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 aimed to evaluate the therapeutic effect of 1% povidone-iodine mouthwash combined with scaling and root planing in patients with stage Ⅰ/Ⅱ class A/B periodontitis, and to provide a basis for the clinical application of povidone-iodine mouthwash. METHODS: Seventy-five subjects were included in this trial and randomly divided into three groups. After full-mouth ultrasonic supragingival cleansing, scaling and root planing, the placebo group was treated with sodium chloride injection (NaCl group), the control group was treated with compound chlorhexidine mouthwash (CHX group), and the experimental group was treated with 1% povidone-iodine mouthwash (PVP-I group), and rinsed their mouths for 1 week, respectively. Subjects were tested at 1, 4, and 12 weeks after dosing for clinical indicators, microbial composition of supragingival plaque, gingival crevicular fluid inflammatory marker levels, and patient-reported outcomes. RESULTS: Sixty-three subjects completed the follow-up. After treatment, the clinical indicators, microbial indicators, and inflammatory indicators were all significantly improved (P<0.05). Comparisons among the groups showed that one week after treatment, the bleeding index and plaque index of the CHX group and the PVP-I group were lower than those of the NaCl group, and the plaque index of the CHX group was lower than that of the PVP-I group (P<0.05). There were no statistically significant differences in the other clinical indicators among the groups (P>0.05). Twelve weeks after treatment, the Shannon index of the CHX group was lower than that of the NaCl group (P<0.05), and there were no statistically significant differences in the other microbial indicators among the groups (P>0.05). Twelve weeks after treatment, the interleukin-10 concentration of the CHX group was higher than that of the NaCl group (P<0.05), and there were no statistically significant differences in the other inflammatory indicators among the groups (P>0.05). The PVP-I group had the highest scores in terms of taste and oral odor. There was no obvious staining on the tooth surfaces and mucosa in all three groups. CONCLUSIONS 1% PVP-I mouthwash combined with scaling and root planing can effectively reduce gingival inflammation and dental plaque, improve clinical symptoms in the short term. While its efficacy is not significantly inferior to that of chlorhexidine, PVP-I mouthwash is more acceptable to patients than chlorhexidine.
Humans ; Povidone-Iodine/administration & dosage* ; Mouthwashes/therapeutic use* ; Dental Scaling ; Root Planing ; Periodontitis/microbiology* ; Gingival Crevicular Fluid/chemistry* ; Anti-Infective Agents, Local/therapeutic use* ; Female ; Male ; Chlorhexidine/therapeutic use* ; Dental Plaque/microbiology* ; Middle Aged ; 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

ABSTRACT Root resorption is a shortening of root dentine which occurs physiologically in deciduous teeth. The present study aimed to quantify dentine sialophosphoprotein (DSPP), aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) in gingival crevicular fluid (GCF) during the physiological process of root resorption of deciduous teeth. A cross-sectional study was conducted with 25 children aged between 4 and 10 years old. GCF was collected from the gingival sulcus using periopaper strips from the upper first deciduous molar (n = 45). The samples were divided equally into three groups, no resorption (R0), moderate resorption (RM) and severe resorption (RS), based on the existing radiographs taken. The GCF samples were then analysed using an enzyme-linked immunosorbent assay (ELISA) kit to determine the DSPP concentration levels and BioAssays System kit for AST and LDH. One-way ANOVA was used to determine the statistical differences between the means of the DSPP, AST and LDH concentration level in the three groups. A difference was considered significant when p < 0.05. High concentration levels of DSPP were significantly noted in RS (p < 0.05), compared to RM and R0. AST also portrayed significant high activity level (p < 0.05) similar to DSPP but LDH showed no significant changes between groups (p > 0.05). The high quantification of DSPP and AST levels in the severe and moderately resorbed roots indicated the potential use of this protein as a biomarker for detecting moderate-severe stages of root resorption.
Root Resorption ; Gingival Crevicular Fluid ; Dentin ; Aspartate Aminotransferases ; Lactate Dehydrogenases

OBJECTIVES: This study aimed to investigate the association between periodontal indexes and biomarkers in gingival crevicular fluid (GCF) and preterm birth (PTB) in pregnancy, as well as to assess the clinical value of these indexes as predictors of PTB. METHODS: A nested case-control study was conducted. A total of 300 systematically healthy pregnant women were selected within 36 weeks of gestation and grouped according to the enrolled weeks. Periodontal indexes, including probing depth (PD), bleeding index (BI), gingival index (GI), and five biomarkers in GCF, including interleukin (IL)-1β, IL-6, tumor necrosis factor-α (TNF-α), prostaglandin E2 (PGE2), and 8-hydroxy-2-deoxyguanosine (8-OHdG) were measured at the enrolled date. The detailed birth outcome was recorded. RESULTS: Only women at 24-28 weeks of gestation per PTB case (four full-term births) were selected as controls subjects, PTB displayed significantly greater GI, BI, and 8-OHdG ( CONCLUSIONS Increased BI and 8-OHdG at 24-28 weeks of gestation are risk factors for PTB. Their combined detection may have some value in the prediction of PTB, but further studies with a larger sample size are needed to explore it and thus provide experiment evidence for establishing an early warning system for PTB in pregnant women with periodontal disease.
Biomarkers ; Case-Control Studies ; Female ; Gingival Crevicular Fluid ; Humans ; Infant, Newborn ; Periodontal Index ; Pregnancy ; Premature Birth

gingival crevicular fluid (GCF). Data were collected and analyzed using Stata software (version 11).RESULTS: The results revealed the significant effect of both PCOS and advanced CP on visfatin levels in the GCF (P < 0.05).CONCLUSION: According to the results of this study, the visfatin level in the GCF could be the probable link of association between PCOS and advanced CP.]]>
Case-Control Studies ; Chronic Periodontitis ; Enzyme-Linked Immunosorbent Assay ; Female ; Gingival Crevicular Fluid ; Healthy Volunteers ; Humans ; Nicotinamide Phosphoribosyltransferase ; Periodontal Diseases ; Polycystic Ovary Syndrome

OBJECTIVE: This study aimed to compare the differences of B cells, plasma cells, and related cytokines expression in gingival tissues between periodontitis and periodontal healthy subjects. METHODS: Gingival tissues were collected from periodontal healthy subjects (periodontal healthy group, n=12) and periodontitis patients (periodontitis group, n=15). Hematoxylin-eosin (HE) staining was used for histopathological examination. Immunohistochemical staining (CD19, CD38, and CD138) was applied to detect the expression of B cells and plasma cells. B cell-activating factor (BAFF) and soluble receptor activator of nuclear factor-κB ligand (sRANKL) were detected by enzyme-linked immunosorbent assay. RESULTS: Extensive inflam-matory cell infiltration was found in the gingival tissues of the periodontitis group. The number of CD19(+), CD38(+), and CD138(+) cells of the periodontitis group was significantly higher than that of the periodontal healthy group (P<0.000 1). BAFF and sRANKL levels of the periodontitis group were higher than those of the periodontal healthy group (P<0.01, P<
0.001, respectively). CONCLUSIONS The expression of B cells, plasma cells, and their related BAFF and sRANKL cytokines were significantly higher in periodon-titis patients than those in the periodontal healthy subjects, sug-gesting that B cells and plasma cells may be involved in the development of periodontitis.
B-Lymphocytes ; Cytokines ; Gingival Crevicular Fluid ; Healthy Volunteers ; Humans ; Periodontitis ; Plasma Cells

BACKGROUND: Smoking exerts an adverse effect on the periodontal tissue by reorganizing the ecosystem of oral microorganisms and is considered to be an important factor in the development of periodontal disease. Although cross-sectional studies on smokers and non-smokers have been attempted to investigate the microbial differences in periodontal oral cavity, only few studies have been conducted to investigate the changes in oral microorganisms during smoking cessation. The purpose of this study was to investigate the changes of bacteria in saliva and gingival crevicular fluid (GCF) over a period of one year among 11 smokers trying to quit smoking.METHODS: Eleven smokers trying to quit smoking visited the clinic at baseline, two weeks, two months, four months, six months, and 12 months to give saliva and GCF samples. The amounts of 16S rRNA, Porphyromonas gingivalis, Treponema denticola, Prevotella intermedia, Fusobacterium nucleatum subsp. nucleatum, Streptococcus mutans, and Streptococcus sobrinus in saliva and GCF were quantified using real-time polymerase chain reaction TaqMan probe assay. The results were analyzed by nonparametric statistical analysis using Friedman test and Spearman correlation coefficient.RESULTS: After cessation of smoking, the amounts of 16S rRNA corresponding to P. gingivalis, F. nucleatum, P. intermedia, and T. denticola in saliva decreased and then again increased significantly. The amount of F. nucleatum 16S rRNA in GCF decreased significantly after smoking cessation. Positive correlations were observed between 16S rRNA and F. nucleatum and between F. nucleatum and T. denticola in saliva and GCF.CONCLUSION: Even if the number of subjects in this study was small, we suggest that smoking cessation may reduce the total bacterial amount and F. nucleatum in GCF. However, the results regarding changes in the microbial ecosystem due to smoking or smoking cessation were inconsistent. Therefore, further in-depth studies need to be carried out.
Bacteria ; Bacterial Load ; Cross-Sectional Studies ; Ecosystem ; Fusobacterium nucleatum ; Gingival Crevicular Fluid ; Mouth ; Periodontal Diseases ; Porphyromonas gingivalis ; Prevotella intermedia ; Real-Time Polymerase Chain Reaction ; Saliva ; Smoke ; Smoking Cessation ; Smoking ; Streptococcus mutans ; Streptococcus sobrinus ; Treponema denticola

OBJECTIVE: Cytokines produced by various cells are strong local mediators of inflammation. Interleukin-1beta (IL-1β) and C-reactive protein (CRP) play essential roles in the development and progression of diabetes mellitus (DM). Thus periodontal diseases could be related to DM via the same mediators of inflammation. To evaluate plasma and gingival crevicular fluid (GCF) levels of IL-1β and CRP in adolescents with DM to further investigate whether DM has an impact on the levels of inflammation factors at an early stage, and to analyze the risk of developing periodontal diseases in adolescents with DM. METHODS: A total of 121 adolescents aged from ten to sixteen years were enrolled, 41 adolescents diagnosed with diabetes mellitus were collected in the DM group, and 80 nondiabetic adolescents as the control group. The periodontal indices of each individual were recorded, including plaque index (PLI), modified bleeding index (mBI), probing depth (PD) and attachment loss (AL). GCF and intravenous blood samples were collected, and CRP and IL-1β levels were detected by enzyme-linked immunosorbent assay (ELISA). RESULTS: (1) PLI of DM group and control group were 1.23±0.05 and 0.95±0.04 separately, with significant difference (P=0.001). DM group and control group had mBI of 0.80±0.08 and 0.51±0.06 separately, with significant difference (P=0.003). Attachment loss was found in none of the subjects. PDs of DM group and control group were (2.37±0.51) mm and (2.31±0.05) mm separately, and there was no significant difference. (2) CRP in GCF was only detectable in partial of the individuals, with a detectable rate of 22.9% (11/48) in total. The detectable rate of CRP in GCF was significantly higher in DM group (38.5%) than that in control group (4.5%, P=0.006). The plasma level of CRP in DM group [0.23 (0.15, 1.89) mg/L] was higher than that in control group [0.19 (0.12, 4.18) mg/L], but without significance (P=0.776). (3) The plasma levels of IL-1β in DM group and control group were (14.11±0.57) ng/L and (14.71±0.50) ng/L separately, but there was no significance (P=0.456). GCF levels of IL-1β in DM group and control group were (12.91±1.95) μg/L and (17.68±3.07) μg/L, without significant difference (P=0.185). CONCLUSION Periodontitis was not observed in adolescents with DM at an early stage. However, the rising levels of periodontal indices and CRP in GCF, might indicate that adolescents with DM have a higher risk of developing periodontal diseases in the future.
Adolescent ; C-Reactive Protein/analysis* ; Dental Plaque Index ; Diabetes Mellitus, Type 2 ; Disease Progression ; Enzyme-Linked Immunosorbent Assay ; Female ; Gingival Crevicular Fluid/chemistry* ; Humans ; Interleukin-1beta/analysis* ; Male ; Periodontal Diseases ; Periodontal Index ; Periodontitis ; Plasma

OBJECTIVES: Chronic periodontitis is a common inflammatory disease of the oral cavity that causes destruction of periodontal tissues and bone around the teeth. Sclerostin is a protein encoded by the SOST gene. In this study, gingival crevicular fluid (GCF) levels of sclerostin in patients with chronic periodontitis were compared with those of healthy subjects. MATERIALS AND METHODS: In this case-control study, a total of 40 subjects were enrolled and divided into the healthy group (n=23) and chronic periodontitis group (n=17). GCF samples were collected, and the concentration of sclerostin was evaluated using enzyme-linked immunosorbent assay. Comparison of significance between groups was assessed using Mann-Whitney U test. RESULTS: Sclerostin concentration was significantly higher in the chronic periodontitis group compared with the healthy group (P < 0.005). CONCLUSION: Despite the limitations of this study, sclerostin can be a possible marker for assessment of periodontal health status.
Case-Control Studies ; Chronic Periodontitis* ; Enzyme-Linked Immunosorbent Assay ; Gingival Crevicular Fluid* ; Healthy Volunteers* ; Humans ; Mouth ; Periodontitis ; Tooth