Gingival crevicular fluid glycosaminoglycan levels in patients with chronic adult periodontitis

Abstract This study investigated levels of hyaluronan and chondroitin-4-sulphate in the crevicular fluid of patients with chronic adult periodontitis at diseased and healthy sites before and after treatment. The relationship between clinical diagnostic parameters and levels of glycosaminoglycans in gingival crevicular fluid were also analysed. Within each patient. 4 sites either mesial or distal and on single rooted teeth were classified as diseased or healthy using a modified gingival index, pocket depth and attachment loss. Crevicular fluid was collected from each site using glass micropipettes and analysed for glycosaminoglycan content by cellulose acetate electrophoresis. Significantly higher levels of chondroitin-4-sulphate were detected at diseased sites prior to treatment correlating with increased pocket depth or attachment levels. Following a period of treatment consisting of oral hygiene instruction and root planing, the patients were reassessed for their response to treatment by measuring the modified gingival index, pocket depth, attachment loss and levels of glycosaminoglycans. Analysis of glycosaminoglycan levels at diseased sites that demonstrated a poor response to treatment also demonstrated significantly higher levels of chondroitin-4-sulphate than those sites that responded well to treatment. Hyaluronan levels were less significantly associated with clinically succesful treatment. This study confirmed the use of the sulphated glycosaminoglycan chondroitin-4–sulphate as a potential diagnostic aid of periodontal tissue destruction; however, further longitudinal studies are required to assess their performance.     

Gingival crevicular fluid osteocalcin in adult periodontitis

This study aimed to detect the levels of osteocalcin in gingival crevicular fluid (GCF) from healthy (< or =3 mm sulcus depth and non-bleeding) and diseased sites (> or =6 mm probing depth and bleeding) in subjects with adult periodontitis, in order to further investigate its potential as a possible marker of the disease process. Periodontal probing depths, attachment levels and gingival indices were recorded from one healthy and one diseased site in each of 20 subjects with adult periodontitis. Both GCF accumulated in the periodontal pocket or sulci and GCF flowing into the periodontal pocket or sulci over a three-minute interval were sampled. The amounts of osteocalcin in each GCF sample was determined using immunoassays. A mean of 2.34 ng/site (2.7 microg/ml) osteocalcin was found at diseased sites and a mean of 2.47 ng/site (5.47 microg/ml) was found at healthy sites for the accumulated GCF collection method. A mean of 0.17 ng/ site (2.17 microg/ml) osteocalcin was found at diseased sites and a mean of 0.14 ng/ site (1.85 microg/ml) at healthy sites for the flow method of GCF collection. There were no statistically significant differences between osteocalcin levels in diseased and healthy sites in subjects with adult periodontitis.     

Gingival crevicular fluid monocyte chemoattractant protein-1 and RANTES levels in patients with generalized aggressive periodontitis

BACKGROUND: Local and systemic inflammatory and immune mechanisms may be implicated in the pathogenesis of the aggressive forms of periodontal disease. Chemokines, monocyte chemoattractant protein-1 (MCP-1) and regulated on activation, normal T cells expressed and secreted RANTES (regulated on activation, normal T cells expressed and secreted), are involved in the activation and recruitment of inflammatory and immune cells to the infected sites and thereby mediating a variety of pathophysiological conditions. The aim of the present study was to examine the gingival crevicular fluid (GCF) levels of MCP-1 and RANTES in patients with generalized agressive periodontitis (G-AgP). METHODS: MCP-1 and RANTES levels were investigated in GCF samples of 10 patients with G-AgP and 10 periodontally healthy subjects. Periodontal status was evaluated by measuring probing depth, clinical attachment loss, presence of bleeding on probing and plaque. In the G-AgP group, GCF samples were collected from the two approximal sites; from one single-rooted tooth and from one first molar tooth with > or =6 mm probing depth. In the healthy group, GCF samples were collected from one of the single-rooted teeth. GCF MCP-1 and RANTES levels were quantified by enzyme immunoassay. RESULTS: The G-AgP patients had significantly higher GCF MCP-1 and RANTES levels compared to the healthy group (p<0.05). GCF MCP-1 and RANTES levels were positively correlated with both probing depth and clinical attachment loss (p<0.05). There was no correlation between GCF MCP-1 and RANTES levels and the percentage of sites with bleeding (p>0.05). CONCLUSIONS: The results of the present study suggest that MCP-1 and RANTES could play key roles in both activation and recruitment of inflammatory and immune cells in periodontal environment of G-AgP patients. In conclusion, these CC chemokines may be considered in the biological mechanism underlying the pathogenesis and progression of G-AgP.     

Variations in crevicular fluid elastase levels in periodontitis patients on long-term maintenance

Granulocyte elastase was determined in the gingival crevicular fluid (GCF) of 18 periodontitis patients. They initially had similar severity of disease but had responded differently to 5-yr maintenance, 13 responders and 5 non-responders. A total of 102 sites were investigated and categorized as: i) consistently healthy, ii) healthy after treatment, iii) gingivitis, and iiii) periodontitis, according to clinical criteria. GCF elastase activity was determined with a granulocyte-specific substrate. The sites from non-responders had consistently higher elastase levels than the corresponding category of sites from responders, despite similar gingival inflammation and periodontal destruction, with the exception of consistently healthy sites. Within the non-responders, the periodontitis sites had higher elastase levels than the gingivitis sites commensurate with probing depth, while no difference existed between gingivitis sites and sites healthy after treatment, despite a difference in probing depth. In contrast, in the responders similar elastase levels were found at the periodontitis sites and gingivitis sites despite difference in probing depth, while both diseased sites had higher elastase levels than the sites healthy after treatment, commensurate with probing depth. This study suggests that increased granulocyte-specific elastase levels in GCF may serve as a diagnostic marker for refractory periodontitis patients.     

Gingival crevicular fluid myeloperoxidase at periodontitis sites

This investigation was undertaken to determine the relationship between the amount of the polymorphoruclear leukocye (PMN) cnzyme mycloperoxidasc (MPO) in gingival crevicular fluid (GCF) collected from periodontius sites before and after root planning and (1) gingival drsease status described by clinical indices. (2) the quantity of GCF. and (3) differential dark-field coums of Subgingival plaque microorganisms. Similar measurements except dark-field microbial counts, were made at sites with minimal signs of periodontal disease. Higher levels of GCF MPO occurred at periodontals sites. Reduced enzyme activity fooled instrumentation therapy. The reduced enzyme activity, however, was not specifically associated with decreases in clinical indices. GCF MPO was only weakly dependent of GCF volume Consequently a major portion of the greater GCF MPO observed before root planning reflected a characteristic of the lesion other than a difference in GCF volume. Before therapy more GCF MPO was noted at sites with a greater number of subgingival plaque microorganisms. However, there was no relationship between GCF MPO and the proportion of spirochetes, mobile and rod forms in the samples. These data support the conclusion that the quantity of GCF MPO reflects factors other than those associated with clinical parameters commonly used to assess inflammatory periodontal disease.     

Gingival tissue and crevicular fluid co-operation in adult periodontitis

Activated matrix metalloproteinase-3 (MMP-3) can contribute to periodontal ligament destruction in adult periodontitis. Since MMP-3 has been reported to activate proMMP-8 and -9, it was speculated that gingival tissue fibroblast-derived MMP-3 might, in periodontitis, be responsible for activation of gingival crevicular fluid (GCF) neutrophil-derived proMMP-8 and -9. Immunohistochemistry disclosed MMP-3 in gingival fibroblasts in periodontitis. Cultured gingival fibroblasts released only pro-MMP-3 when stimulated with tumor necrosis factor-alpha. However, Western blot revealed partially activated MMP-3, MMP-8, and MMP-9 in periodontitis GCF. Active MMP-8 (p < 0.05) and MMP-9 (p < 0.05) correlated with the presence of active MMP-3. It seems that resident gingival fibroblasts produce pro-MMP-3 in GCF, where it becomes activated, probably by cathepsin G or elastase released by neutrophils. Active MMP-3 then activates neutrophil-derived pro-MMP-8 and -9. Different tissue compartments/cells exert co-operative actions in mutual local MMP activation cascades.     

Gingival crevicular fluid lactoferrin levels in adult per iodontitis patients

The present study was designed to determine in a cross-sectional study whether there was any relationship between the levels of lactoferrin in gingival crevicular fluid and clinical periodontal parameters. Crevicular fluid was collected from individual sites using standardized filter paper strips (clinically healthy sites, N=23; periodontitis sites, n=66) and evaluated for lactoferrin by enzyme-linked immunosorbent assay. The data showed that: (1) the total amounts of lactoferrin were 0.003-0.021 ng (30 second sample) (average 0.009±0.005 ng) in a clinically healthy periodontium group and 0.016-3.847 ng (30 second sample) (average 0.575±0.069 ng) in adult periodontitis patients (statistically significantly higher in adult periodontitis patients); and (2) the total amounts of lactoferrin were significantly correlated with clinical parameters,     

Gingival crevicular fluid levels of calprotectin and myeloperoxidase during therapy for generalized aggressive periodontitis

BACKGROUND: Levels of the inflammation marker calprotectin in gingival crevicular fluid correspond to clinical and biochemical parameters of periodontal inflammation. Neutrophil granulocytes (polymorphonuclear neutrophils: PMNs) are supposed to be the main source of calprotectin in gingival crevicular fluid, but evidence is still lacking. The influence of periodontal therapy on gingival crevicular fluid levels of calprotectin has not yet been determined. OBJECTIVES: Gingival crevicular fluid levels of calprotectin were monitored during therapy for generalized aggressive periodontitis. Interrelations between calprotectin and the PMN marker myeloperoxidase (MPO) were evaluated. MATERIAL AND METHODS: Gingival crevicular fluid samples were collected from 23 patients with generalized aggressive periodontitis before and 3 months after non-surgical therapy with an adjunctive antimicrobial medication. Clinical parameters were recorded with a pressure-calibrated electronic probe. Levels of calprotectin and MPO in gingival crevicular fluid were analysed by enzyme-linked immunosorbent assay (ELISA) procedures. RESULTS: At baseline, levels of calprotectin and MPO were highly correlated. Bleeding and suppurating sites showed significantly higher levels of calprotectin and MPO than non-bleeding, non-suppurating sites. Therapy significantly decreased levels of both biomarkers. These changes of calprotectin and MPO were highly correlated and also related to probing-depth reduction. Three months after therapy, the levels of both markers still showed significant correlations in initially deep sites, whereas in initially shallow sites no significant correlation was found. After therapy, levels of markers in bleeding and non-bleeding sites were comparable. CONCLUSION: The correlations between calprotectin and MPO indicate that PMNs are a major contributor to the calprotectin content in gingival crevicular fluid of severely affected sites. Calprotectin levels in gingival crevicular fluid and their changes reflect periodontal inflammation as well as the clinical treatment outcome. A prognostic potential of this marker substance remains to be determined.     

牙周炎患者龈沟液中乳酸脱氢酶水平分析

目的：探讨龈沟液中乳酸脱氢酶（ＬＤＨ）水平对于慢性成人牙周炎患者诊断及预后观察的意义。方法：酶动力学方法。结果：患病部位和健康部位龈沟液中ＬＤＨ水平有非常显著差异（Ｐ〈０．００１）。探诊深度和龈沟液中ＬＤＨ水平呈正相关（Ｐ〈０．０５）。附着丧失水平和龈沟液中ＬＤＨ水平呈正相关（Ｐ〈０．０５）。结论：龈沟液中ＬＤＨ水平对于慢性成人牙周炎的诊断和疗效监测具有一定的临床意义。     

Gingival crevicular fluid and plasma oxidative stress markers and TGM-2 levels in chronic periodontitis

ObjectiveThis study was aimed to evaluate the gingival crevicular fluid (GCF) and plasma transglutaminase-2 (TGM-2), total antioxidant capacity (TAC), total oxidant status (TOS), ferric reducing antioxidant power (FRAP) and thiobarbituric acid reactive substances (TBARS) in patients with chronic periodontal disease.Materials and methodsTwenty patients with chronic periodontitis (CP), 20 patients with gingivitis and 20 healthy subjects were enrolled in the study. Clinical periodontal parameters including probing depth, clinical attachment level, plaque index and papillary bleeding index were recorded. GCF and plasma levels of TGM-2, TAC, TOS, TBARS and FRAP were analyzed.ResultsGCF TGM-2 was significantly lower in CP group than in gingivitis patients (P = 0.006). GCF FRAP in CP and gingivitis groups was significantly lower than in healthy subjects (P < 0.001). Plasma FRAP level was lower in gingivitis group when compared to healthy subjects (P = 0.003). There was no significant difference in GCF and plasma TAC, TOS, TBARS and plasma TGM-2 levels among the study groups (P > 0.05). GCF TGM-2 level was positively correlated with GCF TAC and negatively correlated with CAL.ConclusionsDecreased FRAP in GCF and plasma indicating lower antioxidant status of CP patients might suggest the role of oxidative stress in periodontitis. GCF TGM-2 data might suggest that TGM2 is associated with stabilization of the extracellular matrix and wound healing in periodontium rather than gingival inflammation.     

Gingival crevicular fluid levels of interleukin-1beta and glycemic control in patients with chronic periodontitis and type 2 diabetes

BACKGROUND: Patients with diabetes have increased incidence and severity of periodontal disease not accounted for by differences in the subgingival microbial infection. Poor glycemic control has been consistently associated with periodontal disease severity. Also, recent evidence suggests that hyperglycemia may induce inflammatory cytokine production. Few studies, however, have examined local biochemical measures of periodontal inflammation in patients with type 2 diabetes. The aim of this study was to determine whether glycemic control was related to gingival crevicular fluid (GCF) levels of interleukin-1beta (IL-1beta). METHODs: GCF samples were collected from 45 patients with type 2 diabetes and untreated chronic periodontitis. Plaque index (PI), bleeding on probing (BOP), probing depth (PD), and attachment level (AL) were recorded at six sites per tooth. IL-1beta levels were determined from individual GCF samples by enzyme-linked immunoabsorbent assay (ELISA). Individual site and mean patient values were calculated. Glycated hemoglobin (HbA1c) levels were measured from anticoagulated whole blood using an automated affinity chromatography system. Serum glucose was also determined. RESULTS: Clinical periodontal measures (PD, AL, BOP) and measures of glycemic control (HbA1c, random glucose) were significantly correlated with GCF IL-1beta. Patients with greater than 8% HbA1c had significantly higher mean GCF IL-1beta levels than patients with less than 8% HbA1c. In a multivariate model adjusting for age, gender, PD, AL, BOP, and PI, HbA1c and random glucose were independent predictors of high GCF IL-1beta. CONCLUSIONS: Poor glycemic control is associated with elevated GCF IL-1beta. These data are consistent with the hypothesis that hyperglycemia contributes to an heightened inflammatory response, and suggests a mechanism to account for the association between poor glycemic control and periodontal destruction.     

Serum and gingival crevicular fluid levels of ciprofloxacin in patients with periodontitis

BACKGROUND: The use of antibiotics as an adjunctive therapy in the treatment of periodontal diseases is of special interest to dental practitioners. In addition to using an appropriate antibacterial agent, clinicians may find it useful to determine the local and systemic concentrations of antibiotics in infected periodontal sites to reduce the levels of bacteria. The purpose of this study was to determine the serum and gingival crevicular fluid, or GCF, concentrations of systemic ciprofloxacin in patients with periodontitis. METHODS: Ten subjects with chronic periodontitis received ciprofloxacin (500 milligrams) twice daily for five days. The authors collected GCF and serum samples immediately after administering the first dose (baseline = 0 hours) and at consecutive time points. The orifice method was used for GCF sampling, and 5 milliliters of venous blood was drawn for serum analysis. The authors used high-performance liquid chromatography to determine ciprofloxacin concentrations in GCF and serum. RESULTS: The authors found that ciprofloxacin concentrations in GCF were significantly higher than concentrations in serum at two, four, seven, 24 and 120 hours. Ciprofloxacin reached the maximum concentration, or Cmax (3.72 micrograms/ mL), in GCF two hours after the initial dose was administered. The concentration decreased to 2.06 microg/mL 24 hours after the initial administration of the drug. Serum Cmax was 2.58 microg/mL at 1.5 hours, and the concentration decreased to 0.26 microg/mL at 24 hours. CONCLUSION: The results of this clinical study show that ciprofloxacin is found in GCF and its concentration in GCF is significantly higher than that in serum. CLINICAL IMPLICATIONS: Ciprofloxacin may be useful in treating patients with periodontitis because it reaches higher concentrations in GCF than in serum.     

Profiling gingival crevicular fluid from smoking and non-smoking chronic periodontitis patients

BACKGROUND: Smoking is a major risk factor for the development and progression of chronic periodontitis (CP). Gingival crevicular fluid (GCF) derived from these patients contains many proteins that could serve as important diagnostic indicators. A protein chip technology called Surface Enhanced Laser Desorption/Ionization-Time of Flight-Mass Spectrometry (SELDI-TOF-MS) has recently been developed to facilitate protein profiling of complex biological mixtures. The hypothesis to be tested was SELDI-TOF-MS could distinguish between GCF of CP patients by smoking status and pocket depth. METHODS: GCF samples collected before treatment from sixteen CP patients (eight smokers and eight non-smokers) at both shallow and deep sites were evaluated by SELDI-TOF-MS. Spectral fingerprints were constructed for both cohorts and analyzed by a two-way ANOVA according to smoking status and pocket depth. Significance threshold was set at p < 0.05. Mean molecular weight (MW) peaks and intensities were also analyzed. RESULTS: The spectral fingerprints were significantly different between the two cohorts when analyzed by ANOVA according to smoking status (p < 0.0001) but not pocket depth (p = 0.9876). Also, the mean intensity of many individual MW peaks were determined to be significantly different between the two cohorts. Several peaks ranging in MW from 11-14 kDa were only detected in the GCF obtained from smokers. CONCLUSIONS: This study has demonstrated that profiling of GCF by SELDI-TOF-MS can distinguish between CP smokers and non-smokers. Moreover, over-expressed proteins in GCF from smokers may serve as biomarkers for this high risk patient population.     

Gingival crevicular aspartate aminotransferase levels in periodontitis patients before and after periodontal treatment]

Gingival crevicular fluid samples were collected from 296 teeth from 40 subjects, including 19 rapidly progressive periodontitis (RPP), 8 chronic adult periodontitis (CAP), 7 marginal gingivitis (MG) and 6 healthy subjects (H). The activities of aspartate aminotransferase (AST) in each sample were tested. The results were as follows: (1) The two groups with destructive periodontal disease (RPP and CAP) had greater GCF-AST levels than that from the two non-destructive groups (MG and H). (2) The GCF-AST activities showed significant positive correlations with clinical periodontal parameters, such as probing depth, attachment loss, bleeding index and suppuration. (3) Four weeks after thorough full-mouth root planing, both clinical parameters and GCF-AST levels decreased significantly. The present study suggests that GCF-AST activity might be a sensitive and objective marker for detection of periodontal tissue destruction and inflammation.