糜烂型OLP患者唾液Th1、Th2、Th17相关细胞因子及β防御素的表达

目的：检测Th1、Th2、Th17相关细胞因子和HBD－2,－3在糜烂型口腔扁平苔藓（EOLP）患者唾液中的表达,探讨其在EOLP发生发展中的作用。方法：分别纳入EOLP患者30例及按年龄和性别匹配的健康者20例作为实验组和对照组,采用流式微球技术（Cytometric Bead Array,CBA）检测唾液中IL－1β、TNF－α、IL－6、IL－17、IFN－γ、IL－4、IL－10的表达水平,并采用酶联免疫吸附测试法（ELISA）检测唾液中HBD－2和HBD－3。结果：EOLP患者唾液中IL－1β、IL－6、HBD－2、HBD－3水平显著高于对照组（P＜0．05）,TNF－α、IL－17、IFN－γ、IL－4、IL－10与对照组比较差异无显著性（P＞0．05）。患者唾液HBD－2的表达与IFN－γ呈正相关、与IL－17呈负相关,HBD－3的表达与IL－17和IL－10均呈负相关。结论：唾液中IL－1β、IL－6、HBD－2和HBD－3的高表达可能与EOLP的发生发展密切相关。     

Association study between salivary levels of interferon (IFN)-gamma,interleukin (IL)-17, IL-21, and IL-22 with chronic periodontitis

Objective

To investigate if the salivary levels of IL-17, IL-21, IL-22, and its ratio regarding salivary IFN-γ may be linked with the periodontal clinical status.

Design

One hundred and five chronic periodontitis (CP) subjects and 44 healthy controls (HC) were recruited. Periodontal status was assessed based on full-mouth clinical periodontal measurements. Cytokine salivary levels were analyzed by ELISA. The association between the analytes with CP was analyzed using a binary logistic regression model.

Results

A statistically significant increase in salivary levels of IFN-γ and IFN-γ/IL-22 ratio in CP group could be detected, but there was no significant domination of any Th17 cytokine that could be of predictive value for health/disease status. Univariate and binary logistic regression analyses revealed a strong and independent association of IFN-γ salivary levels and IFN-γ/IL-22 ratio with disease status. An interaction effect of ageing on IFN-γ levels also could be noted.

Conclusion

While salivary levels of IFN-γ and IFN-γ/IL-22 ratio may act as strong/independent indicators of the amount and extent of periodontal breakdown, the low detection frequency of Th17 cytokines in saliva samples make these determinations useless for the detection of disease presence and/or its severity.     

Expression and secretion levels of Th1 and Th2 cytokines in patients with aggressive periodontitis

The role of Th1 and Th2 cytokines in the pathogenesis of aggressive periodontitis has not been previously examined. The aim of this study was to analyse the expression and production of IL-2, IFN-γ, IL-4 and IL-13 in CD4+ cells from the peripheral blood of patients with aggressive periodontitis (AgP) and periodontally healthy controls. Gene expression was analysed in inactivated and activated CD4+ cells by real-time PCR. Cells were activated for 4, 8 and 24?h with anti-CD3/CD28 antibody, phytohemagglutinin (PHA), and Porphyromonas gingivalis (P.g.) outer membrane protein (OMP). Protein levels were measured in supernatants of activated CD4+ cells by bead-based immunoassay (CBA). Statistics were performed using U test (p?相似文献   

Effects of Calcium Hydroxide Intracanal Medications on T Helper (Th1, Th2, Th9, Th17, and Tfh) and Regulatory T (Treg) Cell Cytokines in Apical Periodontitis: A CONSORT RCT

IntroductionThis Consolidated Standards of Reporting Trials randomized clinical trial investigated T helper (Th1, Th2, Th9, Th17, and Tfh) and regulatory T (Treg) cell–type cytokines and their networks in apical periodontitis (AP). We also assessed the effects of calcium hydroxide [Ca(OH)2] intracanal medications (ICMs) on helper T and Treg cell–type cytokines.MethodsTwenty teeth with primary endodontic infection and apical periodontitis were randomly divided into two groups: Ca(OH)2 + saline solution (n = 10) and Ca (OH)2 + 2% chlorhexidine-gel (n = 10). Samples were collected from the periradicular tissue fluid (PTF) before (PTFs1) and after 14 days of ICMs (PTFs2). The Human High Sensitivity T Cell Panel was used to quantify target T-helper (Th)1: interelukin (IL)-2, IL-12, and interferon-gamma (IFN-γ); Th2: IL-4, IL-5, and IL-13; Th9: IL-9; Th17: IL-17; T follicular helper cells (Tfh): IL-21; and Treg-cell-type cytokine: IL-10.ResultsTh1-type cytokines were higher than Th2-type ones, at PTFs1. Positive (+) associations were found among all Th1-type cytokines and all Th2-type cytokines. There were negative (-) correlations between all Th1- and Th2-type cytokines. Size of radiolucent lesions and symptoms (tenderness to percussion and/or pain on palpation) were positively correlated with Th1-type cytokines, IL-17, and IL-21 but negatively correlated with Th2-type cytokines and IL-10 (all, P < .001). Both ICMs increased Th2-type cytokines and IL-10 (P < .05) and decreased Th1-type cytokines, IL-17, and IL-21 (P < .05), with no differences among them (P > .05).ConclusionsComplex T-cell cytokine networks are involved in AP. Both Ca(OH)2 ICMs effectively increased IL-4, IL-5, IL-10, and IL-13 and lowered IL-2, IL-12, IL-17, IL-21, and IFN-γ.     

口腔扁平苔藓患者Th1／Th2失衡表达的研究

目的：观察口腔扁平苔藓（oral liclaen planus,OLP）患者外周血中Th1、Th2型细胞转录因子T-het和GATA-3以及Thl、Th2型细胞因子的表达,进一步探索OLP的发病机制。方法：通过密度梯度离心法分离20例充血糜烂型,16例光滑型OLP病例和19例正常对照组人群的外周血单个核细胞,逆转录-聚合酶链反应（RT—PCR）法检测各组外周血单个核细胞中T—betmRNA和GATA-3 mRNA的表达;ELISA法检测各组血清中干扰素-γ（interferon gamma,IFN-1）和白细胞介素-4（interleukin-4,IL-4）的表达。结果：充血糜烂型和光滑型OLP患者中T—betmRNA、IFN-γ的表达均低于正常对照组,差异有统计学意义（P〈0．01）;而充血糜烂型和光滑型OEP患者中GATA-3 mRNA、IL-4的表达均高于正常对照组,差异有统计学意义（P〈0．01）。T—bet mRNA和GATA-3 mRNA的表达在充血糜烂型及光滑型OLP患者组间的差异有统计学意义（P〈0．01）;而IFN-γ和IL-4的表达在充血糜烂型及光滑型OLP患者组间的差异无统计学意义（P〉0．05）。结论：Th1／Th2失衡表达与OLP发病机制密切相关,为临床治疗OLP提供参考。     

Porphyromonas gingivalis infection enhances Th17 responses for development of atherosclerosis

Objectives

Porphyromonas gingivalis has been shown to associate with the development of atherosclerosis. Recent studies indicate that IL-17-producing T helper 17 (Th17) cells have been correlated with the emergence of atherosclerosis. Therefore, we investigated whether the Th17 cell response and expression of Th17-related molecules, in contrast with Th1- and Treg cells, are enhanced by P. gingivalis-challenge in Apolipoprotein E knockout (ApoE KO) mice.

Design

Five mice were intravenously injected with P. gingivalis three times a week for 3 weeks and killed at 15 weeks of age. The proximal aorta lesion area, flow cytometry analysis and IL-17, IL-10, IFN-γ, and IL-1β levels in splenic cultures, and expression of Th17-related molecules in spleen and hearts were examined.

Results

P. gingivalis-challenge showed notable accumulation of atherosclerotic plaques by Oil Red O-staining in ApoE KO mice. Intracellular cytokine staining revealed that significantly elevated CD4⁺ interleukin (IL)-17A⁺ T cells and slightly increased CD4⁺ Foxp3⁺ T cells was recognized in spleen cells of P. gingivalis-challenged mice compared with those from non-infected mice. P. gingivalis-challenge significantly increased IL-17 and IL-1β production and RORγt expression in splenic cells. Furthermore, the expression of Th17-related genes such as IL-6, TGF-β, RORγt and STAT3 were elevated in splenic cells as well as heart tissue of P. gingivalis-challenged mice.

Conclusion

These results suggest that P. gingivalis infection may enhance pro-inflammatory Th17 cell responses in lesion areas and spleen, thereby accelerating atherosclerosis.     

Analysis of Th-cell subsets in local and systemic environments from experimental periodontitis rats

Objectives : The objective of this study was to explore the effect of periodontitis on Th-cell subsets in local and systemic environments. Methods : A total of 32 male Sprague‒Dawley rats were randomly divided into periodontitis and control groups. Silk ligatures were applied to the mandibular first (M1) molars in the periodontitis group. Inflammation and alveolar bone loss around the M1 molars were analyzed by histological staining and microcomputed tomography. The mRNA expression of interferon-γ (IFN-γ), interleukin 4 (IL-4), IL-17, and IL-10 in the gingiva was measured by qRT-PCR. The proportions of Th1, Th2, Th17, and Treg cells in the submandibular lymph nodes, peripheral blood, and jaw bone marrow were tested using flow cytometry. Results : More inflammatory cells and alveolar bone resorption were found in the periodontitis group, with upregulated mRNA expression of IFN-γ, IL-17, and IL-10. The proportion of Th1 and Th17 cells was significantly elevated in submandibular lymph nodes, and the proportion of Th1, Th2, and Th17 cells was significantly elevated in peripheral blood, while the proportion of Th1, Th17, and Treg cells was significantly elevated in jaw bone marrow in the periodontitis group. Conclusion : This study suggests that periodontitis affects the differentiation of Th-cell subsets in both local and systemic environments, resulting in an increased proportion of proinflammatory cells.     

Th1／Th17细胞相关因子在根管治疗后疾病患牙根尖组织中的表达

目的研究Th1／Th17细胞相关因子在根管治疗后疾病（post—treatment endodontic disease,PED）患牙根尖组织中的表达。方法收集12例根管治疗失败患牙的根尖组织样本,制作组织学切片后,以免疫组化方法检测根尖组织样本中Th1／Th17细胞相关因子白细胞介素-17、干扰素-γ蛋白的表达,用ImageProPlus图像分析软件测量各样本中阳性染色区域的平均光密度值,并进行统计学分析。结果12例样本病理检查确诊2例根尖肉芽肿,10例根尖囊肿,通过免疫组化检验证实12例病变组织均检测出白细胞介素-17、干扰素-γ蛋白的表达,且自细胞介素-17表达高于干扰素-γ（t=6．720,P〈0．05）。结论Th1／Th17细胞相关因子均参与了根管治疗失败患牙根尖炎症过程,而Th17型炎症反应可能在PED发生中发挥主要作用。     

实验性牙周炎大鼠体内外周血及牙周组织中Th17/Treg的检测

目的初步探讨实验性牙周炎大鼠外周血及炎症牙周组织中辅助性T细胞17（Th17）和调节性T细胞（Treg）的表达。 方法采用丝线结扎并牙龈卟啉单胞菌菌液涂抹法建立SD大鼠牙周炎模型,采集外周血、龈沟液以及颌骨样本。体视显微镜及苏木精-伊红染色法观察牙周组织病理改变,流式细胞术检测外周血Th17细胞与Treg细胞的比例,酶联免疫吸附法检测外周血及龈沟液中白细胞介素17（IL-17）、IL-10、转化生长因子β（TGF-β）表达水平,实时荧光定量聚合酶链反应（PCR）法检测牙周组织中RORγt、Foxp3 mRNA的表达。应用SPSS 22.0软件独立样本t检验及方差分析对数据进行统计学分析。 结果实验大鼠牙周组织病理改变符合牙周炎病理改变特征,牙槽骨吸收明显。实验组外周血Th17（F=30.88,P=0.00）、Treg细胞（F=147.03,P=0.00）比例以及Th17/Treg比率较对照组均显著上升（F=219.53,P=0.00）;实验组牙周组织中RORγt（F=35.61,P=0.04）和Foxp3 mRNA（F=216.60,P=0.01）表达较对照组表达显著上升;RORγt/Foxp3 mRNA比率较对照组升高,但差异无统计学意义（F=0.63,P=0.44）;实验组外周血血清中IL-10浓度较对照组显著上升（F=6.41,P=0.02）,而IL-17（F=0.08,P=0.78）、TGF-β（F=3.48,P=0.06）浓度与对照组差异无统计学意义;实验组龈沟液中IL-17（F=9.03,P=0.01）较对照组显著上升;IL-10（F=5.85,P=0.08）及TGF-β含量（F=9.28,P=0.06）含量较对照组升高,但差异无统计学意义。 结论实验性牙周炎大鼠外周血及牙周炎症组织中Th17与Treg表达均显著上调,可能与牙周炎症组织病理改变以及相关全身系统性疾病存在相关性。     

Porphyromonas gingivalis induces myocarditis and/or myocardial infarction in mice and IL-17A is involved in pathogenesis of these diseases

ObjectivesAlthough an association between periodontitis and cardiovascular diseases has been suggested, the role of Porphyromonas gingivalis in cardiovascular diseases is not clear. In this study, we examined whether experimental bacteremia of P. gingivalis causes cardiovascular diseases and investigated the mechanism of pathogenesis of cardiovascular diseases induced by P. gingivalis.DesignC57BL/6 mice were intravenously inoculated with 2.0 × 10⁸ CFU of P. gingivalis A7436 strain. Mice were sacrificed at specified days and their hearts were collected. The collected organs were divided into two halves and used for histological evaluation and cytokine analysis. IL-17A^?/?, IFN-γ^?/? and TNF-α^?/? mice were also intravenously inoculated and the histological changes of hearts in mice were examined.ResultsMyocarditis and/or myocardial infarction were observed in mice injected with P. gingivalis. The levels of IL1-β, IL-6, IL-17A, IL-18, TNF-α and IFN-γ mRNA increased significantly after P. gingivalis injection. In particular, high levels of IL-17A and IFN-γ mRNA expression were observed in hearts of mice after P. gingivalis injection in comparison with these levels before injection. Furthermore, the production of IL-17A was detected in hearts of wild-type mice after P. gingivalis injection. In wild-type, TNF-α^?/? and IFN-γ^?/? mice, moderate infiltration of neutrophils and monocytes was observed in hearts at 5 days after injection. In contrast, no inflammatory findings were observed in hearts of IL-17A^?/? mice.ConclusionWe have demonstrated that an experimental bacteremia of P. gingivalis could induce myocarditis and/or myocardial infarction in mice, and IL-17A plays an important role in the pathogenesis of these diseases.     

Th1/Th2/Th17/Treg Balance in Apical Periodontitis of Normoglycemic and Diabetic Rats

IntroductionThe aim of this study was to evaluate the inflammatory profile of T helper (Th) cells in normoglycemic (N) and diabetic rats with apical periodontitis (AP).MethodsTwenty male Wistar rats were divided in 2 groups: N rats and rats with diabetes mellitus (DM). DM was induced using streptozotocin, and AP was induced by dental pulp exposure of the first mandibular molar to the oral environment. After 30 days, the mandibles were removed and processed for histologic analysis, bacterial analysis, and immunochemical assays for interleukin (IL)-6, tumor necrosis factor alpha, IL-17, IL-23, interferon gamma, and IL-10. The Mann-Whitney U test and Student t test were used for statistical analysis (P < .05).ResultsThe DM group showed more intense inflammatory infiltrate with larger sizes of bone reabsorption and a greater presence of bacteria than the N group (P < .05). Proinflammatory cytokine levels in the DM group were also greater than those in the N group (P < .05). However, interferon gamma was more intense in the N group than in the DM group (P < .05).ConclusionsThe inflammatory profile of AP in DM is different from that in the N group, suggesting that Th1 is a secondary strain and the Th17 strain is predominant in DM.     

Th1/Th2 balance in mouse delayed-type hypersensitivity model with mercuric chloride via skin and oral mucosa

In order to compare delayed-type hypersensitivity (DTH) among different exposure sites, we evaluated the sensitization potency of mercuric chloride (HgCl(2)) via exposure to the skin, or oral or esophageal mucosa using the mouse ear swelling test. Furthermore, we investigated in vitro splenocyte proliferation reaction and cytokine profile in HgCl(2)-exposed and control mice. Sensitization with HgCl(2) was established via the skin and oral mucosa but not via the esophageal mucosa. The splenocyte proliferation reaction was significantly enhanced to a similar degree in skin and oral mucosa-sensitized mice compared with in the control mice. IL-10 levels from cultured splenocytes were significantly increased in skin and oral mucosa-sensitized mice compared with those in control mice, whilst IFN-γ significantly increased only in splenocytes from skin-sensitized mice. These results suggest that exposure of the skin or oral mucosa to HgCl(2) can induce DTH, but that Th1/Th2 balance differs according to the site of antigen exposure.     

Overexpression and varied clinical significance of Th9 versus Th17 cells in distinct subtypes of oral lichen planus

ObjectiveOral lichen planus (OLP) presents with large numbers of T lymphocytes accumulating beneath the epithelium of the oral mucosa; however, its aetiology remains obscure. A potential role for an emerging novel T cell subset, Th9, in OLP has recently been suggested but remains to be clarified. The current aim was to investigate the expression and potential clinical significance of Th9 cells in distinct subtypes of OLP.Materials and methodsPeripheral blood samples were collected from 41 OLP patients and 18 healthy controls (HCs). Flow cytometric analysis was used to detect the CD4+ T helper subset Th9 (IL-9⁺IL-17⁻CD4⁺ Th cells) and Th17 (IL-9⁻IL-17⁺CD4⁺ Th cells) expression levels.ResultsFlow cytometry results showed significantly elevated levels of Th9 cells in reticular and erosive OLP compared to HCs. Th9 expression in erosive OLP was less than in reticular OLP, indicating that Th9 but not Th17 cells may play a predominant role in reticular disease. However, in erosive OLP patients, we found much higher levels of Th17 cells compared to reticular OLP patients and HCs, indicating that Th17 dominates in erosive OLP. Statistical analysis showed positive correlations of Th9 cells and Th17 cells in patients with reticular or erosive OLP but none in HCs.ConclusionsTh9 and Th17 cells may take the predominant roles in reticular and erosive OLP respectively, and their numbers were positively correlated in reticular and erosive OLP patients. Elevated circulating Th9 cells may help maintain immune balance in OLP immunopathogenesis, which requires further investigation.     

Modulatory Roles of Interferon-γ through Indoleamine 2, 3-dioxygenase Induction in Innate Immune Response of Dental Pulp Cells

Introduction

Marked infiltration of inflammatory cells such as activated T cells producing interferon-γ (IFN-γ) is observed in severe pulpitis. However, the roles of IFN-γ in the innate immune response of dental pulp have not been reported. Indoleamine 2, 3-dioxygenase (IDO) is a regulator of immune responses, and the IDO expression is induced by IFN-γ in many cells whose expression in dental pulp is unknown. The purpose of this study was to determine the role of IFN-γ in the immune response through microbial pattern recognition receptors (PRRs) such as Toll-like receptors or nucleotide-binding oligomerization domain–like receptors on the production of proinflammatory cytokines such as CXCL10 and interleukin (IL)-6 and the expression of IDO in cultured human dental pulp cells (HDPCs).

Methods

HDPCs were established from explant cultures of healthy pulp tissues. CXCL10 and IL-6 production was determined using enzyme-linked immunosorbent assay. Confirmation of IDO localization in dental pulp tissues was examined using immunohistochemistry. IDO expression in HDPCs was analyzed by immunoblot.

Results

IFN-γ significantly up-regulated CXCL10 and IL-6 production in the HDPCs stimulated with ligands for PRRs in a concentration-dependent manner. The expression of IDO was detected in inflamed pulp tissue. In addition, IFN-γ in combination with the PRR ligands enhanced IDO expression in HDPCs compared with IFN-γ alone. Moreover, CXCL10 production in IFN-γ–stimulated HDPCs was inhibited by an IDO inhibitor.

Conclusions

This study showed the synergistic effects by IFN-γ on cytokine production and IDO expression in HDPCs, suggesting that IFN-γ may modulate the innate immune response of dental pulp.     

中药养阴益气合剂对口腔扁平苔藓相关细胞因子影响的研究

目的观察中药养阴益气合剂对口腔扁平苔藓患者外周血中相关细胞因子水平的影响,探讨口腔扁平苔藓的免疫应答模式。方法选取阴虚血瘀证型口腔扁平苔藓患者40例,采用中药养阴益气法治疗3个月,检测治疗前、后NF-KB依赖性炎症介质IL-1β、IL-6、IL-8和Th1／Tb2代表性细胞因子IFN-γ、IL-10水平的变化。并选取18例健康志愿者,同时检测IL-1β、IL-6、IL,8、IFN-γ和IL-10水平,作为对照。结果中药养阴益气法治疗阴虚血瘀型口腔扁平苔藓总有效率达87．5％,治疗前,IL-6、IL-8表达水平明显高于对照组（P〈0．05）,治疗后,IL-8、IL-10与治疗前比较差异有显著性（P〈0．05）。结论口腔扁平苔藓的不同阶段患者的免疫状态是变化的,IL-8、IFN-γ/IL-10比值对于监测口腔扁平苔藓的病情可能更加敏感。中药养阴益气合剂可能通过调节Th1／Th2免疫平衡和阻断NF-KB信息通路的激活而发挥其治疗作用。     

牙龈卟啉单胞菌表面相关物质刺激淋巴细胞分化效应T细胞的研究

目的探讨牙龈卟啉单胞菌表面相关物质（SAM）刺激淋巴细胞活化后效应T细胞的表达及意义。方法选取10名全身及牙周组织健康受试者静脉血,分离外周血单核细胞（PBMC）,体外刺激PBMC：实验组加入SAM冻干产物（浓度为25μg/mL）;阳性对照组加入刺激抗原PMA（25μg/mL）＋ionomycin（1μg/mL）;阴性对照组不加任何刺激抗原。流式细胞仪检测T细胞内细胞因子IL-2、IFN-γ、IL-4、IL-10的表达。结果 SAM刺激淋巴细胞后上述四种细胞因子呈低水平表达;SAM诱导IL-4、IL-10表达的能力相对较强。结论 SAM刺激可造成T淋巴细胞免疫缺陷,导致IL-2、IFN-γ等功能障碍,激发TH2细胞活性,可导致机体免疫功能受损和牙周组织的继发性损伤。     

Influences of interferon-gamma on cell proliferation and interleukin-6 production in Down syndrome derived fibroblasts

Objective

Down syndrome, a frequently encountered genetic disorder, is usually associated with medical problems related to infectious disease, such as periodontal diseases and prolonged wound healing. Although affected individuals are considered to have clinical problems related to high interferon (IFN) sensitivity, the molecular mechanisms of IFN activities are not completely understood.

Design

Down syndrome derived fibroblasts, Detroit 539 (D1) and Hs 52.Sk (D2) cells, were used. To analyse the expressions of interferon (IFN) receptors and downstream of IFN-γ, western blotting was performed. Cell proliferation was determined by counting cells following trypan blue staining. Media levels of IL-1β, TNF-α, and IL-6 were quantified using ELISA.

Results

IFN-γ receptor 2 and IFN-α receptor 1, but not IFN-γ receptor 1, were highly expressed in D1 and D2 cells, as compared to the control fibroblast cells. Cell proliferation by D1 and D2 cells was lower than that by the control fibroblasts, further, IFN-γ had a greater effect to inhibit cell proliferation by D1 and D2 cells. In addition, IFN-γ treatment increased the phosphorylation of STAT1 and MAPK in D1 cells as compared to normal fibroblasts. Also, the presence of exogenous IFN-γ in the growth medium significantly induced IL-6, but not IL-1β or TNF-α, in D1 and D2 cells.

Conclusion

Taken together, our results are consistent with hypersensitive reactions to IFN-γ seen in patients with Down syndrome and may provide useful information to elucidate the mechanisms of IFN-γ activities in those individuals.     

Human β-defensin 2 and protease activated receptor-2 expression in patients with chronic periodontitis

Objective

Some previous studies have shown that gingipains, trypsin-like proteases produced by Porphyromonas gingivalis, up-regulate human β defensin-2 (HBD-2) mRNA expression through protease-activated receptor-2 (PAR₂) in gingival epithelial cells. This study aimed at investigating salivary HBD-2 levels and crevicular PAR₂ mRNA expression in human chronic periodontitis and evaluating whether periodontal treatment affected this process.

Methods

Salivary and gingival crevicular fluid (GCF) samples were collected from periodontally healthy (control) and chronic periodontitis patients at baseline and 50 days after non-surgical periodontal treatment. Salivary HBD-2, and GCF TNF-α levels were analysed by ELISA, and PAR₂ mRNA at the GCF was evaluated by RT-PCR.

Results

P. gingivalis was significantly (p < 0.05) more prevalent in patients with chronic periodontitis when compared to controls. This prevalence decreased after periodontal therapy (p < 0.0001). The control group showed statistically significant lower levels of HBD-2, TNF-α, and PAR₂ expression when compared to the chronic periodontitis group. In addition, periodontal treatment significantly reduced PAR₂ expression and HBD-2 levels in chronic periodontitis patients (p < 0.001).

Conclusions

Our results suggest that salivary HBD-2 levels and PAR₂ mRNA expression from GCF are higher in subjects with chronic periodontitis than in healthy subjects, and that periodontal treatment decreases both HBD-2 levels and PAR₂ expression.     

Simultaneous analysis of T helper subsets (Th1, Th2, Th9, Th17, Th22, Tfh,Tr1 and Tregs) markers expression in periapical lesions reveals multiple cytokine clusters accountable for lesions activity and inactivity status

Previous studies demonstrate that the balance between pro- and anti-inflammatory mediators determines the stable or progressive nature of periapical granulomas by modulating the balance of the osteoclastogenic factor RANKL and its antagonist OPG. However, the cytokine networks operating in the development of periapical lesions are quite more complex than what the simple pro- versus anti-inflammatory mediators'' paradigm suggests. Here we simultaneously investigated the patterns of Th1, Th2, Th9, Th17, Th22, Thf, Tr1 and Tregs cytokines/markers expression in human periapical granulomas.

Methods

The expression of TNF-α, IFN-γ, IL-17A, IL23, IL21, IL-33, IL-10, IL-4, IL-9, IL-22, FOXp3 markers (via RealTimePCR array) was accessed in active/progressive (N=40) versus inactive/stable (N=70) periapical granulomas (as determined by RANKL/OPG expression ratio), and also to compare these samples with a panel of control specimens (N=26). A cluster analysis of 13 cytokine levels was performed to examine possible clustering between the cytokines in a total of 110 granulomas.

Results

The expression of all target cytokines was higher in the granulomas than in control samples. TNF-α, IFN-γ, IL-17A and IL-21 mRNA levels were significantly higher in active granulomas, while in inactive lesions the expression levels of IL-4, IL-9, IL-10, IL-22 and FOXp3 were higher than in active granulomas. Five clusters were identified in inactive lesion groups, being the variance in the expression levels of IL-17, IL-10, FOXp3, IFN-γ, IL-9, IL-33 and IL-4 statistically significant (KW p<0.05). Three clusters were identified in active lesions, being the variance in the expression levels of IL-22, IL-10, IFN-γ, IL-17, IL-33, FOXp3, IL-21 and RANKL statistically significant (KW p<0.05).

Conclusion

There is a clear dichotomy in the profile of cytokine expression in inactive and active periapical lesions. While the widespread cytokine expression seems to be a feature of chronic lesions, hierarchical cluster analysis demonstrates the association of TNF-α, IL-21, IL-17 and IFN-γ with lesions activity, and the association of FOXP3, IL-10, IL-9, IL-4 and IL-22 with lesions inactivity.