OBJECTIVETo investigate the effect of human serum albumin (HSA) on cell attachment of human gingival epithelial cells (HGE).

METHODSHGE were primary cultured with keratinocyte serum-free medium (KSFM) and dispase. The cultured cells were immunohistochemically stained by monoclonal anti-pan cytokeratin. MTT test was employed to investigate the influence of HSA on the cell attachment on polystyrene surface. The cell growth curve of HGE which were cultured in KSFM with 50 g/L HSA was observed.

RESULTSThe results showed significant decrease in cell numbers within 8 hours after HGE were inoculated, in which the polystyrene surface was preincubated with 50 g/L HSA. But it did not prove to be the case from 10 hours to 24 hours after HGE were inoculated. There were no significant difference within 24 hours in cell numbers between cultured in KSFM with 50 g/L HSA and control. The cell numbers in cell growth curve of HGE in KSFM with and without 50 g/L HSA did not show significant difference.

CONCLUSIONSHSA preincubation on polystyrene were produce inhibitory effect of HGE attachment in early stage.

Cell Adhesion ; drug effects ; Cell Count ; Cell Division ; drug effects ; Cells, Cultured ; Epithelial Cells ; cytology ; drug effects ; Gingiva ; cytology ; drug effects ; Humans ; Polystyrenes ; Serum Albumin ; pharmacology

OBJECTIVETo investigate the effects of advanced oxidative protein products (AOPP) on the proliferation, apoptosis and matrix metalloproteinase-1 (MMP-1) synthesis of the human gingival fibroblast (HGF). To explore the possible mechanism of the periodontal destruction acceleration in diabetes through AOPP-mediated oxidative stress.

METHODSHGF were isolated by both tissue explant cultivation technique and enzyme digestion method. The culture media with 5, 50, 100 mg/L AOPP-HAS were added into each experimental group, but the culture media in the control group didn't contain AOPP-HAS. MTT colorimetric assay and ELISA were used to measure the changes of HGF proliferation and the levels of MMP-1 protein from HGF at different time periods, respectively. Seventy-two hours after co-culture with 50 mg/L AOPP-HSA, cell apoptosis was detected by flow cytometry with Annexin V/PI staining.

RESULTSCompared to the control group, the growth inhibition rate of HGF in 5, 50, 100 mg/L AOPP-HSA group was significantly different (P < 0.05). The peak value appeared at 48 hours of co-culture [(19.01 +/- 6.28)%, (30.48 +/- 5.75)%, (39.75 +/- 4.60)%, respectively]. There was a dose-dependent relationship between the growth inhibition rate and AOPP-HSA. No significant difference was detected on the apoptotic level between experimental group and the control (P > 0.05). The MMP-1 synthesis in 0.5, 5, 50, 100 mg/L AOPP-HAS group [(55.61 +/- 1.06), (65.78 +/- 4.04), (79.24 +/- 3.09), (89.76 +/- 28.88) mg/L, respectively] was significantly higher than that in the control [(34.90 +/- 3.15) mg/L] after 72 hours co-culture (P < 0.05). There was a dose-dependent relationship between MMP-1 and AOPP-HSA.

CONCLUSIONSAOPP may inhibit the proliferation of HGF and such effect was not achieved through apoptosis. AOPP may increase collagen degradation by promoting MMP-1 synthesis and thus may accelerate periodontal destruction process in diabetes.

Apoptosis ; drug effects ; Blood Proteins ; metabolism ; Cell Proliferation ; drug effects ; Cells, Cultured ; Gingiva ; cytology ; drug effects ; Humans ; Matrix Metalloproteinase 1 ; metabolism ; Oxidation-Reduction ; Oxidative Stress

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 the effect of baicalin on the experimental periodontitis in rats, as well as the expression of MMP-1, MMP-2, MMP-9.

METHODSTwenty-seven adult male Sprague-Dawley rats were divided into three groups, with 9 rats in each group. A nylon thread was placed around the lower first molars of rats, which were sacrificed after 7 days. Baicalin (200 mg/kg) was administered to the experimental group by oral gavage, starting one day before the induction of periodontitis. The negative control group received vehicle (0.5% carboxymethylcellulose) alone. The blank control group did not get induction of periodontitis. The alveolar bone loss (ABL) and the area fraction (AA% ) occupied by collagen fibers were assessed. MMP-1, MMP-2 and MMP-9 protein expressions in the gingiva were detected by immunohistochemistry.

RESULTSBaicalin treatment significantly decreased ABL compared with the negative control group (P = 0.009). AA% of collagen fibers was significantly higher in baicalin-treated group than in the negative control group (P = 0.047). Baicalin treatment significantly down-regulated the protein expression for MMP-1 (P = 0.023) and MMP-9 (P = 0.042) and decreased the expression for MMP-2 (P = 0.099) compared with the negative control group.

CONCLUSIONSBaicalin protects against tissue damage in ligature-induced periodontitis in rats, which might be mediated in part by its inhibitory effect on the expression of MMP-1, MMP-2 and MMP-9.

Animals ; Flavonoids ; pharmacology ; Gingiva ; drug effects ; metabolism ; Male ; Matrix Metalloproteinases ; metabolism ; Periodontitis ; metabolism ; Rats ; Rats, Sprague-Dawley

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

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

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

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

OBJECTIVETo apply the synthesized advanced glycation end products (AGE) to the cultured human gingival fibroblast (HGF) in vitro and then to investigate the effects of AGE on the HGF proliferation and type I collagen synthesis and the potential impact of AGE in the repair of periodontium and its molecular mechanism in diabetes-associated periodontitis.

METHODSThe HGF was obtained from explants of human healthy gingival tissues by using tissue-explant technique. The AGE was prepared and then added to the culture media, its effect on HGF proliferation at different time duration was examined with MTT colorimetric assay. The type I collagen concentrations in cell culture supernatants and intracellular proteins were detected by ELISA, and the type I collagen mRNA expression of HGF was analyzed by real-time RT-PCR.

RESULTS200 mg/L AGE decreased the A value (P < 0.05) and changed the HGF shape. Incubation of HGF with AGE for 72 hours, the quantities of type I collagen were reduced (P < 0.05), and the expression of type I collagen mRNA was down-regulated (P < 0.05).

CONCLUSIONSThe AGE inhibited the HGF proliferation, decreased the synthesis of type I collagen and down-regulated the expression of type I collagen mRNA, impairing the repair of periodontium.

Cell Proliferation ; drug effects ; Cells, Cultured ; Collagen Type I ; biosynthesis ; Fibroblasts ; cytology ; metabolism ; Gingiva ; cytology ; metabolism ; Glycation End Products, Advanced ; pharmacology ; Humans