OBJECTIVESTo study the essence of N-terminal amino acid sequence of 12 000-protein in gingival crevicular fluid (GCF).

METHODSGCF samples from patients with RPP and AP were collected. 12 000-protein was separated by SDS-PAGE and transformed to PVDF by electronic transformation. The aim band was cut to be analyzed in 491 Protein Sequencer.

RESULTSThe first ten of N-terminal amino acid sequence of 12 000-protein in GCF was Met, Leu, Thr, Glu, Leu, Glu, Lys, Ala, Leu, Asn. Through checking up in MS-Edman, the sequence was similar to "Ca binding protein, MRP8" which is the light subunit of Calprotectin.

CONCLUSIONSCalprotectin is a major protein in granulocytes and monocytes, and is related to many inflammatory diseases, maybe served as a effective marker for evaluating the inflammation of periodontium.

Amino Acid Sequence ; Electrophoresis, Polyacrylamide Gel ; Gingival Crevicular Fluid ; chemistry ; Humans ; Leukocyte L1 Antigen Complex ; Periodontium

OBJECTIVETo establish a rapid and sensitive high-performance liquid chromatography (HPLC) method for detecting pazufloxacin concentrations in the saliva, gingival crevicular fluid and serum of healthy adults.

METHODSSamples of saliva, gingival crevicular fluid and serum were obtained from healthy adults receiving intravenous infusion of pazufloxacin. The concentrations of pazufloxacin in the samples were quantified by HPLC equipped with a reversed-phase column (Agilent Zorbax SB-C18 5 µm, 250 mm×4.6 mm). The mobile phase for pazufloxacin was a mixture of acetonitrile and 0.5% phosphoric acid containing 1% triethylamine (155:850), and 20 µl of the resulting solution was injected into the HPLC system at a flow rate of 1.0 ml/min. The detection wavelength was set at 245 nm. The samples were first deproteinized by precipitation with methanol followed by supernatant drying; the residue was reconstituted with the mobile phase and centrifuged, and the supernatants were directly injected into the HPLC system.

RESULTSPazufloxacin in the samples were totally separated without interference by any endogenous substances. The calibration curves showed a good linear regression (r>0.999). The detection limit was 10 ng/ml with within-day and between-day coefficients of variation performance all below 5% and recovery rates all above 91%.

CONCLUSIONHPLC is both sensitive and selective for quantification of pazufloxacin in saliva, gingival crevicular fluid and serum.

Adult ; Chromatography, High Pressure Liquid ; methods ; Female ; Fluoroquinolones ; analysis ; blood ; Gingival Crevicular Fluid ; chemistry ; Humans ; Male ; Oxazines ; analysis ; blood ; Saliva ; chemistry ; Sensitivity and Specificity ; Young Adult

OBJECTIVETo observe the clinical results and the changes of gingival crevicular fluid prostaglandins E(2) (GCF-PGE(2)) levels three months after indomethcin rinsing.

METHODSNineteen periodontal patients who had received periodontal treatment before were chosen and divided into two groups randomly: indomethcin test group and placebo control group. The clinical parameters and gingival crevicular fluid (GCF) samples were obtained respectively at 0, 1, 3 month. The levels of GCF-PGE(2) were measured by radioimmunoassay (RIA).

RESULTSAfter 3 months' rinse treatment, the bleeding index and the amounts of GCF in test group decreased significantly than those of the control group. Attachment level improved in test group as well. The levels of GCF-PGE(2) significantly decreased in test group, which not changed in control group. The percentage value of plaque reduced considerably in both test group and control group after rinsing, but no significant difference was found between these two groups.

CONCLUSIONGingival inflammation and the levels of GCF-PGE(2) are reduced after topical indomethcin administration. The effect is related to the decrease of local PGE(2) levels.

Anti-Inflammatory Agents, Non-Steroidal ; administration & dosage ; Dinoprostone ; analysis ; Double-Blind Method ; Gingival Crevicular Fluid ; chemistry ; Humans ; Indomethacin ; administration & dosage ; Periodontitis ; drug therapy

OBJECTIVE: Cytokines produced by various cells are strong local mediators of inflammation. Interleukin-1beta (IL-1β) and C-reactive protein (CRP) play essential roles in the development and progression of diabetes mellitus (DM). Thus periodontal diseases could be related to DM via the same mediators of inflammation. To evaluate plasma and gingival crevicular fluid (GCF) levels of IL-1β and CRP in adolescents with DM to further investigate whether DM has an impact on the levels of inflammation factors at an early stage, and to analyze the risk of developing periodontal diseases in adolescents with DM. METHODS: A total of 121 adolescents aged from ten to sixteen years were enrolled, 41 adolescents diagnosed with diabetes mellitus were collected in the DM group, and 80 nondiabetic adolescents as the control group. The periodontal indices of each individual were recorded, including plaque index (PLI), modified bleeding index (mBI), probing depth (PD) and attachment loss (AL). GCF and intravenous blood samples were collected, and CRP and IL-1β levels were detected by enzyme-linked immunosorbent assay (ELISA). RESULTS: (1) PLI of DM group and control group were 1.23±0.05 and 0.95±0.04 separately, with significant difference (P=0.001). DM group and control group had mBI of 0.80±0.08 and 0.51±0.06 separately, with significant difference (P=0.003). Attachment loss was found in none of the subjects. PDs of DM group and control group were (2.37±0.51) mm and (2.31±0.05) mm separately, and there was no significant difference. (2) CRP in GCF was only detectable in partial of the individuals, with a detectable rate of 22.9% (11/48) in total. The detectable rate of CRP in GCF was significantly higher in DM group (38.5%) than that in control group (4.5%, P=0.006). The plasma level of CRP in DM group [0.23 (0.15, 1.89) mg/L] was higher than that in control group [0.19 (0.12, 4.18) mg/L], but without significance (P=0.776). (3) The plasma levels of IL-1β in DM group and control group were (14.11±0.57) ng/L and (14.71±0.50) ng/L separately, but there was no significance (P=0.456). GCF levels of IL-1β in DM group and control group were (12.91±1.95) μg/L and (17.68±3.07) μg/L, without significant difference (P=0.185). CONCLUSION Periodontitis was not observed in adolescents with DM at an early stage. However, the rising levels of periodontal indices and CRP in GCF, might indicate that adolescents with DM have a higher risk of developing periodontal diseases in the future.
Adolescent ; C-Reactive Protein/analysis* ; Dental Plaque Index ; Diabetes Mellitus, Type 2 ; Disease Progression ; Enzyme-Linked Immunosorbent Assay ; Female ; Gingival Crevicular Fluid/chemistry* ; Humans ; Interleukin-1beta/analysis* ; Male ; Periodontal Diseases ; Periodontal Index ; Periodontitis ; Plasma

Periodontitis is an inflammatory autoimmune disease. Treatment should alleviate inflammation, regulate the immune reaction and promote periodontal tissue regeneration. Icariin is the main active ingredient of Epimedii Folium, and it is a promising compound for the enhancement of mesenchymal stem cell function, promotion of bone formation, inhibition of bone resorption, alleviation of inflammation and regulation of immunity. The study investigated the effect of icariin on periodontal tissue regeneration in a minipig model of periodontitis. The minipig model of periodontitis was established. Icariin was injected locally. The periodontal clinical assessment index, a computed tomography (CT) scan, histopathology and enzyme-linked immune sorbent assay (ELISA) were used to evaluate the effects of icariin. Quantitative analysis results 12 weeks post-injection demonstrated that probing depth, gingival recession, attachment loss and alveolar bone regeneration values were (3.72 ± 1.18) mm vs. (6.56 ± 1.47) mm, (1.67 ± 0.59) mm vs. (2.38 ± 0.61) mm, (5.56 ± 1.29) mm vs. (8.61 ± 1.72) mm, and (25.65 ± 5.13) mm vs. (9.48 ± 1.78) mm in the icariin group and 0.9% NaCl group, respectively. The clinical assessment, CT scan, and histopathology results demonstrated significant enhancement of periodontal tissue regeneration in the icariin group compared to the 0.9% NaCl group. The ELISA results suggested that the concentration of interleukin-1 beta (IL-1β) in the icariin group was downregulated compared to the 0.9% NaCl group, which indicates that local injection of icariin relieved local inflammation in a minipig model of periodontitis. Local injection of icariin promoted periodontal tissue regeneration and exerted anti-inflammatory and immunomodulatory function. These results support the application of icariin for the clinical treatment of periodontitis.
Animals ; Disease Models, Animal ; Enzyme-Linked Immunosorbent Assay ; Flavonoids ; administration & dosage ; pharmacology ; Gingival Crevicular Fluid ; chemistry ; Inflammation ; drug therapy ; Periodontitis ; diagnostic imaging ; drug therapy ; Swine ; Swine, Miniature ; Tomography, X-Ray Computed

AIMThe aim of this study was to measure the level of Oncostatin M (OSM) a gp130 cytokine in the gingival crevicular fluid (GCF) and serum of chronic periodontitis patients and to find any correlation between them before and after periodontal therapy (scaling and root planing, SRP).

METHODOLOGY60 subjects (age 25-50 years) were enrolled into three groups (n=20 per group), group I (healthy), group II (gingivitis) and group III (chronic periodontitis). Group III subjects were followed for 6-8 weeks after the initial periodontal therapy (SRP) as the group IV (after periodontal therapy). Clinical parameters were assessed as gingival index (GI), probing depth (PD), clinical attachment level (CAL), and radiographic evidence of bone loss. GCF and serum levels of OSM were measured by using Enzyme Linked Immunosorbent Assay (ELISA).

RESULTSIt was found that mean OSM levels had been elevated in both the GCF and serum of chronic periodontitis subjects (726.65 +/- 283.56 and 65.59 +/- 12.37 pg mL(-1), respectively) and these levels were decreased proportionally after the periodontal therapy (95.50 +/- 38.85 and 39.98 +/- 16.69 pg mL(-1) respectively). However, OSM was detected in GCF of healthy subjects (66.15 +/- 28.10 pg mL(-1)) and gingivitis-suffering subjects (128.33 +/- 22.96 pg mL(-1)) and was found as below the detectable limit (approximately equal 0.0 pg mL(-1)) in the serum of same subjects. Significant correlation has been found between clinical parameters and GCF-serum levels of OSM.

CONCLUSIONIncreased OSM level both in the GCF and serum, and the decreased levels after initial periodontal therapy (SRP) may suggest a use as an inflammatory biomarker in the periodontal disease.

Adult ; Analysis of Variance ; Case-Control Studies ; Chronic Periodontitis ; blood ; metabolism ; therapy ; Dental Scaling ; Female ; Gingival Crevicular Fluid ; chemistry ; Gingivitis ; blood ; metabolism ; therapy ; Humans ; Male ; Middle Aged ; Oncostatin M ; analysis ; blood ; metabolism ; Periodontal Index ; Statistics, Nonparametric