慢性牙周炎病人基础治疗前后Th17型细胞因子及RORC的表达变化

目的:研究慢性牙周炎(CP)病人牙周基础治疗前后龈沟液(GCF)中的Th17型细胞因子及其外周血特异性转录因子RORC的表达变化规律。方法:选择CP病人30例,使用ELISA检测牙周基础治疗前后GCF中Th17型细胞因子(IL-17、IL-21)的水平变化,并利用荧光定量PCR检测牙周基础治疗前后外周血CD4+T细胞中特异性转录因子RORC的表达水平。结果:牙周基础治疗后龈沟液中的Th17型细胞因子IL-17、IL-21水平以及外周血CD4+T细胞中特异性转录因子RORC表达水平均较治疗前显著下降,差异均有统计学意义(P<0.05)。结论:Th17型细胞因子在慢性牙周炎发生发展中发挥重要的致炎作用。     

ICAM-1-expressing pocket epithelium, LFA-1-expressing T cells in gingival tissue and gingival crevicular fluid as features characterizing inflammatory cell invasion and exudation in adult periodontitis

Activated T lymphocytes constitute a major component of inflammatory cells in the early periodontal lesion, and also appear in the gingival crevicular fluid. In an attempt to clarify the relationship between the ICAM-1 (CD54) expression of pocket epithelium in gingiva and the infiltrating lymphocyte population, we carried out an analysis of CD11a⁺ (LFA-lα), CD25⁺ (IL-2Rα) and CD4⁺ (Th) cells subjacent to ICAM-1-expressing pocket epithelia and CD11a⁺ CD25⁺CD4⁺ cells in gingival crevicular fluid (GCF). GCF was collected by crevicular washing from 16 patients with periodontitis (P group) and 3 subjects with healthy gingiva (H group). Peripheral blood (PB) was collected at the same time. Mononuclear cells were isolated by Ficoll-paque gradient centrifugation from GCF and PB. Monoclonal antibodies (mAb) to CD11a, CD25, and CD4 were used for three-color flow cytometry. Gingival biopsies were obtained from 7 patients in P group and 3 subjects in H group. Serial cryostat sections (6 μm in thickness) were prepared from each biopsy, on which a double staining was performed. The number of CD11a⁺ CD25⁺ CD4⁺ cells and the fluorescence intensity of FITC conjugated anti-CD 11a were significantly higher in GCF than in PB (p≤0.001 to p≤0.01). CD11a⁺ CD25⁺CD4⁺ cells were not detected in GCF in H group. The pocket epithelia expressed CD54 in P group, but not in H group. The number of CD11a⁺, CD25⁺ and CD4⁺cells infiltrating the connective tissue subjacent to the upper, middle and lower parts of the CD54 positive pocket epithelium (n=16) was 141±26, 38±13, 144±29 (cells/0.04 mm²), respectively, whereas in the CD54 negative pocket epithelium, it was (n=5) 9±2, 3±1, 8±3. In P group, the CD11a⁺CD25⁺CD4⁺cell number in GCF correlated with CD25⁺, CD11a⁺cells in the connective tissue subjacent to the CD54⁺pocket epithelium. These results indicate that expression of ICAM-1 in pocket epithelium is relevant to the migration of CD11a, CD25, CD4 positive cells in connective tissue subjacent to the pocket epithelium into the periodontal pocket. Assessing the relationship of our findings and other adhesion molecules would offer important clues to the understanding of T cell migration in affected gingiva.     

Cytokine profile in the gingival crevicular fluid of periodontitis patients with and without type 2 diabetes: a literature review

Background: Periodontitis may occur in patients with and without type 2 diabetes (T2D). It may be hypothesized that the gingival crevicular fluid (GCF) cytokine profile in patients with periodontitis with poorly controlled T2D may differ from the GCF cytokine profile in medically healthy individuals with periodontitis. The aim was to review the cytokine profiles in the GCF of patients with periodontitis with and without T2D. Methods: Databases were searched from 1988 to August 2011 using different combinations of various keywords. Titles and abstracts of articles that satisfied the eligibility criteria were screened by the authors and checked for agreement. Only articles published in English were included. Results: Ten studies were included. Two studies reported GCF concentrations of interleukin (IL)‐6 to be higher in patients with periodontitis with T2D compared to medically healthy patients with periodontitis. Two studies showed GCF IL‐6 levels to be higher in periodontitis with T2D compared to medically healthy subjects without periodontitis. In one study GCF levels of IL‐17, IL‐23, and interferon‐γ were higher in patients with periodontitis with T2D compared to medically healthy patients with periodontitis. In one study, GCF concentrations of IL‐8 were significantly higher in patients with periodontitis with T2D compared to medically healthy individuals with periodontitis. Three studies reported GCF levels of IL‐1α to be significantly higher in patients with periodontitis with T2D compared to medically healthy individuals with periodontitis. Conclusion: The GCF cytokine profile in patients with and without T2D seems to be governed by the intensity of periodontal inflammation and the role of T2D in this regard is rather secondary.     

Levels of interleukin-21 in patients with untreated chronic periodontitis

Background: A growing body of evidence suggested that interleukin (IL)‐21 enhances the effector phase during T‐cell responses. The aim of our study is to determine the levels of IL‐21 in periodontal sites from patients with chronic periodontitis and controls. Methods: The population studied consisted of 34 patients (15 with chronic periodontitis and 19 healthy patients). Twenty samples (10 gingival crevicular fluid [GCF] and 10 gingival biopsies) were collected from each group before the patients with periodontitis received periodontal treatment. Total protein concentrations were measured in all samples; the presence of IL‐21 was confirmed by immunohistochemistry and Western blot, and IL‐21 levels were quantified through an enzyme‐linked immunosorbent assay. Statistical analyses were performed using statistical software. Data were expressed as patient means ± SDs or medians (interquartile ranges) by using the χ², Student t, and Mann‐Whitney U tests. Results: GCF IL‐21 was mainly detected in patients with chronic periodontitis (P <0.05). Levels of IL‐21 in gingival tissues were significantly higher in patients with chronic periodontitis compared to healthy individuals (P <0.05). The Western blot and immunohistochemical staining confirmed the presence of IL‐21 in periodontal tissues and GCF. Conclusion: IL‐21 was highly expressed in patients with chronic periodontitis, especially in gingival biopsies; therefore, IL‐21 might play a role in the T‐cell response.     

Presence of activated eosinophils, high IgE and sCD23 titers in gingival crevicular fluid of patients with adult periodontitis

Our previous studies showed that the expression of CD23 on polymorphonuclear leukocytes (PMNLs) in gingival crevicular fluid (GCF) from adult periodontitis (AP) patients was higher than in autologous peripheral blood (PB). Percentages of eosinophils in GCF PMNLs ranged between 6 and 14%. The purpose of the present studies was to increase understanding of the potential role of eosinophils and their products, including CD23, in periodontal disease. We analysed the eosinophil fraction in GCF and PB by flow cytometry using monoclonal antibodies to CD23b (BB10), eosinophil cationic protein (ECP) in stored and secretory forms (EG1 and EG2), and CD67 (80H3). Simultaneously, we measured IgE and soluble CD23 titer and GCF and serum by ELISA. Flow cytometric analysis of BB10, EG2 and 80H3 binding showed that GCF eosinophils from AP were activated. A large BB10⁺ EG2⁺ cellular fraction was detected in GCF from AP whereas it was very low in autologous serum (9.30±2.460 vs 0.16±0.10, p>0.001). GCF from gingivitis patients exhibited no flow cytometric evidence for the presence of BB10+ EG2+ cells. BB10+ EG1+ cells, or inactivated eosinophils rated lower in GCF than in PB both in gingivitis and periodontitis patients (0.45±0.63 vs 1.83±0.96 and 0.15±0.30 vs 1.30±0.20, p>0.05. respectively). IgE titer in AP patients reached 1208.1 ±421.2 IU/ml in GCF while only 49.1 ±50.4 in sera. Soluble CD23 in GCF reached 236.1 ±81.3 ng/ml in GCF and 5.6±1.8 ng/ml in sera. GCF of gingivitis patients, however, contained no detectable sCD23. Thus. GCF from AP patients displayed a high rate of activated eosinophils secreting ECP. while GCF of gingivitis patients did not. These results suggest that ECP-secreting activated eosinophils are relevant to the pathology of adult periodontitis.     

Increased Expression of Interleukin (IL)‐35 and IL‐17, But Not IL‐27, in Gingival Tissues With Chronic Periodontitis

Background: Interleukin (IL)‐35 plays an important role in immune regulation through the suppression of effector T‐cell populations, including T‐helper 17 (Th17) cells. Although Th17 cells and IL‐17 are involved in the pathogenesis of periodontitis, the level of IL‐35 in inflamed periodontal tissues is unclear. Here, IL‐35, IL‐17, and IL‐27 production/expression in gingival crevicular fluid (GCF) and human gingival tissue were investigated. Methods: GCF samples were collected from buccal (mesial, center, and distal) sites of teeth from patients with chronic periodontitis (CP) and healthy controls and were analyzed by enzyme‐linked immunosorbent assay for IL‐35 (periodontitis, n = 36; healthy, n = 30) and IL‐17 (periodontitis, n = 16; healthy, n = 13). Gingival tissue, including sulcus/pocket epithelium and underlying connective tissue, was collected from an additional 10 healthy participants and 10 patients with CP and were analyzed by quantitative polymerase chain reaction (qPCR) for Epstein Barr virus‐induced gene 3 (EBI3), IL12A, and IL17A. IL27p28 was also tested by qPCR. Results: IL‐35 and IL‐17 were significantly higher in GCF from patients with periodontitis than healthy participants (P <0.01, P <0.05, respectively). In both healthy participants and those with periodontitis, positive correlations were found among IL‐35 and probing depth and clinical attachment level (CAL) as well as between IL‐17 and CAL. EBI3, IL12A (components of IL‐35), and IL17A messenger RNA expression levels were significantly higher in inflamed gingival tissue than in healthy control tissues (P <0.05). IL27p28 was not detected in any sample, suggesting that IL‐27 is not produced in large quantities in periodontal tissue. Conclusion: IL‐35 and IL‐17, but not IL‐27, may play important roles in the pathogenesis of periodontitis.     

CD9 and HLA-DR expression by crevicular epithelial cells and polymorphonuclear neutrophils in periodontal disease

BACKGROUND, AIMS: The composition of gingival crevicular fluid (GCF) is likely to reflect inflammatory modifications that take place in the gingiva during periodontal diseases. METHOD: In this study, GCF was collected at 3 different sites from 23 periodontal patients. The sites were assessed to be healthy, presenting gingivitis or periodontitis. 10 healthy individuals without any form of periodontal disease formed the control group and were sampled at one site each. The cell content of GCF was collected using Durapore Millipore strips, and 2 types of cells were studied: epithelial cells (EC) and polymorphonuclear neutrophils (PMN). The expression of CD9 and HLA-DR within or on the surface of these cells was studied in immunofluorescence on cytospin smears. RESULTS: Both CD9 and HLA-DR expression on EC differed significantly from control subjects, and the latter decreased according to the severity of the pathology. None of the PMN found in controls expressed CD9 or HLA-DR. However, in periodontal patients, the expression of HLA-DR within PMNs was detectable and increased according to the severity of lesions. CD9 expression on PMNs also increased with inflammation. CONCLUSION: This study shows that clinically healthy sites of periodontal patients already present signs of immunological activation characterised by a down modulation of HLA-DR expression on EC and an upregulation of these 2 molecules in PMN.     

慢性牙周炎患者治疗前后龈沟液中IL-23、IL-17表达水平的变化

目的研究慢性牙周炎患者牙周基础治疗前后龈沟液(gingival crevicular fluid,GCF)中IL-17、IL-23表达水平的变化,探讨IL-23/IL-17轴在慢性牙周炎发生、发展过程中的作用。方法选取来我科就诊的慢性牙周炎患者50例为研究对象。于牙周基础治疗前后,采集龈沟液并测量体积,运用夹心酶联免疫吸附试验测定IL-17和IL-23的质量浓度以及测量两者之间的关系。结果牙周基础治疗后龈沟液中的IL-17、IL-23表达水平均较治疗前显著下降,差异均有统计学意义(P<0.05)。牙周基础治疗前后龈沟液中的IL-17、IL-23表达水平与牙周临床指标呈正相关。结论 IL-17、IL-23在慢性牙周炎发生发展过程中起重要作用。     

Interleukin‐1 β, interleukin‐12 and interleukin‐18 levels in gingival fluid and serum of patients with gingivitis and periodontitis

Introduction: Cytokines are of major importance in periodontal disease progression. Interleukin‐12 (IL‐12) stimulates interferon‐γ production by T helper type 1 (Th1) cells while IL‐18 induces Th1 responses when present with IL‐12 but Th2 responses in the absence of IL‐12. IL‐1β has been correlated with periodontal disease destruction. This study determined the local concentrations of these cytokines in sites of gingivitis and periodontitis. Methods: Gingival crevicular fluid was collected from two sites in each of 10 gingivitis patients and from two gingivitis sites and two periodontitis sites from each of 10 periodontitis patients. Serum samples were also collected. IL‐1β, biologically active IL‐12 p70, the IL‐12 p40 subunit and IL‐18 concentrations were determined by enzyme‐linked immunoabsorbent assay. Results: IL‐1β and IL‐18 concentrations were higher in the gingival crevicular fluid from periodontitis patients than in that from gingivitis patients; IL‐18 concentrations were higher than those of IL‐1β. Very little IL‐12, either p40 or p70, was detected in the gingival crevicular fluid samples. In the serum, very low levels of cytokines were found. The level of serum IL‐12 p40, however, was higher than in the fluid from periodontitis sites of periodontitis patients. Conclusion: The local production of IL‐1β and IL‐18 in the gingival crevicular fluid increased with increasing inflammation and IL‐18 was the predominant cytokine at both gingivitis and periodontitis sites. Very little IL‐12 was detected with levels decreasing with increasing inflammation. These results suggest that there is an association between severity of periodontal disease and levels of IL‐1, IL‐12 and IL‐18.     

Detection of, and anti-collagen antibody produced by, CD5-positive B cells in inflamed gingival tissues

This study was performed to investigate the frequency and distribution of CD5-positive (CD5+) B cells in inflamed gingival tissues using flow cytometric and immunohistochemical analyses. The ability of CD5+ B cells to produce anti-type I collagen antibody was also examined. CD5+ B cells expressed "low" fluorescence intensity in the peripheral blood of both healthy subjects and patients with adult periodontitis. However, in inflamed gingival tissues the intensity of this surface marker was high. The percentage of B cells bearing CD5 surface marker was statistically higher in gingiva than in peripheral blood obtained from both the patients and healthy subjects. These CD5+ B cells were observed in gingival subepithelial connective tissues from the bottom to the middle of the periodontal pocket. This area showed destruction of collagen fibers and dense cell infiltrations. Anti-collagen IgG antibody level in patients' gingival crevicular fluids (GCF) was higher than that in sera from healthy subjects, and slightly higher than in autologous sera. IgM anti-collagen antibody in GCF was lower than in autologous sera and in sera from healthy subjects. EBV-transformed CD5+ B cells produced considerably more IgM and IgG antibody to collagen than CD5- B cells. Therefore CD5+ B cells may contribute to the pathogenesis of inflamed gingival tissues.     

Levels of interleukin-1 beta, -8, and -10 and RANTES in gingival crevicular fluid and cell populations in adult periodontitis patients and the effect of periodontal treatment

BACKGROUND: Various cytokines have been identified at sites of chronic inflammation such as periodontitis. Cytokines are synthesized in response to bacteria and their products, inducing and maintaining an inflammatory response in the periodontium. The purpose of the present study was to investigate the involvement of interleukin-1 beta (IL-1 beta), IL-8, and IL-10 and RANTES (regulated on activation, normally T cell expressed and secreted) and the cell populations associated with the immune response in destructive periodontitis, as well as the effect of periodontal therapy on cytokine levels in gingival crevicular fluid (GCF). METHODS: Data were obtained from 12 patients with moderate to advanced periodontitis and 6 healthy controls. Patients presenting at least 2 sites with > or =2 mm clinical attachment loss were included in the destructive periodontitis group. After monitoring for 4 months, only 6 patients showed destructive periodontitis and GCF samples and soft tissues biopsies were collected from these patients. GCF samples and biopsies were collected both from active (12 CGF samples and 6 biopsies) and inactive (12 CGF samples and 6 biopsies) sites. The comparison with healthy controls was carried out by collecting GCF samples from 6 healthy volunteers (12 samples) and biopsies during the surgical removal of wisdom teeth. In periodontal patients, clinical data and GCF samples were obtained prior to periodontal treatment (72 samples) and 2 months after periodontal therapy (72 samples). GCF was collected using a paper strip; eluted and enzyme-linked immunoabsorbent assays (ELISA) were performed to determine cytokine levels. The inflammatory infiltrate was analyzed by immunohistochemistry of gingival biopsy samples with monoclonal antibodies against CD3, CD8, CD4, CD11c, and CD19 antigens. RESULTS: Cellular components of the inflammatory infiltrate include B and T lymphocytes and monocyte/macrophages. Active sites contained a higher number of B lymphocytes and macrophages. IL-8 and IL-1 beta and RANTES in GCF were detected in the majority of sites from periodontal patients (100%, 94% and 87%, respectively); IL-10 was found in only 43%. IL-8 was the only cytokine detected in the GCF (75%) of the control group. Moreover, IL-1 beta levels were significantly higher in active sites versus inactive sites (P <0.05). IL-8 and IL-10 and RANTES were increased in active sites; however, differences were not significant (P>0.05). A positive correlation between the IL-8 and RANTES (r = 0.677, P<0.05) was observed in periodontitis patients. Periodontal therapy reduced the total amount of IL-1 beta, IL-8, and IL-10 and RANTES. Data showed a weak correlation between the clinical parameters and the total amount of cytokines in periodontitis. CONCLUSIONS: These data suggest that the amount of crevicular IL-1 beta, IL-8, and IL-10 and RANTES is associated with periodontal status. Removal of the bacterial plaque reduces the antigenic stimuli and consequently could modulate the chemokines present in GCF. We propose that the dynamic interactions between cytokines, their production rates, and their quantity could represent factors controlling the induction, perpetuation, and collapse of the cytokine network present in the periodontal disease.     

正常人、牙龈炎和牙周炎患者龈沟液内一氧化氮含量的检测

目的：检测正常人和牙周病患者龈沟液中NO含量,探讨NO在牙周病发病过程中的作用。方法：选择牙周健康组20例,牙龈炎组22例,慢性牙周炎组32例,分别采集龈沟液标本,免疫荧光法检测龈沟液内NO的含量。结果：慢性牙周炎患者和牙龈炎患者龈沟液内NO含量与牙周健康组相比均有高度显著性差异（P〈0．01）,慢性牙周炎患者龈沟液内NO含量与牙龈炎组相比有高度显著性差异（P〈0．01）。结论：牙周健康者、牙龈炎患者、慢性牙周炎患者龈沟液中能检测出NO的存在,NO参与了慢性牙周炎的发展过程,龈沟液内NO含量与慢性牙周炎炎症程度密切相关。     

牙周炎治疗对患者龈沟液IL-8水平的影响

目的　研究牙周炎基础治疗前后IL 8水平的变化。方法　采用双抗夹心ABC ELISA法测定成人牙周炎患者 (AP)、健康对照以及牙周炎治疗前后患者的龈沟液中IL 8浓度和总量。结果　在疾病组和健康组之间以及牙周病治疗前后对照 ,IL 8浓度无统计学差异 (P >0 0 5 ) ,而IL 8总量、GCF量均存在着统计学差异 (P <0 0 5 ,P <0 0 0 1) ,并且发现治疗对IL 8水平有着明显影响。结论　IL 8总量在成人牙周炎病程中显示动态性改变 ,检验GCF中IL 8的水平对评价牙周炎的治疗是有价值的。?     

Expression of HMGB1 and HMGN2 in gingival tissues, GCF and PICF of periodontitis patients and peri-implantitis

Background and objective

High mobility group chromosomal protein B1 (HMGB1) and N2 (HMGN2), two members of high mobility group (HMG) family, play important role in inflammation. The purpose of this study was to investigate the expression of HMGB1 and HMGN2 in periodontistis.

Materials and methods

The expression of HMGB1 and HMGN2 mRNA in gingival tissues and gingival crevicular fluid (GCF) in chronic periodontitis (CP), generalised aggressive periodontitis (G-AgP) patients and healthy subjects was detected by real-time PCR. The protein level of HMGB1 and HMGN2 in peri-implant crevicular fluid (PICF), peri-implant crevicular fluid of peri-implantitis (PI-PICF) and normal patients was determined by Western blotting. Furthermore, IL-1β, IL-6, IL-8, TNF-α and HMGB1 levels in GCF, PI-PICF and healthy-PICF samples from different groups were determined by ELISA.

Results

HMGN2 expression was increased in inflamed gingival tissues and GCF from CP and G-ApG groups compared to control group. HMGB1 expression was the highest in the gingival tissues and GCF from CP patients and was accompanied by increased concentrations of IL-1β, IL-6, IL-8 proinflammaory cytokines.

Conclusion

To our knowledge, this is the first study reporting that the expression of HMGB1 and HMGN2 was increased in the gingival tissues and GCF in CP and G-AgP and the PICF in PICF. Our data suggest that HMGB1 may be a potential target for the therapy of periodontitis and PI.     

Levels of platelet-activating factor in gingival crevicular fluid and gingival tissue in specific periodontal diseases

BACKGROUND: Platelet-activating factor (PAF), a potent phospholipid mediator of inflammatory and immune reactions, is involved in a variety of biological responses seen in periodontal diseases. The aim of the present study was to examine the role of PAF in the pathogenesis of specific periodontal diseases. METHODS: PAF levels were investigated in gingival crevicular fluid (GCF) and gingival tissue (GT) samples of 12 patients with generalized aggressive periodontitis (GAgP), 6 patients with localized aggressive periodontitis (LAgP), 10 patients with chronic periodontitis (CP), 6 with gingivitis (G), and 6 periodontally healthy subjects (H). Periodontal status was evaluated by measuring probing depth, gingival index, papillary bleeding index, and plaque index. PAF was extracted from GCF samples passing through amberlit resin columns, purified by high performance liquid chromatographic method, and then analyzed by radioimmunoassay. RESULTS: GAgP, LAgP, and CP groups had significantly higher GCF PAF levels compared to the H group (P<0.005). Although statistically not significant, GCF PAF levels were also higher in the G group than those of the H group (P = 0.0784). GAgP, LAgP, and CP groups had similar GCF PAF levels (P>0.005). These groups had higher levels of GCF PAF than those of the G group, but the difference was significant only for the GAgP group (P<0.005). When the data were expressed as concentration, GAgP, LAgP, and CP groups were found to have higher concentrations of GCF PAF compared to the H group (P<0.005). GCF PAF concentration was similar in patient groups (P>0.005). All patient groups had significantly higher GT PAF levels compared to the H group (P<0.005). GAgP, LAgP, and CP groups had similar amounts of GCF and GT PAF (P>0.005). GAgP, LAgP, and CP groups had higher GT PAF levels than those of the G group, but the differences were only significant for LAgP and CP groups (P<0.005). No significant correlation was found between GCF and GT PAF levels and clinical parameters. CONCLUSIONS: The results of the present study indicate that PAF is likely to be an important mediator in regulating inflammatory responses in the human periodontal tissues. To our knowledge, this is the first report investigating PAF levels in GCF and GT in specific periodontal diseases. We believe that this potent phospholipid mediator may need to be considered in the pathogenesis of periodontal diseases.     

Interleukin-33 levels in gingival crevicular fluid, saliva, or plasma do not differentiate chronic periodontitis

Background: This study investigates whether gingival crevicular fluid (GCF), saliva, and plasma levels of interleukin‐33 (IL‐33) can differentiate individuals with chronic periodontitis from individuals with healthy periodontium. Methods: GCF, whole saliva, and plasma samples together with full‐mouth clinical periodontal recordings were obtained from 32 otherwise healthy, non‐smoker chronic periodontitis individuals and 25 systemically and periodontally healthy, non‐smoker individuals. IL‐33 levels in the biofluid samples were determined by enzyme‐linked immunosorbent assay. Data were tested statistically by Mann‐Whitney U test. Results: The GCF concentrations of IL‐33 were significantly lower in chronic periodontitis individuals than in healthy individuals (P <0.0001), whereas the total amounts in GCF samples were similar (P >0.05). The salivary and plasma contrations of IL‐33 were indifferent in the two study groups (P >0.05). Conclusions: According to the present findings, the GCF, saliva or plasma levels of IL‐33 could not differentiate chronic periodontitis individuals and periodontally healthy individuals. Larger‐scale intervention studies may better clarify this issue.     

辅助性T17细胞在牙周炎小鼠中的免疫状态研究

目的 研究辅助性T（Th）17细胞在牙周炎小鼠中的免疫状态。方法 将7周龄C57BL/6雌性小鼠随机分为牙周炎组和对照组,每组4只。牙周炎组采用口腔涂抹牙龈卟啉单胞菌（P. gingivalis）的方法建立牙周炎动物模型。对照组涂抹PBS液。在涂抹结束后的第4周取材,流式细胞仪检测CD4+维甲酸相关核孤儿受体（ROR）γτ+（Th17）细胞;酶联免疫吸附（ELISA）检测Th17细胞相关的细胞因子白细胞介素（IL）-17A的蛋白表达。结果 牙周炎小鼠牙龈组织、颈部淋巴结和外周血中CD4+ RORγτ+（Th17）细胞在总CD4+ T细胞中的比例和细胞数量显著高于对照组（P<0.01）。与对照组相比,牙周炎组IL-17A的表达增加（P<0.05）。结论 在牙周炎的发生发展中,Th17细胞介导的细胞免疫应答增强,牙龈组织、颈部淋巴结和外周血可能是Th17细胞介导免疫应答的主要场所。     

Levels of leukotriene B4 in gingival crevicular fluid and gingival tissue in specific periodontal diseases

BACKGROUND: Leukotriene B4 (LTB4), a product of the lipoxygenase pathway of arachidonic acid metabolism, exhibits numerous activities that can account for most of the features of host responses seen in periodontal diseases. The aim of the present study was to examine the role of LTB4 in the pathogenesis of specific periodontal diseases. METHODS: LTB4 levels were investigated in gingival crevicular fluid (GCF) and gingival tissue (GT) samples of 10 patients with chronic periodontitis (CP), 12 patients with generalized aggressive periodontitis (GAgP), 6 patients with localized aggressive periodontitis (LAgP), 6 patients with gingivitis (G), and 6 periodontally healthy subjects (H). Periodontal status was evaluated by measuring probing depth, gingival index, papillary bleeding index, and plaque index. LTB4 was extracted from the samples by solid-phase method using C18 cartridge and was purified by high performance liquid chromatographic method and then analyzed by radioimmunoassay. RESULTS: All patient groups had significantly higher levels of GCF and GT LTB4 compared to the control group (P<0.005). The CP patients had the highest LTB4 levels compared to those in other patient groups (P<0.005). GAgP, LAgP, and G groups had similar amounts of GCF and GT LTB4 (P>0.005). When the data were expressed as concentration, the CP group was found to have higher concentration of LTB4, compared to that of control group (P<0.005). GAgP, LAgP, and G groups had similar LTB4 concentration compared to that of control group (P>0.005). No significant difference was found between GAgP, LAgP, and G groups (P>0.005). The CP group had higher LTB4 concentration compared to both GAgP and LAgP groups (P<0.005). Although the CP group had a higher GCF LTB4 concentration compared to G group, this difference did not reach significance (P>0.005). No significant correlation was found between GCF and GT LTB4 levels and clinical parameters. CONCLUSIONS: The results of the present study indicate that LTB4 is likely to be an important mediator in regulating inflammatory responses in the human periodontal tissues. This lipid mediator may play an important role in the pathophysiology of periodontal disease.