The utility of gingival crevicular fluid matrix metalloproteinase-8 response patterns in prediction of site-level clinical treatment outcome.

BACKGROUND: Different gingival crevicular fluid (GCF) matrix metalloproteinase (MMP)-8 response patterns were studied among non-smoking and smoking patients with chronic periodontitis (CP) and generalized aggressive periodontitis (GAgP) to test the utility of GCF MMP-8 levels predicting the site-level treatment outcome. METHODS: Data from four independent longitudinal studies were combined. Altogether, the studies included 158 periodontal sites from 67 patients with CP and 32 patients with GAgP, and GCF samples were collected at baseline, after the treatment, and during the 6-month maintenance period. All GCF samples were analyzed by immunofluorometric assay for MMP-8. Different site-level MMP-8 response patterns were explored by the cluster analysis. Most optimal MMP-8 cutoff levels were searched with receiver operating characteristic analyses, and the predictive utility of defined levels was tested. RESULTS: Distinct types of MMP-8 response patterns were found in both smokers and non-smokers. MMP-8 levels exceeding the optimal cutoff levels separately defined for smokers and non-smokers indicated increased risk for compromised treatment outcome at baseline and during the maintenance period. Seventy-one percent of non-smokers (positive likelihood ratio of 4.22) and 88% of smokers (positive likelihood ratio of 5.00) with positive test results at both baseline and the maintenance period had compromised treatment outcome. The double-positive result indicated 46% and 39% point risk increase for the compromised outcome, respectively. CONCLUSION: GCF MMP-8 analysis with defined cutoff levels could be used to predict the site-level treatment outcome and for longitudinal monitoring of the disease status during the maintenance period.

Gingival crevicular fluid matrix metalloproteinase-8 levels predict treatment outcome among smokers with chronic periodontitis.

BACKGROUND: Molecular biomarkers are needed for diagnostic use in periodontal diseases. The aim of this study is to explore different gingival crevicular fluid (GCF) matrix metalloproteinase-8 (MMP-8) patterns in smokers and non-smokers with chronic periodontitis (CP) and test the utility of baseline GCF MMP-8 levels in predicting categorically assessed treatment outcomes. METHODS: The study population comprised 15 patients with CP (five non-smokers and 10 smokers). GCF sampling of five to seven periodontal sites per patient was done at baseline, post-treatment, and bimonthly during the maintenance period from 8 to 12 months. GCF MMP-8 levels were measured with an immunofluorometric assay. MMP-8 response patterns were explored by cluster analysis. The ability of baseline MMP-8 levels to predict categorical treatment outcomes was analyzed with receiver operating characteristic curves. RESULTS: GCF MMP-8 response patterns could be clustered into two different site profiles among both smokers and non-smokers. Smoker site profiles 1 and 2 had significantly different clinical attachment level and gingival recession changes by the end of the maintenance period. In smoker sites, baseline MMP-8 levels significantly predicted the categorical treatment outcome. CONCLUSIONS: Baseline GCF MMP-8 levels strongly predict how MMP-8 levels behave during the maintenance period. In smoker sites, high baseline MMP-8 levels indicate weak treatment response.

Differential expression of mitogen activating protein kinases in periodontitis.

AIM: Following toll-like receptor (TLR) engagement, lipopolysaccharide (LPS) can stimulate the expression of pro-inflammatory cytokines thus activating the innate immune response. The production of inflammatory cytokines results, in part, from the activation of kinase-induced signalling cascades and transcriptional factors. Of the four distinct classes of mitogen-activated protein kinases (MAPK) described in mammals, p38, c-Jun N-terminal activated kinases (JNK1-3) and extracellular activated kinases (ERK1,2) are the best studied. Previous data have established that p38 MAPK signalling is required for inflammation and bone loss in periodontal disease pre-clinical animal models. MATERIALS & METHODS: In this study, we obtained healthy and diseased periodontal tissues along with clinical parameters and microbiological parameters. Excised fixed tissues were immunostained with total and phospho-specific antibodies against p38, JNK and ERK kinases. RESULTS: Intensity scoring from immunostained tissues was correlated with clinical periodontal parameters. Rank correlations with clinical indices were statistically significantly positive (p-value < 0.05) for total p38 (correlations ranging 0.49-0.68), phospho-p38 (range 0.44-0.56), and total ERK (range 0.52-0.59) levels, and correlations with JNK levels also supported association (range 0.42-0.59). Phospho-JNK and phospho-ERK showed no significant positive correlation with clinical parameters of disease. CONCLUSION: These data strongly implicate p38 MAPK as a major MAPK involved in human periodontal inflammation and severity.

Cyclooxygenase-2 expression in gingival biopsies from periodontal patients is correlated with connective tissue loss.

BACKGROUND: The objective of this study is to compare cyclooxygenase-2 (COX-2) protein expression in gingival biopsies from patients with chronic periodontitis (CP), patients with gingivitis (GV), and individuals with no periodontal disease (control group) and to establish its relationship with clinical variables and connective tissue loss in the lamina propria. METHODS: A cross-sectional and analytic study was conducted in 108 gingival biopsies from 52 patients with CP, 39 with GV, and 17 controls. All biopsies were processed for conventional histopathologic study, immunohistochemical determination of COX-2 protein expression, and automatic quantification of connective tissue by image analysis. RESULTS: The protein expression of COX-2, mainly produced by plasma cells and monocytes, was significantly related to the presence of periodontal disease, bleeding index, intensity of inflammatory infiltrate, and loss of connective tissue in the lamina propria of gingival biopsies (P <0.01, Spearman test). COX-2 expression was also directly correlated with attachment loss (P <0.05, Spearman test). CONCLUSIONS: COX-2 protein expression is higher in patients with GV and CP than in individuals without periodontal disease and is inversely correlated with the amount of connective tissue in the lamina propria as determined by image analysis. This finding suggests that COX-2 participates in mechanisms and pathway signaling related to the destruction of fibrillar support structures of the periodontium.

Protease inhibitor levels in periodontal health and disease.

BACKGROUND AND OBJECTIVE: Our previous study showed that protease inhibitors were attenuated by the periodontal pathogen Porphyromonas gingivalis in cultured gingival epithelial cells. We hypothesize that fewer protease inhibitors would be present in more advanced periodontal disease sites, where the level of P. gingivalis may be high. The goal of this study was to investigate the relationship between the protease inhibitor [secretory leukocyte protease inhibitor (SLPI), elastase-specific inhibitor (ELAFIN) and squamous cell carcinoma antigen (SCCA)] levels in gingival crevicular fluid and the number of P. gingivalis micro-organisms in subgingival plaque. MATERIAL AND METHODS: Plaque samples from subjects without (n = 18) and with moderate to advanced periodontitis (n = 41) were used to quantify P. gingivalis using real-time PCR. Protease inhibitor levels in the gingival crevicular fluid of all the subjects were determined by ELISA. RESULTS: P. gingivalis was detected in 68.3% of patients with periodontitis, while 16.7% of subjects without periodontitis had a detectable level of P. gingivalis. Patients with periodontitis and P. gingivalis in their plaque exhibited lower SLPI and ELAFIN levels (p < 0.001) compared with control subjects without periodontitis. Secretory leukocyte protease inhibitor was also reduced (p < 0.05) in gingival crevicular fluid of periodontitis patients without a detectable level of P. gingivalis. Periodontitis patients with high vs. low levels of P. gingivalis exhibited reciprocal mean levels of SLPI and ELAFIN concentrations. CONCLUSION: The reduced concentrations of SLPI and ELAFIN may contribute to the loss of host protective capacity and increase susceptibility to breakdown from chronic infection. The work of this investigation may aid in finding diagnostic and prognostic markers in periodontal health and disease and may also help in finding pharmacological targets directed against periodontal inflammation.

The impact of smoking status on antioxidant enzyme activity and malondialdehyde levels in chronic periodontitis.

BACKGROUND: The aim of this study is to investigate the impact of smoking status on the systemic and local superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) activities and malondialdehyde (MDA) levels in subjects with chronic periodontitis (CP). METHODS: Sixty-five CP patients (23 smokers [CP-S], 23 former smokers [CP-FS], and 19 non-smokers [CP-NS]) and 20 periodontally healthy non-smoker controls (PH-NS) were included in the study. After the clinical measurements, serum and gingival tissue samples were collected. SOD, GSH-Px, and CAT activities and MDA levels in hemolysates and gingival tissue samples were spectrophotometrically assayed. RESULTS: Blood MDA levels in all the periodontitis groups were higher than in the PH-NS group but only the difference between CP-FS and PH-NS groups was significant (P <0.01). Gingival tissue MDA levels in the periodontitis groups were significantly higher than that in the control group (P <0.01). However, the control group had the highest gingival SOD, GSH-Px, and CAT activities compared with all the periodontitis groups (P <0.01). The CP-S group had the highest gingival MDA levels and SOD, GSH-Px, and CAT activities among the periodontitis groups, whereas the lowest values were observed in the CP-NS group (P <0.01). The blood and gingival MDA levels in the CP-FS group were similar in the CP-NS group, whereas they were lower than in the CP-S group. CONCLUSIONS: Systemic and local MDA levels are increased by smoking in addition to the impact of periodontitis. The decreased local SOD, GSH-Px, and CAT activities observed in periodontitis patients may increase with smoking.

Crevicular fluid matrix metalloproteinase-8, -13, and TIMP-1 levels in type 2 diabetics.

OBJECTIVES: To evaluate whether type 2 diabetes mellitus (DM) enlarged and if so the quantum of such increase in the gingival crevicular fluid (GCF) levels of matrix metalloproteinase-8 (MMP-8), MMP-13 and tissue inhibitor of metalloproteinases-1 (TIMP-1). METHODS: Subjects (n = 73) were divided into five groups as follows: 12 DM patients with gingivitis (DM-G), 12 DM patients with periodontitis (DM-P), 12 systemically healthy patients with gingivitis (H-G), 13 systemically healthy patients with periodontitis (H-P) and 24 periodontally, systemically healthy volunteer subjects (H-C). Full-mouth clinical periodontal measurements were performed at six sites per tooth. Gingival crevicular fluid samples were obtained from two sites representing the clinical periodontal diagnosis in single-rooted teeth. Gingival crevicular fluid levels of MMP-8, MMP-13 and TIMP-1 were analysed by immunofluorometric MMP assay (IFMA), enzyme-linked immunosorbent assay (ELISA). Data were tested statistically by parametric tests. RESULTS: All clinical periodontal measurements were similar in both diabetic and systemically healthy patients with periodontal disease (all P > 0.05). Total amounts of MMP-8 in GCF samples were significantly lower in H-C group than DM-G, DM-P, H-P groups (all P < 0.05). Matrix metalloproteinase-13, TIMP-1 total amounts were similar in study groups (P > 0.05). Diabetes mellitus patients exhibited similar levels of MMP-8, MMP-13, TIMP-1 with systemically healthy gingivitis/periodontitis patients (P > 0.05). CONCLUSIONS: Within the limits of this study, DM does not seem to significantly affect GCF levels of MMP-8, MMP-13, TIMP-1 or clinical periodontal status.

Alanine aminopeptidase and dipeptidyl peptidase IV in saliva of chronic periodontitis patients.

BACKGROUND: Alanine aminopeptidase (ALAP) and dipeptidyl peptidase IV (DPPIV) are ectopeptidases that play a role in collagen degradation and are thought to be involved in the destruction of periodontal tissue. This study compared the activities of salivary ALAP and DPPIV in patients with periodontitis and periodontally healthy subjects. The correlations of enzyme activities with clinical variables and the presence of Porphyromonas gingivalis were also evaluated. METHODS: Whole saliva was collected from 30 periodontally healthy subjects, 30 localized chronic periodontitis (LCP) patients, and 30 generalized chronic periodontitis (GCP) patients to determine the activities of ALAP and DPPIV. The presence of P. gingivalis in subgingival plaque was detected by polymerase chain reaction. Periodontal clinical assessments included probing depth, clinical attachment level, and bleeding on probing. RESULTS: The activities of DPPIV in the LCP and GCP groups were not significantly different from one another, but both groups had significantly higher enzyme activities than the periodontally healthy group (P = 0.001). DPPIV activity was positively correlated with all clinical parameters and the prevalence of P. gingivalis. The ALAP activities were not significantly different among the three study groups. There was no significant correlation of ALAP activity with any of the clinical and bacterial parameters. CONCLUSION: DPPIV, but not ALAP, activity is associated with periodontitis and the presence of P. gingivalis.

Maternal periodontal disease and soluble fms-like tyrosine kinase-1 expression.

BACKGROUND: This study was conducted to examine the relationship between maternal periodontal disease and plasma angiogenic factor expression of soluble fms-like tyrosine kinase (sFlt)-1. METHODS: This was a nested case-control study of 220 women, including 45 healthy women with evidence of active periodontal disease, 98 women without evidence of active periodontal disease, 13 women with fetal exposure to oral pathogens, and 64 women without fetal exposure to oral pathogens. Active periodontal disease was defined as the presence of moderate/severe periodontal disease and evidence of periodontal disease progression. Fetal exposure to oral pathogens was determined by fetal immunoglobulin M (IgM) umbilical cord seropositivity. Maternal plasma was collected at <26 weeks of gestation; umbilical cord blood was collected at delivery. sFlt-1 was measured with an immunoradiometric assay. Demographic and medical data were chart abstracted. Maternal variables and sFlt-1 concentrations were compared between cases and controls using the Student t and chi(2) tests and analysis of variance. RESULTS: The median sFlt-1 concentration at the time of enrollment for all women was 2,374 pg/ml (interquartile range [IQR]: 1,504 to 3,194 pg/ml). Women with evidence of fetal exposure to oral pathogens had significantly higher sFlt-1 concentrations compared to IgM-negative fetuses (3,383 pg/ml [IQR: 2,610 to 4,244 pg/ml] versus 2,123 pg/ml [IQR: 1,456 to 3,011 pg/ml]; P = 0.03). CONCLUSION: Fetal exposure to oral pathogens was associated with increased plasma concentrations of sFlt-1 early in pregnancy.

Differential gene and protein expression of tissue inhibitors of metalloproteinases (TIMP)-3 and TIMP-4 in gingival tissues from drug induced gingival overgrowth.

OBJECTIVES: The purpose of this study was to analyse mRNA expression and protein localization of tissue inhibitors of metalloproteinases (TIMP)-3 and TIMP-4 in gingival tissues removed from drug (calcium-channel blocker) induced gingival overgrowth and periodontitis patients. DESIGN: Employing RT-PCR, we evaluated TIMP-3 and TIMP-4 mRNA levels of 20 human gingival tissue samples taken from patients suffering gingival overgrowth (GO) and periodontitis (P). Then, using immunohistochemistry we investigated the TIMP-3 and TIMP-4 protein localization of five sample tissues from each group. RESULTS: TIMP-4 mRNA levels in GO-gingiva tended to be lower than in P-gingiva but the results differed little (p = 0.22). Varying degrees of inflammation in the protein localization of TIMP-3 and TIMP-4 were found. TIMP-4 immunoreactivity (IR) was weak in the endothelial cells, fibroblasts, epithelial basal and parabasal cells while the degree of inflammation differed as well. TIMP-3 and TIMP-4 IR in inflammatory cells, including lymphocytes, plasma cells, and macrophages, were faint and intense respectively. For P-gingiva, both TIMP-3 and TIMP-4 IR expression was weak in the endothelial cells, fibroblasts, basal and parabasal epithelial layers. Expression of TIMP-3 was faint in the inflammatory cells, whereas TIMP-4 IR was strong. CONCLUSION: Our findings suggest that TIMP-3 and TIMP-4 expression differs in GO and P-gingival tissues, both of which are potentially involved in pathogenesis.

Matrix metalloproteinase-8 concentration in shallow crevices associated with the extent of periodontal disease.

OBJECTIVE: The aim of this study was to analyse the association between matrix metalloproteinase-8 (MMP-8) concentration in shallow, mostly non-bleeding gingival crevices, and the extent of periodontal disease. MATERIAL AND METHODS: Plaque, bleeding on probing (BOP), probing pocket depth (PPD) and attachment level (AL) were assessed clinically in 48 patients with chronic periodontitis. MMP-8 concentrations in gingival crevicular fluid (GCF) from four shallow (PPDor=4 mm (p=0.028) and AL>or=6 mm (p<0.001). CONCLUSION: The above association between MMP-8 concentration in shallow crevices and attachment loss provides a new aspect to future studies of MMP-8 as a prognostic marker for periodontal disease.

Tumor necrosis factor-alpha-converting enzyme (TACE) levels in periodontal diseases.

Tumor necrosis factor-alpha-converting enzyme (TACE) is a metalloprotease which can shed several cytokines from the cell membrane, including receptor activator of NF-kappaB ligand (RANKL). This study aimed to investigate the hypothesis that TACE would be elevated in the gingival crevicular fluid (GCF) of persons with periodontitis. Total TACE amounts in GCF were higher in persons with chronic and aggressive periodontitis than in those with gingivitis or in healthy persons. TACE concentrations in GCF were higher in persons with chronic and aggressive periodontitis than in those with gingivitis, although not significantly higher than in healthy persons. Persons with chronic periodontitis receiving immunosuppressive treatment exhibited over 10-fold lower TACE levels than the other periodontitis groups. TACE was positively correlated with probing pocket depth, clinical attachment levels, and RANKL concentrations in GCF. In conclusion, the increased GCF TACE levels in persons with periodontitis and their positive correlation with RANKL may indicate an association of this enzyme with alveolar bone loss, and may warrant special attention in future therapeutic approaches.

MMP-13 and TIMP-1 determinations in progressive chronic periodontitis.

UNLABELLED: Matrix metalloproteinase (MMP)-13 is a collagenase involved in extracellular matrix degradation either by its direct degradative effects or by processing bioactive substrates. The aim of this study was to determine the levels of MMP-13 and tissue inhibitor of metalloproteinase (TIMP)-1 in gingival crevicular fluid (GCF) and gingival biopsies obtained from active and inactive sites during chronic periodontitis progression. MATERIALS AND METHODS: This was a longitudinal study in which chronic periodontitis patients with moderate to severe disease were included and followed until they developed progression determined by the tolerance method. GCF samples were obtained from periodontitis, active, inactive and healthy sites and additional gingival biopsies were taken from active and inactive sites. MMP-13 and TIMP-1 determinations were carried out by immunodot blots and immunowestern blots. RESULTS: In progressive periodontitis, MMP-13 and TIMP-1 remained unchanged between active and inactive sites, but as the TIMP-1 relative levels increased together with MMP-13 elevation in inactive samples, an inverse correlation was observed in active sites. Besides, MMP-13 was undetectable in healthy controls. CONCLUSION: Chronic periodontitis is characterized by increased MMP-13 expression. During disease progression, active sites tended to decrease TIMP-1 levels in association with MMP-13 elevation.

Expression of cathepsin-K in gingival crevicular fluid of patients with periodontitis.

Cathepsin-K is a highly expressed cysteine protease, and it plays a key role in bone remodeling and cartilage breakdown in bone. Cathepsin-K is used as a well-known marker of osteoclast activity, because this enzyme is mainly derived from osteoclasts. The receptor activator for NF-kappaB ligand (RANKL) plays an important role in osteoclast formation. Although a recent study suggests the involvement of RANKL in the pathogenesis of periodontal disease, no one has previously examined the level of cathepsin-K in the body fluid of human subjects. If the presence of cathepsin-K, as well as RANKL, can be detected in body fluids, it would be indirect proof of the differentiation and/or activation of osteoclasts in the tissues bathed by these fluids. This communication reports on the in vivo concentrations of cathepsin-K and RANKL in the gingival crevicular fluid (GCF) of normal subjects and those patients with severe, moderate, and mild forms of the disease. Increased concentrations of cathepsin-K and RANKL were detected in the GCF from patients with periodontitis (P<0.005 versus control subjects). Also, there was a positive correlation between cathepsin-K and RANKL levels (r=0.726), suggesting that both of them contribute to osteoclastic bone destruction in periodontal disease.

The associations between gingival crevice fluid matrix metalloproteinase-9, tissue inhibitor of metalloproteinase-1 and periodontitis in human immunodeficiency virus-positive patients.

BACKGROUND AND OBJECTIVE: The study aimed to determine whether matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in gingival crevice fluid could serve as prognostic factors for the progression of periodontitis in human immunodeficiency virus (HIV) -positive patients. Activated inflammatory cells produce inflammatory mediators, which stimulate the production of MMPs and their inhibitors. It is likely that the compromised immune system contributes to the pathogenesis of periodontitis in HIV-positive patients. METHODS: Clinical measurements including gingival index, plaque index, bleeding index, probing depth, attachment loss, and gingival crevice fluid samples were taken from two healthy sites (including sites with gingival recession, gingival index = 0; probing depth < or = 3 mm; attachment loss < or = 2 mm), three gingivitis sites (gingival index > 0; probing depth < or = 3 mm; attachment loss = 0) and three periodontitis sites (gingival index > 0; probing depth > or = 5 mm; attachment loss > or = 3 mm) of each of the 35 patients at baseline visits and 6-month visits by means of paper strips. Gingival crevice fluid levels of MMP-9 and TIMP-1 were determined by sandwich enzyme-linked immunosorbent assays. RESULTS: The mean amounts of MMP-9 and TIMP-1 in the gingivitis and periodontitis sites sites were significantly higher than in the healthy sites (P < 0.0001). The progressing site was defined as a site that had 2 mm or more attachment loss during the 6-month study period. Gingival crevice fluid levels of MMP-9 were significantly correlated with probing depth, attachment loss, TIMP-1, age, smoking pack years, and viral load values at baseline and 6-month visits (0.0001 < P < 0.001). TIMP-1 levels were only correlated with CD4, viral load, attachment loss, and MMP-9 (0.001 < P < 0.01). Repeated measures analysis of 11 active sites vs. 269 inactive sites indicated that MMP-9 and TIMP-1 levels were significantly higher in active sites than in inactive sites (P < 0.0001). These data indicate that sites with high ginigval crevice fluid levels of MMP-9 and TIMP-1 in HIV-positive patients are at significantly greater risk for progression of periodontitis.

Genetic polymorphisms in the MMP-1 and MMP-3 gene may contribute to chronic periodontitis in a Brazilian population.

OBJECTIVES: Matrix metalloproteinase-1 and -3 (MMP-1, MMP-3) represent proteinases that degrade macromolecules of the extracellular matrix. These enzymes play a fundamental role during destruction of periodontal tissues. Genetic polymorphisms were characterized in the promoter region of the MMP-1 and MMP-3 genes. The aim of this study was to investigate the relationship between these genetic variations with chronic periodontitis in a Brazilian population. MATERIAL AND METHODS: Non-smoking subjects (n = 114) exhibiting sites > or = 5 mm clinical attachment loss were recruited for study. Control subjects (n = 109) should not exhibit clinical signals of periodontitis. MMP-1 (-1607 1G/2G, -519 A/G) and MMP-3 (-1612 5A/6A) gene promoter polymorphisms were genotyped using PCR-RFLP methods. RESULTS: Analysis of polymorphisms showed no differences in distribution of the -1607 1G/2G and -519 A/G variants in the MMP-1 gene between the healthy and periodontitis group (p > 0.05). However, the distribution of genotype frequencies of the -1612 5A/6A polymorphism in the MMP-3 gene showed that the 5A/5A genotype was significantly more frequent in the periodontitis group (p = 0.008). The same was not observed in the 5A/6A genotype once only one 5A allele is carried. We also observed a trend to increase the frequency of the MMP-1/MMP-3 haplotype (2G/5A) in the periodontitis group (p = 0.08). CONCLUSION: On the basis of the results, no significant association is found for the MMP-1 polymorphisms with susceptibility of periodontitis, while the MMP-3 gene polymorphism may contribute to periodontal tissue destruction during periodontitis in Brazilian subjects.

Gingival crevicular fluid alkaline phosphatase levels in postmenopausal women: effects of phase I periodontal treatment.

BACKGROUND: The aim of this study was to determine how estrogen status may possibly influence gingival crevicular fluid (GCF) alkaline phosphatase (ALP) levels in estrogen-deficient (ED) and -sufficient (ES) postmenopausal women at baseline (BL) and 1 year after periodontal phase I treatment (AT). METHODS: Thirty-six postmenopausal women on estrogen supplements (mean serum estradiol levels >30 pg/ml; estrogen sufficient) and 37 postmenopausal women not on estrogen supplements (mean serum estradiol levels <30 pg/ml; ED) were divided into two subgroups as chronic periodontitis and clinically healthy controls after clinical and radiographic examination. The ES group consisted of 19 control (ES/C) and 17 chronic periodontitis (ES/P) patients, and the ED group consisted of 20 control (ED/C) and 17 chronic periodontitis (ED/P) patients. Plaque (PI) and gingival (GI) indices, bleeding on probing (BOP), probing depths (PD), clinical attachment loss (CAL) scores, and GCF samples were recorded at BL and AT. The levels of ALP in the GCF were measured photometrically. The paired samples Student t and Wilcoxon tests were used to compare the ALP levels and clinical parameters between BL and AT. The correlation among the ALP and clinical parameters was analyzed using the Pearson correlation. RESULTS: The mean of all clinical parameters (PI, GI, BOP, PD, and CAL) was significantly (P <0.05) higher in periodontitis groups (ES/P and ED/P) than controls (ES/C and ED/C). Periodontitis groups (ES/P and ED/P) demonstrated significantly increased GCF volumes and GCF ALP levels (P <0.05) compared to controls (ES/C and ED/C). There were no significant differences in the concentrations of ALP between periodontitis and control groups (P >0.05). The BL GCF ALP total levels of the ED/P group were significantly higher than the ES/P group (P <0.05). The BL and AT serum ALP levels of the ED/P group were not significantly but were numerically higher than the ES/P group. One year after periodontal treatment, the GCF volume, GCF ALP total, and concentrations decreased significantly in both periodontitis groups (P <0.05). However, the GCF ALP levels were still numerically higher in the ED/P group. A positive statistical correlation was found between total ALP levels and PD (r = 0.621; P <0.05). CONCLUSION: These data suggest that the presence of ALP in GCF is not simply a reflection of the local inflammation state and that a patient's estrogen status may possibly influence local ALP levels in GCF.

Analysis of superoxide dismutase activity levels in gingiva and gingival crevicular fluid in patients with chronic periodontitis and periodontally healthy controls.

OBJECTIVES: Superoxide dismutase (SOD) is an antioxidant enzyme that acts against superoxide, an oxygen radical, released in inflammatory pathways and causes connective tissue breakdown. In this study, SOD activities in gingiva and gingival crevicular fluid (GCF) from patients with chronic periodontitis (CP) and periodontally healthy controls were compared. MATERIAL AND METHODS: Twenty-six CP patients and 18 controls were studied. In patients, teeth with moderate-to-severe periodontal breakdown and > or =5 mm pockets that required full-thickness flap surgery in the right or left maxillary quadrant, and in controls, teeth scheduled for extraction for orthodontic reasons were studied. After the clinical measurements (probing depth, clinical attachment level, gingival index, gingival bleeding index, plaque index), GCF samples were collected. Tissue samples were harvested from the same teeth, during flap operation in patients and immediately after tooth extraction in controls. SOD activities were spectrophotometrically assayed. The results were statistically analysed. RESULTS: Gingival SOD activity was significantly higher in the CP group than in controls (p<0.05). No significant difference was found in GCF SOD activity between the groups (p>0.05). Correlations between gingival and GCF SOD activities were not statistically significant in CP and control groups (p>0.05). CONCLUSION: In CP, SOD activity seems to increase in gingiva, probably as a result of a higher need for SOD activity and protection in gingiva in CP than in periodontal health, while not significantly changing in GCF, suggesting a weak SOD activity in GCF in periodontal disease state. The weak correlation between gingival and GCF SOD activities suggests distinct actions of these SODs.

Beta-glucuronidase activity in saliva: relationship to clinical periodontal parameters.

BACKGROUND: Beta-glucuronidase (betaG) in gingival crevicular fluid (GCF), a marker of neutrophil influx into the crevicular environment, has previously been shown to be correlated with periodontal clinical parameters at individual sites (probing depth and clinical attachment level). Furthermore, elevated levels of betaG were found to be a risk factor for periodontal attachment loss. Analysis of betaG in saliva may be a measure of crevicular neutrophil influx for the whole mouth. The purpose of this cross-sectional study was to evaluate the relationship between betaG activity in saliva and periodontal clinical parameters in subjects demonstrating various levels of periodontal disease. METHODS: The study population consisted of 380 subjects (108 males and 272 females). A sample of unstimulated whole saliva and a venous blood sample were obtained from each subject, and a periodontal examination, which included measurement of probing depth (PD), attachment level (AL), and gingival index (GI) was performed. The unmodified saliva samples were frozen at -20 degrees C and analyzed for betaG activity. The blood samples were analyzed for number of white blood cells, neutrophils, monocytes, lymphocytes, and platelets. Statistical analysis was conducted to determine the association between salivary betaG activity and the whole-mouth clinical periodontal parameters, complete blood count, smoking status, and age. RESULTS: Highly significant correlations between salivary betaG activity and mean probing depth (MPD), mean gingival index (MGI), and the number of sites with probing depth > or = 5 mm were found. When subjects were divided into tertiles based on their MPD and MGI, elevated salivary betaG activity was detected in subjects in the 2 upper tertiles. Logistic regression modeling was used to determine which of the clinical or laboratory parameters were able to identify patients with at least 4 sites with PD > or = 5 mm. Salivary betaG activity > or = 100 produced an odds ratio (OR) of 3.77. In comparison, current and former smokers had an OR of 3.15 and 2.29, respectively. CONLCUSIONS: The results suggest that a significant association exists between periodontal clinical parameters and salivary betaG activity. Due to the non-invasive and simple nature of saliva collection, this association should be studied to determine its usefulness as a screening test for periodontitis, and a means of monitoring the response to treatment.

Nitric oxide synthesis and severity of human periodontal disease.

The expression of the inducible nitric oxide synthase enzyme (iNOS) is a response to an inflammatory stimulus and produces a large amount of nitric oxide (NO), which may act as a cytotoxic molecule against the invading microorganism and may be related to both harmful and beneficial effects to tissues. OBJECTIVE AND MATERIAL AND METHODS: In order to further characterize the presence of NO in human periodontal disease, we undertook a quantitative study of iNOS positive cells in samples of clinically healthy gingival tissues, plaque-induced gingivitis and localized chronic periodontitis using immunohistochemistry. RESULTS: A significant increase in the number of iNOS+ cells mm-2 was found in the samples of the gingivitis and periodontitis compared with those of the control. In all groups most of the polymorphonuclear cells showed intense immunoreactivity for iNOS independent of the disease stage, and the percentage of iNOS+ polymorphonuclear cells increased significantly in periodontal disease when compared with the control. CONCLUSION: Our results indicate that iNOS increases in the presence of periodontal disease. In addition, our findings suggest that polymorphonuclear cells present an additional activation pathway in periodontal disease, expressing significant iNOS and probably representing an important source of NO in human periodontal disease that has not been previously reported.