RESUMO
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.
Assuntos
Humanos , Pessoa de Meia-Idade , Doenças Periodontais/diagnóstico , Infecções por HIV/imunologia , Líquido do Sulco Gengival/imunologia , Periodontite Crônica/classificação , Brasil , Citocinas/análise , Líquido do Sulco Gengival/química , Síndrome da Imunodeficiência Adquirida , Interleucina-6/análise , Interferon-alfa , Estatísticas não Paramétricas , Comitês de Ética em PesquisaRESUMO
Abstract The presence of neopterin in gingival crevicular fluid (GCF) is a marker for local and acute immune activation, and the presence of vascular cell adhesion molecule (VCAM-1) in GCF is accepted as a marker for chronic vascular inflammation. Objectives This study aimed to evaluate effects of periodontal treatment on GCF levels of neopterin and VCAM-1 in patients with chronic periodontitis (CP) with acute myocardial infarction (AMI) compared with systemically healthy CP patients. Material and methods Sixty subjects (20 CP patients with AMI, 20 healthy CP patients, and 20 healthy controls) were included. GCF samples were analyzed at baseline and after 3 and 6 months, and the probing pocket depth (PD), clinical attachment level (CAL), bleeding on probing, gingival (GI) and plaque (PI) indices were recorded. We determined neopterin and VCAM-1 levels (concentration and total amount) using enzyme-linked immunosorbent assay (ELISA). No significant differences were seen between the AMI+CP and CP groups for PI, GI, GCF levels of neopterin and VCAM-1 at baseline. Results The number of teeth with 5 mm≤CAL<7 mm and CAL≥7 mm were significantly increased in the AMI+CP group at baseline. There were no significant differences between the AMI+CP and CP for PI, CAL, GCF volumes, and the AMI+CP group had the highest clinical improvement in the number of teeth with 5 mm≤CAL<7 mm at the sixth month. There were significant positive correlations between clinical periodontal inflammation and the presence of neopterin and VCAM-1 in GCF prior to and following periodontal treatment, and between the GCF volume and clinical parameters. Conclusions Data suggest that the total amount and concentration of neopterin and VCAM-1 in GCF seemed to be closely associated with periodontal disease severity in CP patients with AMI. Moreover, the results of our study demonstrate that the past periodontal status is potentially correlated between groups, with similar periodontal disease severity.
Assuntos
Humanos , Masculino , Feminino , Adulto , Idoso , Líquido do Sulco Gengival/química , Molécula 1 de Adesão de Célula Vascular/análise , Neopterina/análise , Periodontite Crônica/patologia , Periodontite Crônica/terapia , Infarto do Miocárdio/patologia , Valores de Referência , Fatores de Tempo , Índice de Gravidade de Doença , Ensaio de Imunoadsorção Enzimática , Estudos de Casos e Controles , Índice Periodontal , Índice de Placa Dentária , Análise de Variância , Resultado do Tratamento , Perda da Inserção Periodontal , Estatísticas não Paramétricas , Medição de Risco/métodos , Periodontite Crônica/complicações , Pessoa de Meia-Idade , Infarto do Miocárdio/etiologiaRESUMO
Abstract Recent studies investigating protease-activated receptor type 2 (PAR-2) suggest an association between the receptor and periodontal inflammation. It is known that gingipain, a bacterial protease secreted by the important periodontopathogen Porphyromonas gingivalis can activate PAR-2. Previous studies by our group found that PAR-2 is overexpressed in the gingival crevicular fluid (GCF) of patients with moderate chronic periodontitis (MP). The present study aimed at evaluating whether PAR-2 expression is associated with chronic periodontitis severity. GCF samples and clinical parameters, including plaque and bleeding on probing indices, probing pocket depth and clinical attachment level, were collected from the control group (n = 19) at baseline, and from MP patients (n = 19) and severe chronic periodontitis (SP) (n = 19) patients before and 6 weeks after periodontal non-surgical treatment. PAR-2 and gingipain messenger RNA (mRNA) in the GCF of 4 periodontal sites per patient were evaluated by Reverse Transcription Polymerase Chain Reaction (RT-qPCR). PAR-2 and gingipain expressions were greater in periodontitis patients than in control group patients. In addition, the SP group presented increased PAR-2 and gingipain mRNA levels, compared with the MP group. Furthermore, periodontal treatment significantly reduced (p <0.05) PAR-2 expression in patients with periodontitis. In conclusion, PAR-2 is associated with chronic periodontitis severity and with gingipain levels in the periodontal pocket, thus suggesting that PAR-2 expression in the GCF reflects the severity of destruction during periodontal infection.
Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Líquido do Sulco Gengival/química , Receptor PAR-2/análise , Periodontite Crônica/patologia , Valores de Referência , Índice de Gravidade de Doença , Cisteína Endopeptidases/análise , Biomarcadores/análise , Estudos de Casos e Controles , Expressão Gênica , Índice Periodontal , Índice de Placa Dentária , Perda da Inserção Periodontal , Porphyromonas gingivalis , Estatísticas não Paramétricas , Adesinas Bacterianas/análiseRESUMO
O objetivo dessa tese foi avaliar a expressão de citocinas Th1 (IL-12 e INFγ), citocinas Th2 (IL-4, IL-6 e IL-10) e das citocinas pró-inflamatórias IL-18, IL-1β e TNFα no fluido gengival de pacientes com periodontite crônica portadores da doença de Crohn (DC), de retocolite ulcerativa idiopática (RCUI) e em indivíduos saudáveis (o grupo controle, GC). Como objetivo secundário, avaliamos a função dos neutrófilos no fluido gengival desses pacientes através da mensuração das metaloproteinases da matriz -8, -9 (MMP-8 e MMP-9) e da atividade da elastase. Quinze pacientes com DC (idade média 38.2 ± 11.4 anos), 15 pacientes com RCUI (idade média 45.0 ± 10.5 anos) e 15 pacientes saudáveis (idade média 42.1 ± 7.8 anos) participaram desse estudo. Todos os dentes presentes, com exceção dos terceiros molares, foram examinados. Profundidade de bolsa (PB), nível de inserção clínica (NI), presença de placa e de sangramento a sondagem foram avaliados em seis sítios por dente. Em cada paciente, o fluido de 4 sítios com periodontite (PB ≥ 5 mm e NI ≥ 3mm) e de 4 sítios com gengivite (PB ≤ 3 mm e NI ≤ 1 mm) foram coletados através de pontas de papel absorvente pré-fabricadas. O sistema LUMINEX® foi utilizado na mensuração das IL-1β, IL-4, IL-6, IL-10, IL-12p70, TNFα, INFγ, MMP-8 e MMP-9. A IL-18 foi analisada através do ensaio ELISA e a atividade de elastase através de uma reação enzimática. O soro desses pacientes também foi analisado e o coeficiente de correlação de Pearson foi utilizado na análise da correlação entre as citocinas no soro e no fluido gengival. Nos sítios com gengivite, a quantidade total de IL-4 foi significativamente menor no grupo RCUI do que no grupo GC (p=0.016). Nos sítios com periodontite, a quantidade total de IL-4 foi significativamente menor no grupo DC do que no grupo GC (p=0.029)...
The aim of this thesis was to evaluate the expression of Th1 cytokines (IL-12 and INF-γ), Th2 cytokines (IL-4, IL-6 and IL-10) and the pro-inflammatory cytokines IL-18, IL-1β and TNF-α in the gingival crevicular fluid (GCF) from Crohn's disease (CD) patients, ulcerative colitis (UC) patients and healthy individuals (control group, CG) who had chronic periodontitis. Besides, we measured elastase activity, matrix metalloproteinase -8 and -9 (MMP-8 and -9) to address the neutrophil function in the GCF. Fifteen CD patients (mean age 38.2 ± 11.4 years), 15 UC patients (mean age 45.0 ± 10.5 years) and 15 systemically healthy controls (mean age 42.1 ± 7.8 years) were enrolled in this study. All the present teeth, except for the third molars were examined. Probing pocket depth (PPD), clinical attachment loss (CAL), presence of plaque and presence of bleeding on probing were assessed in six sites per tooth. In every subject, GCF from 4 gingivitis sites (PPD ≤ 3mm and CAL ≤ 1mm) and from 4 periodontitis sites (PPD ≥ 5mm and CAL ≥ 3mm) were collected with filter strips. The data were reported as total amount and concentration. IL-1β, IL-4, IL-6, IL-10, IL-12p70, TNFα, INFγ, MMP-8 and MMP-9 were analyzed by the Luminex® analyzer. IL-18 was analyzed using a commercially available ELISA assay and the elastase activity by an enzymatic reaction. The serum was also analysed and the correlations between the cytokines in the GCF and in the serum were calculated by Pearson correlation analysis. In gingivitis sites, the total amount of IL-4 was significantly lower in the UC group than in the CG group (p=0.016). In periodontitis sites, the total amount of IL-4 was significantly lower in CD group than in the CG group (p=0.029). The total amount of IL-4 was lower in UC group than in CD group (p=0.077)...
Assuntos
Humanos , Citocinas/química , Citocinese/imunologia , Líquido do Sulco Gengival/química , Linfócitos/química , Periodontite Crônica/enzimologia , Estudos de Casos e Controles , Doença de Crohn , Doenças Inflamatórias Intestinais , Elastase de Leucócito , Metaloproteinases da Matriz/química , ProctocoliteRESUMO
The objectives of this study were to determine arginine and glutamate levels in the gingival crevicular fluid (GCF) of adult chronic periodontitis patients versus periodontally healthy controls, and to compare two kinds of microdialysis probes: normal and U-shaped probes. The analysis of GCF components was developed to improve the diagnosis of periodontal disease (PD). Proteolysis in the periodontal tissues increases the concentration of amino acids (aa) in the GCF and the levels of these aa may reveal PD features and stages. GCF samples were collected by microdialysis in situ from 5 periodontally affected sites (probing depth >5 mm, clinical attachment loss >3 mm) in 14 adult chronic periodontitis patients and from 14 adult periodontally healthy controls. Capillary zone electrophoresis coupled to laser induced fluorescence detection was used to measure concentration of arginine and glutamate in the GCF. Data were analyzed statistically by ANOVA and Tukey's post-hoc tests (?=0.05). Arginine concentration was increased (p<0.001) and glutamate concentration was decreased (p<0.001) in chronic periodontitis patients as compared to controls. There were no significant differences (p=0.069) between the normal and U-shaped probes. In conclusion, the increase of arginine and decrease of glutamate concentration in GCF were associated to the presence of periodontitis, and might be used as markers to recognize periodontally susceptible subjects as well as to evaluate the treatment course.
Os objetivos deste estudo foram determinar os níveis de arginina e glutamato no fluido gengival crevicular (FGC) em pacientes com periodontite crônica contra controles saudáveis e comparar dois tipos de cânulas de microdiálise: normais e em forma de U. A análise dos componentes do FGC desenvolveu-se para melhorar o diagnóstico da doença periodontal (DP). A proteólise dos tecidos periodontais aumenta a concentração de aminoácidos (aa) no FGC e os níveis destes aa podem revelar as características e estágios da DP. Amostras de FGC foram obtidas pela técnica de microdiálise in situ de cinco zonas com o periodonto afetado (profundidade de sondagem >5 mm, perda da inserção clínica >3 mm) em 14 pacientes adultos com periodontite crônica e 14 controles saudáveis. Para medir a concentração de arginina e glutamato no GFC, usou-se a técnica de eletroforese capilar com detecção de fluorescência induzida por laser. Nos pacientes com periodontite crônica, a concentração de arginina aumentou significantemente (p<0.001), enquanto a de glutamato diminuiu significantemente (p<0.001) em comparação aos controles. Não houve diferenças significantes (p=0.069) entre as cânulas normais e as cânulas em forma de U. Conclui-se que o aumento da concentração de arginina e diminuição de glutamato no FGC estavam associados à presença de periodontite, e podem ser usados como marcadores para identificar pacientes suscetíveis à periodontite bem como avaliar a evolução do tratamento.