OBJECTIVES: This study aimed to observe the effects of initial periodontal therapy on the level of neutrophil extracellular traps (NETs) in gingival crevicular fluid (GCF) of patients with severe periodontitis and to analyze the factors related to the formation of NETs. METHODS: Thirty-one patients with stage Ⅲ-Ⅳ periodontitis were recruited. Clinical periodontal parameters, including plaque index (PLI), gingival index (GI), probing depth (PD), and clinical atta-chment loss (CAL), were recorded before and 6-8 weeks after initial periodontal therapy. Levels of NETs in GCF were detected by immunofluorescence staining. Quantities of total bacteria, Porphyromonas gingivalis (P. gingivalis), Aggregatibacter actinomycetemcomitans (A. actionomycetemcomitans) and Prevotella intermedia (P. intermedia)in unattached subgingival plaque were determined by real-time quantitative PCR, and levels of tumor necrosis factor-α (TNF-α) and interleukin-8 (IL-8) in GCF were explored by enzyme-linked immunosorbent assay. In addition, the correlations between the level of NETs and the above indicators were analyzed. RESULTS: After initial periodontal therapy, the level of NETs in GCF, PLI, GI, PD, and CAL; quantities of total bacteria, P. gingivalis, A. actinomycetemcomitans, and P. itermedia; and levels of IL-8 and TNF-α significantly decreased (P<0.05). We observed strong positive correlations between the level of NETs and PLI, GI, PD, CAL, the amount of total bacteria, P. gingivalis, TNF-α, and IL-8 (P<0.05). CONCLUSIONS Initial periodontal therapy might decrease the level of NETs in GCF from patients with severe periodontitis, which might be positively correlated with the quantities of P. gingivalis andthe levels of TNF-α and IL-8 in GCF.
Humans ; Gingival Crevicular Fluid ; Extracellular Traps/metabolism* ; Porphyromonas gingivalis/isolation & purification* ; Aggregatibacter actinomycetemcomitans/isolation & purification* ; Periodontitis/metabolism* ; Tumor Necrosis Factor-alpha/analysis* ; Prevotella intermedia/isolation & purification* ; Interleukin-8/analysis* ; Male ; Female ; Middle Aged ; Periodontal Index ; Adult

OBJECTIVES: This study aims to explore the potential relationships of cell-free DNA (cfDNA) in gingival crevicular fluid (GCF) with periodontal clinical indicators and the expression of DNA receptor pathway cyclic guanosine phosphate-adenosine phosphate synthase (cGAS)-stimulator of interferon genes (STING) in gingival tissues and human gingival fibroblasts (HGFs). METHODS: GCF and gingival tissue samples were collected from periodontally healthy individuals and patients diagnosed with periodontitis. Periodontal clinical indicators were recorded, including plaque index (PLT), bleeding index (BI), probing depth (PD), and clinical attachment level (CAL). The concentration of cfDNA in GCF was quantified, and the correlation between GCF and periodontal clinical indicators was analyzed. Immunofluorescence and reverse transcription-quantitative polymerase chain reaction (RT-qPCR) were used to assess the distribution of cGAS, STING, and p-STING in gingival tissues. Additionally, the mRNA expression levels of the key components of the cGAS-STING signaling pathway, namely, cGAS, STING, inhibitory of kappa-B kinase (IKK), nuclear factor kappa-B p65 (NF-κB p65), interleukin (IL)-1β, IL-6, and tumor necrosis factor-α (TNF-α), were measured. Furthermore, cfDNA extracted from GCF was employed to stimulate HGFs in the healthy control and periodontitis groups, and the mRNA expression levels of the key molecules of cGAS-STING signaling pathway were detected through Western blot and RT-qPCR. RESULTS: The concentration of cfDNA in GCF was found to be significantly elevated in the periodontitis group compared with the control group. Moreover, cfDNA concentration demonstrated a strong positive correlation with the periodontal clinical indicators. Immunofluorescence analysis revealed considerably increased percentage of fluorescence co-localization of cGAS, STING, and p-STING with the gingival fibroblast FSP-1 marker in the gingival tissues of the periodontitis group. The mRNA expression levels of cGAS, STING, IKK, NF-κB p65, IL-1β, IL-6,and TNF-α were significantly higher in the periodontitis group. In vitro stimulation of HGFs with GCF-derived cfDNA resulted in increased protein expression of cGAS and p-STING and considerably upregulated the mRNA expression levels of cGAS, STING, IKK, NF-κB p65, IL-1β, IL-6, and TNF-α in the healthy and periodontitis groups compared with the blank group. Correlation analysis showed that the concentration of cfDNA at the sampling site was positively correlated with the mRNA expression levels of cGAS, STING, NF-κB p65, and IL-6 in gingival tissues. CONCLUSIONS cfDNA concentrations in the GCF of patients with periodontitis are considerably elevated, and are associated with the activation of the cGAS-STING signaling pathway in HGFs. These findings suggest that cfDNA contributes to the progression of periodontitis.
Humans ; Gingival Crevicular Fluid/metabolism* ; Signal Transduction ; Gingiva/cytology* ; Nucleotidyltransferases/genetics* ; Membrane Proteins/genetics* ; Cell-Free Nucleic Acids/analysis* ; Fibroblasts/metabolism* ; Tumor Necrosis Factor-alpha/metabolism* ; Periodontitis/metabolism* ; Interleukin-1beta/metabolism* ; Interleukin-6/metabolism* ; Adult ; RNA, Messenger/metabolism* ; Male ; Female

This study is to prepare the in situ forming sustained-release injection which can perform sustained release behavior at the periodontal site for 7 days and to evaluate its in vitro and in vivo properties. After preparation of in situ forming sustained-release injection the in situ time was studied. And the surface of the solid injection was characterized by SEM. The rheological curve at 0 degrees C, 25 degrees C, 37 degrees C was determined and the impact of the temperature on the viscosity was examined. The in vitro release behavior was investigated. At last, rabbit periodontitis model was established to study its pharmacokinetics. The injection was stable, hard to stratify and decompose. The in situ forming time was about 6 seconds. It can easily adhere into periodontal pockets. There were lots of holes on the surface of the solid injection for the drug to diffuse. The drug releasing curves could be fit by Korsmeyer-Peppas equation. The drug smoothly released for 7 days at pH 7.4 PBS buffer with a very slight burst release and maintained a certain concentration. In vivo pharmacokinetics results indicated that after administration with the in situ forming injection, achievement of tinidazole (TNZ) concentration in gingival crevicular fluid (GCF) was more comparable and long-lasting than usual solution of TNZ management and relatively constant TNZ levels were attained until 168 h. All these results supported the prospect of tinidazole in situ forming sustained-release injection in clinical applications.
Animals ; Antitrichomonal Agents ; administration & dosage ; pharmacokinetics ; Delayed-Action Preparations ; Drug Carriers ; Drug Compounding ; methods ; Endotoxins ; Gingival Crevicular Fluid ; metabolism ; Injections ; Periodontal Pocket ; metabolism ; Periodontitis ; chemically induced ; metabolism ; Polyesters ; chemical synthesis ; pharmacokinetics ; Polyethylene Glycols ; chemical synthesis ; pharmacokinetics ; Rabbits ; Random Allocation ; Rheology ; Tinidazole ; administration & dosage ; pharmacokinetics

OBJECTIVETo observe the effect of periodontal initial therapy together with Chinese herbal compound for clearing heat and detoxification (Abbr. TCM) on the levels of tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 beta (IL-1 beta) in gingival crevicular fluid (GCF) of patients with chronic periodontitis (CPD).

METHODSThirty-four teeth of 24 patients with CPD were randomly assigned to the control group (17 teeth of 11 patients) and the treated group (17 teeth of 13 patients), they were treated for 8 weeks with periodontal initial therapy, but TCM was given additionally to the treated group. The GCF of the observed teeth was collected at the time before initial therapy (datum line) and at 2 and 8 weeks after treatment. At the same time, the clinical parameters, including probing depth (PD), clinical attachment loss (CAL), and bleeding on probing (BOP) were recorded, and the levels of TNF-alpha and IL-1 beta in GCF were measured by radioimmunoassay.

RESULTSThe levels of TNF-alpha and IL-1 beta in GCF after treatment were significantly lowered in all patients as compared with those before treatment (P<0.01), while PD, CAL and BOP showed significant reduction (P<0.01). After treatment, TNF-alpha and IL-1 beta levels in the treated group were lower than those in the control group (TNF-alpha: 2.5551 +/- 1.0157 microg/L vs 3.4341 +/- 1.0762 microg/L, IL-1 beta: 0.1335 +/- 0.0216 microg/L vs 0.1471 +/- 0.0169 microg/L; P<0.05).

CONCLUSIONTCM could enhance the effect of periodontal initial therapy in lowering TNF-alpha and IL-1 beta levels, which was attributed to reducing the inflammatory reaction of periodontal tissues.

Adult ; Chronic Periodontitis ; metabolism ; therapy ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Gingival Crevicular Fluid ; metabolism ; Humans ; Interleukin-1beta ; metabolism ; Male ; Middle Aged ; Phytotherapy ; Tumor Necrosis Factor-alpha ; metabolism

AIMThe aim of this study was to measure the level of Oncostatin M (OSM) a gp130 cytokine in the gingival crevicular fluid (GCF) and serum of chronic periodontitis patients and to find any correlation between them before and after periodontal therapy (scaling and root planing, SRP).

METHODOLOGY60 subjects (age 25-50 years) were enrolled into three groups (n=20 per group), group I (healthy), group II (gingivitis) and group III (chronic periodontitis). Group III subjects were followed for 6-8 weeks after the initial periodontal therapy (SRP) as the group IV (after periodontal therapy). Clinical parameters were assessed as gingival index (GI), probing depth (PD), clinical attachment level (CAL), and radiographic evidence of bone loss. GCF and serum levels of OSM were measured by using Enzyme Linked Immunosorbent Assay (ELISA).

RESULTSIt was found that mean OSM levels had been elevated in both the GCF and serum of chronic periodontitis subjects (726.65 +/- 283.56 and 65.59 +/- 12.37 pg mL(-1), respectively) and these levels were decreased proportionally after the periodontal therapy (95.50 +/- 38.85 and 39.98 +/- 16.69 pg mL(-1) respectively). However, OSM was detected in GCF of healthy subjects (66.15 +/- 28.10 pg mL(-1)) and gingivitis-suffering subjects (128.33 +/- 22.96 pg mL(-1)) and was found as below the detectable limit (approximately equal 0.0 pg mL(-1)) in the serum of same subjects. Significant correlation has been found between clinical parameters and GCF-serum levels of OSM.

CONCLUSIONIncreased OSM level both in the GCF and serum, and the decreased levels after initial periodontal therapy (SRP) may suggest a use as an inflammatory biomarker in the periodontal disease.

Adult ; Analysis of Variance ; Case-Control Studies ; Chronic Periodontitis ; blood ; metabolism ; therapy ; Dental Scaling ; Female ; Gingival Crevicular Fluid ; chemistry ; Gingivitis ; blood ; metabolism ; therapy ; Humans ; Male ; Middle Aged ; Oncostatin M ; analysis ; blood ; metabolism ; Periodontal Index ; Statistics, Nonparametric

PURPOSE: The aim of this preliminary study was to determine the possible relationship between alkaline phosphatase (ALP) levels in the gingival crevicular fluid (GCF) and periodontal disease in men with hypergonadotropic hypogonadism (HH). MATERIALS AND METHODS: A total of 41 patients were divided into four groups. 9 with HH and periodontitis (P/HH), 11 with HH and gingivitis (G/HH), 12 with systemically healthy and periodontally healthy (H/C) and 9 with systemically healthy and periodontitis (P/C). The clinical evaluation of patients was based on the following parameters; the plaque index (PI), gingival index (GI), probing depths (PD) and attachment level (AL). The levels of ALP in the GCF were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: No significant difference could be detected in the mean clinical parameter data between the P/HH and P/C groups (p > 0.05). The periodontitis patients in both groups (P/C and P/HH) had higher mean probing depths than the H/C and G/HH patients (p < 0.001). The concentrations and total amounts of ALP in the GCF were significantly higher in both periodontitis groups compared to healthy and gingivitis groups (p < 0.01). The serum ALP levels were significantly higher in the P/HH group when compared to the other groups (p < 0.001). CONCLUSION: The findings of this study suggested that HH could be implicated as a contributing factor to the progress of periodontal disease.
Adolescent ; Adult ; Alkaline Phosphatase/*metabolism ; Gingival Crevicular Fluid/*enzymology ; Humans ; Hypogonadism/diagnosis/*enzymology ; Male ; Periodontium/*enzymology

OBJECTIVETo investigate the volatile fatty acids in gingival crevicular fluid (GCF) and to analyze the relationship between the levels of the volatile fatty acids and chronic periodontitis.

METHODSGCF samples taken from 37 patients with chronic periodontitis and 16 volunteers with healthy periodontal status were analyzed by capillary electrophoresis.

RESULTSThe detection frequencies and concentrations of succinic acid, butyric acid and valeric acid were significantly higher in GCF of chronic periodontitis than in that of healthy group. The detection frequencies of propionic acid had no statistic difference between the two groups, but the concentrations of it was significantly higher in inflammation group. We also found that the concentrations of succinic acid, propionic acid and butyric acid were significantly lower in shallow pockets than that in deep pockets.

CONCLUSIONSThe volatile fatty acids, especially succinic acid, propionic acid, butyric acid and valeric acid were associated significantly with the severity and inflammation of periodontal disease. The levels of succinic acid, propionic acid and butyric acid in GCF were related to pocket depth.

Adult ; Aged ; Butyrates ; analysis ; Case-Control Studies ; Chronic Periodontitis ; metabolism ; Electrophoresis, Capillary ; Fatty Acids, Volatile ; analysis ; Female ; Gingival Crevicular Fluid ; metabolism ; Humans ; Male ; Middle Aged ; Periodontium ; metabolism ; Propionates ; analysis ; Succinic Acid ; analysis

OBJECTIVETo study the effect of smoking on the periodontium and level of aspartate aminotransferase in gingival cervical fluid (GCF-AST).

METHODSA proportion matched case-control filtration was performed in examined population, college students, outpatients from 1999 May to 2001 March 115 smokers aged from 23 to 65 years (102 men and 13 women) and 90 non-smokers aged from 25 to 70 years (80 men and 10 women) were chosen. Debris index (DI), calculus index (CI), periodontal disease index (PDI), GCF-AST were measured.

RESULTSNo obvious differences were observed in DI in smokers (0 degrees, 27.2%; 5 degrees, 5.0%) and non-smokers (0 degrees, 27.8%; 5 degrees, 4.2%),whereas obvious differences were found in CI in smokers and non-smokers. The percentages of patients without calculus were lower in smokers (9.8%) than in non-smokers (20.0%). The percentages of patients with weighty calculus were higher in smokers (25.4%) than in non-smokers (12.8%). The PDI values in smokers were higher than in non-smokers. The percentages of their normal periodontium were lower in smokers (9.6%) than in non-smokers (20.8%). The percentages of their periodontitis were higher in smokers (38.5%) than in non-smokers (25.8%). The smoking quantity were positively related to periodontitis (P < 0.001). No obvious differences were found in the level of GCF-AST with same PDI (P > 0.05).

CONCLUSIONSmoking is considered as one of the risk factors in the prevalence of periodontal disease, but may not have any direct effect on GCF-AST.

Adult ; Aged ; Aspartate Aminotransferases ; metabolism ; Case-Control Studies ; Female ; Gingival Crevicular Fluid ; enzymology ; Humans ; Male ; Middle Aged ; Periodontitis ; etiology ; Periodontium ; enzymology ; Smoking ; adverse effects

OBJECTIVETo investigate the relationship between smoking and gingival crevicular fluid volume (GCF), level of elastase (EA) in 37 severe periodontitis patients before and after 1 month periodontal initial treatment.

METHODSThe GCF samples were collected from 122 sites in 22 heavy smokers (>or= 20 cigarettes/day) and 90 sites in 15 non-smokers before and after 1 month periodontal initial treatment. There is no difference (P > 0.05) on pocket depth between smoking sites (5.6 +/- 1.2) mm and non-smoking sites (5.4 +/- 1.2) mm at baseline. The volume of each GCF sample was measured by Periotron 6000 and the elastase in GCF were determined by substrate (meosuc-als-als-pro-val-NA) method.

RESULTSAfter non-surgical treatment both GCF volume and elastase level were significantly decreased (P < 0.001) in both smokers and non-smokers. But the decrease of GCF volume (91 sites, 74.6%) and elastase level (70 sites, 76.1%) in smokers were significant lower (P < 0.01) than non-smokers (GCF, 88 sites, 97.8%; EA, 56 sites, 93.3%).

CONCLUSIONThese findings suggest that smoking has effect on gingival crevicular fluid volume and elastase level of patients with periodontitis.

Adult ; Gingival Crevicular Fluid ; enzymology ; physiology ; Humans ; Male ; Middle Aged ; Pancreatic Elastase ; analysis ; Periodontitis ; metabolism ; therapy ; Smoking ; adverse effects