Macrophage Migration Inhibitory Factor Levels in Gingival Crevicular Fluid, Saliva, and Serum of Chronic Periodontitis Patients.

Chronic periodontitis (CP) is an infection that affects the teeth supporting structure. Macrophage migration inhibitory factor (MIF) is an important effector cytokine of the innate immune system. Due to its functional characteristics, MIF may be involved in the immunopathology of CP. The aim of the present study was to evaluate MIF levels in gingival crevicular fluid (GCF), saliva, and serum of CP patients. A cross-sectional study was conducted on 60 subjects divided into two groups: subjects with CP (n= 30) and periodontally healthy subjects without CP (n=30). MIF was quantified in GCF, saliva, and serum of all participants by enzyme-linked immunosorbent assay. MIF concentrations were higher in GCF, saliva, and serum in the group with CP compared with the group without CP and a higher MIF concentration was observed in GCF (p=0.001) and saliva (p=0.009) in the group with CP. MIF intragroup comparisons between fluids demonstrated significant high levels of MIF in saliva compared with GCF and serum in both study groups (p<0.05). A positive correlation was found between clinical signs and MIF concentration in GCF (p<0.05). There is an association between the MIF and the clinical signs of the disease. Therefore, MIF could have an important role in the pathology and progression of CP.

Interleukin-6 and Interferon-α Levels in Gingival Crevicular Fluid in HIV-1 Patients with Chronic Periodontitis / Niveles de Interleukin-6 e Interferon-α de Fluido Crevicular Gingival en Pacientes VIH-1 con Periodontitis Crónica

ABSTRACT: The influence of cytokine on the progression of chronic periodontitis in human immunodeficiency virus (HIV) patients is still controversial and poorly investigated. This study aimed to analyze and compare IL-6 and IFN-α levels in the gingival crevicular fluid of HIV-1-positive and HIV-1-negative patients with chronic periodontitis and different grades of tissue destruction and inflammation. Samples from the gingival crevicular sulcus were obtained from 35 HIV-1-positive individuals with chronic periodontitis and 35 seronegative patients with chronic periodontitis. Probing depth and clinical attachment level, as well as the results of the Enzyme-Linked Immunosorbent Assay for confirmation of patient diagnostics, were evaluated. Statistical analyses were performed using Student t, Mann-Whitney and Spearman tests. IL-6 levels were significantly lower, while IFN-α levels were significantly higher in HIV-1 patients. Clinical attachment level was directly associated with IFN-α levels in HIV-1 carriers, connected to probing depth in these patients. Clinical data in association with gingival crevicular fluid cytokine levels may reveal a localized immunological response pattern, which may contribute to the understanding of periodontitis pathogenesis in HIV-1 carriers.

RESUMEN: La influencia de la citocina en la progresión de la periodontitis crónica en pacientes con el virus de la inmunodeficiencia humana (VIH) sigue siendo controvertida y poco investigada. Este estudio tuvo como objetivo analizar y comparar los niveles de interleuquina-6 (IL6) e interferón-α (IFN-α) en el líquido crevicular gingival de pacientes VIH-1-positivos y VIH-1-negativos con periodontitis crónica y diferentes grados de destrucción e inflamación tisular. Se obtuvieron muestras del surco crevicular gingival de 35 individuos VIH-1 positivos con periodontitis crónica y 35 pacientes seronegativos con periodontitis crónica. Se evaluaron la profundidad de sondeo y el nivel de inserción clínica, así como los resultados del Ensayo Inmunoabsorbente Ligado a Enzimas para la confirmación del diagnóstico del paciente. Los análisis estadísticos se realizaron utilizando pruebas t de Student, Mann-Whitney y Spearman. Los niveles de IL-6 fueron significativamente más bajos, mientras que los niveles de IFN-a fueron significativamente más altos en los pacientes con VIH-1. El nivel de inserción clínica se asoció directamente con los niveles de IFN-α en los portadores del VIH1, conectados a la profundidad del sondaje en estos pacientes. Los datos clínicos en asociación con los niveles de citoquinas de los fluidos creviculares gingivales pueden revelar un patrón de respuesta inmunológica localizado, que puede contribuir a la comprensión de la patogénesis de la periodontitis en los portadores del VIH-1.

Gingival crevicular fluid levels of human beta-defensin 1 in individuals with and without chronic periodontitis.

BACKGROUND AND OBJECTIVE: Human beta-defensins (hBDs) contribute to innate immunity antimicrobial activity. They are also effective in the adaptive immune response and may play a crucial role in the susceptibility to diseases of the oral cavity. This study aimed to evaluate the levels of hBD-1 in the gingival crevicular fluid of individuals with and without chronic periodontitis. MATERIAL AND METHODS: Twenty periodontally healthy individuals (H) and 20 individuals with chronic periodontitis were recruited. Gingival crevicular fluid samples were collected from: healthy sites (Hh) from periodontally healthy individuals; and healthy sites (Ph), sites with gingivitis (Pg), and sites with periodontitis (Pp) from individuals with periodontitis. The levels of hBD-1 (pg/mL) were measured using enzyme-linked immunosorbent assay. Comparisons of hBD-1 between individuals (H and chronic periodontitis) and among sites (Hh, Ph, Pg, Pp) were performed through hierarchical linear modeling. RESULTS: Gingival crevicular fluid levels of hBD-1 were: Hh = 229.52 ± 138.96 (median 199.26), Ph = 53.88 ± 58.17 (median 35.75), Pg = 57.11 ± 40.18 (median 39.90) and Pp = 55.31 ± 37.28 (median 54.19). No influence of site diagnosis (level 1; health/gingivitis/periodontitis) was observed; however, individual diagnosis (level 2; health/periodontitis) influenced the levels of hBD-1 (P < .001). CONCLUSION: Periodontally healthy individuals showed higher gingival crevicular fluid levels of hBD-1 when compared to individuals with chronic periodontitis. This suggests a potential protective role of hBD-1 in the susceptibility to chronic periodontitis.

Cytokines and MMPs levels in gingival crevicular fluid from patients with chronic periodontitis before and after non-surgical periodontal therapy

The study of host response in periodontal disease may provide a mechanism to monitor disease progression. the purpose of the present research was to determine the levels of IL-1alfa, IL-1beta, TNF-alfa, IL-6, IL-6sR, IL-8, IL-10, MMP-3 and MMP-8 in gingival crevicular fluid (GCF), before and after non-surgical periodontal treatment (NSPT) in order to evaluate therapy response. methodology: eleven patients diagnosed with chronic periodontitis and eleven healthy subjects were selected for this study. clinical measurements, including probing depth (PD) and clinical attachment loss (CAL) were carried out in patients diagnosed with chronic periodontitis and periodontal healthy controls. the clinical indexes evaluated were: gingival index (GI) and plaque index (PI). samples of GCF were taken from one tooth per quadrant before and 45 days after NSPT. the levels of inflammatory mediators were measured by ELISA. results: the values of all clinical parameters decressed significsntly after treatment. the concentration levels of all cytokines and MMP-3 and MMP-8 in the GCF sample were higher in patients diagnosed with chronic periodontitis compared to the healthy group. all inflammatory mediators decreased after therapy, but did not reach control values; IL-6, Il-6sR, IL-10 and TNF-alfa, attained the highest reduction (70 percent -54 percent); the vales of MMP3, IL-1alfa, IL-1beta and IL-8 were reduced between 50 percent ­ 34 percent; and MMP-8 showed the lowest decrease (28 percent). conclusion: all clinical parameters and cytokines levels decreased after NSPT. the mediators TNF-alfa IL-6, IL-6sR, and IL-10 showed the largest variation between before and after NSPT and could thus be used to evaluate therapy response.

Matrix metalloproteinase-8 levels in periodontal disease patients: A systematic review.

Periodontal disease is characterized as a disorder of the oral microbiota resulting in an immune response which, in turn, leads to the destruction of periodontal tissue. Matrix metalloproteinase-8 (MMP-8) has been reported as the major metalloproteinase involved in periodontal disease, being present at high levels in gingival crevicular fluid and salivary fluid (SF). The aim of this systematic review was to evaluate the scientific literature regarding the expression of MMP-8 in gingival crevicular fluid and SF in patients with periodontal disease, analyzing its validity as a possible biomarker in the diagnosis of periodontal disease. A systematic review of the literature was performed using the PubMed/Medline, CENTRAL and Science Direct databases. Studies concerning the use of MMP-8 in the diagnosis of periodontal disease that evaluated its effectiveness as a biomarker for periodontal disease were selected. The search strategy provided a total of 6483 studies. After selection, six articles met all the inclusion criteria and were included in the present systematic review. The studies demonstrated significantly higher concentrations of MMP-8 in patients with periodontal disease compared with controls, as well as in patients presenting more advanced stages of periodontal disease. The findings on higher MMP-8 concentrations in patients with periodontal disease compared with controls imply the potential adjunctive use of MMP-8 in the diagnosis of periodontal disease.

Análise clínica e de marcadores biológicos no fluido do sulco peri-implantar correlacionando-os à mucosa ceratinizada ao redor de implantes dentários / Clinical and biological markers analysis in the peri-implant groove fluid correlating them to the keratinized mucosa around dental implants

Uma faixa adequada de mucosa ceratinizada (MC) é importante para garantir condições mínimas necessárias para o estabelecimento da homeostasia do periodonto de proteção. Frente à infecção bacteriana, os tecidos periodontais e peri-implantares desenvolvem uma resposta imune inflamatória local, resultando na produção e liberação de diversos mediadores inflamatórios que podem ser encontrados no fluido do sulco gengival e peri-implantar. Entretanto, é escassa a literatura acerca dos níveis desses mediadores em sítios peri-implantares considerando a faixa de MC. Assim, o objetivo deste trabalho foi avaliar a associação entre a quantidade e qualidade da MC peri-implantar e parâmetros clínicos e a qualidade da resposta imune através da análise da concentração de mediadores inflamatórios (IL-1, IL-4, IL-6, IL-8, MIP-1, TNF- e VEGF) presentes no fluido peri-implantar humano antes (T1) e depois (T2) da raspagem subgengival, através de imunoensaio. Parâmetros clínicos avaliaram índice de placa (IP), supuração a sondagem (S), profundidade de sondagem (mesial-PSM, centro-PSC e distal-PSD), índice de sangramento (mesial-ISM, centro-ISC e distal-ISD), nível de inserção relativo (NIR), largura (LMC) e espessura (EMC) da MC na face vestibular. Amostras de fluido sulcular foram coletadas e analisadas. Os implantes foram divididos em grupos de acordo com a faixa de MC (G12mm e G2>2mm) e espessura de MC (GA11mm e GB1>1mm; GA21,5mm e GB2>1,5mm). Foram avaliados 20 pacientes (11 homens e 9 mulheres) com idade entre 40 e 80 anos (53,45±10,32), que apresentaram 42 implantes (G1=25 e G2=17). Os resultados clínicos demonstraram diferença estatística significativa apenas entre T1 e T2 dentro do G1 para IP (T1=56% e T2=16%) e ISM (T1=68% e T2=40%). Foi observada diferença estatística entre G1 e G2 apenas para IL-1 em T2 (G1=9,77pg/ml±12,44 e G2=30,13pg/ml±32,29). Intra-grupos, todas as citocinas aumentaram significativamente, mas apenas no G2, demonstrando diferença de reatividade entre grupos. Quanto à espessura da MC (GA1=6 e GB1=36), resultados clínicos revelaram diferença inter-grupos para ISC em T2 (GA1=16,67% e GB1=61,11%) e intra-grupos para IP no GB1 (T1=52,78% e T2=27,78%). Houve aumento significativo no GB1 para todas as citocinas, exceto VEGF, assim como para IL-1 no GA1. Quando a amostra foi redistribuída em GA2=24 e GB2=18, os resultados clínicos indicaram diferença estatística inter-grupos para PSC em T2 (GA2=2,58mm±1,06 e GB2=3,11mm±1,02) e intra-grupos para IP (T1=62,5% e T2=20,83%) e PSC (T1=2,92mm±1,18 e T2=2,58mm±1,06) no GA2 e para ISM (T1=55,56% e T2=27,78%) no GB2. Intra-grupos observou-se aumento significativo para todas as citocinas no GA2 exceto VEGF, assim como IL-8 no GB2. Conclui-se que as diferenças clínicas apresentadas tenderam a evidenciar a importância da MC principalmente após o preparo inicial e, além disso, uma faixa de MC maior que 2mm influenciou os níveis dos mediadores inflamatórios avaliados após a raspagem subgengival. Adicionalmente, a falta de diferença estatística significativa na comparação entre grupos com diferentes espessuras de MC, bem como tal diferença ora no grupo espesso ora no grupo fino quando se adotam diferentes valores de corte (1mm ou 1,5mm respectivamente), demonstra resultados inconclusivos, ressaltando a importância de novas pesquisas para responder esta questão.(AU)

An adequate keratinized mucosa (KM) width is important to ensure minimal conditions necessary to establish protect periodontium homeostasis. When a bacterial infection occurs, periodontal and peri-implant tissues develop a local inflammatory immune response that results in production and release of several inflammatory mediators that may be found in gingival crevicular and in peri-implant fluids. However, there is a lack of literature concerning about the levels of these mediators in peri-implant sites considering KM width. The aim of this study was to evaluate the association between KM peri-implant quantity and quality and clinical parameters and immune response quality by analyzing the inflammatory mediators concentration (IL-1, IL-4, IL-6, IL-8, MIP-1, TNF- and VEGF) present in human peri-implant fluid before (T1) and after (T2) subgingival scaling, by immunoassay. Clinical parameters evaluated plate index (PI), probing suppuration (S), probing depth (mesial-PDM, center-PDC and distal-PDD), bleeding index (mesial-BIM, center-BIC and distal-BID), relative attachment level (RAL), keratinized mucosa width (KMW) and thickness (KMT) on the buccal face. Sulcular fluid samples were collected and analyzed. The implants were divided in groups according KM width (G12mm and G2>2mm) and KM thickness (GA11mm and GB1>1mm, GA21,5mm and GB2>1,5mm). Twenty patients (11 men and 9 women) aged 40 to 80 years (53,45±10,32) were evaluated, with 42 implants (G1=25 and G2=17). Clinical results showed a significant statistical difference only between T1 and T2 within G1 for PI (T1=56% and T2=16%) and BIM (T1=68% and T2=40%). Statistical difference was observed between G1 and G2 only for IL-1 in T2 (G1=9,77pg/ml±12,44 and G2=30,13pg/ml±32,29). Intra-groups, all cytokines increased significantly, but only in G2, showing reactivity difference between groups. As to KM thickness (GA1=6 and GB1=36), clinical results revealed intergroup differences for BIC in T2 (GA1=16,67% and GB1=61,11%) and intra-groups for PI in GB1 (T1=52,78% and T2=27,78%). There was a significant increase in GB1 for all cytokines except VEGF, as well as for IL-1 in GA1. When the sample was redistributed in GA2=24 and GB2=18, clinical results indicated statistical inter-group differences for PDC in T2 (GA2=2,58mm±1,06 and GB2=3,11mm±1,02) and intra-groups for PI (T1=62,5% and T2=20,83%) and PDC (T1=2,92mm±1,18 and T2=2,58mm±1,06) in GA2 and for BIM (T1=55,56% and T2=27,78%) in GB2. Intra-groups were observed significantly increase for all cytokines in GA2 except VEGF, as well as IL-8 in GB2. Concluded that clinical differences presented tended to show the KM importance principally after the initial preparation and, in addition, KM width greater than 2mm influenced the inflammatory mediators levels evaluated after subgingival scaling. Additionally, the absence of significant statistic difference between groups when comparing the keratinized mucosa thickness, as well as this difference sometimes in the thick group or in the thin group when different court values was adopted (1mm or 1,5mm respectively), show inconclusive results, emphasizing the importance of new research that may answer this question.(AU)

Cytokine measurements in gingival crevicular fluid and periodontal ligament: Are they correlated?

INTRODUCTION: Mechanical stress can induce molecular changes in gingival crevicular fluid (GCF) and the periodontal ligament (PDL). It is still not clear whether changes in the PDL and GCF are linked. In this study, we aimed to analyze the expression of cytokines in GCF and PDL after mechanical stress. METHODS: Twenty-three healthy patients were included. The experimental group consisted of premolars subjected to a force of 0.980 N for 1, 3, 7, 14, 21, or 28 days. The contralateral teeth were the controls. GCF and PDL samples were collected at the same time points for analysis of cytokines using the cytometric bead array. RESULTS: Interleukin (IL)-6 (IL-6) production was significantly elevated in the PDL on day 1 after force application. Significantly strong positive correlations between GCF and PDL in experimental group were seen on days 3 (interferon-gamma), 7 (IL-10), 14 (IL-17A), and 28 (IL-17A, tumor necrosis factor-alpha), and significantly strong negative correlation were seen on days 14 (interferon-gamma) and 21 (IL-2, IL-10). CONCLUSIONS: Different patterns of IL-6 expression were seen in the PDL and GCF after mechanical stress. Despite occasional correlations between GCF and PDL, the molecular contributions of the PDL to the GCF changes could not be clearly defined by our model.

Alcohol Consumption and Periodontitis: Quantification of Periodontal Pathogens and Cytokines.

BACKGROUND: There are few studies on periodontal status related to microbiologic and immunologic profiles among individuals not or occasionally using alcohol and those with alcohol dependence. The aim of this study is to determine the effect of alcohol consumption on the levels of subgingival periodontal pathogens and proinflammatory cytokines (interleukin [IL]-1ß and tumor necrosis factor [TNF]-α) in the gingival fluid among individuals with and without periodontitis. METHODS: This observational analytic study includes 88 volunteers allocated in four groups (n = 22): individuals with alcohol dependence and periodontitis (ADP), individuals with alcohol dependence and without periodontitis (ADNP), individuals not or occasionally using alcohol with periodontitis (NAP), and individuals not or occasionally using alcohol without periodontitis (NANP). Levels of Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Prevotella intermedia, Eikenella corrodens, and Fusobacterium nucleatum were determined by real-time polymerase chain reaction on the basis of the subgingival biofilm, and IL-1ß and TNF-α were quantified by enzyme-linked immunosorbent assay in gingival fluid samples. RESULTS: Individuals with alcohol dependence showed worse periodontal status and higher levels of P. intermedia, E. corrodens, F. nucleatum, and IL-1ß than non-users. No significant correlations between TNF-α and bacterial levels were observed. However, in the ADP group, higher levels of E. corrodens were correlated with higher levels of IL-1ß. CONCLUSION: A negative influence of alcohol consumption was observed on clinical and microbiologic periodontal parameters, as well as a slight influence on immunologic parameters, signaling the need for additional studies.

Clinical, microbial, and immune responses observed in patients with diabetes after treatment for gingivitis: a three-month randomized clinical trial.

BACKGROUND: Although patients with diabetes are frequently affected by periodontitis, only a few investigations have focused on gingivitis in this at-risk population. This randomized placebo-controlled clinical trial compared the response to a gingivitis treatment protocol that combined mechanical procedures and daily use of an essential oil (EO) mouthrinse between patients with and without diabetes. METHODS: The whole-mouth periodontal probing depth (PD), gingival index (GI), and plaque index (PI) were monitored in gingivitis cases among systemically healthy patients (n = 60) or those with diabetes (n = 60) at baseline and 3 months after treatment. Levels of Porphyromonas gingivalis, Tannerella forsythia, Aggregatibacter actinomycetemcomitans, and total bacterial load were determined by a real-time polymerase chain reaction in intrasulci plaque samples. The volume of gingival crevicular fluid (GCF) was quantified, and interleukin-1ß (IL-1ß) levels were determined in GCF samples. After a full-mouth ultrasonic debridement, patients were randomly assigned to an EO or a placebo rinse for 90 days (40 mL/day). The data were analyzed through repeated-measures analysis of variance and multiple comparisons Tukey tests (P <0.05). RESULTS: GI was more severe in the diabetes group. Diabetes impaired GI and reduced GCF volume. PD, bacterial levels, and IL-1ß improved similarly in both systemic conditions. The adjunctive use of EO provided greater reductions of PI, GI, total bacterial load, T. forsythia, A. actinomycetemcomitans, and GCF volume. CONCLUSIONS: Response to gingivitis treatment in patients with diabetes can slightly differ from that in patients without diabetes. Daily use of an EO mouthrinse after ultrasonic debridement benefited patients with and without diabetes.

Antimicrobial photodynamic therapy as an adjunct to non-surgical treatment of aggressive periodontitis: a split-mouth randomized controlled trial.

BACKGROUND: The management of aggressive periodontitis (AgP) represents a challenge for clinicians because there are no standardized protocols for an efficient control of the disease. This randomized controlled clinical trial evaluated the effects of repeated applications of antimicrobial photodynamic therapy (aPDT) adjunctive to scaling and root planing (SRP) in patients with AgP. METHODS: Using a split-mouth design, 20 patients with generalized AgP were treated with aPDT + SRP (test group) or SRP only (control group). aPDT was applied at four periods. All patients were monitored for 90 days. Clinical, microbiologic, and immunologic parameters were statistically analyzed. RESULTS: In deep periodontal pocket analysis (probing depth [PD] ≥ 7 mm at baseline), the test group presented a decrease in PD and a clinical attachment gain significantly higher than the control group at 90 days (P < 0.05). The test group also demonstrated significantly less periodontal pathogens of red and orange complexes and a lower interleukin-1ß/interleukin-10 ratio than the control group (P < 0.05). CONCLUSION: The application of four sessions of aPDT, adjunctive to SRP, promotes additional clinical, microbiologic, and immunologic benefits in the treatment of deep periodontal pockets in single-rooted teeth in patients with AgP.

Do subjects with aggressive and chronic periodontitis exhibit a different cytokine/chemokine profile in the gingival crevicular fluid? A systematic review.

BACKGROUND AND OBJECTIVE: Microbiological and immunological hypotheses have been raised to explain the differences in the clinical manifestations of aggressive periodontitis and chronic periodontitis. However, studies comparing the cytokine/chemokine profiles in gingival crevicular fluid between these two clinical conditions have so far not been compiled. This systematic review aimed to answer the following question: "Do subjects with aggressive periodontitis and chronic periodontitis have a different profile of cytokines/chemokines in the gingival crevicular fluid?" MATERIAL AND METHODS: An electronic database search of MEDLINE/PubMed and Embase was performed from 1990 up to and including August 2013, using MeSH terms and other keywords. Titles and abstracts were screened and the papers that satisfied eligibility criteria were assessed. RESULTS: Of 1954 titles, 17 studies reporting the levels of 21 different cytokines/chemokines were included. Most studies did not find any significant differences in the gingival crevicular fluid levels of cytokines/chemokines between aggressive periodontitis and chronic periodontitis. Some studies demonstrated that the levels of specific proinflammatory and anti-inflammatory cytokines/chemokines were higher (n = 5) and lower (n = 3), respectively, in aggressive periodontitis than in chronic periodontitis. The studies differed in the manner in which they reported the results (e.g. concentrations or total amounts). It was not clear in some studies whether the sample sites from both groups were matched for disease severity. Some studies did not take into account confounders, such as smoking. CONCLUSION: The current weight of evidence is not sufficient to prove that there are distinct gingival crevicular fluid cytokine/chemokine profiles for patients with aggressive periodontitis and chronic periodontitis.

O efeito adjunto de uma suplementação alimentar à base de ômega-3 no tratamento da periodontite: um estudo clínico e imunológico / The adjunctive effects of Omega-3 dietary supplementation in the treatment of periodontitis: a clinical and immunological study

O objetivo foi avaliar se a suplementação com ômega (ω)-3 no tratamento não cirúrgico da periodontite melhora os resultados clínicos e imunológicos. Avaliar os níveis séricos dos ω-3 e ω-6 na doença periodontal, investigar se o tratamento periodontal associado ou não a suplementação com ω-3 afeta os níveis séricos destes. Assim como avaliar a expressão de um painel de citocinas relacionadas a osteoclatogênese no fluido gengival (FG) de sítios dos pacientes com gengivite e periodontite, adicionalmente relacionar estas citocinas a densidade óptica do osso alveolar destes sítios. Por fim, investigar se as terapias propostas neste estudo afetam os parâmetros clínicos periodontais, a densidade óptica do osso alveolar e a expressão das citocinas no FG. Para isso foi realizado um ensaio clínico piloto, randomizado, duplo-cego, controlado-placebo, com vinte e um pacientes com periodontite e dezesseis com gengivite. Estes foram investigados para os níveis séricos dos ácidos graxos poli-insaturados de cadeia longa (AGPI-CL), ácido eicosapentaenóico (EPA), ácido docosahexaenóico (DHA), ácidos docosapentaenóico (DPA), o ácido araquidônico (AA) usando cromatografia gasosa. O FG dos sítios controle-gengivite, controle-periodontite e destruído de pacientes com periodontite foi avaliado para Receptor ativador do fator nuclear kappa-B ligante (RANK-L), osteoprotegerina (OPG), a osteocalcina (OC), fator de necrose tumoral (TNF)-α, interferon (IFN)-γ, interleucina (IL) -1ß, IL-4, IL-6 e IL-10, utilizando imunoensaio multiplex. Pacientes com periodontite foram subagrupados em grupos com e sem Síndrome Metabólica para avaliação dos AGPI-CL. Sítios dos pacientes com periodontite foram subagrupados em sítios sem e com considerável desmineralização ópticas baseado na avaliação da densidade óptica alveolar representada pelos valores de pixels das radiografias intraorais digitais para avaliação das citocinas no FG. Pacientes com periodontite foram randomizados em raspagem e alisamento radicular mais suplementação com ω-3 (RAR+ω-3) ou placebo (RAR+placebo), e foram reavaliados depois de quatro meses para AGPI-CL no soro, citocinas no FG e densidade alveolar óptica. Os níveis significativamente maiores dos AGPI-CLs foram observados em pacientes com periodontite em comparação com gengivite. A profundidade de bolsa mostrou uma correlação positiva significativa com DHA, DPA e AA. Após RAR+ω-3, DPA, AA, AA/EPA e AA/DHA reduziram significativamente, e depois da RAR+placebo todos os níveis dos AGPI-CLs reduziram significativamente. Os níveis das citocinas, IFN-γ, IL-4, IL-10, RANK-L e OC, no FG de sítios controle-gengivite foram menores comparados aos sítios controle-periodontite e destruído. O mesmo resultado foi observado para os níveis da IL-1ß, IL-6 e OPG em comparação com sítios destruídos. Pacientes com periodontite apresentaram níveis mais baixos de IL-1ß e OPG em sítios controle-periodontite comparado aos destruídos. Após RAR+ω-3, o nível da IL-4 aumentou e os níveis de OC, IFN-γ e IL-10 diminuíram. Não houve diferenças significativas entre os grupos experimentais para a densidade alveolar óptica e parâmetros. Em conclusão, os níveis séricos dos AGPI-CLs são afetados pela gravidade de doença periodontal e diferem com a presença da síndrome metabólica. Citocinas relacionadas a osteoclatogênese no FG refletem a gravidade periodontal e diferem com densidade óptica alveolar alterada. A suplementação com ω-3 não melhora clinica ou imunologicamente o resultado do tratamento periodontal.

The aim was to test whether omega (ω)-3 supplementation associated to non-surgical treatment of periodontitis improves clinical and immunological results. Evaluate the serum levels of ω-3 and ω-6 in periodontal disease, hence, to investigate whether periodontal treatment with or without ω-3 supplementation affects serum levels of these. Therefore, to evaluate the expression of a panel of cytokines related to osteoclastogenesis in the gingival crevicular fluid (GCF) sites of patients with gingivitis and periodontitis, additionally relate these cytokines with optical alveolar bone density in these sites. Finally, to investigate whether the therapies proposed in this study affect the clinical periodontal parameter, the optical density of the alveolar bone and the expression of cytokines in GCF. Then was conducted a pilot clinical trial, randomized, double-blind, placebo-controlled, with twenty-one patients with periodontitis and sixteen with gingivitis. These were investigated for blood levels of long chain polyunsaturated fatty acids (LC-PUFA), eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), docosapentaenoic acid (DPA), arachidonic acid (AA) using gas chromatography. The GCF of the sites of control-gingivitis, control-periodontitis and destroyed from patients periodontitis were evaluated for receptor activator of kappa-B ligand nuclear factor (RANK-L), osteoprotegerin (OPG), osteocalcin (OC), tumor necrosis factor (TNF) -α, interferon (IFN) -γ, interleukin (IL) -1ß, IL-4, IL-6 and IL-10 using multiplex immunoassay. Patients with periodontitis were subgrouped in groups with and without metabolic syndrome to evaluate the LC-PUFA. Sites of patients with periodontitis were subgrouped at sites with and without considerable optical demineralization based on the evaluation of alveolar optical density represented by the pixel values of the digital intraoral radiographs for evaluation of cytokines in GCF. Patients were randomized into the periodontal scaling and root planing more ω-3 supplementation (ω-3 SRP +) or placebo (SRP + placebo), and were assessed after four months to LC-PUFA in serum, cytokines in GCF and alveolar optical density. The significantly higher levels of LC-PUFA were observed in patients with periodontitis compared to gingivitis. The pocket depth showed a significant positive correlation with DHA, DPA and AA. After RAR + ω-3, APD, AA, AA / AA and EPA / DHA reduced significantly, and, after SRP + placebo, all levels of LC-PUFA reduced significantly. The levels of cytokines, IFN-γ, IL-4, IL-10, and OC RANK-L, from controlgingivitis sites GCF were lower compared to control-periodontitis and destroyed sites. The same result was observed for levels of IL-1ß, IL-6 and OPG compared to destroyed sites. Periodontitis patients had lower levels of IL-1ß and OPG in control-periodontitis compared to destroyed sites. After SRP + ω-3, the level of IL-4 and OC increased and levels of IFN-γ and IL-10 decreased. There were no significant differences between the experimental groups to optical density and clinical parameters. In conclusion, serum levels of LC-PUFA are affected by periodontal disease severity and differ in the presence of the metabolic syndrome. Cytokines related to osteoclastogenesis in GCF reflect the periodontal severity and differ with impaired alveolar optical density. Ω-3 supplementation does not improve clinical or immune result of non-surgical periodontal treatment.

A double-blind randomised clinical trial of two techniques for gingival displacement.

Knowledge about security and the potential damage originated by the gingival displacement techniques has not been described through randomised clinical studies. This crossover, double-blind, randomised clinical trial evaluated clinical and immunological factors related to conventional and cordless gingival displacement (GD) techniques, and patients' perceptions in 12 subjects with the employment of 2 GD techniques: conventional (gingival cord + 25% AlCl3 astringent gel) and cordless (15% AlCl3 astringent-based paste). In each subject, two anterior teeth were treated and a 10-day wash-out period separated the two treatments. Periodontal indices were evaluated before (baseline) and 1 and 10 days after GD. Interleukin 1ß, interleukin 6 and tumour necrosis factor α concentrations in gingival crevicular fluid were measured before and 1 day after GD. Subjective parameters (pain, unpleasant taste and stress) were also evaluated. Data were analysed by one-way repeated-measures analysis of variance and Tukey's test (immunological factors), the Friedman test (periodontal parameters) and Fisher's exact or chi-squared test (subjective parameters), with a significance level of 95%. Gingival bleeding index, probing depth and plaque index values did not differ significantly between groups at any timepoint. Neither technique resulted in worse periodontal indices. Both techniques yielded similar results for pain and unpleasant taste, but conventional GD was significantly more stressful than cordless GD for volunteers. Both treatments significantly increased mean concentrations of the three cytokines, with the conventional technique producing the highest cytokine levels. Cordless GD is less stressful for patients and results in lower post-treatment levels of inflammatory cytokines compared with conventional GD.

Clinical outcomes of periodontal therapy are not influenced by the ATC/TTC haplotype in the IL8 gene.

BACKGROUND AND OBJECTIVE: Although chronic periodontitis (CP) is a multifactorial condition, few studies have investigated the potential association of gene variants with the outcome of periodontal therapy. In a previous study, we reported that variants in the interleukin-8 (IL8) gene were associated with CP in a Brazilian population. The aim of this nonrandomized study was to investigate whether genetic susceptibility to CP, conferred by the presence of the IL8 ATC/TTC haplotype, influences the clinical outcomes of nonsurgical periodontal therapy and the IL-8 protein levels in the gingival crevicular fluid. MATERIAL AND METHODS: Forty-one individuals were grouped according to the presence (susceptible to CP; n = 21) or absence (not susceptible to CP; n = 20) of the IL8 ATC/TTC haplotype. These individuals received nonsurgical periodontal therapy from one periodontist, who was blinded to the genetic status of each patient, and follow up continued for 45 d. The clinical parameters and gingival crevicular fluid samples were collected at baseline and on day 45. The IL-8 levels were determined by an ELISA. The data were subjected to the Mann-Whitney U-test, Wilcoxon and Spearman tests and to multiple logistic-regression analysis. RESULTS: No significant differences between patients with or without the IL8 ATC/TTC haplotype were found for the outcome of nonsurgical periodontal therapy and IL-8 levels. The multiple logistic-regression analysis did not show a statistically significant association between the IL8 haplotype and the variables studied. CONCLUSION: In this longitudinal clinical study, we observed that neither the outcome of nonsurgical periodontal therapy nor the IL-8 levels were influenced by the IL8 ATC/TTC CP-susceptibility haplotype. Additional studies of CP patients from other ethnic populations are necessary to confirm these results.

Local levels of inflammatory mediators in uncontrolled type 2 diabetic subjects with chronic periodontitis.

AIM: The aim of this study was to evaluate the levels of a wide panel of cyto/chemokines in the gingival crevicular fluid (GCF) of uncontrolled type 2 diabetic subjects as compared with non-diabetic subjects with periodontitis. METHODS: Twenty-six uncontrolled type 2 diabetic subjects (glycated haemoglobin levels >7.5%) and 20 non-diabetic subjects with chronic periodontitis were enrolled in this study. The levels of 14 cyto/chemokines were measured in the GCF of healthy and diseased sites of the diabetic and non-diabetic subjects using multiplex bead immunoassays. RESULTS: The concentrations of eotaxin, macrophage inflammatory protein-1α, granulocyte-macrophage colony-stimulating factor, interleukin (IL)-6, tumour necrosis factor-α and IL-12 were higher in healthy and diseased sites of diabetic than non-diabetic subjects, after adjustment for multiple comparisons (p < 0.0035). CONCLUSION: Uncontrolled type 2 diabetes mellitus modulated the local levels of several cyto/chemokines at both healthy and diseased periodontal sites in favour of a proinflammatory profile, which may partially explain the greater susceptibility of diabetic subjects to periodontal breakdown.

Effect of periodontal therapy on metabolic control and levels of IL-6 in the gingival crevicular fluid in type 2 diabetes mellitus.

BACKGROUND: The aim of this study was to compare the efficacy of metabolic control and levels of interleukin 6 (IL-6) in gingival crevicular fluid after periodontal therapy in type 2 diabetes mellitus (DM) and nondiabetic (NDM) patients. METHODS: This study was performed in 20 subjects (10 type 2 DM and 10 NDM patients with generalized chronic periodontal disease. Both groups were recorded for clinical parameters (plaque index (PI), gingival index (GI), probing depth (PD), gingival recession (GR) and clinical attachment level (CAL)), metabolic control (fasting glucose levels, glycated a-hemoglobin (HbA1c), low-density lipoprotein (LDL), high-density lipoprotein (HDL), triglycerides (TR)), and IL-6 levels at baseline and 3 months after periodontal treatment. RESULTS: DM and NDM patients revealed significant statistical reductions for clinical parameters (P < 0.05, RM ANOVA) after 3 months. DM group had improvements on HbA1c and had significant statistical increased of TRG (P < 0.05, RM ANOVA) after 3 months. No differences of LDL/HDL and IL-6 levels were found after 3 months in both groups. CONCLUSION: DM group presented a significant reduction of HbA1c levels after periodontal therapy. However, TRG levels increased after 3 months, which suggest more confirmatory studies to investigate if these results will be repeated in other studies.

Interleukins 1ß and 10 expressions in the periimplant crevicular fluid from patients with untreated periimplant disease.

PURPOSE: To analyze the interleukin (IL)-1ß and IL-10 expressions in periimplant crevicular fluid (PICF) in healthy and diseased regions to elucidate the inflammatory process around implants and its influence on clinical diagnosis. MATERIALS AND METHODS: PICF samples from 30 patients were analyzed for IL-1ß and IL-10 concentrations by enzyme-linked immunosorbent assay. Patients were divided in Groups A (health), B (mucositis), and C (periimplantitis). Plaque accumulation, periodontal phenotype (PP), depth on probing, and history of periodontitis (HP) were evaluated. RESULTS: IL-1ß levels were lower in healthy group compared with Groups B (P < 0.0005) and C (P < 0.001). IL-10 levels were higher in Groups A compared with B (P = 0.033) and C (P = 0.0001). Patients with HP and thin PP had 9 and 4.5 times more chance of presenting disease, respectively. CONCLUSIONS: Lower IL-1ß and higher IL-10 levels characterized healthy periimplant conditions, which demonstrate the anti-inflammatory predominance in sites without disease signs. IL-10 levels decrease significantly according to increase of disease status. Therefore, its levels can differentiate healthy, mucositis, and periimplantitis. Thin PP and HP are associated with periimplant disease. These findings suggest the use of ILs as a biochemical marker for early diagnosis of periimplant disease.

Effectiveness of non-surgical treatment to reduce il-18 levels in the gingival crevicular fluid of patients with periodontal disease.

The aim of this study was to evaluate the effectiveness of the non-surgical periodontal treatment in reducing the gingival crevicular fluid (GCF) levels of IL-18 from inflamed periodontal sites. Fourteen patients with periodontal disease were included, being 9 patients with chronic periodontitis (mean age: 48.8 SD ± 7.4 years) and 5 patients with gingivitis (mean age: 43.6 SD ± 11.8). The patients were divided in the following groups: gingivitis sites from periodontitis patients (sites GP), periodontitis sites from periodontitis patients (sites PP), and gingivitis sites from gingivitis patients (sites GG). Probing pocket depth (PPD), probing attachment level (AL), plaque index (PI) and gingival index (GI) were recorded, and gingival fluid samples were collected. The subjects received non-surgical treatment and were re-evaluated 30 days after treatment (day 30 AT). There was a significant reduction in PI in GG (1.0 ± 0.4 to 0.5 ± 0.2), GP (1.2 ± 0.3 to 0.5 ± 0.3), and in PP (1.3 ± 0.4 to 0.7 ± 0.3) 30 AT. There was also a significant reduction in the GI in GG (1.3 ± 0.3 to 0.7 ± 0.4). PPD reduced significantly in GG (2.4 ± 0.6 to 1.9 ± 0.1), and PP (6.7 ± 1.1 to 5.2 ± 0.9) 30 AT. When all the samples were analyzed together, there was a significant reduction in IL-18 (12.9 ± 7.2 to 10.0 ± 3.1). This study showed that non-surgical treatment was effective in reducing GCF levels of IL-18 from inflamed periodontal sites.

The combination of amoxicillin and metronidazole improves clinical and microbiologic results of one-stage, full-mouth, ultrasonic debridement in aggressive periodontitis treatment.

BACKGROUND: The aim of the present study is to assess clinical, microbiologic, and immunologic benefits of amoxicillin/metronidazole (AM) when performing full-mouth ultrasonic debridement (FMUD) in generalized aggressive periodontitis (GAgP) treatment. METHODS: Twenty-four GAgP patients were divided into two groups: the FMUD group (n = 12), which received FMUD plus placebo, and the FMUD+AM group (n = 12), which received FMUD and 375 mg amoxicillin plus 250 mg metronidazole for 7 days. The following clinical outcomes were tested: plaque and bleeding on probing indices, pocket probing depth (PD), relative gingival margin position (GMP), and relative clinical attachment level (CAL). Total amount of Porphyromonas gingivalis (Pg), Aggregatibacter actinomycetemcomitans (Aa), Tannerella forsythia (Tf), and gingival crevicular fluid (GCF) concentration of interleukin (IL)-10 and IL-1ß were also determined. All clinical, microbiologic, and immunologic parameters were assessed at baseline and at 3 and 6 months post-therapy. The ANOVA/Tukey test was used for statistical analysis (α = 5%). RESULTS: Amoxicillin/metronidazole used as an adjunct to the FMUD protocol added clinical and microbiologic benefits to GAgP treatment (P <0.05). FMUD+AM groups presented an additional PD reduction in initially deep PDs at the 3-month follow-up (3.99 ± 1.16 mm and 3.09 ± 0.78 mm for FMUD+AM and FMUD, respectively; P <0.05), a lower number of residual pockets at the 3- and 6-month follow-ups, and a statistical reduction in amounts of Aa (P <0.05). Analysis of Tf and Pg amounts, as well as IL-10 and IL-1ß GCF concentrations failed to demonstrate a difference between the groups (P >0.05). CONCLUSION: It may be concluded that amoxicillin/metronidazole improves clinical and microbiologic results of FMUD in GAgP treatment.

Partial- and full-mouth scaling and root planing in type 2 diabetic subjects: a 12-mo follow-up of clinical parameters and levels of cytokines and osteoclastogenesis-related factors.

BACKGROUND AND OBJECTIVE: The aim of this study was to evaluate the effects of full-mouth scaling and root planing (FMSRP) and partial-mouth scaling and root planing (PMSRP), up to 12 mo after treatment, on clinical parameters, and levels of cytokines and osteoclastogenesis-related factors in type 2 diabetic subjects with chronic periodontitis. MATERIAL AND METHODS: Thirty-four subjects received FMSRP (n = 17) or PMSRP (n = 17) within 24 h or in multiple sessions, respectively. Clinical parameters and local levels of tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), interleukin (IL)-17, IL-23, IL-4, receptor activator of NF-ß ligand and osteoprotegerin were assessed at baseline, and 3, 6 and 12 mo after therapies. RESULTS: Clinical parameters improved after both therapies (p < 0.05), and no between-group differences were observed at any time-point (p > 0.05). Overall, there were no considerable differences in the local levels of the biomarkers studied between groups (p > 0.05). The IL-23 concentration and total amount of IFN-γ increased in the FMSRP group and decreased in the PMSRP group from baseline to 3 mo and from baseline to 6 mo, respectively (p < 0.05). CONCLUSION: Both PMSRP and FMSRP promoted benefits in clinical parameters and showed a similar modulation of cytokines and osteoclastogenesis-related factors at 12 mo in type 2 diabetic subjects.