Increased levels of soluble Fcγ receptor III in gingival fluid from periodontal lesions

Enzyme-linked immunosorbent assay was used for determination of the concentration of soluble Fc gamma receptor III (Fc gamma RIII) in 40 samples of gingival fluid obtained from periodontal pockets in 30 patients with periodontitis. The assay was based on a monoclonal immobilized antibody binding Fc gamma RIII and a polyclonal Fc gamma RIII rabbit antibody for its quantification. The results indicate a substantially increased concentration of soluble Fc gamma RIII in gingival fluid as compared to the serum level. This increased concentration of soluble Fc gamma RIII may interfere with phagocytosis and immune homeostasis in the periodontal lesions.     

Soluble Fcγ receptors in periodontal lesions

Soluble Fcγ-binding components were detected in gingival fluid from periodontal lesions by incubation with biotinylated human Fcγ fragments. FcγIII receptor was identified by incubation of gingival fluid with monoclonal antibody. Sodium dodecyl sulfate-polyacrylamide gel electophoresis and Western transfer showed that most of the Fcγ-binding components had minimal mobility in a 4–15% gradient gel under nonreducing conditions. Under reducing conditions, the main band of Fcγ-binding components in gingival fluid migrated corresponding to protein A of 49 kDa. The pattern of Fcγ-binding components was similar in serum and gingival fluid except for the observation in gingival fluid of Fcγ-binding components migrating like standard proteins of 19 to 20 kDa, a size that corresponds to the polypeptide part of FcγII receptor and FcγIII receptor.     

Serum IgG2 level, Gm(23) allotype and FcγRIIa and FcγRIIIb receptors in refractory periodontal disease

Abstract. The purpose of this investigation was to compare the levels of serum IgG2, the frequency of detection of Gm(23)-negative allotype and frequency of detection of FcγRIIa and FcγRIIIb receptor haplotypes in 32 refractory, 54 successfully treated and 27 periodontally healthy individuals. Refractory subjects showed mean full mouth attachment loss and/or > 3 sites with attachment loss > 2.5 mm within 1 year after both scaling and root planing, and surgery plus systemically administered tetracycline. Successfully treated subjects showed mean attachment level gain and no sites with attachment loss > 2.5 mm 1 year post-therapy. Periodontally healthy subjects exhibited no pocket depth or attachment level > 3 mm. and no evidence of progressing disease during 1 year of monitoring. Blood was obtained from each subject al baseline. Serum IgG2 and Gm(23) allotype were determined using radial immunodiffusion. DNA was extracted from whole blood and the FcγR genotypes determined using PCR and allele specific oligonucleotide probes. Significance of differences among clinical groups were sought using the Kruskal-Wallis or chi-square tests. Associations between 2 or more variables were tested using regression analysis. Refractory subjects exhibited higher mean attachment loss and pocket depth than successfully treated or periodontally healthy subjects. Smoking status did not differ significantly among groups. No significant differences in serum IgG2 levels and frequency of detection of Gm(23)-negative allotype were observed among the clinical groups. Serum IgG2 level was positively associated with the number of serum antibody responses to subgingival species (r-0.51.P<0.001). Subjects with the Gm(23)- negative allotype exhibited lower mean levels of serum lgG2 (3.06±0.3 versus 3.9±0.2. p<0.01) and mean number of serum antibodies to subgingival species (17.7±1.7 versus 23.3±1.4, p<0.05) than allotype positive individuals. No significant differences in FcγR haplotype distribution were observed among the 3 clinical groups. Associations of serum IgG2 level. Gm(23) allotype. FcγRIIa and FcγRIIIb receptor haplotypes and smoking status were weakly related or not related to clinical status. This lack of relationship may have been due to a reality of no relationship, or the inadvertent pooling of subjects where these factors were of primary importance with subjects in whom these factors played a less important role.     

Epithelial expression of HLA class II antigens and Feγ receptors in patients with adult periodontitis

Abstract The distribution of HLA class II (DR, DP, DQ) and FcγR (I, II, III) was analyzed in the epithelia of patients with advanced marginal periodontitis using cryostat sections incubated with monoclonal antibodies (MoAb) against the Langerhans cell (LC) (CDla) and various subtypes of HLA class II and FcγR, and the indirect immunofluorescence technique. In the oral gingival epithelium (OGE), LC were concentrated subjacent to the connective tissue papillae, while in the pocket epithelium (PE), they were most abundant at the gingival margin. HLA-DP, DQ, and DR stained LC in both OGE and PE. HLA-DQ+ LC were significantly fewer than DP+ and DR+ LC. HLA-DR also stained keratinocytes (KC) in the whole extension of both OGE and PE. HLA-DP was also observed on KC, but not HLA-DQ. FcγR II stained both LC and focal areas of KC. In PE FCγR II+ LC were concentrated near the bottom of the pocket, while in the OGE, they were concentrated at the gingival margin. FcγR III was present only on KC, especially in the basal and suprabasal layer. The results indicate that the epithelial cells are actively involved in the development and maintenance of the inflammation of periodontal disease.     

Feγ-binding bacteria in periodontal lesions

The proportion of bacteria exhibiting surface Feγ-binding proteins was determined in periodontal pockets of 20 patients diagnosed with periodontal disease and in subgingival areas of 20 patients without periodontal lesions. Bacterial smears were examined by fluorescence microscopy based on DNA staining (Hoechst 33256) and staining of Feγ-binding proteins by human biotin-labelled Feγ and Texas red-conjugated streptavidin. Feγ-binding proteins were observed in all smears from the patients diagnosed with periodontitis, and in a majority of the smears high proportions of the bacteria were positive for Feγ-binding proteins. In contrast, most smears from patients without periodontal lesions included low or undetectable proportions of bacteria with Feγ-binding proteins.     

Fcgamma receptor polymorphisms and periodontal status: a prospective follow-up study

AIMS: The aims of this study were to assess: (i) the distribution of Fcgamma receptor polymorphisms among patients with chronic periodontitis ("cases") and control subjects with no/minimal loss of periodontal tissue support in a Caucasian population; (ii) whether these polymorphisms can serve as severity markers for periodontitis; and (iii) whether they have any bearing on the response to periodontal therapy. METHODS: The study sample consisted of 132 cases and 73 controls of comparable age and gender. Full-mouth periodontal status was assessed. Subgingival plaque (PL) samples and blood samples were obtained and analysed with respect to 19 bacterial species and homologous serum immunoglobulin G titres. Polymorphisms in the Fcgamma receptor IIa (131R/H) and IIIb (NA1/NA2) were assessed by polymerase chain reaction. Patients underwent periodontal therapy and were followed up at 4 and 30 months. RESULTS: Neither polymorphism showed a skewed distribution among cases and controls. At baseline, periodontitis patients with Fcgamma RIIa-H/H131 genotype had more PL and deeper pockets than patients in other genotype groups (p < 0.05). Both bacterial levels and antibody titres were unrelated to genotype. The longitudinal analysis failed to detect an association between genotype and response to periodontal therapy. CONCLUSIONS: The present data failed to demonstrate a clinically relevant relationship between the Fcgamma receptor IIa (131R/H) or IIIb (NA1/NA2) polymorphism and periodontal status.     

Hyper-reactive peripheral neutrophils in adult periodontitis: generation of chemiluminescence and intracellular hydrogen peroxide after in vitro priming and FcγR-stimulation

Abstract. We have earlier reported a higher Fcγ-receptor (FcγR)-mediated generation of reactive oxygen species, measured as luminol-enhanced chemiluminescence (CL) from peripheral neutrophils in adult periodontitis patients. The aims of this study were to confirm our previous results and lo elucidate the mechanism of this phenomenon by measuring CL in parallel with the intracellular production of hydrogen peroxide, after stimulation with opsonized bacteria. To determine whether the higher CL was associated with altered responsiveness to priming, the cells were preincubated with tumor necrosis factor α (TNFα) and lipopolysaccharide (LPS). While CL was significantly higher in subjects with periodontitis, there was no difference in hydrogen peroxide production between the patients and the controls, indicating that the hyperreactivity is related to the generation of other oxygen species than H₂O₂ and/or to processes in the outer cell membrane. The responsiveness to priming with LPS on CL was slightly but not significantly higher in the periodontitis group, suggesting that priming could be of value for distinguishing subjects with periodontitis. When assaying intracellular production of H₂O₂, TNFα and LPS had both a priming and an activating effect. There were no significant differences between the two groups. In conclusion, this study shows a higher FcγR-mediated CL of peripheral neutrophils from adult patients with periodontitis, thus confirming our earlier results. The hyperreactivity seems to be related to the outer cell membrane or to oxygen species other than H₂O₂.     

Longitudinal evaluation of GCF IFN-gamma levels and periodontal status in HIV+ patients

BACKGROUND/AIM: Loss of periodontal support and related tooth loss is a common finding among HIV+ patients. The etiology of this destruction may be an increase in the levels of pro-inflammatory cytokines and subsequent increase in periodontal disease activity. The purpose of this study was to investigate the associations between gingival crevicular fluid interferon gamma (GCF IFN-gamma) and clinical measures of periodontal disease in HIV+ individuals. We monitored GCF IFN-gamma and periodontal status of selected sites in 33 HIV+ subjects over a 6-month period. METHOD: Clinical measurements including gingival index, plaque index, bleeding on probing, probing depth, attachment loss (AL), and GCF samples were taken from four lower incisors and the upper right posterior sextant of each patient at baseline and 6-month visits by means of sterile paper strips. GCF levels of IFN-gamma were determined by sandwich ELISA assays. A progressing site was defined as a site that had 2 mm or more AL during the 6-month study period. RESULTS: Twenty-five of the 264 examination sites showed 2 mm or more clinical AL during the 6-month study period. Significantly higher GCF levels of IFN-gamma were found at progressing sites than in nonprogressing sites (p < 0.001). GCF levels of IFN-gamma were highly correlated with clinical measurements taken at baseline and 6-month visits (0.001相似文献   

Changes in amount of gingival crevicular fluid after a single episode of periodontal treatment

A total of 51 periodontal sites from 6 adults with no systemic diseases or medication were selected for the study. All sites showed radiologic bone loss and pockets of 4 mm or more. Crevicular fluid (CF) was collected by inserting filter paper strips into periodontal pockets for 5 s and was measured by Periotron. Samples were collected before and 2, 5, 10, 20, and 40 days after a single episode of periodontal treatment (scaling, root planing and curettage). Plaque Index (PII), Papilla Bleeding Index (PBI) and pocket depth (PD) were measured before and 40 days after treatment. The amount of bone loss was estimated from orthopantomograms taken immediately before the trial. Two days after treatment an increase in the amount of CF was seen. After this the amount of CF decreased, reaching the pretreatment level on day 5 after treatment and a level clearly below pretreatment level on day 10 after treatment. Forty days after treatment a slight increase in the amount of CF was seen. The difference between pretreatment values and values at days 2, 10, 20, and 40 was highly significant. In pretreatment samples, positive correlations were found between the amount of CF and PD, PBI and bone loss and, in samples collected 40 days after treatment between CF and PD. CF measurements made before treatment were of no value in predicting the changes in clinical parameters after treatment.     

Gingival crevicular fluid of patients with gingivitis or periodontal disease: evaluation of elastase-α, proteinase inhibitor complex

Elastase-alpha 1 proteinase inhibitor (E alpha 1PI) concentrations were assessed in gingival crevicular fluids and evaluated in relation to the clinical signs of periodontal disease. 7 gingivitis patients (group G), 38 patients with adult periodontitis and clinically stable lesions (group AP), 21 patients with rapidly progressive periodontitis and clinically stable lesions (group RPP) and 11 patients with either adult periodontitis or rapidly progressive periodontitis and clinically progressive lesions (group Pr) were studied. 6 healthy subjects served as the control group (group H). Significantly differences were observed in the E alpha 1PI concentration between the healthy, gingivitis, clinically stable periodontitis and clinically progressive periodontitis group. In the control group, no E alpha 1PI was detected. Groups G, AP and RPP showed mean E alpha 1PI concentrations of 10.95 +/- 4.96 micrograms/ml, 35.55 +/- 18.64 micrograms/ml and 38.56 +/- 20.89 micrograms/ml, respectively. In these groups, high enzyme levels were correlated with clinical signs of inflammation. The highest E alpha 1PI levels were observed in the clinically progressive lesions. However, they were not necessarily associated with bleeding on probing or clinical evidence of inflammation. These data suggest that a significant increase in crevicular E alpha 1PI levels may be an early manifestation of a progressive or potentially progressive periodontal lesion.     

Collagen III aminoterminal propeptide in gingival crevicular fluid before and after periodontal treatment

The amount of procollagen III aminoterminal propeptide (PIIINP) in crevicular fluid (CF) was measured from three periodontitis patients before treatment (scaling, root planing, and curettage) and 2, 5, 10, 20, and 40 days after treatment. CF was collected by placing two paper strips in each pocket for 5 s, after which the amount of fluid was measured. Control samples were collected from three subjects with minimal gingival inflammation. PIIINP was extracted into saline solution and determined by a radioimmunological method. Plaque Index, Papilla Bleeding Index, and pocket depth were recorded before and 40 days after treatment. Forty days after treatment clinical parameters indicated healing. The CF PIIINP mean concentration was 162 μg/1 (range 0–430) in the pretreatment samples. Ten days after treatment, PIIINP mean concentration was 1400 μg/1 (range 1000–9000). After this, the concentration gradually decreased, reaching the pretreatment level 40 days after treatment. Most of the control samples showed undetectable amounts of PIIINP. It was suggested that elevated PIIINP concentrations in CF after periodontal treatment reflected increased type III collagen synthesis in gingiva. PIIINP concentrations before treatment reflected the rate of type III collagen turnover in inflamed periodontal tissues. It was further suggested that CF PIIINP has clinical value as an indicator of the healing process of inflamed gingiva.     

Increased levels of soluble Fcγ receptor III in gingival fluid from periodontal lesions

Enzyme-linked immunosorbent assay was used for determination of the concentration of soluble Fcγ receptor III (FcγRIII) in 40 samples of gingival fluid obtained from periodontal pockets in 30 patients with periodontitis. The assay was based on a monoclonal immobilized antibody binding FcγRIII and a polyclonal FcγRIII rabbit antibody for its quantification. The results indicate a substantially increased concentration of soluble FcγRIII in gingival fluid as compared to the serum level. This increased concentration of soluble FcγRIII may interfere with phagocytosis and immune homeostasis in the periodontal lesions.     

Calprotectin in gingival crevicular fluid correlates with clinical and biochemical markers of periodontal disease

Clinical and biochemical markers of periodontal disease have been used for precise objective diagnosis of periodontal inflammation. Interleukin 1beta (IL-1beta) and prostaglandin E2 (PGE2), inflammatory factors, levels in gingival crevicular fluid (GCF) of patients with periodontal disease are elevated and have been studied as biochemical markers. The levels of calprotectin, a leukocyte protein, in body fluids of patients with some inflammatory diseases are raised. Recently, we detected calprotectin in GCF and its concentrations in periodontal pockets were higher than those in healthy gingival crevices. In this study, we investigated the correlations between GCF calprotectin levels and clinical indicators (probing depth and bleeding on probing, BOP), and the IL-1beta or PGE2 levels in GCE Probing depth and BOP at 130 sites of 110 subjects with periodontal or other oral diseases were examined, then GCF samples were collected and their calprotectin, IL-1beta and PGE2 were determined by ELISA. The calprotectin level correlated positively with the probing depth and was significantly higher at BOP-positive than BOP-negative sites. There were significant, positive correlations between the calprotectin and IL-1beta or PGE2 concentrations. These results indicate that the calprotectin level in GCF correlates well with clinical and biochemical markers of periodontal disease and suggest that calprotectin may be useful for evaluating the extent of periodontal inflammation.     

龈沟液中与牙周病有关的细胞因子的研究进展

牙周病时龈沟液内含有多种可作为诊断指标的细胞因子,由于取样简单无创,且能重复取样,易为患者所接受。因此,近年来学者们对龈沟液内的细胞因子在牙周炎活动期的诊断﹑治疗及疗效评价中作用的研究很多。本文对龈沟液中与牙周病有关的细胞因子的研究进展作一综述。     

Interleukin-1 receptor antagonist and interleukin-4 in gingival crevicular fluid of patients with inflammatory periodontal disease

Interleukin-1 receptor antagonist (IL-1ra) and interleukin-4 (IL-4) were detected in gingival crevicular fluid (GCF) by an immunochemical method. However, we could not detect IL-4 in GCF from severe inflammation sites. In addition, we sought to detect which cells had produced cytokines in moderately inflamed gingival tissues by means of immunohistochemistry. The cell types expressing CD 68 were identified as monocytes/macrophages and stained positively for IL-1ra. The helper T cells identified by immunostaining for CD 4 stained positively for IL-4. These results suggest that IL-4 is one of the mediators regulating the degree of local inflammation in periodontal disease.     

Gingival crevicular fluid laminin-5 gamma2-chain levels in periodontal disease

AIM: Our study aimed to examine the molecular forms and gingival crevicular fluid (GCF) levels of laminin-5 gamma2-chain in patients with different periodontal disease, and compare the effects of P.gingivalis trypsin-like proteinase on intact laminin-5 gamma2-chain species. METHODS: Eighteen patients with generalized aggressive periodontitis (G-AgP), 29 patients with chronic periodontitis (CP), 20 with gingivitis and 20 periodontally healthy subjects were included. Probing depth, clinical attachment loss, presence of bleeding on probing and plaque were recorded. Molecular forms and GCF laminin-5 gamma2-chain levels and the effects of P. gingivalis trypsin-like proteinase on intact laminin-5 gamma2-chain were analysed by computer-quantitated Western immunoblotting. RESULTS: Laminin-5 gamma2-chain 40 and 70 kDa fragments could be detected in all groups, in varying levels.The CP group had elevated GCF laminin-5 gamma2-chain fragment levels compared with the gingivitis and healthy groups (p<0.008).The G-AgP group had GCF laminin-5 gamma2-chain fragment levels similar to the gingivitis and healthy groups (p>0.008). GCF laminin-5 gamma2-chain fragments differed clearly from the multiple lower molecular size fragments of P.gingivalis trypsin-laminin-5 gamma2-chain proteinases. CONCLUSION: Increased GCF laminin-5 gamma2-chain fragments in periodontitis sites with deep periodontal pocket suggest that these cleaved 40 and 70 kDa fragments could reflect the extent of the inflammatory reaction in CP.     

Plasma and crevicular fluid osteopontin levels in periodontal health and disease

BACKGROUND AND OBJECTIVE: The level of osteopontin in gingival crevicular fluid has been found to correlate with clinical measures of periodontal disease. The present study was designed to assess the relationship between clinical parameters and osteopontin levels of the gingival crevicular fluid from inflamed gingivae, periodontitis sites and after treatment of periodontitis sites, and to correlate them to the osteopontin levels of the plasma. MATERIAL AND METHODS: Thirty, gender-matched subjects were divided into three groups--healthy, gingivitis and chronic periodontitis--based on modified gingival index scores and clinical attachment loss. The fourth group consisted of 10 subjects in the periodontitis group, 6-8 wk after initial therapy. Plasma and gingival crevicular fluid samples were collected and quantified for osteopontin using an enzyme immunoassay. RESULTS: The highest mean gingival crevicular fluid and plasma osteopontin concentrations were observed in the periodontitis group (1575.01 and 1273.21 ng/mL, respectively) and the lowest in the healthy group (1194.80 and 476.35 ng/mL, respectively). After treatment of the periodontitis group, the level of osteopontin decreased to 1416.15 in gingival crevicular fluid and to 1051.68 ng/mL in plasma. In all groups the gingival crevicular fluid osteopontin levels showed a statistically significant positive correlation with that of plasma and clinical attachment loss. CONCLUSION: Osteopontin levels were highest in the gingival crevicular fluid from sites with periodontal destruction; however, periodontal treatment resulted in the reduction of osteopontin levels. Gingival crevicular fluid and plasma osteopontin levels showed a positive correlation in all of the groups.