Gingival crevicular fluid levels of calprotectin and myeloperoxidase during therapy for generalized aggressive periodontitis

BACKGROUND: Levels of the inflammation marker calprotectin in gingival crevicular fluid correspond to clinical and biochemical parameters of periodontal inflammation. Neutrophil granulocytes (polymorphonuclear neutrophils: PMNs) are supposed to be the main source of calprotectin in gingival crevicular fluid, but evidence is still lacking. The influence of periodontal therapy on gingival crevicular fluid levels of calprotectin has not yet been determined. OBJECTIVES: Gingival crevicular fluid levels of calprotectin were monitored during therapy for generalized aggressive periodontitis. Interrelations between calprotectin and the PMN marker myeloperoxidase (MPO) were evaluated. MATERIAL AND METHODS: Gingival crevicular fluid samples were collected from 23 patients with generalized aggressive periodontitis before and 3 months after non-surgical therapy with an adjunctive antimicrobial medication. Clinical parameters were recorded with a pressure-calibrated electronic probe. Levels of calprotectin and MPO in gingival crevicular fluid were analysed by enzyme-linked immunosorbent assay (ELISA) procedures. RESULTS: At baseline, levels of calprotectin and MPO were highly correlated. Bleeding and suppurating sites showed significantly higher levels of calprotectin and MPO than non-bleeding, non-suppurating sites. Therapy significantly decreased levels of both biomarkers. These changes of calprotectin and MPO were highly correlated and also related to probing-depth reduction. Three months after therapy, the levels of both markers still showed significant correlations in initially deep sites, whereas in initially shallow sites no significant correlation was found. After therapy, levels of markers in bleeding and non-bleeding sites were comparable. CONCLUSION: The correlations between calprotectin and MPO indicate that PMNs are a major contributor to the calprotectin content in gingival crevicular fluid of severely affected sites. Calprotectin levels in gingival crevicular fluid and their changes reflect periodontal inflammation as well as the clinical treatment outcome. A prognostic potential of this marker substance remains to be determined.     

广泛型侵袭性牙周炎患者龈沟液中白介素-17的检测

目的 比较广泛型侵袭性牙周炎患者与健康人龈沟液中白介素-17(IL-17)的表达水平及与临床指标的关系。方法 选择广泛型侵袭性牙周炎患者18例和健康人16例,共68颗牙,记录临床牙周检查指标,用滤纸条法收集龈沟液。采用抗体夹心ELISA法测定广泛型侵袭性牙周炎患者治疗前、治疗后3个月及健康对照者龈沟液中IL-17总量和浓度。结果 广泛型侵袭性牙周炎患牙治疗前龈沟液中IL-17的总量和浓度高于牙周健康牙,基础治疗3个月后广泛型侵袭性牙周炎患牙龈沟液中IL-17的总量和浓度较治疗前下降。广泛型侵袭性牙周炎患牙龈沟液中IL-17浓度与探诊深度、附着丧失和出血指数呈正相关关系,而IL-17总量仅与探诊深度呈正相关关系。结论 龈沟液中IL-17浓度可能与广泛型侵袭性牙周炎牙周破坏及炎症的严重程度相关。?     

Intracytoplasmic enzymes in gingival crevicular fluid of patients with aggressive periodontitis

Castro CE, Koss MA, López ME. Intracytoplasmic enzymes in gingival crevicular fluid of patients with aggressive periodontitis. J Periodont Res 2011; 46: 522–527. © 2011 John Wiley & Sons A/S Background and Objective: Biochemical parameters of crevicular fluid could provide evidence of periodontal tissue disease. The aim of this study was to analyze enzymes in crevicular fluid in aggressive localized and generalized periodontitis. Material and Methods: One hundred and twenty‐four subjects were classified as having localized (n = 36) or generalized aggressive periodontitis (n = 38) and subclassified into moderate and severe groups. Controls were 50 periodontitis‐free subjects. Activities of the enzymes lactate dehydrogenase, neutrophil elastase, alkaline phosphatase and aspartate aminotransferase were determined. Data were analyzed using one‐way ANOVA and Tukey’s test. Results: Among the subjects with localized aggressive periodontitis, values of lactate dehydrogenase and alkaline phosphatase increased notably in moderate and severe periodontitis compared with control subjects. Values for aspartate aminotransferase increased with the severity of the disease, and neutrophil elastase was increased in the moderate and severe states. In generalized aggressive periodontitis, lactate dehydrogenase showed higher values than in control subjects in both periodontal subgroups. Alkaline phosphatase and neutrophil elastase showed higher significant differences between moderate and severe periodontitis compared with the control group. Aspartate aminotransferase showed differences between the severe and moderate periodontitis groups compared with the control group. Of all the enzymes analyzed, only lactate dehydrogenase showed higher values in localized than in generalized aggressive periodontitis. Conclusion: Lactate dehydrogenase may distinguish localized and generalized aggressive periodontitis. Alkaline phosphatase increases from moderate to severe states in both types of periodontitis. Aspartate aminotransferase and neutrophil elastase only increase with strong evidence of periodontal destruction.     

Calprotectin levels in gingival crevicular fluid predict disease activity in patients treated for generalized aggressive periodontitis

Kaner D, Bernimoulin J‐P, Dietrich T, Kleber B‐M, Friedmann A. Calprotectin levels in gingival crevicular fluid predict disease activity in patients treated for generalized aggressive periodontitis. J Periodont Res 2011; 46: 417–426. © 2011 John Wiley & Sons A/S Background and Objective: Clinical parameters such as probing depth and bleeding on probing are commonly used for monitoring after periodontal treatment. However, these parameters have poor prognostic utility. The biomarker calprotectin is used to monitor conditions such as inflammatory bowel disease because of its ability to predict disease activity. Levels of calprotectin in gingival crevicular fluid correlate with periodontal disease severity and treatment outcome. The validity of calprotectin as predictor for future periodontal disease activity has not yet been investigated. Material and Methods: Thirty‐six subjects with generalized aggressive periodontitis were treated with scaling and root planing (SRP), and with adjunctive antimicrobial medications. Probing depth, clinical attachment level and bleeding on probing were assessed at baseline, and 3 and 6 mo after SRP. A gingival crevicular fluid sample was collected from the initially deepest site in each patient 3 mo after SRP and analysed for calprotectin levels. Activity was defined as a probing depth increase of > 0.5 mm between 3 and 6 mo at the sample site. The ability of individual parameters to predict activity was analysed by construction of receiver operating characteristic curves. Results: Nine active sites were identified. Clinical attachment level, probing depth, bleeding on probing and gingival crevicular fluid volume showed no predictive utility [area under the curve (AUC) < 0.6, p > 0.05]. However, calprotectin concentration (AUC = 0.793, p = 0.01) and the total amount/sample of calprotectin (AUC = 0.776, p = 0.02) significantly predicted activity. Patients with calprotectin levels above calculated cut‐off values had significantly more active sites than patients with negative results. Conclusion: Calprotectin levels were predictors of disease activity at both site and subject levels. The calculated cut‐off values provide a dichotomous basis for prospective evaluation of calprotectin as a diagnostic marker for monitoring periodontal treatment.     

TNF-alpha and IL-4 levels in generalized aggressive periodontitis subjects

Objectives:  The aim of this study was to evaluate tumor necrosis factor (TNF)-α and interleukin (IL)-4 levels in healthy sites and sites exhibiting signs of moderate and advanced generalized aggressive periodontitis (GAgP) in the same subject.
Methods:  The following sites were selected for crevicular fluid sampling in the same AgP subject ( n  = 14): Healthy sites (HS): no marginal bleeding or bleeding on probing (BOP) and probing depth (PD) ≤ 3 mm; Moderate sites (MS): BOP and PD between 4 and 6 mm; Advanced sites (AS): BOP and PD ≥ 7 mm. One site from periodontally healthy subjects ( n  = 13) was sampled for use as a control. TNF-α and IL-4 levels were measured using ELISA.
Results:  The total amount of TNF-α was lower for control sites, while there were no differences among healthy and diseased sites from GAgP subjects ( P  < 0.05). The concentration of TNF-α was higher in HS, in relation to the other sites ( P  < 0.05). There were no significant differences among the groups regarding total amounts of IL-4 ( P  > 0.05), while IL-4 concentration was significantly higher in control sites, when compared with sites from GAgP subjects ( P  < 0.05).
Conclusion:  In conclusion, high levels of TNF-α and low levels of IL-4 were observed in both healthy and diseased sites within the same generalized AgP individuals.     

Relationship of neutrophil phagocytosis and oxidative burst with the subgingival microbiota of generalized aggressive periodontitis

Introduction: Polymorphonuclear neutrophil (PMN) dysfunctions have been associated with severe forms of periodontitis. This study evaluated the correlation between PMN phagocytosis and oxidative burst with the subgingival microbiota of patients with generalized aggressive periodontitis (GAgP). Methods: Heparinized peripheral blood samples were obtained from 18 GAgP patients and 11 periodontally healthy (PH) subjects, and PMNs were isolated on a Ficoll–Hypaque gradient. For phagocytosis analysis, PMNs were incubated with fluorescein‐labeled Staphylococcus aureus. The oxidative burst was evaluated by incubation of PMNs with dihydroethidium and activation by S. aureus. The assays were examined using flow cytometry. Subgingival biofilm samples were obtained from periodontal sites with and without periodontitis and 24 species were detected by checkerboard. Results: A significantly lower phagocytosis rate was observed for patients with GAgP compared with PH subjects over time (P < 0.05). No differences between groups were found for superoxide production. GAgP patients presented significantly higher prevalence and levels of Porphyromonas gingivalis, Tannerella forsythia, and Aggregatibacter actinomycetemcomitans serotype b than controls (P < 0.05). Significant negative correlations between T. forsythia and P. gingivalis and PMN functions were observed. Conclusions: GAgP subjects presented diminished phagocytic activity of peripheral PMNs and high prevalence and levels of classical periodontal pathogens.     

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。     

Subgingival microbiota of periodontally untreated Mexican subjects with generalized aggressive periodontitis

BACKGROUND AND AIM: Specific microbial profiles that may distinguish between generalized aggressive-periodontitis (GAgP) and generalized chronic-periodontitis (GCP) have, to date, not been described. The purpose of the present study was to describe the subgingival microbial composition of Mexican subjects with GAgP and compare it with that of individuals with GCP and periodontal health (PH). MATERIAL AND METHODS: Seventy-seven subjects with GAgP (n=19), GCP (n=39) and PH (n=19) were selected. Clinical measurements included plaque accumulation, gingival erythema, bleeding on probing, suppuration, pocket depth and attachment level. Up to 28 subgingival plaque samples were obtained from each subject and analysed using the checkerboard DNA-DNA hybridization technique. RESULTS: GAgP and GCP subjects harboured significantly higher levels and/or proportion of Porphyromonas gingivalis, Tannerella forsythia (levels: p<0.001, proportion: p<0.01), Prevotella nigrescens (p<0.05 levels) and "red" complex species (p<0.001 proportion) than PH subjects. All GAgP subjects were carriers of P. gingivalis and P. nigrescens. No significant differences in any of the 40 microbial species tested were detected between GAgP and GCP subjects. CONCLUSIONS: Our results revealed that the microbial differences between GAgP and GCP subjects were only discrete and none of the bacterial species tested seemed to specifically differentiate the subgingival microbial profile of either periodontitis group.     

Effect of azithromycin, as an adjunct to nonsurgical periodontal treatment, on microbiological parameters and gingival crevicular fluid biomarkers in generalized aggressive periodontitis

Emingil G, Han B, Özdemir G, Tervahartiala T, Vural C, Atilla G, Baylas H, Sorsa T. The effect of azithromycin, as an adjunct to nonsurgical periodontal treatment, on microbiological parameters and gingival crevicular fluid biomarkers in generalized aggressive periodontitis. J Periodont Res 2012; 47: 729–739. © 2012 John Wiley & Sons A/S Background and Objective: To study the effectiveness of azithromycin in combination with nonsurgical periodontal therapy on clinical and microbiological parameters, and on the MMP‐8 and TIMP‐1 levels in gingival crevicular fluid, over a 6‐mo time‐period in patients with generalized aggressive periodontitis. Material and Methods: Thirty‐two patients with generalized aggressive periodontitis were included in this randomized, double‐blind, placebo‐controlled, parallel‐arm study. They were randomly assigned to azithromycin or placebo groups (500 mg once daily for 3 d). Probing depth, clinical attachment levels, presence of bleeding on probing and plaque were recorded. Gingival crevicular fluid samples were obtained from one single‐rooted tooth, while microbiological samples were obtained from two single‐rooted teeth, all with a probing depth of ≥ 6 mm. Microbiological parameters were analyzed by quantitative real‐time PCR for Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, Fusobacterium nucleatum, Prevotella intermedia and total bacteria. Gingival crevicular fluid biomarkers were determined by immunofluorometric assay and ELISA. Results: All clinical parameters improved, and microbiological parameters and gingival crevicular fluid MMP‐8 levels significantly decreased, over the 6‐mo period (p < 0.05); both groups demonstrated similar improvements. The azithromycin group presented a higher percentage of deep pockets resolved (probing depth reduction of ≥ 3 mm from baseline) compared with the placebo group at 1 mo (p < 0.05). Conclusion: Adjunctive azithromycin therapy provides no additional benefit over nonsurgical periodontal treatment on clinical parameters, microbiological parameters and gingival crevicular fluid biochemical markers investigated in patients with generalized aggressive periodontitis.     

侵袭性牙周炎和慢性牙周炎的龈下优势菌分析

目的 :分析侵袭性牙周炎 (aggressiveperiodontitis ,AgP)与慢性牙周炎 (chronicperiodontitis ,CP)的龈下优势菌群 ,为探讨牙周炎分类、病因和诊断提供实验依据。方法 :将中学生流调筛选 (16例 )及牙周病专科就诊(2 4例 )的AgP和CP患者 ,采集龈下菌斑样本 ,在厌氧菌基础培养基 (CDC)和选择性培养基 (TSBV)上培养分析。结果 :局限型AgP患者的伴放线放线杆菌 (Actinobacillusactinomycetemcomitans ,Aa)及兼性厌氧菌的检出率显著高于中度CP患者 (P <0 .0 5 ,P <0 .0 1) ,而广泛型AgP和重度CP患者的厌氧菌总数较局限型AgP和中度CP患者显著增加 (P <0 .0 5 )。结论 :局限型AgP和中度CP的龈下优势菌有明显差别 ,Aa是一个重要的危险因子。     

基础治疗对侵袭性牙周炎患者龈沟液骨钙素水平的影响

目的观察牙周基础治疗对侵袭性牙周炎患者的牙周临床指标及龈沟液骨钙素水平的影响。方法检测20例侵袭性牙周炎患者60颗牙齿牙周治疗前和治疗后4周的牙周临床指标,采用电化学发光法检测治疗前后龈沟液中的骨钙素水平。并与20例牙周健康对照组的60颗牙齿对照。结果实验组龈沟液骨钙素水平明显高于对照组;牙周治疗后实验组骨钙素较治疗前明显降低(P<0.05)。结论牙周基础治疗能明显改善侵袭性牙周炎的临床指数,降低龈沟液骨钙素含量。     

慢性牙周炎龈沟液中白细胞介素-8与硫化物的相关关系

目的：探讨慢性牙周炎龈沟液（gingival crevicular fluid，GCF）中白细胞介素-8（interleukin-8，IL-8）与硫化物（suleus sulphide level，SUL）的关系。方法：采用双抗体夹心ELISA法、金刚牙周诊断仪对龈沟液中IL-8和硫化物的含量和临床指标进行测定。23个牙周健康牙作正常对照组（C），12个慢性牙周炎健康牙作实验组1（T1），30个慢性牙周炎患牙作实验组2（T2）。用标准化滤纸条采集观察牙位GCF样本，记录相应位点30’硫化物浓度，同时记录龈沟出血指数（SBI）、牙周袋探诊深度（PPD）、牙周附着丧失水平（CAL）。结果：①受检位点龈沟液中IL-8总量存在显著性差别，其中慢性牙周炎炎症牙位组IL-8总量高于健康牙位组、正常对照组（p〈0.05），但龈沟液中IL-8浓度无显著性差别。龈沟液中IL-8总量与SBI、PPD、CAL之间有明显相关关系（P〈0.05），但IL-8的浓度与临床指标间相关关系不明显。②慢性牙周炎炎症牙位组的硫化物浓度与健康牙位组、正常牙位组之间均有显著性差别（P〈0.05），慢性牙周炎健康牙位组与正常牙位组之间无显著性差别。③慢性牙周炎炎症牙位组硫化物浓度与临床指标间具正相关关系（P〈0.05），而健康牙位组和正常对照组硫化物浓度与临床指标间无相关关系。GCF中慢性牙周炎炎症牙位组的硫化物浓度与IL-8总量具负相关关系（P〈0．05），而慢性牙周炎健康牙位组和正常对照组的硫化物浓度与IL-8总量之间无相关关系。结论：慢性牙周炎患牙龈沟液中IL-8和硫化物的含量与临床指标之间有相关性。龈沟液中细菌代谢产物所产生的硫化物对IL-8的含量存在一定抑制作用。     

牙龈卟啉单胞菌与龈沟液中白细胞介素8关系的研究

目的:探讨龈沟液(GCF)中白细胞介素8(IL -8)在慢性牙周炎病程中的变化、IL- 8与牙周临床检测指标以及同龈下菌斑中细菌含量的相关关系。方法: 5名非牙周炎患者的10颗牙周健康牙和30名慢性牙周炎(CP)患者的70颗牙(其中10颗为健康牙, 60颗为牙周炎患牙)纳入本研究。采集观察牙的GCF样本、龈下菌斑样本,同时记录所有牙的牙龈指数(GI)、牙周袋探诊深度(PPD)、临床附着丧失水平(CAL)。用ELISA法检测样本中IL -8的水平,用厌氧菌培养技术培养菌斑标本,用PCR法检测菌斑中的牙龈卟啉单胞菌(Pg),结果:慢性牙周炎组的GCF中IL -8的总量高于临床牙周健康组,而IL -8的质量浓度在各组中无差异。GCF中IL- 8浓度与GI、PPD、CAL无相关关系,而IL -8的总量与GI、PPD、CAL呈正相关关系。龈下菌斑中Pg的检出量与GCF中IL- 8的含量间未发现有相关关系。结论:慢性牙周炎患者龈沟液中IL- -8量增加与龈沟液分泌增加有关而与牙龈卟啉单胞菌检出CFU无关。