Cytokine expression in gingival and intestinal tissues of patients with periodontitis and inflammatory bowel disease: An exploratory study.

OBJECTIVE: To evaluate the expression of the cytokines IFN-γ, IL-1ß, IL-4, IL-6, IL-10, IL-21, IL-22, IL-23, IL-25, IL-31, IL-33, IL-17A, IL-17F, sCD40L, and TNF-α in gingival tissue and intestinal mucosa of patients having both periodontitis and inflammatory bowel disease (IBD) and assess how they cluster in both tissues. METHODS: This cross-sectional study selected 28 patients with periodontitis (18 with Crohn's disease and 10 with ulcerative colitis) from the IBD gastroenterology outpatient clinic at the Pedro Ernesto University Hospital. Patients were assessed using questionnaire, medical chart check and periodontal examination. Gingival and intestinal biopsies were collected and homogenized using a cell disruptor. Cytokines expression was evaluated through multiplex technology. Cluster analysis was performed based on cytokinés correlation strength and presented in dendrograms. RESULTS: Crohn's disease and ulcerative colitis patients exhibited no significant difference between them in cytokine levels (p>0.05), so they were analysed together. Significantly higher levels of IL-17A, IL-17F, IL-22, IL-25, IL-33, IL-10, and INF-γ were found in gingival tissues in comparison with intestinal mucosa (p<0.05). In gingival tissue, cytokines formed the following clusters: IL-25/IL-10/IL-33 (r=0.775), IL-22/IL-23/IL-6 (r=0.681) and IL-6/IL-25/IL-33/IL-10 (r=0.660). In intestinal mucosa, the following clusters were formed: IL-6/IL-21/IL-10 (r=0.880), IL-17A/IL-6/IL-21/IL-10 (r=0.826), IL-I7F/IL-33/IL-25 (r=0.813) and IL-23/IL-2/IL-17A/IL-6/IL-21/IL-10 (r=0.785). CONCLUSION: Expression of IL-17A, IL-17F, IL-22, IL-25, IL-33, IL-10, and INF-γ was significantly increased in gingival tissue in comparison with intestinal mucosa of patients with periodontitis and IBD. The cytokine clustering pattern was different in gingival and intestinal tissues.

Presença do polimorfismo do gene TNFA em pacientes com doença de crohn e com periodontite crônica / Presence of TNFA gene polymorphism in patients with crohn's disease and chronic periodontitis

Indivíduos com doença inflamatória intestinal (DII) apresentam uma maior prevalência de periodontite. O objetivo desse estudo piloto foi investigar a presença do polimorfismo do gene do TNF-A -308 G>A em indivíduos com DII e periodontite comparando com indivíduos sistemicamente saudáveis com periodontite. Nossa hipótese é que nos indivíduos com DII e periodontite, a presença do polimorfismo do fator de necrose tumoral (TNFA) -308 G>A seja mais prevalente. Dez pacientes com Doença de Crohn (DC) e periodontite, 6 pacientes com retocolite ulcerativa idiopática (RCUI) e periodontite e 8 pacientes sem DII com periodontite participaram desse estudo. Foram considerados com periodontite indivíduos com perda de inserção clínica maior ou igual a 3 mm em pelo menos 4 sítios em diferentes dentes. Células epiteliais da mucosa oral foram coletadas com cotonetes esterilizados. O DNA foi extraído através de um kit comercial. O DNA obtido foi utilizado como molde em reações de amplificação das regiões genômicas de interesse através da técnica de reação em cadeia polimerase - PCR, utilizando-se oligonucleotídeos específicos. A comparação entre grupos foi feita com o teste ANOVA e com o teste T. O teste x² foi utilizado para analisar a presença do polimorfismo. O nível de significância foi determinado em 5% (p< 0,05). Nos pacientes com DC, 60% apresentavam 1 alelo polimórfico. Nos com RCUI, não foi encontrado o alelo polimórfico. Nos saudáveis com periodontite, 25% apresentaram o alelo polimórfico. Esse estudo demonstrou que pacientes com DC e periodontite possuíam uma maior prevalência de um alelo polimórfico do TNFA -308 G> A. Esse resultado sugere que apesar de não necessariamente determinar a ocorrência da DII e nem da periodontite, o polimorfismo do TNFA -308 G>A pode exercer um papel na modificação do fenótipo de pacientes com DII e periodontite

Subjects with inflammatory bowel disease (IBD) have a higher prevalence of peridontitis. The aim of this pilot study was to assess the polymorphism of TNFA -308 G>A in IBD patients who had periodontitis and to compare to systemically health individuals who have periodontitis. Our hypothesis is that in subjects with IBD and periodontitis the occurrence of a genetic polymorphism of tumor necrosis factor alpha (TNFA) -308 G>Ais more prevalent. Ten patients with Crohn 's Disease (CD), 6 patients with ulcerative colitis (UC) and 8individuals systemically health otherwise periodontitis were included in this study. Periodontitis was defined by clinical attachment loss equal or higher 3 mm in at least 4 sites in different teeth. Epithelial cells from the oral mucosa were obtained through a sterilized swab. The DNA extraction was realized through a commercial kit. The obtained DNA was used for the aimed region in polymerase chain reaction –PCR using specific oligonucleotides. Comparisons between the groups were analyzed by ANOVA and T tests. To analyze thepresence of the polymorphism x² test was used. Significance was set at 5% (p< 0.05). Sixty per cent of CD patients had one polymorphicalele. None of UC patients had the polymorphic allele. Twenty five per cent of systemically health individuals had the polymorphic allele. This study showed that patientswith Crohn 's Disease who have periodontitis present a higher prevalance of the polymorphic allele of TNA-308. This preliminary result suggests that although this polymorphism is not a risk for occurrence of IBD or periodontitis, this polymorphism may have a role in modifying the phenotype of IBD patients who have periodontitis.

Níveis elevados de IL-6 no fluido gengival de pacientes com periodontite crônica e retrocolite ulcerativa idiopática / Elevated levels of IL-6 in the gingival crevicular fluid from patients with chronic periodontitis and inflammatory bowel disease

O nosso objetivo foi mensurar os níveis de Interleucina-6 (IL-6) no fluido gengival de pacientes com periodontite e doença inflamatória intestinal (DII), comparando-os com pacientes sistemicamente saudáveis, com periodontite. Como objetivo secundário será avaliada a IL-6 no soro desses pacientes. Foram selecionados 15 pacientes com doença de Crohn (DC, idade média 38.2, DP ± 11.4 anos), 15 com retrocolite ulcerativa idiopática (RCUI, 45.0 ± 10.5 anos) e 15 pacientes saudáveis (C, 42.1 ± 7.8 anos). A Profundidade de bolsa (PB), nível de inserção clínica (NI), presença de placa e de sangramento a sondagem foram avaliados em seis sítios por dente. O fluido gengival foi coletado de quatro sítios com periodontite (PP: PB ≥ 5mm, NI ≥ 3mm) e quatro sítios com gengivite (GP: PB ≤ 3mm e NI≤ 1mm), em dentes diferentes, com pontas de papel absorvente pré-fabricadas. O soro destes pacientes também foi coletado. A análise da IL-6 foi realizada pelo LUMINEX®. A quantidade total e concentração da IL-6 estavam significantemente maiores no fluido gengival dos sítios PP do grupo RCUI quando comparados aos sítios PP do grupo controle (p=0.028; p=0.044, respectivamente). O grupo DC apresentou a quantidade total de IL-6 significantemente maior no sítio PP do que no GP (p=0.028). Já no soro, a IL-6 não diferiu entre os grupos. Sendo assim, pode-se concluir que os indivíduos com retrocolite ulcerativa idiopática apresentavam níveis mais altos de IL-6 nos sítios com periodontite, o que pode indicar um importante papel dessa citocina no estabelecimento e progressão da doença periodontal nesses pacientes.

Our aim was to measure the levels of interleukin-6 in gingival crevicular fluid (GCF) from patients with periodontitis and inflammatory bowel disease (IBD), and compare with systemically healthy controls with periodontitis. Besides, was measured IL-6 level in serum. Fifteen patients with Crohn’s disease (CD, mean age 38.2 ± 11.4 years), 15 with ulcerative colitis (RCUI, 45.0 ± 10.5) and 15 controls (C, 42.1 ± 7.8) participated in this study. Probing pocket depth (PPD), attachment loss (CAL), presence of plaque and presence of bleeding on probing, were assessed in six sites per tooth. In each subject, GCF from four sites with gingivitis (GP- PPD≤ 3mm and CAL ≤ 1mm) and from 4 sites with periodontitis (PP- PPD ≥ 5mm and CAL ≥ 3mm), on different teeth, were collected with filter strips. The serum of those patients was also collected. The IL-6 was analyzed in the Luminex®. The total amount and the concentration of IL-6 in GCF was significantly higher in PP sites from RCUI than in controls (p=0.028 e p=0.044, respectively). The total amount of IL-6 was significantly higher in PP than GP sites in the DC group (p=0.028). In serum, IL-6 does not differ between groups. Therefore was concluded that subjects with ulcerative colitis showed higher levels of IL-6 in periodontitis sites, which might indicate an important role of this cytokine in the onset and progression of periodontal disease in such patients.