Gingival Crevicular Fluid Of Patients Research Articles

Involvement of TSP1 and MMP9/NGAL in angiogenesis during orthodontic periodontal remodeling.

In the present study the aim was to measure the levels of Thrombospondin-1 (TSP1) and Lipocalin-2/matrix metalloproteinase 9 (MMP9/NGAL) complex in gingival crevicular fluid (GCF) at different time points of orthodontic treatment, to determine the relationship between these values and those of total-matrix metalloproteinase 9 (MMP9) and theirs implication in angiogenesis balance, in the situation of a good control of the bacterial plaque, emphasizing the role of TSP1 and MMP9/NGAL complex. GCF samples were collected from 16 young orthodontic patients requiring upper canine distalization (test tooth) with first premolar extraction. The contralateral canine (control tooth) was free from orthodontic force. For the orthodontic appliance, brackets Roth 0.018 inch with 0.012 inch NiTi archwire and a laceback were used. TSP1, MMP9/NGAL, and MMP9 increased from 1 hour before activation of orthodontic appliance to a maximum at 8 hours for MMP9 and 72 hours for MMP9/NGAL and TSP1. The results show a change in time of TSP1, MMP9/NGAL, and MMP9 levels in GCF of patients with this method of orthodontic treatment. The powerful correlation of MMP9/NGAL with TSP1 suggests their stronger involvement in angiogenesis processes in PDL during orthodontic periodontal remodeling, in the situation of a healthy periodontium and a good control of the bacterial plaque.

Effect of periodontal therapy on C-reactive protein levels in gingival crevicular fluid of patients with gingivitis and chronic periodontitis: A clinical and biochemical study.

Background and Objectives:C-reactive protein (CRP) is a type I acute phase reactant. A number of studies have reported elevated serum CRP levels in periodontitis subjects, which decrease following periodontal therapy. However, the data of interventional studies on gingival crevicular fluid (GCF) levels of CRP is very scarce. The aim of the present study was to evaluate the effect of periodontal therapy on GCF CRP levels in patients with gingivitis and chronic periodontitis.Materials and Methods:A total of 60 subjects were included in the study with 20 subjects each in following groups: I-Healthy, II-Gingivitis, III-Mild periodontitis based on community periodontal index scores. Periodontal therapy was performed on Group II and Group III patients. GCF was collected from each subject at baseline and 3 months after periodontal therapy. The collected sample was subjected to biochemical analysis to detect CRP levels by using commercially available chemiluminescence immunoassay kit.Results:The present study demonstrated that the periodontitis group had a higher mean CRP level (2.49 ± 0.47 ng/ml) when compared with the Gingivitis group (1.40 ± 0.32 ng/ml) and Healthy group (0.56 ± 0.20 ng/ml). The mean CRP values after periodontal therapy were found to be reduced to 0.44 ± 0.23 ng/ml in Group II and 1.30 ± 0.36 ng/ml in Group III patients.Interpretation and Conclusion:Within the limitations of this study, it can be concluded that GCF CRP level progressively increases from periodontal health to disease. It can also be stated that there is a decrease in GCF CRP levels with periodontal treatment.

Diabetes‐Associated Periodontitis Molecular Features in Infrared Spectra of Gingival Crevicular Fluid

It has been established previously that infrared spectroscopy (IRS) can be used to identify periodontitis-specific molecular signatures in gingival crevicular fluid (GCF) and to confirm clinical diagnoses. This follow-up study is designed to assess whether this novel technique is also able to differentiate diseased from healthy sites in patients with diabetes mellitus (DM) by analyzing the molecular fingerprints embedded in the GCF. A total of 65 patients with DM with moderate-to-severe chronic periodontitis (CP) was recruited, and 15 individuals without DM (65 sites) without periodontal diseases were used as control. Clinical examination and GCF samples were taken from a total of 351 sites, including periodontitis (109), gingivitis (115), and healthy (127) sites. Corresponding absorption spectra of GCF samples were acquired and processed, and the relative contributions of key functional groups in the infrared spectra were identified and analyzed. The qualitative assessment of clinical relevance of these GCF spectra was interpreted with multivariate statistical analysis: linear discriminant analysis (LDA). Spectral analysis revealed several molecular signatures representing vibrations in protein (amide I and II), lipid ester, and sugar moieties in the GCF of patients with DM with CP and non-DM controls. The diagnostic accuracy for distinction between healthy and CP sites in patients with DM determined by LDA of GCF spectra was 95.3% for the training set of samples and 87.5% for the validation set. Additional LDA of GCF spectra from healthy sites of non-DM controls and patients with DM revealed 100% diagnostic accuracy for the training set and 86.7% for the validation set. The regions robotically selected by LDA for the two analyses were slightly different in that first LDA identified major regions clustered with the side chain vibrations originating from protein and DNA contents, whereas the second was predominantly the glycation and protein components. IRS is a feasible method to differentiate disease-specific molecular signatures in GCF in the presence of DM and to generate a complex biochemical profile of GCF to identify DM-specific spectral features.

Oxidized-low density lipoprotein in gingival crevicular fluid of patients with chronic periodontitis: a possible link to atherogenesis

Objective. To investigate a possible link between periodontitis and atherogenesis by examining the levels of anti-oxidized low density lipoprotien (ox LDL) and low density lipoprotien (LDL) in gingival crevicular fluid (GCF) and serum of healthy subjects and chronic periodontitis patients. Methods. Sixty male subjects (35–55 years) were grouped into 30 healthy individuals and 30 subjects with chronic periodontitis. Serum and GCF samples were obtained from each subject and were assessed for anti-ox LDL and LDL levels. Results. A significant difference (p < 0.001) was found between the anti-ox-LDL levels in GCF of healthy vs chronic periodontitis groups. Also the ratio of GCF anti-ox LDL to GCF LDL was significantly higher (p < 0.001) in chronic periodontitis patients as compared to the healthy group. Conclusions. A significant rise in ox LDL level in otherwise systemically healthy chronic periodontitis patients may put these subjects at an increased risk of developing atherosclerosis.

Elevated Levels of a Disintegrin and Metalloproteinase 8 in Gingival Crevicular Fluid of Patients With Periodontal Diseases

A disintegrin and metalloproteinase 8 (ADAM8) is involved in inflammation and is essential for osteoclastogenesis. Elevated ADAM8 levels are detected in human serum and other body fluids in several inflammatory conditions. Therefore, we hypothesized that ADAM8 levels are also raised in gingival crevicular fluid (GCF) of patients with periodontal diseases. Forty-five patients with periodontal diseases (n = 15 for each group: the group of patients with gingivitis, the group with aggressive periodontitis [AgP], and the group with chronic periodontitis [CP]) and 15 volunteers who exhibited healthy gingiva were recruited. Four periodontal parameters, gingival index, plaque index, probing depth, and clinical attachment level, were recorded before GCF collection. The presence of ADAM8 in GCF was shown by immunoblotting using anti-human ADAM8 polyclonal antibody against its prodomain, and the ADAM8 levels were measured by an enzyme-linked immunosorbent assay. Four immunoreactive bands at 120, 70, 50, and <30 kDa were detected in the groups of patients with periodontitis, whose intensities were stronger than those in the group of patients with gingivitis, consistent with significantly greater ADAM8 levels in both groups of patients, with either CP or AgP, than those in the group of patients with gingivitis and in the group that was healthy (P <0.001). Moreover, the ADAM8 levels correlated significantly with the four periodontal parameters (P <0.001), indicating that ADAM8 levels are positively associated with the degree of periodontal tissue inflammation and destruction. The ADAM8 levels are elevated in the GCF of patients with periodontal diseases, including gingivitis, CP, and AgP, in comparison to control participants who are healthy, and they correlate with four clinical parameters that reflect the degree of disease severity.

Expression of human beta defensins (HBDs) 1, 2 and 3 in gingival crevicular fluid of patients affected by localized aggressive periodontitis

To study the effect of nonsurgical periodontal therapy on the expression frequencies of human beta-defensin (HBD)-1, -2, and -3 in the gingival crevicular fluid (GCF) of patients affected by localized aggressive periodontitis. Twenty patients affected by localized aggressive periodontitis (age range, 20-35years) and 20 healthy subjects (age range, 21-37years) were examined with clinical periodontal parameters and radiographic examination with the long-cone parallel technique. All periodontitis patients underwent nonsurgical periodontal therapy combined with doxycycline treatment and a maintenance program (including brushing with regular toothpaste). GCF samples were collected from patients and healthy control subjects at baseline as well as 3months after periodontal therapy for the patient group. In the patient group, the expression frequencies of HBD-1, -2, and -3 mRNA at baseline were 30%, 85%, and 35%, respectively, which changed after periodontal therapy to 80%, 45%, and 85%, respectively (all P<0.001). In the healthy control subjects, the expression frequencies were 95%, 40%, and 95% for HBD-1, -2, and -3, respectively, which were different from those of diseased patients at baseline (all P<0.001). The appropriate expression of HBD peptides in health and disease may contribute to the maintenance of periodontal homeostasis, possibly through its antimicrobial effects and the promotion of adaptive immune responses.

RP-HPLC assay of doxycycline in human saliva and gingival crevicular fluid in patients with chronic periodontal disease

A reversed-phased HPLC method with fluorescence detection was optimized and validated for determination of DOXY in human saliva and gingival crevicular fluid (GCF) with tetracycline as internal standard. Single step extraction with acetonitrile for both types of samples was performed. The separation was achieved at Zorbax Extend-C18 analytical column at 30°C. Mobile phase was consisted of an aqueous phase containing magnesium acetate, ammonium acetate, Na2EDTA, triethyl-ammonium acetate buffered to pH 7.5 with ammonium hydroxide solution and acetonitrile. The volume ratio of the buffered water mixture of salts and acetonitrile was 86:14. Fluorescence detector was set at λex=380nm and λem=520nm. Under the optimized experimental conditions, good linearity was found in the range of 5.0–250.0ng/mL for GCF with LOD of 1.63ng/mL and LOQ of 4.93ng/mL and 20.0–500.0ng/mL for saliva with LOD of 6.36ng/mL and LOQ of 19.28ng/mL. This method was successfully applied for determination of DOXY in saliva and GCF obtained from patients with chronic periodontal disease.

Gingival crevicular fluid levels of human beta‐defensins 1 and 3 in subjects with periodontitis and/or type 2 diabetes mellitus: a cross‐sectional study

Human β-defensins (hBDs) have a strong antibacterial action against various microorganisms, especially periodontal pathogens. The aim of this study was to compare the total levels of hBD-1 and hBD-3 in the gingival crevicular fluid of healthy patients with gingivitis (HG), healthy patients with chronic periodontitis (HP), patients with type 2 diabetes mellitus (DM) and gingivitis (DM2G) and patients with type 2 DM and chronic periodontitis (DM2P). A total of 80 patients were included: 20 HG, 20 HP, 20 DM2G and 20 DM2P. The levels of hBD-1 and hBD-3 in gingival crevicular fluid were measured using ELISA. The DM2P group had significantly higher periodontal clinical parameters at sites from which gingival crevicular fluid was collected compared with the other groups. The HG group had significantly lower periodontal clinical parameters within the gingival crevicular fluid-collected sites than did the HP, DM2G and DM2P groups. The gingival crevicular fluid of the DM2P group had significantly higher levels of total hBD-1 and hBD-3 than did that of the other groups; the hBD-1 and hBD-3 levels were significantly higher in the gingival crevicular fluid of the DM2G group than in that of the the non-DM type 2 groups (HG and HP). The gingival crevicular fluid of the HP group had significantly higher levels of total hBD-1 and hBD-3 in comparison with that of the HG group. As a result of the observed vascular and cell activity changes that occur within patients diagnosed with DM, periodontal diseases become more severe. These changes hinder the migration and the ability of chemotactic factors and leukocytes to protect periodontal tissues from the effects of microorganisms. In order to eliminate microorganisms, the epithelial cells in patients with DM may release more hBD-1 and hBD-3 into the gingival crevicular fluid. Determining the amount of hBD-1 and hBD-3 in the gingival crevicular fluid of patients with and without DM will help to elucidate the relationship among hBD-1, hBD-3, DM and periodontal disease.

A comparative analysis of Superoxide dismutase activity levels in gingiva and gingival crevicular fluid in patients with chronic Periodontitis and periodontally healthy controls

Background and Objectives: Superoxide dismutase (SOD), a metalloenzyme is a powerful antioxidant in the body that acts against Superoxide, released in inflammatory pathways and causes extracellular structure degradation. In this study, SOD activities in gingiva and gingival crevicular fluid (GCF) from patients with chronic periodontitis (CP) and periodontally healthy controls were compared. Materials and Methods: Thirty patients, involving teeth with moderate-to-severe periodontal breakdown and ≥ 5 mm pockets that required full-thickness flap surgery in the maxillary quadrants, and in controls, thirty patients with teeth scheduled for extraction for reasons other than periodontal destruction were studied. After clinical measurements, GCF samples were collected. Tissue samples were harvested from the same teeth, during flap operation in CP patients and immediately after tooth extraction in controls. SOD activities were spectrophotometrically assayed. The results were statistically analyzed. Results: Gingival SOD activity was significantly higher in the CP group than in the controls (p<0.001). Significant difference was found in GCF SOD activity between the groups (p = 0.012). Correlations between gingival and GCF SOD were statistically highly significant in CP and control groups (p = 0.00). Conclusion: In chronic periodontitis, superoxide dismutase activity seems to increase in gingiva, probably as a result of a higher need for superoxide dismutase activity and protection in gingiva in CP, while a lower activity seen in GCF which could be due to the low amounts of superoxide dismutase in the extracellular fluid. The strong correlation between the gingival and GCF SOD activities suggests distinct actions of these superoxide dismutases.

Haplotypes of susceptibility to chronic periodontitis in the Interleukin 8 gene do not influence protein level in the gingival crevicular fluid

ObjectivePreviously, we identified that the ATC/TTC haplotype formed by polymorphisms in the Interleukin-(IL)8 gene conferred susceptibility to chronic periodontitis (CP). The aim of the study was to investigate whether the IL8 haplotype ATC/TTC was associated with the volume of gingival crevicular fluid (GCF), the concentration of interleukin IL-8 in the GCF, as well as periodontal conditions in patients with CP in comparison to controls without CP. MethodsSeventy-nine individuals (CP: n=41, controls: n=38) were grouped according to the presence (susceptible for CP) or absence (not susceptible for CP) of the IL8 ATC/TTC haplotype. After periodontal clinical evaluation, they were subdivided by the presence or absence of CP. GCF was collected from each patient and the IL-8 levels were determined by ELISA. The GCF volume of each subject was measured by means of a calibrated electronic device. Comparisons of means between carriers and non-carriers of the ATC/TTC haplotype were evaluated using the Mann–Whitney test. Linear regression and stepwise linear regression analysis were used to analyse the association of the GCF volume with potential covariates and their contribution for the phenotype. ResultsWe did not find significant differences of both periodontal conditions and IL-8 concentration in the GCF of patients with the presence or absence of the IL8 ATC/TTC haplotype. However, the GCF volume was significantly higher amongst the patients affected by CP that are absent for the IL8 ATC/TTC haplotype. In addition, linear regression analysis showed a statistically significant association between GCF volume and CP, IL8 haplotype ATC/TTC and IL-8 concentration. ConclusionsThe IL8 haplotype of susceptibility to CP was neither associated with IL-8 cytokine levels nor with clinical periodontal parameters. Also, CP, IL8 haplotype and IL-8 concentration showed a positive association with the GCF volume levels in the studied patients.

Longitudinal study of volatile fatty acids in the gingival crevicular fluid of patients with periodontitis before and after nonsurgical therapy

Short-chain fatty acids (SCFAs) are important metabolic products of subgingival organisms and their concentrations are associated with the status of inflammation. The purpose of this study was to observe and analyze the change in concentration of SCFAs in the gingival crevicular fluid of patients with chronic periodontitis before and after periodontal treatment. Gingival crevicular fluid samples were taken from 21 patients with chronic periodontitis before periodontal treatment and 2 wk, and 2, 4 and 6 mo after treatment. The concentrations of six different SCFAs in the gingival crevicular fluid were measured using high-performance capillary electrophoresis. The presence of porphyromonas gingivalis in the same pretreatment gingival crevicular fluid samples used to measure SCFAs was analyzed using PCR amplification. Two weeks after periodontal treatment, the concentrations of lactic acid, propionic acid, butyric acid and isovaleric acid in the gingival crevicular fluid of patients with chronic periodontitis had decreased to the levels found in the healthy control group. However, the concentration of formic acid had increased. Statistically significant differences were found in the levels of these SCFAs before and after treatment. In the longitudinal observation, the concentrations of butyric acid and isovaleric acid in the gingival crevicular fluid had increased to a high level 2 mo after treatment. At the last two study time-points (4 and 6 mo after treatment), butyric acid and isovaleric acid were still present at a high level and showed a tendency to continue to increase. In contrast, the concentration of formic acid in gingival crevicular fluid showed a gradual decrease over the study period. The concentration of formic acid in the gingival crevicular fluid has an inverse relationship with the severity of periodontitis, whereas the increased concentrations of butyric and isovelaric acids during the long-term observation period after therapy may indicate the status of recolonization of periodontal pathogens and reflect the subgingival ecology. These two fatty acids could be used as indicators for the development and progression of periodontitis.

Cytokine Profile in the Gingival Crevicular Fluid of Periodontitis Patients With and Without Type 2 Diabetes: A Literature Review

Periodontitis may occur in patients with and without type 2 diabetes (T2D). It may be hypothesized that the gingival crevicular fluid (GCF) cytokine profile in patients with periodontitis with poorly controlled T2D may differ from the GCF cytokine profile in medically healthy individuals with periodontitis. The aim was to review the cytokine profiles in the GCF of patients with periodontitis with and without T2D. Databases were searched from 1988 to August 2011 using different combinations of various keywords. Titles and abstracts of articles that satisfied the eligibility criteria were screened by the authors and checked for agreement. Only articles published in English were included. Ten studies were included. Two studies reported GCF concentrations of interleukin (IL)-6 to be higher in patients with periodontitis with T2D compared to medically healthy patients with periodontitis. Two studies showed GCF IL-6 levels to be higher in periodontitis with T2D compared to medically healthy subjects without periodontitis. In one study GCF levels of IL-17, IL-23, and interferon-γ were higher in patients with periodontitis with T2D compared to medically healthy patients with periodontitis. In one study, GCF concentrations of IL-8 were significantly higher in patients with periodontitis with T2D compared to medically healthy individuals with periodontitis. Three studies reported GCF levels of IL-1α to be significantly higher in patients with periodontitis with T2D compared to medically healthy individuals with periodontitis. The GCF cytokine profile in patients with and without T2D seems to be governed by the intensity of periodontal inflammation and the role of T2D in this regard is rather secondary.

Effect of the emodin thermosensitive hydrogel on coilagenase in gingival crevicular fluid of patients with chronic periodontitis

Objective To observe the effect of the emodin thermosensitive hydrogel on collagenase in gingival crevicular fluid of patients with chronic periodontitis and to evaluate the clinical efficacy.Methods Forty patients with chronic periodontitis were selected.For each patient after scaring and root planning,the one teeth on one side in a person was assigned at the test group and treated with the emodin thermosensitive hydrogel in the pocket once a week for four weeks.The teeth on the other side were assigned as the control group,just with the primary periodontal treatment.The PLI、SBI、PD、AL、MD and content of collagenase in gingival crevicular fluid were examined at baseline,before administration and after administration four weeks.The COL-II level in GCF was detected using ELISA method.Results There were no significant differences in PLI、SBI、PD、AL、MD and content of collagenase in gingival crevicular fluid before the treatment(P ＞ 0.05).While there are some evidences that periodontal indexes dropped down and the type Ⅱ collagenase level in gingival crevicular fluid also declined after using the emodin thermosensitive hydrogel than the control group(t =3.46,4.02,4.18,3.03,2.79,4.29,all P ＜ 0.05).Conclusion Ultrasonic scaling using the emodin thermosensitive hydrogelas was more effective in reducing the type Ⅱ collagenase level in gingival crevicular fluid.And improving clinical parameters associated with periodontal health in patients with chronic periodontitis. Key words: Collagen type Ⅱ ; Emodin; Silicone gels; Periodontitis; Fluids and secretions

Detection of IgG, IgA and IgM antibodies against Porphyromonas gingivalis in gingival crevicular fluid and saliva in patients with chronic periodontitis

Humoral immune response is an important host mechanism against periodontopathogenic microorganisms, such as Porphyromonas gingivalis. P. gingivalis is the most frequent causative agent of chronic periodontitis in Mexico, but its detection presents difficulties. The aim of this study was to identify antibodies against semi-purified culture-secreted proteins of P. gingivalis in saliva and gingival crevicular fluid of patients with chronic periodontitis. Semi-purified culture-secreted proteins of P. gingivalis were immobilized in Enzyme-Linked Immunosorbent Assay (ELISA) plaque to detect IgA, IgG and IgM antibodies in saliva and gingival crevicular fluid from chronic periodontitis patients. IgG antibody against proteins from P. gingivalis was detected in 65% of the samples of patients, 27% of patients had IgA antibodies, while IgM was detected in 45% of patients. IgG, IgA and IgM antibodies were detected in saliva and gingival crevicular fluid mainly in patients with severe chronic periodontitis, confirming that P. gingivalis could be the causative agent of the infection. Key words: Antibodies, gingival crevicular fluid, Porphyromonas gingivalis, and saliva.

Effectiveness of non-surgical treatment to reduce il-18 levels in the gingival crevicular fluid of patients with periodontal disease

The aim of this study was to evaluate the effectiveness of the non-surgical periodontal treatment in reducing the gingival crevicular fluid (GCF) levels of IL-18 from inflamed periodontal sites. Fourteen patients with periodontal disease were included, being 9 patients with chronic periodontitis (mean age: 48.8 SD ± 7.4 years) and 5 patients with gingivitis (mean age: 43.6 SD ± 11.8). The patients were divided in the following groups: gingivitis sites from periodontitis patients (sites GP), periodontitis sites from periodontitis patients (sites PP), and gingivitis sites from gingivitis patients (sites GG). Probing pocket depth (PPD), probing attachment level (AL), plaque index (PI) and gingival index (GI) were recorded, and gingival fluid samples were collected. The subjects received non-surgical treatment and were re-evaluated 30 days after treatment (day 30 AT). There was a significant reduction in PI in GG (1.0 ± 0.4 to 0.5 ± 0.2), GP (1.2 ± 0.3 to 0.5 ± 0.3), and in PP (1.3 ± 0.4 to 0.7 ± 0.3) 30 AT. There was also a significant reduction in the GI in GG (1.3 ± 0.3 to 0.7 ± 0.4). PPD reduced significantly in GG (2.4 ± 0.6 to 1.9 ± 0.1), and PP (6.7 ± 1.1 to 5.2 ± 0.9) 30 AT. When all the samples were analyzed together, there was a significant reduction in IL-18 (12.9 ± 7.2 to 10.0 ± 3.1). This study showed that non-surgical treatment was effective in reducing GCF levels of IL-18 from inflamed periodontal sites.

Cytokines and Bone‐Related Factors in Systemically Healthy Patients With Chronic Periodontitis and Patients With Type 2 Diabetes and Chronic Periodontitis

This study compares the levels of cytokines and bone-related factors in the gingival crevicular fluid (GCF) of systemically healthy patients with chronic periodontitis (CP); and better-controlled, and poorly controlled patients with type 2 diabetes and CP. Thirty-seven patients with type 2 diabetes and CP and 20 systemically healthy patients with CP were enrolled in this study. The patients with diabetes mellitus were categorized as better-controlled (n = 17; HbA(1c) levels ≤8%) or poorly controlled (n = 20; glycated hemoglobin values >8%). Levels of tumor necrosis factor-α, interleukin (IL)-4, interferon (IFN)-γ, IL-23, IL-17, soluble receptor activator of nuclear factor-kappa B ligand (sRANKL), and osteoprotegerin (OPG) in GCF of diseased sites were analyzed by enzyme-linked immunosorbent assay. Type 2 diabetes mellitus, as a whole, upregulates the levels of OPG, sRANKL, IFN-γ, IL-17, and IL-23 and downregulates the production of IL-4 in sites with CP (P <0.05). Better-controlled individuals exhibited the highest levels of IFN-γ, whereas poorly controlled patients presented the highest levels of IL-17 (P <0.05). There were no differences in the levels of tumor necrosis factor-α, OPG, and IL-23 among systemically healthy, better-controlled, and poorly controlled patients with diabetes (P >0.05). Increased levels of proinflammatory cytokines and RANKL were observed in the GCF of patients with type 2 diabetes with CP, compared to patients without diabetes. In addition, poor or good glycemic status seems to modulate osteo-immunoinflammatory mediators in a different manner.

Nitric oxide in gingival crevicular fluid and nitric oxide synthase expression in the gingiva of patients with sickle cell disease.

This study aimed to biochemically measure the production of nitric oxide in gingival crevicular fluid and immunohistochemically measure the expression of inducible nitric oxide synthase in the gingiva of patients with sickle cell disease. Additionally, we aimed to obtain insight into the immunopathology of sickle cell disease by comparing inducible nitric oxide synthase levels in patients with sickle cell disease and controls using gingiva and gingival crevicular fluid. The study included 20 sickle cell disease patients and 20 healthy controls. Immunohistochemical analysis was used to measure inducible nitric oxide synthase expression in gingiva and nitric oxide levels in gingival crevicular fluid were spectrophotometrically measured. Nitric oxide levels in the patients and controls did not differ significantly (21.2±4.5 and 23.1±2.3 μM L-1, respectively, p>0.05). There weren't any statistically significant differences in infiltrated inflammatory cells, density of inflammatory cells that stained with inducible nitric oxide synthase, or nitric oxide expression in gingiva between the patient and control groups (p>0.05). To the best of our knowledge this is the first study to examine the expression of inducible nitric oxide synthase in the gingiva and gingival crevicular fluid in patients with sickle cell disease. Using the gingiva and gingival crevicular fluid we were unable to observe sickle cell disease-associated inducible nitric oxide synthase expression and a difference in nitric oxide levels.

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.

Effect of surgical flap on IL-1β and TGF-β concentrations in the gingival crevicular fluid of patients with moderate to severe chronic periodontitis.

Growth factors play a major part in wound healing in many tissues including the periodontium. Transforming growth factor-β1 (TGF-β1) is one of these factors present in the gingival crevicular fluid. In addition, it is considered as one of the most important anti-inflammatory cytokines. Interleukin-1β is a proinflammatory cytokine that presents itself in gingival inflammation and the GCF. Such factors might be of value as prognostic markers of wound healing activity and the therapeutic progress following flap surgery. The aim of this study was to evaluate the effect of surgical flap on the concentration of IL-1β and TGF-β in the GCF of patients with moderate to severe chronic periodontitis. The GCF samples were collected, using the Perio-Paper strip at phase 1 (pre-surgery), phase 2 (4th week post surgery) and phase 3 (12th week post surgery) from 20 sites of 10 patients undergoing flap surgery. After the elution, IL-1β and TGF-β concentrations were measured by enzyme-linked immunosorbent assay (ELISA). The mean TGF-β and IL-1β concentration decreased from phase 1 to phase 3 (p<0.05). There were no significant statistical correlations between IL-1β and TGF-β1 concentrations in the 3 assessment phases. There was a significant statistical correlation between TGF-β1 concentrations and the Plaque Index (PI) in phase 2 (p<0.05). There was a significant statistical correlation (p<0.05) between IL-1β and TGF-β1 concentration and the probing pocket depth (PPD). The flap surgery has a significant effect on decreasing IL-1β concentration. In the case of TGF-β1, probably the decrease in the concentration after treatment might be due to the removal of the inflammatory stimulants.

Receptor activator of nuclear factor kappa B ligand (RANKL) levels in peri-implant crevicular fluid.

Receptor activator of nuclear factor kappa B ligand (RANKL) is one of the key cytokines in the induction of osteoclastogenesis both in vitro and in vivo. Several reports indicated the presence of sRANKL in gingival crevicular fluid of patients with periodontal diseases. To determine the presence of RANKL in peri-implant crevicular fluid samples of implants with peri-implantitis, peri-implant mucositis and healthy controls. In this study, 40 implants were categorized as clinically healthy, peri-implant mucositis and peri-implantitis according to the clinical and radiographic findings. Filter paper strips were used to collect peri-implant crevicular fluid for 30 seconds in the base of the crevice/pocket. Peri-implant crevicular fluid (PICF) samples were obtained from buccal and lingual aspects of implants. Plaque index, probing depth, gingival index and bleeding on probing were recorded at six sites per implant. Enzyme-linked immunosorbent assay (ELISA) was performed to determine the PICF levels of sRANKL. There were no statistically significant differences in sRANKL concentration between healthy group, peri-implant mucositis and periimplantitis (p=0.12). There were also no statistical correlation between the concentration of sRANKL and probing pocket depth (R=0.051, p=0.65), or any of the other clinical regarding (p>0.05). No differences between the mean sRANKL concentration in the buccal and lingual sites were found (p=0.693). Our results may suggest that peri-implant crevicular fluid analysis of sRANKL in conjunction with some other osteoclastogenic mediators could be further investigated in further well-designed prospective longitudinal studies on a larger-scale sample size in the evaluation of dental implants.