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.     

Differential expression of chemokines and chemokine receptors in inflammatory periapical diseases

BACKGROUND: Periapical lesions are thought to be the result of a local inflammatory response mediated by inflammatory cell infiltration and production of inflammatory mediators. Although chemokines are strongly implicated in the migration and activation of leukocytes in different inflammatory diseases and experimental models, little is known regarding the expression of chemokines and their receptors in human apical periodontitis. OBJECTIVE AND METHODS: The objective of this study was to determine the expression of chemokines and their receptors by real-time polymerase chain reaction in samples obtained from healthy gingiva, periapical granulomas, and inflammatory periradicular cysts. The inflammatory infiltrate was characterized by immunohistochemistry. RESULTS: Comparing cysts and granulomas, an increase in CD4+ and CD8+ cells was observed in granulomas, despite the similar numbers of CD45RO-positive cells detected in both lesions. The analysis of mRNA expression revealed increased levels of CCR1, CCR2, CCR3, CCR5, CXCR1, and CXCR3 in both types of lesion compared with controls. Cysts exhibited a higher expression of CCR3, CCR5, CXCR1, and CXCR3 compared to granulomas. A significantly higher expression of RANTES, IP-10, and MCP-1 was detected in cysts compared with controls or granulomas. The expression of interleukin-8, MIP-1alpha, and MIP-1beta was not different in the three experimental groups. CONCLUSIONS: The increase in Th1 type (CCR1, CCR5, and CXCR3) and Th2 type (CCR2 and CCR3) receptors in both periapical lesions suggests the concomitant occurrence of Th1 and Th2 responses. Furthermore, the prevalent expression of the receptors CCR3, CCR5, CXCR1, and CXCR3 and of the chemokines RANTES, IP-10, and MCP-1 in cysts may point to a role in the progression of granulomas to cysts.     

Gingival crevicular fluid monocyte chemoattractant protein-1 and RANTES levels in patients with generalized aggressive periodontitis

BACKGROUND: Local and systemic inflammatory and immune mechanisms may be implicated in the pathogenesis of the aggressive forms of periodontal disease. Chemokines, monocyte chemoattractant protein-1 (MCP-1) and regulated on activation, normal T cells expressed and secreted RANTES (regulated on activation, normal T cells expressed and secreted), are involved in the activation and recruitment of inflammatory and immune cells to the infected sites and thereby mediating a variety of pathophysiological conditions. The aim of the present study was to examine the gingival crevicular fluid (GCF) levels of MCP-1 and RANTES in patients with generalized agressive periodontitis (G-AgP). METHODS: MCP-1 and RANTES levels were investigated in GCF samples of 10 patients with G-AgP and 10 periodontally healthy subjects. Periodontal status was evaluated by measuring probing depth, clinical attachment loss, presence of bleeding on probing and plaque. In the G-AgP group, GCF samples were collected from the two approximal sites; from one single-rooted tooth and from one first molar tooth with > or =6 mm probing depth. In the healthy group, GCF samples were collected from one of the single-rooted teeth. GCF MCP-1 and RANTES levels were quantified by enzyme immunoassay. RESULTS: The G-AgP patients had significantly higher GCF MCP-1 and RANTES levels compared to the healthy group (p<0.05). GCF MCP-1 and RANTES levels were positively correlated with both probing depth and clinical attachment loss (p<0.05). There was no correlation between GCF MCP-1 and RANTES levels and the percentage of sites with bleeding (p>0.05). CONCLUSIONS: The results of the present study suggest that MCP-1 and RANTES could play key roles in both activation and recruitment of inflammatory and immune cells in periodontal environment of G-AgP patients. In conclusion, these CC chemokines may be considered in the biological mechanism underlying the pathogenesis and progression of G-AgP.     

Chemokine RANTES in gingival crevicular fluid of adult patients with periodontitis

Background, aims: This study presents the first evidence on the presence of the chemokine RANTES in the gingival fluid crevicular (GCF) of patients with periodontitis. RANTES is a chemokine that selectively attracts and activates macrophages and lymphocytes. Leucocytes play a critical rôle in the host response to the subgingival microflora. Method: In this study, the presence de RANTES in GCF was determined in samples obtained from adult patients with periodontitis and from control subjects with clinically healthy gingiva. GCF was collected from different probing depths (<3 mm, 4–6 mm, >6 mm) (n=72); and active (n=12) and inactive sites (n=12). An active site was defined as attachment loss >2 mm, as determined by sequential probing and the tolerance method. GFC was collected for 30 s using Periopaper^® strips, and RANTES was quantified by ELISA. Results: The presence of RANTES was detected exclusively in the group of patients with periodontitis, presenting a total amount of 40.43±16 pg and a concentration 67.80±41 pg/μl. RANTES concentration was significantly higher in probing depth <3 mm than in probing depth >6 mm (87.24 versus 51.87, p=0.014). Total amount and concentration in the GCF samples from active sites were higher that in inactive sites (p>0.05). Conclusions: The finding that RANTES is found only in patients with periodontitis, may represent a general feature of chronic inflammatory in periodontal diseases. Finally, RANTES may be implicated in the biological mechanisms underlying the pathogenesis and progression of periodontal disease.     

The influence of diabetes on gingival crevicular fluid beta-glucuronidase and interleukin-8

OBJECTIVES: Polymorphonuclear neutrophil (PMN) dysfunction is associated with diabetes. We examined the gingival crevicular fluid (GCF) beta-glucuronidase (BG) and interleukin-8 (IL-8) levels of periodontitis patients with and without type 2 diabetes mellitus (DM). MATERIAL AND METHODS: Forty five adults with type 2 DM and 32 adults without DM, both with chronic periodontitis were enrolled. GCF was collected from eight posterior sites in each quadrant, and periodontal parameters were recorded. GCF was assayed for IL-8 by ELISA and BG by a fluorometric assay. RESULTS: GCF IL-8 was positively correlated with probing depth (PD), and GCF BG but not clinical attachment level (CAL), bleeding on probing (BOP), or plaque index (PI). In contrast, GCF BG was strongly correlated with each of the clinical measures of periodontal disease. Subjects with DM significantly lower levels of both BG (73.0+/-44.8 versus 121.9+/-84.6 pg/sample; p=0.002) and IL-8 (32.1+/-33.1 versus 90.8+/-83.2 pg/sample; p<0.0001) even after adjustments for age, gender, PD, CAL, BOP, and PI. Neither BG nor IL-8 was correlated with HbA1c levels in subjects with DM. CONCLUSION: These data suggest that an inadequate local response by PMN, partially explained by an altered chemokine gradient, may contribute to periodontal disease in patients with type 2 DM.     

白细胞介素-21在不同牙周状态龈沟液中的表达及相关性分析

目的检测白细胞介素(Interleukin,IL)-21在不同牙周状态下龈沟液中的表达,探讨IL-21在牙周免疫中的可能作用。方法本研究共纳入47例患者,分为健康对照组(15例),轻度牙周炎组(10例),中重度牙周炎组(22例);记录患者一般信息、用Florida牙周探针检查牙周袋探诊深度(PD)和临床附着丧失(CAL)情况;收集牙周治疗前的龈沟液样本,采用酶联免疫吸附法(ELISA)测定不同牙周条件下龈沟液中IL-21的水平;并进行Pearson秩相关检验统计IL-21的表达与相关临床指标的关系。结果 3组龈沟液中IL-21平均浓度分别为(107.20±2.54)、(218.90±5.11)、(367.80±7.27)pg/m L,轻度牙周炎组及中重度牙周炎组龈沟液中IL-21水平显著高于对照组(P<0.05),其中以中、重度牙周炎组增高最为显著;中、重度牙周炎组龈沟液中IL-21水平分别与PD、CAL呈正相关关系(P<0.001)。结论 IL-21在牙周疾病中有较高的表达,尤其是以中重度牙周炎患者最为显著;由此推测IL-21在牙周疾病的发生发展中占有重要地位,与牙周组织的破坏有密切关系。     

Expression of RANTES and CCR1 in oral lichen planus and association with mast cell migration

BACKGROUND: T lymphocytes and mast cells infiltrate the lamina propria in oral lichen planus (OLP). Chemokines and their receptors are involved in T cell and mast cell migration and accumulation during the inflammatory process. METHODS: In the present study, we investigated the role of RANTES and its receptors in OLP using immunohistochemistry, RT-PCR and an in vitro chemotaxis assay. RESULTS: RANTES and CCR1 were expressed on T cells and mast cells in OLP, while OLP lesional T cell supernatants stimulated CCR1 mRNA expression in a human leukemia mast cell line (HMC-1). TNF-alpha stimulated CCR1, CCR4 and CCR5 mRNA expression in the same cell line. OLP lesional T cell supernatants stimulated HMC-1 migration, which was partly inhibited by anti-RANTES antibody. CONCLUSIONS: The present study shows, for the first time, the distribution of RANTES and CCR1 in OLP. It is hypothesized that RANTES and CCR1 may play important roles in mast cell trafficking and related events in OLP.     

Effect of scaling and root planing on gingival crevicular fluid cytokine/chemokine levels in smokers with chronic periodontitis: A systematic review

In the present study, the impact of scaling and root planing (SRP) on gingival crevicular fluid (GCF) cytokine/chemokine levels in smokers with chronic periodontitis was assessed. The PICO (population, intervention, comparison, outcome) question was: In smokers with chronic periodontitis (population), what is the effect of SRP (intervention) in comparison to SRP in non‐smokers with chronic periodontitis (comparison) on the GCF cytokine/chemokine level (outcome)? Indexed databases were searched up to September 2017. Of 4330 titles, nine studies reporting the levels of 13 different cytokines/chemokines were included. Eight studies had a moderate risk of bias, while one study had a high risk of bias. Almost all cytokines/chemokines were pro‐inflammatory cytokines. Five cytokines/chemokines studied in four clinical studies were decreased in the smoker‐chronic periodontitis group following SRP. One study observed that the GCF levels of interleukin‐17 increased, while anti‐inflammatory osteoprotegerin was reduced in both the SCP and non‐smoker‐chronic periodontitis groups at follow up. However, the majority of cytokines/chemokines did not change in the SCP groups at follow up. The current weight of evidence is not sufficient to prove that SRP has an impact on GCF cytokine/chemokine profile in smokers with chronic periodontitis. Evaluation of wide panels of pro‐inflammatory cytokines/chemokines related to collagen degradation and alveolar bone destruction in future studies are warranted.     

CCR5 mediates pro-osteoclastic and osteoclastogenic leukocyte chemoattraction

Periodontal disease (PD) progression involves the selective leukocyte infiltration into periodontium, supposedly mediated by the chemokine/chemokine receptor system. In this study, we investigated the role of chemokine receptor CCR5 in the immunoregulation of experimental PD in C57BL/6 (WT) and CCR5KO mice. Aggregatibacter actinomycetem comitans infection triggered the chemoattraction of distinct CCR5+ leukocyte subpopulations (determined by flow cytometry): CCR5+F4/80+ leukocytes, which co-express CD14 , CCR2, TNF-α, and IL-1β, indicative of activated macrophages; and CCR5+CD4+ cells, which co-express CXCR3, IFN-γ, and RANKL, indicative of Th1 lymphocytes, therefore comprising pro-osteoclastic and osteoclastogenic cell subsets, respectively. CCR5KO mice presented a lower PD severity (lower inflammation and alveolar bone loss) when compared with the WT strain, since the migration of F4/80+, TNF-α+, CD4+, and RANKL+ cells specifically decreased due to the lack of CCR5. Also, ELISA analysis demonstrated that the production of TNF-α, IL-1β, IL-6, IFN-γ, and RANKL in periodontal tissues was significantly decreased in the CCR5KO strain. The periodontal bacterial load and antimicrobial patterns were unaltered in CCR5KO mice. Our results demonstrate that the chemokine receptor is involved in the migration of distinct leukocyte subpopulations throughout experimental PD, being a potential target for therapeutic intervention in PD.     

Interleukin-1 beta levels in gingival crevicular fluid in type 2 diabetes mellitus and adult periodontitis.

Interleukin-1 beta (IL-1beta) is a potent bone-resorptive cytokine that also mediates soft-tissue destruction by stimulating prostaglandin production and inducing collagenase and other protease activity. The literature suggests that this substance may be an important mediator of attachment loss in human periodontitis, and indicates that IL-1beta may be useful for locating sites of periodontal disease activity. There is some evidence that IL-1beta is produced by cells of the periodontium, and that it can be detected in gingival crevicular fluid (GCF). Many factors are known to contribute to the destruction of periodontal tissue. One of the most important is immune deficiency in diabetes. The aim of this study was to measure and compare the concentration of IL-1beta in the GCF of patients with non-insulin-dependent diabetes mellitus (Type 2 DM), otherwise healthy adults with periodontitis, and individuals with no periodontal disease in order to assess whether diabetes alters IL-1beta levels. We also examined relationships between GCF levels and the clinical parameters of pocket depth, plaque index, and bleeding index in each group. Seventeen patients with Type 2 DM, 17 adult periodontitis patients (AP), and 17 healthy controls were selected. The levels of IL-1beta in the GCF were quantified by ELISA. The mean IL-1beta concentrations in the Type 2 DM, AP, and control groups were 200.1 +/- 65.34 pg/microl, 131.35 +/- 67.66 pg/microl, and 80.0 +/- 36.08 pg/microl, respectively. The levels in the diabetic patients were significantly higher than those in the AP and control subjects. There were no significant correlations between IL-1beta level and any of the clinical data parameters for each group. We believe that the macrophages may over produce IL-beta in Type 2 DM and increased IL-1beta levels in diabetic patients could be linked to altered immune function.     

慢性牙周炎龈沟液中硫离子水平与临床相关性研究

目的:分析慢性牙周炎(CP)患者龈沟液中硫离子(su lfides)水平的变化与临床牙周指数的相关关系及其对诊断预后的意义。方法:采用金刚牙周诊断仪进行龈沟液硫离子和牙周临床指标测定。选定实验组(T):36例慢性牙周炎患者,57颗牙位,共342个位点。其中健康牙位(T1)21颗,位点126个;炎症牙位(T2)36颗,位点216。对照组(C):全身及牙周健康者8例,16颗牙位,共96个位点。测定所选位点龈沟液(GCF)中硫化物水平(su lcussu lph ide level,SUL),牙周袋探诊深度(prob ing depth,PD),牙周临床附着丧失水平(c lin ical attachm ent level,CAL),龈沟出血指数(su lcus b leed ing index,SB I)。所有统计结果均采用SPSS11.0进行统计学分析。结果:1)牙周健康对照组(C)GCF中硫离子SUL的浓度均值为(0.0648±0.0169)pg/mL,明显低于慢性牙周炎炎症牙位组(T2)(0.3249±0.0489)pg/mL及慢性牙周炎健康牙位组(T1)(0.1160±0.0271)pg/mL;慢性牙周炎炎症牙位组(T2)GCF中硫离子(SUL)的浓度均值均高于正常对照组及慢性牙周炎健康牙位组(T1)。2)经相关性分析,慢性牙周炎炎症牙位组(T1)GCF中SUL的浓度均值与PD、SB I和CAL均呈正相关关系。而慢性牙周炎健康牙位组(T1)及正常对照组(C)GCF中SUL的浓度均值与PD、SB I及CAL间无相关性。结论:慢性牙周炎(CP)炎症牙位组龈沟液中硫离子(SUL)的浓度均值与牙周临床指标之间具有相关关系,其水平的高低变化可客观反映牙周组织的炎症状态。     

Ⅱ型糖尿病合并牙周病患者龈沟液中细胞因子/趋化因子的表达水平

目的 探讨Ⅱ型糖尿病合并牙周病患者与单纯牙周病患者龈沟液（gingival crevicular fluid,GCF）中细胞因子/趋化因子的表达水平。 方法 选取伴Ⅱ型糖尿病的牙周病患者52例,单纯牙周病患者40例,用Luminex FLEXMAP3D仪和Human Cytokine/Chemokine试剂盒检测GCF中14种细胞因子/趋化因子的表达水平。 结果 牙周病部位：嗜酸性粒细胞趋化因子、巨噬细胞炎症蛋白-1α、粒细胞-巨噬细胞集落刺激、白介素-6、肿瘤坏死因子-α和白介素-12的浓度,糖尿病组受试者高于非糖尿病组受试者（P<0.0035）。 结论 糖尿病可影响牙周病部位细胞因子/趋化因子的表达,糖尿病可能是牙周病的促进因素。     

The new concept of periodontal disease pathogenesis requires new and novel therapeutic strategies

In this issue Bostanci et al. (2007) demonstrate that receptor activator of NF-kappaB ligand (RANKL) and osteoprotegerin (OPG) were oppositely regulated in gingival crevice fluid (GCF) from periodontitis patients. RANKL, RANK and OPG are key molecules that regulate osteoclast recruitment, differentiation and activation. New concepts of the pathogenesis of periodontitis have implicated inflammation triggered by host immune response to periodontal biofilm microorganisms(s) in disease. Host response to bacteria involves activation of T and B cells in the inflammatory infiltrate which bear abundant RANKL that promotes osteoclastic bone resorption. Periodontal tissue destruction can be ameliorated by immunobiological interference with immune cell RANKL expression or function. The new disease concepts provide a foundation to build biological approaches to target RANKL production in periodontal lesions.     

Interleukin-8 and β-glucuronidase in gingival crevicular fluid

Abstract Polymorphonuclear leukocytes (PMN) play a critical role in the host's response to the subgingival microflora. Interleukin-8 (IL-8) is a potent chemotactic and activating factor for PMN. In this study, the presence of IL-8 in gingival crevicular fluid (GCF) was examined in relation to the PMN indicator β-glucuronidase (βG), as well as clinical parameters of chronic inflammatory periodontal disease. Data was obtained from 30 patients with periodontitis and 14 healthy controls. For the control group, GCF and clinical data were obtained only once. For the periodontitis patients, clinical data and GCF samples were collected prior to treatment, and GCF samples were again collected 2 weeks after scaling and root planing. Comparing control and periodontitis patients prior to treatment, IL-8 concentration was lower in the patients with periodontitis. Scaling and root planing resulted in either an increase or a decrease in total IL-8 and IL-8 concentration GCF. A reduction in total IL-8 or IL–8 concentration was accompanied by a corresponding reduction in βG activity. An increase in total IL-8 or IL-8 concentration after scaling and root planing was associated with an increase in βG activity in some patients and a reduction in βG activity in other patients. The periodontitis patients who did not demonstrate a linkage between IL-8 and βG activity in GCF were those individuals with the highest βG activity prior to treatment. As elevated βG activity in GCF has been associated with an increased risk for probing attachment loss, the absence of a direct relationship between IL-8 in GCF and PMN recruitment into the gingival crevice may characterize individuals at risk for progression of periodontitis.     

Interleukin-1 and IL-1 receptor antagonist in gingival crevicular fluid

BACKGROUND/AIMS: This study aimed to investigate the cytokine IL-1beta and its receptor antagonist IL-1ra in gingival crevicular fluid (GCF), in patients with adult periodontitis. METHOD: A total of 40 GCF samples were harvested from 10 subjects with moderate to severe adult periodontitis and 10 healthy controls. Subjects were selected from both genders, with all the upper anterior teeth present, and with no relevant systemic illness, pregnancy or recent medication. All subjects were non-smokers and had not received any periodontal therapy within the preceding 3 months. Deep bleeding sites, deep non-bleeding sites and healthy sites were investigated in relation to upper anterior teeth. Clinical measurements were recorded for each site, after obtaining a GCF sample. IL-1beta and IL-1ra were quantified using new commercially available ELISA kits (Quantikine), and could be detected in all samples. RESULTS: The mean concentration for IL-1beta was 0.11 (SD 0.14) pg/microl for bleeding periodontitis sites, 0.04 (0.05) pg/microl for non-bleeding periodontitis sites, and 0.01 (0.03) pg/microl for healthy sites (p<0.001). In contrast, the mean concentration for IL-1ra was 6.99 (9.78) pg/microl for healthy sites, 0.59 (0.44) pg/microl for non-bleeding periodontitis sites, and 0.44 (0.36) pg/microl for bleeding periodontitis sites (p<0.001, except for comparisons between bleeding and non-bleeding periodontitis sites, p>0.05). For healthy sites, a strong inverse relationship was found between IL-1beta and IL-1ra levels in GCE. CONCLUSIONS: The results suggest a strong relationship between the severity of adult periodontitis and the increasing GCF levels of IL-1beta and decreasing levels of IL-1ra.     

Profiling the cytokines in gingival crevicular fluid using a cytokine antibody array

BACKGROUND: Various compounds have been detected in gingival crevicular fluid (GCF) as indicators of periodontal disease activity. Therefore, the analysis of GCF may be especially beneficial for diagnosing current periodontal status and addressing the effects of treatment. Moreover, the identification of new markers in GCF may also contribute to elucidating novel mechanisms involved in periodontal disease. This study sought novel marker proteins specific to chronic periodontitis by profiling cytokines in GCF using a cytokine antibody array system. METHODS: Human cytokine array V, which detects 79 cytokines on one membrane, was used to determine the profile of cytokines in GCF from seven subjects with chronic periodontitis and seven subjects with healthy periodontia. The profile was exposed to x-ray film and quantified using image analysis software. Healthy and diseased sites were compared statistically. RESULTS: We detected 10 cytokines in periodontally healthy sites and 36 cytokines in periodontally diseased sites. Interleukin-8 (IL-8) and transforming growth factor-beta 2 (TGF-beta2) were detected at high levels in healthy and diseased subjects. There were significant differences between healthy and diseased subjects in the levels of tissue inhibitor of metalloproteinases-2 (TIMP-2), tumor necrosis factor-beta (TNF-beta), growth-related oncogene (GRO), interferon-inducible protein-10 (IP-10), angiogenin (Ang), vascular endothelial growth factor (VEGF), insulin-like growth factor binding protein-3 (IGFBP-3), osteoprotegerin (OPG), epidermal growth factor (EGF), glial-derived neurotrophic factor (GDNF), pulmonary and activation-regulated chemokine (PARC), oncostatin M (OSM), fibroblast growth factor-4 (FGF-4), IL-16, homologous to lymphotoxins (LIGHT), and placenta growth factor (PlGF). Of these, the newly detected cytokines were GRO, Ang, IGFBP-3, GDNF, PARC, OSM, FGF-4, IL-16, LIGHT, and PlGF. CONCLUSIONS: In this study, we detected several cytokines in GCF using a cytokine antibody array system, including both inflammatory cytokines and various growth factors. Therefore, periodontal disease may participate in the wound healing process and in tissue destruction via the inflammatory process. Our results suggest that the quantification of these cytokines in GCF provides useful information for the diagnosis of periodontal disease status.     

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。     

Identification of the osteoprotegerin/receptor activator of nuclear factor-kappa B ligand system in gingival crevicular fluid and tissue of patients with chronic periodontitis

BACKGROUND AND OBJECTIVE: Recent findings have suggested that osteoclastogenesis is directly regulated by receptor activator of nuclear factor-kappa B ligand (RANKL) and its decoy receptor, osteoprotegerin (OPG). However, no studies have described interactions of OPG/RANKL and the gp130 cytokine family in periodontal disease. This study aimed to identify and quantify OPG/RANKL in the gingival crevicular fluid (GCF) and connective tissue of patients with periodontitis, and to clarify possible correlations with disease severity and interleukin-6 (IL-6) cytokines. MATERIAL AND METHODS: Ninety-five sites in 20 patients with generalized chronic periodontitis were divided into four groups by site based on probing depth (PD) and bleeding on probing (BOP). In periodontitis patients, GCF was obtained using sterile paper strips from clinically healthy sites (PD 6 mm with BOP, n = 27). Fourteen clinically healthy sites from four periodontally healthy individuals were used as the control group. The levels of OPG, RANKL and two gp130 cytokines - IL-6 and oncostatin M (OSM) - in the GCF were determined by an enzyme-linked immunosorbent assay (ELISA) and are expressed as total amounts (pg/site). Immunohistochemical localization of OPG- and RANKL-positive cells was also performed on gingival connective tissues harvested from patients with periodontitis (inflammatory group, n = 8 biopsies) and from non-diseased individuals (healthy group, n = 8 biopsies). RESULTS: GCF RANKL, but not OPG, was elevated in diseased sites of patients with periodontitis. However, the expressions of OPG and RANKL showed no correlation with disease severity (r = 0.174 and 0.056, respectively), but the content of RANKL in the GCF was significantly positively correlated with those of IL-6 (r = 0.207) and OSM (r = 0.231) (p < 0.01). Immunohistochemical staining showed that RANKL-positive cells were significantly distributed in the inflammatory connective tissue zone of diseased gingiva, compared with those of samples from non-diseased persons (p < 0.01). However, few OPG-positive cells were found in connective tissue zones of either the diseased gingiva or healthy biopsies. CONCLUSION: These findings imply that in this cross-sectional study of GCF, RANKL, IL-6 and OSM were all prominent in periodontitis sites, whereas OPG was inconsistently found in a few samples of diseased sites but was undetectable in any of the control sites. The results also imply that the expression of RANKL was positively correlated with IL-6 and OSM in the GCF.