OBJECTIVETo investigate the expression of interleukin-10 (IL-10) mRNA in gingival tissue of active and stable stage in patients with adult periodontitis.

METHODS12 patients with acute abscesses of the periodontium, 12 patients after periodontal initial treatment and 6 periodontal healthy patients having extraction of impacted wisdom tooth were randomly divided into group A (active stage group), group B (stable stage group) and the control group. Biopsies of gingival tissues were collected from every subject of three groups. Technique of in situ hybridization was applied to observe the expression of IL-10 mRNA in the biopsies from three groups semi-quantitatively.

RESULTSIL-10 mRNA was positively expressed in lymphocytes, macrophages and fibroblasts. The quantity of IL-10 mRNA of group A was the lowest in the three groups and was significantly lower than that of control group and group B respectively (P < 0.01). The quantity of IL-10 mRNA of group B was the highest in the three groups and was significantly higher compared with the control group and group A (P < 0.01).

CONCLUSIONThe quantities of IL-10 mRNA expression are closely related with various clinical states of periodontitis.

Case-Control Studies ; Chronic Periodontitis ; metabolism ; Gingiva ; metabolism ; Humans ; Interleukin-10 ; metabolism ; RNA, Messenger ; metabolism

OBJECTIVETo investigate the expression of cytokeratins (CK) in the normal human gingival epithelium and to explore the difference between junctional epithelium (JE), oral epithelium (OE) and sulcular epithelium (SE).

METHODSTeeth specimens with gingival tissue were collected from 5 people. Paraffin-embedded sections were stained with monoclonal antibodies responded respectively to human CK5/6, 7, 8/18, 10/13, 16, 17, 19, 20.

RESULTSCK7 and 17 was not expressed in all strata of JE, OE and SE. CK5/6 and 20 were weekly or moderately expressed in the suprabasal, and not expressed in the basal layer of all three epithelia. CK10/13 and 16 were positive in all strata of JE and in the suprabasal layers of OE and SE. CK10/13 was moderately to strongly expressed and CK16 was weekly to moderately expressed. The staining for CK19 was intense in all strata of JE and the basal layer of OE and SE. There was a remarkable demarcation between JE and SE. The pattern of CK8/18 expression was similar to that of CK19, but was weaker. Besides the basal layer, some suprabasal layers close to the basal layer were stained.

CONCLUSIONSJE is an unique non-differentiated stratified epithelium different from OE and SE. CK19 would be a histological marker and CK10/13, 16 would be the cellular markers to differentiate JE from OE and SE.

Epithelial Attachment ; cytology ; metabolism ; Gingiva ; metabolism ; Humans ; Keratins ; metabolism ; Mouth Mucosa ; metabolism

OBJECTIVES: This study was performed to clarify the effects of sitagliptin on METHODS: Healthy gingival samples were collected from the donors. HGFs were isolated with enzymic digestion method and identified. The effects of LPS and sitagliptin on cell viability were detected by cell-counting kit-8 (CCK8). The mRNA levels of inflammatory cytokines, namely, interleukin (IL)-6, IL-8, C-C motif ligand 2 (CCL2), and superoxide dismutase 2 (SOD2), were evaluated by quantity real-time polymerase chain reaction (qRT-PCR) and enzyme-linked immune sorbent assay (ELISA) was used to measure the secretion protein levels of IL-6, IL-8, and CCL2. Western blot analysis was used to further investigate the activation of nuclear factor (NF)-κB signaling pathway. The effect of NF-κB pathway inhibitor BAY11-7082 on LPS-induced HGF inflammatory cytokines at the gene level was verified by qRT-PCR. RESULTS: Low concentrations of sitagliptin (0.1, 0.25, and 0.5 µmol·L CONCLUSIONS Sitagliptin could significantly inhibit LPS-induced HGF inflammatory response by blocking the NF-κB signaling pathway activation.
Fibroblasts ; Gingiva/metabolism* ; Humans ; Lipopolysaccharides ; NF-kappa B/metabolism* ; Signal Transduction ; Sitagliptin Phosphate

OBJECTIVETo study the effects and the therapeutic mechanism of hyperbaric oxygen (HBO) on prostaglandin E(2) (PGE(2)) in alveolar bone and gingiva of experimental periodontitis in animal.

METHODSExperimental periodontitis was produced by silk thread sutures combined with high content sugar diet. For HBO therapy, they were exposed to a pressure of 0.25 MPa (2.5ATA), breathing pure oxygen one session a day for 60 min. The treatment course was 2 weeks. The value of PGE(2) in gingiva and alveolar bone was analyzed by enzyme immunoassay (EIA).

RESULTSThe value of PGE(2) in gingiva of control group was 3.21 ng/g, and that of PGE(2) in alveolar bone was 3.22 ng/g. The contents of PGE(2) in gingiva (13.96 ng/g) and alveolar bone (13.32 ng/g) of periodontitis group increased markedly than control group (P < 0.01). The contents of PGE(2) in gingiva (5.21 ng/g) of HBO group were 62.7% which was lower than that of periodontitis group, and the value of PGE(2) in alveolar bone (4.05 ng/g) were 69.6% lower than that of periodontitis group. The difference of PGE(2) in gingiva or alveolar bone was significant for the HBO group and periodontitis group (P < 0.01).

CONCLUSIONSThe contents of PGE(2) in alveolar bone and gingiva increased markedly when experimental periodontitis has formed. The value of PGE(2) in alveolar bone and gingiva reduce markedly after HBO exposure, and the decreased rate of PGE(2) in alveolar bone is more evident than that of PGE(2) in gingiva after HBO therapy.

Alveolar Process ; metabolism ; Animals ; Dinoprostone ; metabolism ; Disease Models, Animal ; Female ; Gingiva ; metabolism ; Guinea Pigs ; Hyperbaric Oxygenation ; Male ; Periodontitis ; metabolism

OBJECTIVE: To observe the effects of xipayi mouth rinse on the DNA synthesis and change of cell cycles of human gingival fibroblast (HGF) induced by lipopolysaccharide(LPS). METHODS: HGF was stimulated with LPS at 25 mg/L. Flow cytometry was used to examine the effect of xipayi mouth rinse at 25 mg/L on the DNA synthesis and change of HGF cell cycles. RESULTS: The percentage of HGF in G( 1) phase increased after the cells were induced by LPS, while the percentage of HGF in S phase decreased. Xipayi mouth rinse could ameliorate this phenomenon. CONCLUSION Xipayi mouth rinse can significantly ameliorate the inhibitory effect of LPS on the proliferation of HGF, suggesting the anti-inflammatory effect of xipayi mouth rinse in the treatment and prevention of periodontal diseases.
Cell Cycle ; drug effects ; DNA ; biosynthesis ; Drugs, Chinese Herbal ; pharmacology ; Fibroblasts ; cytology ; metabolism ; Gingiva ; cytology ; Humans ; Lipopolysaccharides ; Mouthwashes ; pharmacology

OBJECTIVETo investigate the effect of alloy leaching liquor of four different types of base metal alloy on the expression of prostaglandin E(2) (PGE(2)) and cyclo-oxygenase-2(COX-2) by human gingival fibroblast(HGF) in vitro.

METHODSNi-Cr, Co-Cr, pure Ti and Au ceramic alloys were incubated in Dulbecco's modified Eagle's medium (DMEM) to prepare alloy leaching liquor, and then added in HGF medium. DMEM was prepared as negative control. Aliquots were taken from exposed media after 1, 6, 12, 24 h. Assays for PGE(2) were carried out by enzyme-linked immunosorbent assay (ELISA).

RESULTSIn 6, 12, 24 h, the expression of PGE(2) in Ni-Cr and Co-Cr alloy groups (Ni-Cr: 45.568 ± 0.926, 60.538 ± 0.988, 73.754 ± 0.507; Co-Cr: 40.496 ± 0.693, 53.216 ± 0.327, 65.470 ± 1.086) were significantly higher than those in other experimental groups (Ti: 31.564 ± 0.719, 31.998 ± 0.856, 32.066 ± 0.513; Au alloy: 31.540 ± 0.821, 31.136 ± 0.518, 31.340 ± 0.443) and control group (31.122 ± 0.642, 31.230 ± 0.634, 30.980 ± 0.746) (P < 0.05). No significant difference were found in the expression of PGE(2) among pure Ti, Au alloy groups and the control group (P > 0.05). Immunofluorescence showed dark and uniform COX-2 stain in Ni-Cr and Co-Cr alloy groups, while in pure Ti group, Au alloy group, and negative control group shallow and uneven distribution of COX-2 stain were observed.

CONCLUSIONSOur findings suggested that pure Ti and Au alloy did not cause elevated PGE(2) and COX-2 release from HGF. However, Ni-Cr and Co-Cr alloy caused increase in PGE(2) and COX-2 levels.

Cells, Cultured ; Chromium Alloys ; adverse effects ; Cyclooxygenase 2 ; metabolism ; Dental Alloys ; adverse effects ; Dinoprostone ; metabolism ; Fibroblasts ; cytology ; metabolism ; Gingiva ; cytology ; Gold Alloys ; adverse effects ; Humans ; Titanium ; adverse effects

OBJECTIVEThis study investigates the immune status of T helper (Th) 17 cells in mice with periodontitis.

METHODSSeven-week-old C57BL/6 female mice were randomly divided into the control and periodontitis groups. Each group comprisedfour mice. Experimental periodontitis was induced in mice by oral infection with Porphyromonas gingivalis in the periodontitis group. Phosphate-buffered saline solution was used in the control group. All mice were sacrificed 4 weeks after the last P. gingivalis infection. CD4⁺retinoid-related orphan receptor (ROR) γτ⁺(Th17) cells were analyzed by flow cytometry. The protein expression of Th17 cell-related cytokine interleukin (IL)-17A was detected by enzyme-linked immunosorbentassay (ELISA).

RESULTSFlow cytometry showed that the percentage of CD4⁺RORγτ⁺(Thl7) cells in all CD4⁺ cells and the cell number in gingival tissues, cervical lymph nodes (CLNs), and the peripheral blood obviously increased (P < 0.01) in the periodontitis group. ELISA showed that compared with the control group, the periodontitis group exhibited an obvious increase in the protein expression of IL-17A (P < 0.05).

CONCLUSIONTh17 cell-mediated cell response is enhanced, and the gingival tissues, CLNs, and the peripheral blood are probably the main locations of Thl7 cell-mediated cell response during the development of periodontitis.

Alveolar Bone Loss ; Animals ; Cytokines ; Female ; Flow Cytometry ; Gingiva ; Interleukin-17 ; metabolism ; Mice ; Mice, Inbred C57BL ; Periodontitis ; metabolism ; Porphyromonas gingivalis ; Random Allocation ; T-Lymphocytes, Helper-Inducer

OBJECTIVE: To observe the effects of xipayi mouth rinse of different concentrations on the activities of interleukin-6 (IL-6) from human gingival fibroblast (HGF) induced by lipopolysaccharide (LPS). METHODS: HGF was stimulated with LPS at 25 g/mL, and the radioimmunoassay (RIA) was used to examine the effect of xipayi mouth rinse at 12.5 approximately 200 g/mL on the secretion of IL-6 in the supernatant of the cell culture. RESULTS: IL-6 secreted by human gingival fibroblast was significantly inhibited by xipayi mouth rinse in a dose dependent manner. CONCLUSION Xipayi mouth rinse can inhibit the secretion of IL-6 from HGF induced by LPS, suggesting the anti-inflammatory effect of xipayi mouth rinse to treat and prevent periodontal diseases.
Anti-Inflammatory Agents ; pharmacology ; Cells, Cultured ; Depression, Chemical ; Drugs, Chinese Herbal ; pharmacology ; Fibroblasts ; metabolism ; Gingiva ; cytology ; Humans ; Interleukin-6 ; metabolism ; Lipopolysaccharides ; Mouthwashes

OBJECTIVEThis study aimed at investigating effects of exogenous interleukine-10 (IL-10) on IL-6 and intercellular adhesion molecular (ICAM-1) expression in inflamed gingival tissue.

METHODSThe expression of IL-6 and ICAM-1 was examined using immunohistochemical techniques. Inflammatory gingival tissue treated with IL-10 was taken as the experimental group and the same patient's inflammatory gingival tissue without treatment of IL-10 was included into the control group.

RESULTSIL-6 expression was found mainly in monocytes, macrophages, lymphocytes, endotheliocytes and fibroblasts. The expression of ICAM-1 was found mainly in epithelial cells, monocot-macrophages, lymphocytes, endotheliocytes and fibroblasts. The immunohistochemical optical density (IOD) of the expression of IL-6 and ICAM-1 was detected by using Image-Proplus software, and the results showed that the expression in the experimental group differed significantly from that in the control group.

CONCLUSIONThe exogenous IL-10 may down-regulate IL-6 and ICAM-1 expression in inflammatory gingival tissue.

Adult ; Down-Regulation ; Female ; Gingiva ; metabolism ; Gingivitis ; metabolism ; Humans ; Intercellular Adhesion Molecule-1 ; biosynthesis ; genetics ; Interleukin-10 ; pharmacology ; Interleukin-6 ; biosynthesis ; genetics ; Male ; Middle Aged

This study used in vivo microdialysis to examine the effects of intragingival application of lipopolysaccharide (LPS) derived from Porphyromonas gingivalis (Pg-LPS) on gingival tumour necrosis factor (TNF)-α and interleukin (IL)-6 levels in rats. A microdialysis probe with an injection needle attached to the surface of the dialysis membrane was implanted into the gingiva of the upper incisor. For comparison, the effects of LPS derived from Escherichia coli (Ec-LPS) on IL-6 and TNF-α levels were also analysed. Pg-LPS (1 μg/1 μL) or Ec-LPS (1 or 6 μg/1 μL) was applied by microsyringe, with gingival dialysates collected every hour. Enzyme-linked immunosorbent assay (ELISA) revealed that gingival dialysates contained approximately 389 pg·mL⁻¹ of IL-6 basally; basal TNF-α levels were lower than the detection limit of the ELISA. Pg-LPS failed to alter IL-6 levels but markedly increased TNF-α levels, which remained elevated for 2 h after treatment. Neither IL-6 nor TNF-α were affected by Ec-LPS. Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis revealed that the gingiva expresses Toll-like receptor (TLR) 2 and TLR4 mRNA. Immunohistochemical examination showed that TLR2 and TLR4 are expressed by gingival epithelial cells. The present study provides in vivo evidence that locally applied Pg-LPS, but not Ec-LPS, into the gingiva transiently increases gingival TNF-α without affecting IL-6. The present results suggest that TLR2 but not TLR4 expressed on gingival epithelial cells may mediate the Pg-LPS-induced increase in gingival TNF-α in rats.
Animals ; Gingiva ; drug effects ; metabolism ; Interleukin-6 ; metabolism ; Lipopolysaccharides ; administration & dosage ; Male ; Porphyromonas gingivalis ; metabolism ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Toll-Like Receptor 2 ; genetics ; metabolism ; Toll-Like Receptor 4 ; genetics ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism