Some non-periodontitis patients had detectable levels of CRP in the GCF. CONCLUSION: CRP in the GCF appears to be of systemic origin, and therefore may be indicative of systemic inflammation from either a periodontal infection or inflammatory disease elsewhere. The correlation between levels of CRP in GCF and serum requires validation in future studies.

[Usefulness of micronutrients in the treatment of periodontitis]. / Bruikbaarheid micronutriënten bij behandeling van parodontitis.

Micronutrients can inhibit oxidation reactions due to micro-organisms in human tissues. In this randomized, double-blind and placebo-controlled pilot study, it was examined whether suppletion of a mixture of selected micronutrients could affect the outcome of conservative treatment of periodontitis. A group of 16 for periodontitis treated patients received a mixture of micronutrients consisting of alpha-tocopherol acetate, ascorbic acid, cholecalcipherol and an extract from green tea, whereas a group of 16 similar patients received a placebo. At baseline and after 3 and 6 months, in every patient the number of teeth with periodontitis, the number of gingival bleedings on probing, and the pocket depths were registered. An evident improvement was defined as 40% reduction of the number of gingival bleedings on probing or of the summated pocket depth. After 6 months, the number of gingival bleedings on probing was significantly more reduced in the experimental group when compared with the placebo group. Also evident improvement was significantly more observed in the experimental group when compared with the placebo group.

In vivo determination of multiple indices of periodontal inflammation by optical spectroscopy.

BACKGROUND AND OBJECTIVE: Visible, near-infrared (optical) spectroscopy can be used to measure regional tissue hemodynamics and edema and therefore may represent an ideal tool with which to study periodontal inflammation in a noninvasive manner. The study objective was to evaluate the ability of optical spectroscopy to determine simultaneously multiple inflammatory indices (tissue oxygenation, total tissue hemoglobin, deoxyhemoglobin, oxygenated hemoglobin and tissue edema) in periodontal tissues in vivo. MATERIAL AND METHODS: Spectra were obtained, processed and evaluated from healthy, gingivitis and periodontitis sites (n = 133) using a portable optical, near-infrared spectrometer. A modified Beer-Lambert unmixing model that incorporates a nonparametric scattering loss function was used to determine the relative contribution of each inflammatory component to the overall spectrum. RESULTS: Optical spectroscopy was harnessed to generate complex inflammatory profiles of periodontal tissues. Tissue oxygenation at periodontitis sites was significantly decreased (p < 0.05) compared to sites with gingivitis and healthy controls. This was largely the result of an increase in deoxyhemoglobin in the periodontitis sites compared with healthy (p < 0.01) and gingivitis (p = 0.05) sites. Tissue water content per se showed no significant difference between the sites, but a water index associated with tissue electrolyte levels and temperature differed significantly between periodontitis sites and both healthy and gingivitis sites (p < 0.03). CONCLUSION: This study established that optical spectroscopy can simultaneously determine multiple inflammatory indices directly in the periodontal tissues in vivo. Visible, near-infrared spectroscopy has the potential to be developed into a simple, reagent-free, user-friendly, chairside, site-specific, diagnostic and prognostic test for periodontitis.

Increased expression of extracellular matrix metalloproteinase inducer is associated with matrix metalloproteinase-1 and -2 in gingival tissues from patients with periodontitis.

BACKGROUND AND OBJECTIVE: Extracellular matrix metalloproteinase inducer (EMMPRIN), an immunoglobulin-like cell surface glycoprotein, could promote collagenolytic balance in favor of the expression and activation of matrix metalloproteinases (MMPs). This study was to investigate the expression of EMMPRIN in gingival tissues from different periodontal conditions and to correlate it with the production of MMP-1 and MMP-2. MATERIAL AND METHODS: Gingival biopsies were collected from 15 patients with untreated advanced chronic periodontitis and 15 patients with aggressive periodontitis (AgP). The control group consisted of 12 subjects diagnosed either as periodontally healthy individuals or as individuals with a gingival index of one (H/G1). The peptides and mRNA of EMMPRIN, MMP-1 and MMP-2 were detected by immunohistochemistry and semi-quantitative reverse transcriptase-polymerase chain reaction, respectively. RESULTS: The expression of EMMPRIN, MMP-1 and MMP-2 peptides in periodontally healthy tissues was mainly confined to the gingival epithelium. The EMMPRIN was strongly expressed in the cell membrane of the basal layer. Immunoreactivity for EMMPRIN was more intensive and more widespread in periodontitis, extended from the epithelial layers to the underlying connective tissues, and was essential in both inflammatory and fibroblast-like cells. In addition, MMP-1 and MMP-2 showed the same localized expression. The chronic periodontitis group had a significantly higher mRNA expression of EMMPRIN and MMP-2 compared with the H/G1 subjects (p < 0.05). Production of MMP-1 and MMP-2 by gingival tissues was correlated with the mRNA level of EMMPRIN (r = 0.463, p = 0.013 for MMP-1 and r = 0.404, p = 0.033 for MMP-2). CONCLUSION: The expression of EMMPRIN in human normal and diseased gingiva might contribute to periodontal physiological and pathological processes; moreover, its increased production might be associated with MMP-1 and MMP-2 expression.

B-1a cells and plasma cells in periodontitis lesions.

BACKGROUND AND OBJECTIVE: Host response mechanisms in periodontal tissues are complex and involve numerous systems of interactions between cells. The B-cell lineage seems to predominate in chronic periodontitis lesions. The aim of the present investigation was to study the correlation between inflammatory cells and some functional markers in gingival lesions obtained from subjects with severe chronic periodontitis. MATERIAL AND METHODS: Thirty-eight Caucasian subjects volunteered to take part in the study. A gingival biopsy from one randomly selected diseased proximal site (probing pocket depth > 6 mm and bleeding on probing positive) was obtained from each patient. Immunohistochemical preparation was used to identify inflammatory cells and functional markers. Correlations between the different percentages of cell markers were analyzed by pairwise correlation. RESULTS: B cells (B-1a and B-2 cells) occurred in larger proportions than T cells and plasma cells. A statistically significant correlation was found between the percentage of B-1a cells and plasma cells and between all B lymphocytes and plasma cells. About 60% of B lymphocytes exhibited autoreactive features. CONCLUSION: It is suggested that B-1a cells constitute a significant part of the host response in periodontitis lesions and that plasma cells may develop from both B-2 and B-1a cells.

Effects of caloric restriction on inflammatory periodontal disease.

OBJECTIVE: Dietary caloric restriction (CR) has been found to reduce systemic markers of inflammation and may attenuate the effects of chronic inflammatory conditions. The purpose of this study was to examine the effects of long-term CR on naturally occurring chronic inflammatory periodontal disease in a nonhuman primate model. METHODS: The effects of long-term CR on extent and severity of naturally occurring chronic periodontal disease, local inflammatory and immune responses, and periodontal microbiology, were evaluated in a cohort of 81 (35 female and 46 male; 13-40 y of age) rhesus monkeys (Macaca mulatta) with no previous exposure to routine oral hygiene. CR monkeys had been subjected to 30% CR for 13-17 y relative to control-fed (CON) animals starting at 3-5 y of age. RESULTS: Same sex CR and CON monkeys exhibited similar levels of plaque, calculus, and bleeding on probing. Among CON animals, males showed significantly greater periodontal breakdown, as reflected by higher mean clinical attachment level and periodontal probing depth scores, than females. CR males exhibited significantly less periodontal pocketing, lower IgG antibody response, and lower IL-8 and ss-glucuronidase levels compared to CON males, whereas CR females showed a lower IgG antibody response but comparable clinical parameters and inflammatory marker levels relative to CON females. Long-term CR had no demonstrable effect on the periodontal microbiota. CONCLUSION: Males demonstrated greater risk for naturally occurring periodontal disease than females. Long-term CR may differentially reduce the production of local inflammatory mediators and risk for inflammatory periodontal disease among males but not females.

Anti-tumor necrosis factor-alpha therapy and periodontal parameters in patients with rheumatoid arthritis.

BACKGROUND: The aim of this study was to evaluate the influence of anti-tumor necrosis factor-alpha (TNF-alpha) therapy on the clinical and immunologic parameters of the periodontium. METHODS: Ten patients with rheumatoid arthritis (RA) who routinely received infusions of infliximab, 200 mg (RA+), 10 patients with RA without anti-TNF-alpha therapy (RA-), and 10 healthy controls (C) were included. Clinical parameters, including the plaque index (PI), gingival index (GI), probing depth (PD), clinical attachment loss (AL), and bleeding on probing (BOP), were assessed, and total gingival crevicular fluid (GCF) TNF-alpha level was determined using enzyme-linked immunosorbent assay. Analysis of variance with Scheffe modification and the Pearson correlation test were used for statistical analysis. RESULTS: The ages of the patients ranged from 22 to 76 years (mean, 50.73 +/- 9.1 years). The mean PI was similar among the groups. However, mean inflammatory parameters in the three groups varied significantly; GI was greater in the RA- group compared to RA+ and C groups (P = 0.0042). The RA+ group exhibited less BOP than RA- and C groups (21.1% +/- 3.0%, 45.9% +/- 6.2%, and 39.1% +/- 7.2%, respectively; P = 0.0146). The mean PD in the RA+ group was shallower than in RA- and C groups (3.22 +/- 0.13 mm, 3.85 +/- 0.22 mm, and 3.77 +/- 0.20 mm, respectively; P = 0.055). Clinical AL in the RA+ group was lower than in RA- and C groups (3.68 +/- 0.11 mm, 4.52 +/- 0.26 mm, and 4.35 +/- 0.24 mm, respectively; P = 0.0273). TNF-alpha levels in the GCF of the RA+ group were the lowest compared to RA- and C groups (0.663, 1.23, and 0.949 ng/site, respectively; P = 0.0401). A significant positive correlation was found between TNF-alpha levels in the GCF and clinical AL (r = 0.448; P = 0.0283). CONCLUSIONS: Patients with RA receiving anti-TNF-alpha medication had lower periodontal indices and GCF TNF-alpha levels. Thus, suppression of proinflammatory cytokines might prove beneficial in suppressing periodontal diseases.

Immunohistochemical localization of Toll-like receptors 1-10 in periodontitis.

BACKGROUND/AIM: In periodontitis, bacteria and pathogen-associated molecular patterns are sensed by Toll-like receptors (TLRs), which initiate intracellular signaling cascades that may lead to host inflammation. In this study, the expression and distribution of TLRs (TLR-1 to TLR-10) were immunohistochemically detected in gingival epithelium and connective tissue. METHODS: Immunohistochemistry was used for the localization of TLRs in gingival tissue samples from 10 patients with chronic periodontitis and 10 healthy controls; these samples were obtained during routine periodontal flap operations and during extraction operations performed for retained wisdom teeth, respectively. For the evaluation, epithelial layers were stratified to basal, spinous, and superficial layers, and the percentages of TLR-positive cells were determined. RESULTS: Both healthy and periodontitis gingival tissues expressed all TLRs except TLR-10. In patients with periodontitis, epithelial cells showed increased TLR expression towards the basal layer. Healthy controls showed more variable cellular TLR expression and distribution between the layers of epithelium. In the connective tissue, consistently higher TLR expression was found within the periodontitis group compared to the healthy group. CONCLUSIONS: For the first time, the cellular expression and distribution of TLR-1 to TLR-10 have been studied in periodontitis, indicating that TLR-1 to TLR-9 are differentially expressed both in connective tissue and epithelial layers. Except for TLR-7 and TLR-8, all the other TLRs showed statistically significant differences between patients with periodontitis and healthy controls, suggesting their involvement in the pathogenesis of periodontitis.

Significance of thrombomodulin release from gingival epithelial cells in periodontitis patients.

BACKGROUND AND OBJECTIVE: Thrombomodulin, a cell transmembrane glycoprotein, binds to thrombin and converts it from a procoagulant protease to an anticoagulant enzyme that activates protein C. Thrombomodulin is very important in regulating the function of thrombin. Elevated soluble thrombomodulin is present in the gingival crevicular fluid of subjects with periodontitis. The objective of the present study was to investigate the mechanisms about the elevated soluble thrombomodulin in gingival crevicular fluid. MATERIAL AND METHODS: Gingival sections from six patients with chronic periodontitis and from three periodontally healthy subjects were immunostained for thrombomodulin detection. Thrombomodulin levels were investigated in the gingival crevicular fluid of 11 subjects with chronic periodontitis. The effects of neutrophil enzymes on thrombomodulin release and on thrombomodulin in the gingival crevicular fluid were examined by an enzyme-linked immunosorbent assay or by Western blotting. RESULTS: The expression of gingival epithelial thrombomodulin was lost or decrease near infiltrating neutrophils. Thrombomodulin was rapidly released from gingival epithelial cells by neutrophil enzymes, and gingival crevicular fluid with periodontitis included the proteolytic cleavage thrombomodulin using immunoblotting analysis. The thrombomodulin release was not caused by rapid cell damage, on lactate dehydrogenase assay. There were significant differences in thrombomodulin content between gingival crevicular fluid samples from healthy and diseased sites, regardless of the degree of probing depth. CONCLUSION: Neutrophil enzymes induced rapid thrombomodulin release from the membrane surface of gingival epithelial cells. This might explain the thrombomodulin increase in gingival crevicular fluid with local diseased gingiva. Elevation of thrombomodulin in gingival crevicular fluid may be a potential marker of epithelial cell membrane injury.