Enzymes In Gingival Crevicular Fluid Research Articles

The mRNA expression of genes encoding selected antioxidant enzymes and thioredoxin, and the concentrations of their protein products in gingival crevicular fluid and saliva during periodontitis.

The activity of antioxidant enzymes in periodontitis is reduced, but results vary between studies and are subject to bias. In turn, the expression of genes encoding antioxidant factors has not been examined yet. This is the first study to evaluate the expression of genes encoding superoxide dismutase 1 (SOD1), glutathione peroxidase 1 (GPX1) and thioredoxin 1 (TXN1) in the saliva and gingival tissue of patients with periodontitis. The activity of the antioxidant enzyme protein products in the unstimulated and stimulated saliva and the gingival crevicular fluid (GCF) of patients with periodontitis was also investigated. The prospective study involved 65 patients with periodontitis, who were divided into groups depending on the disease stage, and a control group of 31 ageand gender-matched healthy patients. We demonstrated that the expression of genes encoding GPX1 and TXN1 in saliva was significantly higher, and the expression of genes encoding SOD1, GPX1 and TXN1 in the gingival tissue was significantly lower in periodontitis patients as compared to the control group. We noted a lower activity of GPX1 in unstimulated saliva, of SOD1 in stimulated saliva and of both antioxidant enzymes in GCF in patients with periodontitis. The GPX1 transcriptome and its activity in the salivary and GCF proteome appear to be dependent on the oxidative stress related to the destructive inflammatory changes in periodontitis.

Evaluation of alkaline phosphatase in gingival crevicular fluid and saliva of patients with periodontitis and healthy individuals.

In recent decades, biomarkers have been used to predict the progression of chronic periodontitis. One of these biomarkers is alkaline phosphatase (ALP). Due to limitations of the performed studies, this study was performed to determine the amount of salivary ALP and gingival crevicular fluid in patients with chronic periodontitis and healthy individuals. Twenty-three patients with severe chronic periodontitis and 23 healthy individuals referred to the Periodontology Department of Ahvaz Jundishapur School of Dentistry were evaluated in this analytical epidemiological study. Salivary ALP and gingival crevicular fluid (GCF) were measured using ALP measuring kit and Hitachi device. Mean (standard deviation) of ALP enzyme was 19.43 (12.5) in GCF of patients with chronic periodontitis and 12 (1.48) in the healthy group, and it was 80.17 (23.9) in the saliva of patients with periodontitis and 24.78 (4.37) units per litre in the healthy group. There was a significant difference in the mean of this enzyme in GCF and saliva of patients with chronic periodontitis and healthy individuals (P < 0.001). The results showed that mean of ALP enzyme is significantly higher in GCV and saliva of patients with chronic periodontitis than in healthy individuals. Therefore, it seems that this parameter can be used as a useful biochemical parameter for the diagnosis of periodontal disease.

Effect of orthodontic forces on levels of enzymes in gingival crevicular fluid (GCF): A systematic review.

Objective: Orthodontic force application releases multiple enzymes in gingival crevicular fluid (GCF) for activation, resorption, reversal, deposition of osseous elements and extracellular matrix degradation. The current systematic review critically evaluated all existing evidence on enzymes in orthodontic tooth movement. Methods: Literature was searched with predetermined search strategy on electronic databases (PubMed, Scopus, Embase), along with hand search. Results: Initial search identified 652 studies, shortlisted to 52 studies based on PRISMA. Quality assessment further led to final inclusion of 48 studies (13 moderately and 35 highly sensitive studies). Primary outcomes are significant upregulation in GCF levels of enzymes-aspartate aminotransferase (AST), alkaline phosphatase (ALP), matrix metalloproteinases (MMPs), lactate dehydrogenase (LDH), β-glucuronidase (βG), tartrate resistant acid phosphatase (TRAP), acid phosphatase (ACP) and down regulation in cathepsin B (Cb). Site specificity is shown by ALP, TRAP, AST, LDH, MMP9 with levels at compression site increasing earlier and in higher quantities compared with tension site. ALP levels are higher at tension site only in retention. A positive correlation of LDH, ALP and AST is also observed with increasing orthodontic force magnitude. Conclusions: A strong evidence of variation in enzymes (ALP, AST, ACP TRAP, LDH, MMPs, Cb) in GCF is found in association with different magnitude, stages and sites of orthodontic force application.

A pilot study of active enzyme levels in gingival crevicular fluid of patients with chronic periodontal disease.

To determine whether combinations of enzymes in gingival crevicular fluid (GCF) can act as improved biomarkers compared with single enzymes for predicting the outcome of treatment and also for diagnosing the clinical status of sites. Thirty subjects with chronic periodontitis were recruited to a 12-month longitudinal pilot study. GCF samples from three representative sites: healthy (≤3mm), deep non-bleeding (NB) (≥6mm) and deep bleeding (DB) (≥6mm) sites and clinical data were collected at baseline, 3months, 6months and 12months following periodontal treatment. Active enzyme levels (MMP-8, cathepsin G, elastase, trypsin-like activity and sialidase) in GCF samples were assessed. The enzyme profiles and clinical data of each site were analysed for correlation and logistic regression was performed to find the predictive value of the active enzyme levels regarding the outcome of treatment. Twenty-two individuals completed the study. All active enzyme levels were significantly higher in diseased sites than healthy sites. Logistic regression showed that the combination of MMP8, elastase and sialidase provided accurate predictions of treatment outcome (88% for NB and 86% for DB), which was significantly better than each enzyme alone (61%). This pilot has suggested that combined active enzyme profiling could provide significant prediction of outcome of treatment.

Periodontal Diagnostik Biyobelirteç Olarak Diş Eti Oluğu Sıvısındaki Konak Kaynaklı Enzimler-Bölüm 1

Bu çalışma, diş eti oluğu sıvısındaki (DOS) periodontal hastalık tanısının konulmasına yardımcı olan konak kaynaklı enzimlere ve onların inhibitörlerine odaklanılan çalışmanın ilk kısmıdır. İnflamatuar bir durum olan periodontit, alveolar kemik rezorpsiyonu, periodontal ataşman kaybı ve bağ dokusunun yıkımı ile karakterizedir. Periodontal cep derinliği, ataşman seviyesi, sondalamada kanama gibi olağan periodontal ölçümler periodontal hastalıkların teşhisi için gereklidir. Ancak hastalığın şiddetinin, prognozunun ve tedaviye verilen yanıtın değerlendirilmesinde yetersiz kalabilirler. DOS, periodontal hastalık ilerlemesi için olası biyolojik belirteç olarak belirtilmiş 65'den fazla bileşenden oluşmaktadır. Bu tür bileşenlerin varlığının incelenmesi, periodontal hastalığın derecesinin ya da periodontal tedavinin sonuçlarının değerlendirilmesinde faydalı olabilir. Bu çalışma, 'Pubmed' veri tabanında periodontal hastalık, diş eti oluğu sıvısı ve enzimler anahtar kelimeleriyle araştırılan, 1970-2015 yılları arasında yayınlanmış makaleler incelenerek hazırlanmıştır. İlk bölümde; aspartat aminotransferazı, alkalen fosfatazı, asit fosfatazı, beta glukuronidazı, nötrofil elastazı, elastaz inhibitörlerini, katepsinleri, tripsin-benzeri enzimleri içeren konak kaynaklı enzimlere ve onların inhibitörlerine odaklanılmıştır. İkinci bölümde de, immünoglobulin parçalayıcı proteazı, glukozidazı, dipeptidil peptidazı, spesifik olmayan nötral proteinazları, metalloproteinazları, metalloproteinaz doku inhibitörünü, miyeloperoksidazı, laktat dehidrogenazı, arilsülfatazı, ß-N-asetil hekzoaminidazı içeren konak kaynaklı enzimlere ve onların inhibitörlerine odaklanılmıştır.

Periodontal Diagnostik Biyobelirteç Olarak Diş Eti Oluğu Sıvısındaki Konak Kaynaklı Enzimler-Bölüm 2

Bu çalışma, diş eti oluğu sıvısındaki (DOS) periodontal hastalık tanısının konulmasına yardımcı olan konak kaynaklı enzimlere ve onların inhibitörlerine odaklanılan çalışmanın ikinci kısmıdır. Bu çalışma

Comparative analysis of the potential effect of phase I therapy on gingival crevicular fluid myeloperoxidase levels in non-diabetic and diabetic patients with periodontitis

Objectives: To evaluate the effect of periodontal therapy on clinical parameters as well as on myeloperoxidase (MPO) activity in the gingival crevicular fluid (GCF) of patients with type 2 diabetes mellitus and of systemically healthy individuals. Materials and Methods: 24 type 2 DM patients subjects, and 21 systemically healthy individuals, both groups with chronic periodontitis, and systemically and periodontally healthy individuals were enrolled. Periodontal clinical parameters, namely periodontal probing depth (PD), plaque index (PI), gingival index (GI), clinical attachment level (CAL), and gingival bleeding time index (GBI), as well as GCF MPO activity, were assessed before and 3 months after non-surgical periodontal therapy. GCF enzyme activity was spectrophotometrically analyzed. Possible correlation between clinical periodontal status and MPO activity was also evaluated. Results: Despite the relatively stable clinical measures and GCF MPO content in the periodontally-healthy subjects, the clinical periodontal status improved, as significant reductions were observed in all of the clinical parameters in periodontitis patients with and without diabetes (p<0.05). GCF MPO activity presented with significant reductions in both of the periodontally-diseased groups after phase I treatment (p<0.05), however it could not reach to the level of periodontal health (p<0.05). Conclusions: Periodontal inflammation is likely to play the major role in the higher MPO activity observed at diseased sites, while diabetes mellitus do not seem to further increase the production of this enzyme. Periodontal treatment can provide successful improvement in clinical periodontal parameters and reduction in enzyme profile of GCF in diabetics.

Erratum to: Evaluation of bleeding on probing and gingival crevicular fluid enzyme activity for detection of periodontally active sites during supportive periodontal therapy

Erratum to: Evaluation of bleeding on probing and gingival crevicular fluid enzyme activity for detection of periodontally active sites during supportive periodontal therapy

Evaluation of bleeding on probing and gingival crevicular fluid enzyme activity for detection of periodontally active sites during supportive periodontal therapy

This study aimed to analyze the enzyme activity in gingival crevicular fluid (GCF) and its association with clinical parameters, especially bleeding on probing (BOP), and thus reconsider the significance and accuracy of recording BOP. A total of 184 patients who had entered supportive periodontal therapy were selected and GCF was collected from 401 sites before recording the clinical parameters, probing pocket depth (PPD), BOP, clinical attachment level, gingival index and plaque index. The enzyme activity of neutrophil elastase and aspartate aminotransferase and amount of protein in GCF were also analyzed. In the clinical parameters for biochemical data, amount of GCF showed the most correlation. A cut-off value for BOP and PPD were determined by the ROC curve and Youden index. Analysis was performed with all clinical parameters and biochemical data. Of the 401 sites, 51 were less than the cut-off value and were BOP-negative. On the other hand, 29 sites had values more than the cut-off value, with 14 BOP-negative sites and 15 BOP-positive sites. A conclusion is as follows: twenty-nine sites with values more than the cut-off value were diagnosed as sites requiring periodontal management, however, 14 of these were BOP-negative. These results suggest that combining other biochemical tests with examination of BOP and PPD may improve the validity of periodontal disease diagnosis. In future studies, it will be essential to find a marker that can precisely detect periodontal disease activity, and to develop a diagnostic tool for chair-side use.

Intracytoplasmic enzymes in gingival crevicular fluid of patients with aggressive periodontitis

Biochemical parameters of crevicular fluid could provide evidence of periodontal tissue disease. The aim of this study was to analyze enzymes in crevicular fluid in aggressive localized and generalized periodontitis. One hundred and twenty-four subjects were classified as having localized (n = 36) or generalized aggressive periodontitis (n = 38) and subclassified into moderate and severe groups. Controls were 50 periodontitis-free subjects. Activities of the enzymes lactate dehydrogenase, neutrophil elastase, alkaline phosphatase and aspartate aminotransferase were determined. Data were analyzed using one-way ANOVA and Tukey's test. Among the subjects with localized aggressive periodontitis, values of lactate dehydrogenase and alkaline phosphatase increased notably in moderate and severe periodontitis compared with control subjects. Values for aspartate aminotransferase increased with the severity of the disease, and neutrophil elastase was increased in the moderate and severe states. In generalized aggressive periodontitis, lactate dehydrogenase showed higher values than in control subjects in both periodontal subgroups. Alkaline phosphatase and neutrophil elastase showed higher significant differences between moderate and severe periodontitis compared with the control group. Aspartate aminotransferase showed differences between the severe and moderate periodontitis groups compared with the control group. Of all the enzymes analyzed, only lactate dehydrogenase showed higher values in localized than in generalized aggressive periodontitis. Lactate dehydrogenase may distinguish localized and generalized aggressive periodontitis. Alkaline phosphatase increases from moderate to severe states in both types of periodontitis. Aspartate aminotransferase and neutrophil elastase only increase with strong evidence of periodontal destruction.

Periodontal Status and Neutrophilic Enzyme Levels in Gingival Crevicular Fluid During Pregnancy and Postpartum

Pregnancy induces or enhances susceptibility to gingivitis; however, the presence and role of neutrophilic enzymes in pregnancy-related gingivitis are not well known. The present study demonstrates the relationship between neutrophilic enzymes in gingival crevicular fluid (GCF) and periodontal status during pregnancy and postpartum. At baseline, 30 periodontally healthy pregnant women (Pr group) and 24 non-pregnant women (N-Pr group) as their controls participated in the study. The Pr group was examined once per each trimester and twice during postpartum and the N-Pr group three times (on successive months). During each visit, GCF samples were collected from all first molars, and clinical measurements (visible plaque index, bleeding on probing [BOP], probing depth [PD], and clinical attachment level) were recorded. The samples were analyzed for matrix metalloproteinase (MMP)-8, polymorphonuclear neutrophil (PMN) elastase, myeloperoxidase (MPO), and tissue inhibitor of matrix metalloproteinase (TIMP)-1. Their levels were compared to the periodontal status at the collection site. In the Pr group, BOP and PD scores significantly increased between the first and second trimester, indicating pregnancy gingivitis. This increased inflammation was not reflected by the enzymes examined in GCF; the amounts of PMN elastase decreased continuously during the follow-up period, and those of MPO and MMP-8 did not increase until delivery, whereas TIMP-1 amounts remained stable throughout the follow-up period. In the N-Pr group, all parameters remained steady. Despite an increased susceptibility to gingivitis during mid-pregnancy, the host response does not seem to activate its own degradative enzymes.

Effect of Systemically Administered Naproxen Sodium on Clinical Parameters and Myeloperoxidase and Elastase‐Like Activity Levels in Gingival Crevicular Fluid

The present study was conducted to determine the possible effect of naproxen sodium on clinical status and the enzymatic profile of gingival crevicular fluid (GCF) when given as adjunct to periodontal treatment. A total of 34 subjects with chronic periodontitis were selected and divided into two groups to receive either naproxen sodium or placebo. At baseline, GCF samples were obtained and probing depths (PD), gingival index (GI), plaque index (PI), and gingival bleeding index (GBI) scores were recorded. In the non-steroidal anti-inflammatory drug (NSAID) group, patients were treated with a protocol consisting of baseline periodontal treatment (scaling, root planing) and naproxen sodium (275 mg) administration daily for 6 weeks. In the placebo group, patients received the same treatment except placebo was given instead of naproxen sodium. At the end of the experimental period, clinical recordings and GCF sampling were repeated. Myeloperoxidase (MPO) and elastase-like enzyme activity (ELA) levels were determined in GCF samples by a spectrophotometric method. GCF enzymatic content was calculated both as total enzyme activity and enzyme concentration. All of the clinical parameters, except mean GBI, were significantly lower in the experimental group (P <0.05). At baseline and at the end of the experimental period, there were no significant differences between the NSAID and placebo groups regarding GCF MPO and ELA levels in either mode of data presentation (P <0.05). However, in the NSAID group, mean ELA concentration (P = 0.002) and mean total ELA (P = 0.003) presented significant decreases with treatment. Also, with treatment, a general reduction in MPO levels was seen; however, this difference was not significant. Although constant and stable correlations between GCF enzyme levels and clinical parameters could not be found, positive and strong correlations were observed between total enzyme activity and enzyme concentrations. Based on the positive clinical effect and the ELA profile of GCF, it can be suggested that NSAIDs given as an adjunct to baseline periodontal treatment could be beneficial in the outcome of treatment.

Association of gingival crevicular fluid and serum intracytoplasmic enzyme levels in periodontally healthy homozygous (major) beta-thalassemia patients.

To assess tissue necrosis in beta-thalassemia major patients, as in other areas of medicine, lactate dehydrogenase (LDH), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) have been widely used. These markers of tissue degradation have also been studied in the gingival crevicular fluid (GCF) in relation to periodontal disease status. The purpose of this study was to investigate whether periodontal diagnostic tests based on these markers of tissue degradation are influenced from the enzymes' levels in serum and, therefore, could be used in the assessment of the patient's periodontal status. Forty-four periodontally healthy, homozygous beta-thalassemia patients were enrolled in the study. GCF and serum samples were obtained and the levels of AST, ALT and LDH were determined; the measurements took place in an automated analyzer (Hitachi 777) using the kits of Roche Company. Lack of correlations between serum and GCF enzyme levels was demonstrated. Serum LDH and serum AST, serum AST and serum ALT are significantly positively correlated. Concerning the GCF, AST and ALT were proved to be significantly positively correlated. Elevated values of LDH, ALT and AST in serum do not constitute a confounding factor in GCF measurements of the respective enzymes.

Crevicular fluid levels of TGFβ1 in drug-induced gingival overgrowth

Objective: To determine if local, gingival crevicular fluid (GCF) levels of TGFβ1 were altered in drug-induced gingival overgrowth. Patients and methods: GCF samples were collected on Periopaper™ strips from 45 renal transplant recipients who had been medicated with cyclosporin or cyclosporin in combination with other putative overgrowth-inducing drugs for a minimum of 6 months. Twenty-two subjects had gingival overgrowth while the other 23 patients showed no signs of gingival changes and constituted the medicated control group. Non-medicated controls consisted 20 periodontally healthy individuals who had never taken overgrowth-inducing drugs. GCF levels of TGFβ1 and alkaline phosphatase, a marker of inflammation, were determined by enhanced chemiluminescence ELISA and enzyme activity assays, respectively. Results: TGFβ1 levels in GCF from overgrowth and non-overgrowth sites in overgrowth sufferers did not differ. However, there were significant differences in median concentration ( P=0.001) and GCF levels of TGFβ1 per sample ( P=0.05) between study groups with overgrowth patients having higher amounts per sample and lower concentrations than medicated and healthy controls. Median levels of alkaline phosphatase per GCF sample differed between site ( P=0.01) with higher levels present at overgrowth sites. Despite this, the concentration of enzyme in GCF did not differ between site or patient group. Conclusions: GCF TGFβ1 detected in overgrowth patients could reflect a higher level of gingival inflammation because of difficulties in plaque control consequent on the development of overgrowth. However, the higher local levels of total TGFβ1 in overgrowth patients could indicate that it is a risk factor for developing gingival overgrowth.

PMN responses in chronic periodontal disease: evaluation by gingival crevicular fluid enzymes and elastase-alpha-1-proteinase inhibitor complex.

In the present trial, the hypothesis was examined that the local PMN responses in untreated and treated chronic periodontitis can be differentiated by gingival crevicular fluid lysosomal enzyme activities and elastase-alpha-1-proteinase inhibitor complex. In nine subjects (average age 49.2 +/- 7.1 years) with chronic periodontitis, clinical parameters and markers of the PMN-derived inflammatory tissue response in gingival crevicular fluid (GCF) were assessed before and 6 months after surgical periodontal therapy. Myeloperoxidase (MPO), beta-N-acetyl-hexosaminidase (beta-NAH) and cathepsin D (CD) were analyzed as indicators of the PMN-associated host tissue destruction, and elastase-alpha-1-proteinase inhibitor complex (alpha-1-EPI) as the major serum protein inactivating PMN elastase. The total activities of the lysosomal enzymes MPO and beta-NAH were evaluated spectrophotometrically, the CD levels by liquid scintillation counting with [14C] hemoglobin as substrate, and the total alpha-1-proteinase inhibitor complex using a sandwich-immunoassay. The clinical parameters revealed a statistical significant decrease at the 6-month reexamination. PD levels dropped from 5.40 to 2.88 mm (change 2.52 +/- 1.04 mm), the CAL scores from 6.67 to 4.43 mm (change 2.24 +/- 0.77 mm). The 30 s GCF volumes dropped from 129.8 to 68.6, displaying a change of 61.1 +/- 18.6, p </= 0.05. The decrease in total MPO, beta-NAH and CD levels (medians: 1.7/0.6 micro U MPO, 0.035/0.020 micro U beta-NAH, 1.3/0.5 ng CD) following therapy was associated with a significant drop in total GCF amounts of alpha-1-EPI from 76.3 ng at baseline to 52.4 ng after 6 months. The clinical healing in chronic periodontal disease is associated with a downregulation of the local PMN responses following periodontal therapy. The reorganization of periodontal tissues is characterized by a decrease of lysosomal enzyme activities and the alpha-1-proteinase inhibitor complex in gingival crevicular fluid.

The importance of data presentation regarding gingival crevicular fluid myeloperoxidase and elastase-like activity in periodontal disease and health status.

The enzymatic profile of gingival crevicular fluid (GCF) is being analyzed with increasing interest, but related studies lack a general consensus on most methodological points, including the appropriate mode of data presentation. GCF myeloperoxidase (MPO) and elastase-like activity (ELA) levels were spectrophotometrically determined on a total of 60 subjects who were divided into three equal subgroups as early-onset periodontitis (EOP), adult periodontitis (AP), and healthy. GCF enzyme levels were calculated and evaluated both as total enzyme activity and enzyme concentration. The correlations between these GCF enzymes and clinical periodontal status were also analyzed. With both modes of data presentation, the results regarding MPO activity were consistent. When presented either as total MPO activity or MPO concentration, the periodontally healthy group showed significantly lower MPO activity than the two patient groups (P<0.05). However, two modes of data presentation did not match when GCF ELA was concerned. When data were reported as total ELA, the healthy group exhibited lower enzyme activity (0.02 +/- 0.001 U) than EOP (0.04 +/- 0.01 U) and AP (0.06 +/- 0.02 U) groups; but when reported as concentration, the highest ELA levels were seen in the healthy group (221 +/- 31.53 nmol/min/ml), followed by AP (98.63 +/- 23.03 nmol/min/ml) and EOP (70.49 +/- 12.02 nmol/min/ml) (P<0.05). A strong-positive and significant correlation existed between GCF MPO and ELA. Correlations with clinical parameters were mostly observed with total activities. The findings of the present study confirm the relationship between GCF ELA and MPO activity and periodontal disease and also support the functional relativity between the two enzymes. Furthermore, based on these findings, it can be suggested that data presentation by use of total activity seems to be more sensitive in both the reflection of the actual enzymatic profile of GCF and also the existing clinical periodontal status. For each GCF component, the validity of different modes of data presentation should be considered.

Assessment of a novel screening test for neutrophil collagenase activity in the diagnosis of periodontal diseases.

Increased levels of active neutrophil collagenase (MMP-8) in the gingival crevicular fluid (GCF) are associated with progressive periodontitis. The measurement of this enzyme in GCF could facilitate diagnosis. However, assays with sufficient sensitivity to detect collagenase in whole-mouth GCF currently use radiolabeled substrates and require several days to complete. To provide more rapid analyses of collagenase activity that are better adapted to clinical studies, we developed and validated a novel assay (soluble biotinylated-collagen assay: SBA) based on chemiluminescent detection of biotinylated collagen digestion products. The concordance of the novel SBA assay with a radioactive collagen substrate assay was assessed by parallel analyses of enzyme from 35 neutrophil preparations and from 41 samples of GCF from periodontitis patients, followed by Pearson correlation analysis. To test whether the assay appropriately measured MMP-8 activity, enzyme activity was assessed after incubation with specific collagenase blockers. We examined the diagnostic utility of the SBA in cross-sectional and longitudinal analyses of 125 patients with adult periodontitis, 5 patients with early-onset periodontitis, 1 edentulous patient, and in 32 control patients without periodontitis. The assay detected <56 pg collagen degraded/hour/microl sample, which is comparable to the most sensitive radioactive assay. The total assay time was 22 hours and reproducibility on replicate measurements was high (r = 0.96). In direct comparisons of MMP-8 activity in GCF with enzyme from peripheral blood neutrophils using the SBA and radioactive assays, there was a high correlation (r = 0.97). As expected, EDTA and TIMP-1 and -2, known inhibitors of MMP-8, completely blocked enzyme activity with this assay. Cross-sectional and longitudinal analyses of GCF showed that MMP-8 activity was >18-fold higher in severe periodontitis than in stable periodontitis and decreased to <25% of pretreatment levels following therapy. Based on measurements of collagenase activity in different disease groups, we estimated a value of 80 nano units as a threshold for severe periodontitis. These results indicate that active MMP-8 is detected in GCF by a novel assay that is specific, simple, rapid, and reproducible and which may facilitate diagnostic discrimination between stable and progressive lesions.

Subgingival temperature: relation to gingival crevicular fluid enzymes, cytokines, and subgingival plaque micro-organisms.

There have been no reports on the relationship of subgingival temperature to specific gingival crevicular fluid (GCF) components. Therefore, the purpose of this cross-sectional study was to determine whether there was any relationship between subgingival temperature and GCF levels of neutrophil elastase (NE), myeloperoxidase (MPO), beta-glucuronidase (BG), interleukin-1 alpha (IL-1), and interferon alpha (IFN). Furthermore, another objective was to confirm an association of subgingival temperature with clinical parameters and specific subgingival plaque micro-organisms as has been reported earlier. 27 human subjects each having healthy (n = 50), gingivitis (n = 59) and periodontitis (n = 53) sites were evaluated. The plaque index (PI), subgingival temperature, probing depth, attachment loss, bleeding index and gingival index were measured. GCF was sampled following the measurement of the PI and removal of the supragingival plaque. GCF samples were assayed for the enzymes NE, BG, MPO and the cytokines IFN-alpha and IL-1 alpha. A sterile Gracey curette was utilized at each sampled site to collect subgingival plaque. The plaque samples were evaluated using an immunoassay. Subgingival temperature was found to directly correlate with all clinical parameters (p < 0.001). Significant, albeit not large, correlations were found between subgingival temperature and NE (r = 0.35, p < 0.001), MPO (r = 0.26, p < 0.001) and BG (r = 0.23, p < 0.01). Temperature was found to correlate positively with E. corrodens (r = 0.33, p < 0.02) and F. nucleatum (r = 0.25, p < 0.05) but not with P. intermedia (r = 0.02, p = 0.9), P. gingivalis (r = 0.20, p = 0.1) and A. actinomycetemcomitans (r = 0.01, p > 0.9). In conclusion, subgingival temperature is correlated with the GCF enzymes, NE, MPO and BG as well as the clinical parameters and specific plaque micro-organisms associated with periodontal disease.

The effects of low dose doxycycline regimen on gingival crevicular fluid enzyme actmty of diabetic patients with periodontitis and adult periodontitis patients

The effects of low dose doxycycline regimen on gingival crevicular fluid enzyme actmty of diabetic patients with periodontitis and adult periodontitis patients

Risk indicators for periodontal disease in a racially diverse urban population.

A cross-sectional study of 117 subjects from a dental clinic serving a diverse population (i.e., Whites, African-Americans, Native-Americans, and Asians) was performed to evaluate risk indicators of periodontal disease. Gingival crevicular fluid (GCF) and subgingival plaque were taken at the same visit from 4 posterior sites of the most diseased sextant in each subject. Age, smoking packyears, beta-glucuronidase (beta G), neutrophil elastase (NE), myeloperoxidase (MPO), Fusobacterium nucleatum (F. nucleatum), and Porphyromonas gingivalis (P. gingivalis) were significantly (p < 0.05-0.005) correlated with attachment loss. Probing depth was significantly correlated with smoking packyears, beta G, NE, MPO, F. nucleatum and Prevotella intermedia (P. intermedia) (p < 0.05-0.005). Mean NE value of Whites was lower than the mean NE values of African-Americans, Native-Americans and Asians (p < 0.05). Whites had a lower mean beta G value compared to African Americans, and a lower mean MPO value compared to African Americans and Native Americans. The %s of patients positive for F. nucleatum, P. intermedia and Eikenella corrodens (E. corrodens) were higher in Native Americans compared to Whites. Step-wise multiple regression analysis was performed to construct models for the estimation of probing depth and attachment loss. The most parsimonious regression models which had the best R2 values included the following variables and accounted for the indicated % of variability: models 1 and 2: beta G, race, and F. nucleatum accounted for 50% of the variability in mean probing depth and 39% of the variability in a single site (first molar) for probing depth, respectively; model 3: age, beta G, and F. nucleatum accounted for 53% of the variability in mean attachment loss; model 4: age, NE, and F. nucleatum explained 35% of the variability in a single site (first molar) for attachment loss. The results suggest that age, race, smoking packyears, beta G, NE, MPO, F. nucleatum, P. gingivalis and P. intermedia are risk indicators for periodontal disease in this racially diverse urban population. Regression models which include multiple variables (i.e., demographic factors, GCF enzymes and periodontopathic bacteria) can be used to estimate periodontal disease status.