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

Oral lichen planus (OLP) is a chronic inflammatory disease that is frequently detected in oral tissues. The aim of our study was to identify the prevalence of the detection of periodontopathogenic microorganisms (Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythia and Treponema denticola in OLP patients and to compare with this prevalence of periodontopathogenic microorganisms in healthy non-OLP patients. Our study included 27 (18 chronic periodontitis (OLPP) and 9 gingivitis (OLPG)) patients diagnosed with OLP along with 26 (13 chronic periodontitis (HP) and 13 gingivitis (HG)) healthy non-OLP patients. The multiplex polymerase chain reaction (PCR) with subsequent reverse hybridization method (micro-IDent) was used for identifying periodontopathogenic microorganisms present in subgingival plaque samples. The percentages of detection for A. actinomycetemcomitans, P. gingivalis, P. intermedia, T. forsythia and T. denticola in subgingival plaque samples taken from OLP patients (OLPG and OLPP) were 18.5%, 85.1%, 81.4%, 88.8% and 74%, respectively. Meanwhile, in the non-OLP patients (HG and HP), these values were 7.6%, 50%, 46.1%, 73% and 57.7%, respectively. Thus, comparing the non-OLP groups with the OLP groups, the periodontopathogens' percentages of detection in the OLP groups were higher than those in the non-OLP groups. According to our study results, OLP patients have higher levels of infection with A. actinomycetemcomitans, P. gingivalis, P. intermedia, T. forsythia and T. denticola than non-OLP patients. We argue that the high percentages in patients with OLP may help identify the importance of periodontopathogenic microorganisms in the progress of periodontal diseases of OLP.
Actinobacillus Infections ; diagnosis ; Adult ; Aggregatibacter actinomycetemcomitans ; isolation & purification ; Bacteroidaceae Infections ; diagnosis ; Bacteroides ; isolation & purification ; Bacteroides Infections ; diagnosis ; Chronic Periodontitis ; microbiology ; Dental Plaque ; microbiology ; Dental Plaque Index ; Female ; Gingivitis ; microbiology ; Gram-Negative Bacteria ; isolation & purification ; Humans ; Lichen Planus, Oral ; microbiology ; Male ; Middle Aged ; Periodontal Attachment Loss ; microbiology ; Periodontal Index ; Periodontal Pocket ; microbiology ; Porphyromonas gingivalis ; isolation & purification ; Prevotella intermedia ; isolation & purification ; Treponema denticola ; isolation & purification ; Treponemal Infections ; diagnosis

OBJECTIVETo investigate an effective purification method for removing endotoxin from Prevotella intermedia.

METHODSThe main protein ingredients of bacteria prepared from ammonium sulfate precipitation were further treated with octyl phenyl polyoxyethylene ether (Triton X-114), and then processed at 4°C, 37°C and 25°C. The obtained aqueous phase after at least two more cycle repeated operations was assayed for endotoxin by Western blotting, LAL-clotting method, in vitro cell stimulation and in vivo animal experiments.

RESULTSWestern blotting and LAL-clotting method demonstrated that the reduction in endotoxin level was greater than 99.99% and recovery of the proteins after endotoxin removal was greater than 90% with Triton X-114 treatment for 3 cycles. The cytokines expression level was lower in both in vitro cell stimulation and in vivo animal experiments than in untreated group (P < 0.05).

CONCLUSIONSThe extraction method provides a new choice for endotoxin removal from large volumes of the oral Prevotella intermedia.

Animals ; Bacterial Proteins ; isolation & purification ; Endotoxins ; isolation & purification ; Female ; HEK293 Cells ; Humans ; Interleukin-1alpha ; blood ; Interleukin-6 ; blood ; Interleukin-8 ; metabolism ; Lipopolysaccharides ; pharmacology ; Mice ; Mice, Inbred C57BL ; Polyethylene Glycols ; chemistry ; Prevotella intermedia ; chemistry ; metabolism ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVETo extract the effective ingredient (crystal I) from effective section (saponin) of Ligustrum Lucidum Ait, identify the chemical structure of crystal I, study the effect of crystal I on P. gingivalis, B. forsythus and P. intermedia.

METHODSIsolated crystal I from saponin using the silica gel column chromatograph. Identified crystal I with IR spectra, (1)H-NMR and (13)C-NMR. Measured the minimal inhibitory concentration (MIC) and the minimal bactericidal concentration (MBC) through micro-liquid dilution. Studied the killing curve of ursolic acid on B. forsythus and P. intermedia.

RESULTSThe crystal I was identified as ursolic acid; its MIC and MBC to P. gingivalis, B. forsythus and P. intermedia were 0.740 and 0.295 microg/L respectively. The killing curve indicated that 0.800 microg/L ursolic acid could kill P. intermedia and B. forsythus in 3 and 6 hours respectively.

CONCLUSIONUrsolic acid has obvious effect to inhibit periodontal pathogen.

Bacteroides ; drug effects ; Cell Division ; drug effects ; Ligustrum ; Magnetic Resonance Spectroscopy ; methods ; Microbial Sensitivity Tests ; Periodontium ; microbiology ; Plant Extracts ; isolation & purification ; pharmacology ; Porphyromonas gingivalis ; drug effects ; Prevotella intermedia ; drug effects ; Triterpenes ; pharmacology