The aim of this study was to investigate the cytotoxicity of modified nonequilibrium plasma with chlorhexidine digluconate (CHX) on human gingival fibroblasts (HGFs), and to evaluate the biosecurity of modified nonequilibrium plasma with 2% CHX as a new method of root canal treatment. Tissue samples taken from human gingiva were primarily cultured and passaged. Cells from passages 3-7 were used. HGFs were treated by modified nonequilibrium plasma with 2% CHX for 0 min (control group), 30 s, 1 min, 1.5 min, 3 min, 5 min, and 10 min, respectively, and then they were incubated for 0, 24, and 48 h. After that, cell counting kit-8 (CCK-8) assay was applied to analyze the cytotoxicity of modified nonequilibrium plasma with 2% CHX on HGFs. There was no significant difference between the 0 h group treated with the modified nonequilibrium plasma for 1 min and the control group (P>0.05). However, there were significant differences between all the other treated groups and the control group (P<0.05). When treated for 1.5 min or shorter, the cell viability was obviously increased; while treated for 3 min or longer, it was obviously reduced. Moreover, when successively cultured for 0, 24, and 48 h, cell viability was decreased at first and then increased in the 3-min-treated and 5-min-treated groups. The modified nonequilibrium plasma with 2% CHX was of no influence on cell viability in 1.5 min treatment, and it could be safely used on root canal treatment.
Adolescent ; Adult ; Anti-Infective Agents, Local ; adverse effects ; toxicity ; Cell Survival ; drug effects ; Cells, Cultured ; Chlorhexidine ; adverse effects ; analogs & derivatives ; toxicity ; Fibroblasts ; drug effects ; Gingiva ; cytology ; Humans ; Plasma ; chemistry ; Root Canal Therapy ; instrumentation ; methods

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

OBJECTIVETo investigate the effect of cyclosporin A (CsA) and tumor necrosis factor-α (TNF-α) on cell proliferation of cultured human gingival fibroblasts (GF), and the relationship between gingival inflammation and drug-induced gingival overgrowth.

METHODSHuman GF were cultured in vitro using tissue culture method. then cells from the 4 - 8 th passage were used in the experiment. The cells were cultured and incubated with various concentrations of CsA and TNF-α (A: blank group, B1: 10 µg/L CsA, B2: 50 µg/L CsA, B3: 250 µg/L CsA, B4: 1250 µg/L CsA, C: 5 µg/L TNF-α, D1: 10 µg/L CsA + 5 µg/L TNF-α, D2: 50 µg/L CsA + 5 µg/L TNF-α, D3: 250 µg/L CsA + 5 µg/L TNF-α, D4: 1250 µg/L CsA + 5 µg/L TNF-α) solution for 3, 5 and 7 days. Methyl thiazolyl tetrazolium assay was used to evaluate the cell proliferation in the culture meidiun.

RESULTSThe proliferation of fibroblasts was inhibited when exposed to different concentration of CsA and A value decreased. There was no significant difference between group B1, B2, B3 and the control group, while the A value of group B4 was significantly higher than that of control group (P < 0.01). Fibroblast proliferation was significantly increased while cultured with 5 µg/L TNF-α. A value increased (P < 0.01). When exposed to CsA + TNF-α, A value of group D1, D2, D3 was much higher than that of group A, but was lower than that of group C (P < 0.05). Cell proliferation in group D4 was significantly increased, and significantly different with that in group C (P < 0.01).

CONCLUSIONSCsA did not stimulate the cell proliferation, and high concentration of CsA inhibited cell proliferation. TNF-α can stimulate the cell proliferation. High-concentration CsA + TNF-α can enhance the fibroblast proliferation, which suggests that CsA in certain concentration have amplification effect on TNF-α to stimulate fibroblast proliferation.

Adult ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cyclosporine ; adverse effects ; pharmacology ; Dose-Response Relationship, Drug ; Fibroblasts ; cytology ; Gingiva ; cytology ; Gingival Overgrowth ; chemically induced ; Humans ; Immunosuppressive Agents ; adverse effects ; pharmacology ; Tumor Necrosis Factor-alpha ; adverse effects ; pharmacology

Acetylcysteine ; pharmacology ; Antioxidants ; pharmacology ; Composite Resins ; toxicity ; Dental Materials ; toxicity ; Fibroblasts ; drug effects ; metabolism ; Gingiva ; cytology ; drug effects ; Humans ; Methacrylates ; toxicity ; Polyethylene Glycols ; toxicity ; Polymethacrylic Acids ; toxicity ; Polymethyl Methacrylate ; toxicity ; Reactive Oxygen Species ; metabolism ; Resins, Synthetic ; toxicity

PURPOSE: The purpose of this study was to examine the effects of using gauze frozen with normal saline or ice on thirst-relief and oral condition of laparoscopic cholecystectomy patients. METHODS: A quasi-experimental nonequivalent control group, pretest-posttest design was used. Participants (n=53) received either gauze frozen with normal saline (n=17), ice (n=18) or wet gauze (n=18) for thirst-relief. The subjective thirst level and oral condition of the participants were assessed before the intervention, 15 min after the first intervention and 15 min after the second intervention. RESULTS: After oral care was provided twice, there were significant differences in thirst level among the groups. When oral care was provided twice, the oral condition of tongue, saliva, mucosal membrane, and gingiva was improved in patients receiving gauze frozen with normal saline or ice. CONCLUSION: Gauze frozen with normal saline and ice can be effective for oral care in reducing the thirst level and improving the condition of the oral cavity.
Adult ; Aged ; *Cholecystectomy, Laparoscopic ; Female ; Freezing ; Gallbladder Diseases/*surgery ; Gingiva/drug effects ; Humans ; *Ice ; Male ; Middle Aged ; Mouth Mucosa/drug effects ; Pilot Projects ; Saline Solution, Hypertonic ; Saliva/physiology ; *Thirst/drug effects ; Tongue/drug effects

OBJECTIVETo observe the effects of Porphyromonas gingivalis (Pg), Actinobacillus actinomycetemcomitans (Aa), Escherichia coli (Ec) lipopolysaccharides (LPS) on the production of IL-11 and IL-6 from healthy human gingival fibroblasts (HGF), and the effects of endogenous prostaglandin on HGF IL-11 and IL-6 production stimulated with the above LPS.

METHODSHGF were stimulated with Pg-, Aa-, Ec-LPS of different concentrations (0.1, 1, 10 mg/L) for 24 h. And HGF were also stimulated with the combinations of 10 mg/L Pg-, Aa-, Ec-LPS and 10(-6) mol/L indomethacin respectively for 24 h. Levels of IL-11 and IL-6 in the supernatants were quantitated by ELISA.

RESULTSLPS from Aa, Ec, at the concentration of 10 mg/L and from Pg at the concentrations 1, 10 mg/L significantly augmented IL-11 production by HGF. IL-6 production was also significantly increased by stimulation with Aa-LPS at concentrations 1, 10 mg/L and with Ec-, Pg-LPS at concentrations 0.1, 1, 10 mg/L. In addition, IL-11 production was lower than IL-6 production by HGF stimulated with LPS. Indomethacin significantly inhibited IL-6 and IL-11 production in LPS-stimulated HGF.

CONCLUSIONSAa-, Pg-, Ec-LPS may significantly increase IL-11 and IL-6 level in the supernatants of HGF, and endogenous prostaglandin may upregulate IL-11 and IL-6 production in LPS-stimulated HGF.

Aggregatibacter actinomycetemcomitans ; chemistry ; Cells, Cultured ; Dose-Response Relationship, Drug ; Escherichia coli ; chemistry ; Fibroblasts ; drug effects ; metabolism ; Gingiva ; cytology ; metabolism ; Humans ; Indomethacin ; pharmacology ; Interleukin-11 ; biosynthesis ; Interleukin-6 ; biosynthesis ; Lipopolysaccharides ; pharmacology ; Porphyromonas gingivalis ; chemistry

OBJECTIVETo evaluate the effect of verapamil on the proliferation of normal gingival fibroblast (NGF) in vitro.

METHODSNGF was isolated and cultured. The 5th passage of NGF was incubated with 0, 0.1, 1, 10, 100 micromol/L verapamil respectively. Methyl thiazolyl tetrazolium (MTT) assay and flow cytometry were used to detect cell proliferation and cell cycles.

RESULTSIncubated with 100 micromol/L verapamil for 66 h, the A value of normal gingival fibroblast was significantly lower than those without verapamil groups (P < 0.01). Incubated with 100 micromol/L verapamil for 18 h, 69% of cells were at the G(0) - G(1) phase, 27% were at the S phase. For control group (without verapamil) 41% of cells were at G(0) - G(1) phase and 49% cells were at S phase. There was significant difference between the two groups (P < 0.001).

CONCLUSIONS100 micromol/L verapamil inhibited proliferation of normal gingival fibroblast by a cell-cycle arrest.

Calcium Channel Blockers ; pharmacology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Dose-Response Relationship, Drug ; Fibroblasts ; drug effects ; Gingiva ; cytology ; Humans ; Verapamil ; pharmacology

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 effects of hydroxyapatite (HA) and porous tricalcium phosphate/hydroxyapatite (TCP/HA)-coating titanium on the adhesion behavior of human gingival fibroblasts (HGFs).

METHODSCoatings of HA and duplex phases TCP/HA on titanium (Ti) were formed by ion beam assisted deposition (IBAD) method. Attachment, spreading, extracellular matrix (ECM) production, and focal adhesion plaque formation of HGFs were investigated on commercially pure (CP) titanium, HA-coated CP titanium and porous TCP/HA-coated CP titanium. After incubation of HGFs on these substrates, the number of attached cell, the area of cell spreading, immunostained ECM including fibronectin (FN) and type I collage, and vinculin (presenting the formation of focal adhesion plaque) were quantified by morphometric analysis using immunofluorescence microscope.

RESULTSTCP/HA and HA coatings exhibited that the attached cell number and cell spreading area were greater than those of CP titanium (P < 0.05), and the formation of focal adhesion plaque was earlier than that of uncoated substrate (P < 0.05). The number of attached cell and the formation of type I collagen on TCP/HA were more than those on Ti and HA. After 24-hour incubation on TCP/HA surface, the number of attached cell was 198.1 +/- 27.7 and the fluorescent intensity of type I collagen was 154.10 +/- 31.56. While under the same condition, the corresponding numbers for the CP titanium were 125.1 +/- 29.9 and 132.63 +/- 35.26. The differences between the two groups were significant (P < 0.05).

CONCLUSIONSIn this study, the porous TCP/HA coating significantly facilitated the adherence of human gingival fibroblasts to Ti surface and could improve the biocompatibility of titanium.

Calcium Phosphates ; pharmacology ; Cell Adhesion ; drug effects ; Cells, Cultured ; Coated Materials, Biocompatible ; chemistry ; Durapatite ; pharmacology ; Fibroblasts ; cytology ; drug effects ; Gingiva ; cytology ; Humans ; Materials Testing ; Titanium ; chemistry

OBJECTIVETo investigate what role IL-11 plays in periodontal disease and to determine the level of IL-11 in HGFs stimulated with IL-1alpha and TNF-alpha.

METHODSHGFs were stimulated with IL-1alpha and TNF-alpha alone or in combination. The production of IL-11 was measured using enzyme-linked immunosorbent assay (ELISA). IL-11 and Glyceraldehyde 3-Phosphate Dehydrogenase (GAPDH) messenger RNA (mRNA) levels in HGFs were determined by real-time reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSIL-1alpha significantly increased the levels of IL-11 in HGFs. TNF-alpha also significantly augmented IL-11 production in HGFs, and synergistically stimulated HGFs to produce IL-11 when combined with IL-1alpha. Indomethacin, an inhibitor of prostaglandin synthesis, significantly reduced IL-11 production by HGFs stimulated with IL-1alpha and TNF-alpha individually or in combination. IL-1alpha alone or combined with TNF-alpha enhanced the ratio of IL-11/GAPDH mRNA expression in HGFs, and the augmentation was abolished by indomethacin after co-incubation for 24 hs.

CONCLUSIONSProduction of IL-11 in HGFs stimulated with IL-1alpha and TNF-alpha was transcriptionally upregulated by the endogenous prostaglandin synthesis. Inhibition of prostaglandin might suppress the osteoclastogenesis by IL-11 in inflammatory periodontal diseases.

Cells, Cultured ; Cyclooxygenase Inhibitors ; pharmacology ; Fibroblasts ; drug effects ; metabolism ; Gingiva ; cytology ; metabolism ; Humans ; Indomethacin ; pharmacology ; Interleukin-11 ; biosynthesis ; Interleukin-1alpha ; pharmacology ; RNA, Messenger ; biosynthesis ; Tumor Necrosis Factor-alpha ; pharmacology