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

Objective To investigate the effect of Echinococcus granulosus cyst fluid(EgCF) on the cytoskeletal rearrangement and phagocytosis and the migration of macrophages induced by lipopolysaccharide(LPS). Methods Peritoneal macrophages of C57BL/6 mice were isolated and cultured in vitro, and divided into control group and LPS group and LPS combined with EgCF group. After 48 hours of treatment, filamentous actin (F-actin) changes were observed with rhodamine-labelled phalloidin staining and fluorescence microscopy; Transwell^TM chamber was used to test cell migration ability and flow cytometry to test cell phagocytosis. After 1 hour of treatment, PI3K and AKT, phosphorylated AKT (p-AKT), Rac1, guanosine triphospho-Rac1 (GTP-Rac1), WASP and Arp2 protein expressions were detected with Western blot analysis. Results Compared with the control group, after LPS stimulation, macrophages were deformed significantly; pseudopodia increased; actin cytoskeleton increased and was more distributed in pseudopodia; the ability of migration and phagocytosis were significantly improved, and the expression of PI3K, p-AKT, GTP-Rac1, WASP and Arp2 proteins significantly increased. EgCF treatment caused cell shrinkage and disappearance of pseudopodia protrusions of LPS-activated cells, and led to the reduced phagocytic and migratory of cells; the protein expression of PI3K, p-AKT, GTP-Rac1, WASP and Arp2 decreased significantly compared with the LPS group. Conclusion LPS induces the migration and enhances phagocytosis of macrophages while EgCF inhibits these effects, which is related to actin cytoskeleton rearrangement.
Mice ; Animals ; Lipopolysaccharides/pharmacology* ; Echinococcus granulosus/metabolism* ; Proto-Oncogene Proteins c-akt ; Cyst Fluid/metabolism* ; Mice, Inbred C57BL ; Macrophages/metabolism* ; Phagocytosis ; Actins/metabolism* ; Phosphatidylinositol 3-Kinases/metabolism* ; Guanosine Triphosphate/pharmacology*

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

Burns often cause the damaged tissue to produce a large amount of exudate and the formation of blisters on the wound. The burn blister fluid contains a large number of molecules related to wound healing, which can reflect the state of local tissue microenvironment of the burn wound. Analyzing relevant information such as cellular components, signal mediators, and protein molecules in burn blister fluid is helpful to understand the local reaction and tissue microenvironment of burn wounds, and then help clinical burn treatment. In this article, by understanding the production mechanism of burn blister fluid, discussing its role in wound evaluation, and integrating the research progress of burn blister fluid in proteomics, metabolomics, cellular components, and pharmacokinetics, we propose our thoughts and prospects on the research of burn blister fluid, in order to provide assistance for clinical evaluation and treatment of burn wounds, and also provide idea for the follow-up study of burn blister fluid.
Humans ; Blister/metabolism* ; Follow-Up Studies ; Burns/metabolism* ; Exudates and Transudates/metabolism* ; Wound Healing

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

After one-month of oral treatment with traditional Chinese medicine decoction, without using other drugs, the lung inflammatory exudate, pulmonary fibrosis and quality of life of a 61-year-old female patient with corona virus disease 2019 (COVID-19) were significantly improved. No recurrence or deterioration of the patient's condition was found within seven weeks of treatment and follow-up, and no adverse events occurred, indicating that oral Chinese medicine decoction was able to improve the pulmonary inflammation and fibrosis in a patient recovering from COVID-19, but further research is still needed.
Administration, Oral ; COVID-19/virology* ; Drugs, Chinese Herbal/therapeutic use* ; Exudates and Transudates ; Female ; Humans ; Inflammation/etiology* ; Lung/pathology* ; Magnoliopsida ; Medicine, Chinese Traditional ; Middle Aged ; Phytotherapy ; Pulmonary Fibrosis/etiology* ; SARS-CoV-2

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

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