Periodontopathogens influence in the expression and production of antimicrobial peptides and inflammatory cytokines in neutrophils from healthy and periodontitis individuals

Orientador: Reginaldo Bruno GonçalvesTese (doutorado) - Universidade Estadual de Campinas. Faculdade de Odontologia de PiracicabaResumo: Dentre os mecanismos da imunidade inata, os quais são importantes para a proteção da cavidade oral, podemos citar os peptídeos antimicrobianos e células fagocíticas como os neutrófilos. Os peptídeos antimicrobianos, LL-37 e ?-defensinas ou peptídeos de neutrófilos humanos (HNP 1-3) são produzidos por neutrófilos e têm ampla atividade contra bactérias, fungos e vírus envelopados, um mecanismo microbicida independente de oxigênio. Contudo, os neutrófilos também executam atividade antimicrobiana através da produção de óxido nítrico (NO) durante a fagocitose. Adicionalmente, estas células produzem diferentes citocinas em resposta aos patógenos. Sendo assim, os neutrófilos tornam-se importantes na resposta contra a doença periodontal, mas, não está bem descrito se há diferenças funcionais entre os neutrófilos de pacientes com periodontite e os de indivíduos saudáveis após estimulação destes com lipopolossacarídeo (LPS) de periodontopatógenos. Dessa forma, avaliamos os níveis de LL-37, HNP1-3, NO e citocinas no sobrenadante das culturas de neutrófilos de voluntários com e sem periodontite e estimulados com LPS de Porphyromonas gingivalis (Pg) e Aggregatibacter actinomycetemcomitans (Aa) e Escherichia coli (Ec), bem como analisamos a expressão de RNAm, para esses peptídeos e citocinas nessas células. Nossos resultados demonstraram que neutrófilos de voluntários com periodontite estimulados com LPS de Aa expressaram maiores níveis de RNAm de LL-37, HNP 1-3 e IFN- ?, bem como produziram maiores quantidades de IL-12 e IFN-_ quando comparados aos saudáveis. LPS de Pg e Ec aumentaram significativamente a expressão de HNP 1-3, IFN-? e IL-8, bem como a produção de LL-37, IL-12 e IFN-?. Em contrapartida, neutrófilos de pacientes com periodontite produziram níveis reduzidos de NO e nenhuma diferença foi observada na expressão de IL-12 e na produção de HNP 1-3 entre ambos os grupos de voluntários estudados. Estes dados permitem concluir que neutrófilos de pacientes com periodontite respondem de forma mais intensa aos estímulos de LPS de bactérias periodontopatogênicas em comparação aos neutrófilos de voluntários com periodonto saudável.Abstract: Among the innate mechanisms, which are essential for the oral cavity protection, we can include antimicrobial peptides and phagocytic cells such as neutrophils, as a major. Antimicrobial peptides, LL-37 and ?-defensins or human neutrophil peptides (HNP 1-3) are produced by neutrophils and have broad activity against bacteria, fungi and enveloped viruses, an oxygen-independent microbicidal mechanism. However, neutrophils also do antimicrobial activity through nitric oxide (NO) production in phagocytosis. Also, neutrophils produce special cytokines against pathogens. Thus, neutrophils are central in the periodontal disease response, but is unclear the functional differences between neutrophils from periodontitis and healthy volunteers pos-stimulation with periodopathogens lipopolossacarideo (LPS). Then, we studied of LL-37, HNP1-3, NO and cytokines levels in culture supernatants of cells from volunteers with or without periodontitis post Porphyromonas gingivalis (Pg), Aggregatibacter actinomycetemcomitans (Aa) and Escherichia coli (Ec) LPS. We analyzed also peptides and cytokines expression in these cells. We show that neutrophils from periodontitis volunteers cultured with Aa LPS expressed higher LL-37, HNP 1-3 and IFN-? mRNA and produced more IL-12 and IFN- ? levels as the controls. Pg and Ec LPS notably improved the HNP1-3, IFN-? and IL-8 expression and the LL-37, IL-12 and IFN- ? secretion. But, neutrophils from periodontitis patients produced low NO levels and similar IL-12 mRNA and HNP 1-3 production in both the volunteers groups studied. Briefly, we can conclude that neutrophils from periodontitis patients respond more strongly than healthy neutrophils when stimulated with LPS periodontopathic bacterias.DoutoradoMicrobiologia e ImunologiaDoutor em Biologia Buco-Denta

Microbiological, lipid and immunological profiles in children with gingivitis and type 1 diabetes mellitus

Objective The aim of this study was to compare the prevalence of periodontal pathogens, systemic inflammatory mediators and lipid profiles in type 1 diabetes children (DM) with those observed in children without diabetes (NDM), both with gingivitis. Material and methods Twenty-four DM children and twenty-seven NDM controls were evaluated. The periodontal status, glycemic and lipid profiles were determined for both groups. Subgingival samples of periodontal sites were collected to determine the prevalence of periodontal microorganisms by PCR. Blood samples were collected for IL-1-β, TNF-α and IL-6 analysis using ELISA kits. Results Periodontal conditions of DM and NDM patients were similar, without statistical differences in periodontal indices. When considering patients with gingivitis, all lipid parameters evaluated were highest in the DM group; Capnocytophaga sputigena and Capnocytophaga ochracea were more prevalent in the periodontal sites of DM children. “Red complex” bacteria were detected in few sites of DM and NDM groups. Fusobacterium nucleatum and Campylobacter rectus were frequently found in both groups. Similar levels of IL-1-β, TNF-α and IL-6 were detected in DM and NDM children. Conclusion Clinical and immunological profiles are similar between DM and NDM children. The presence of Capnocytophaga sputigena and Capnocytophaga ochracea were associated with gingivitis in DM children

Microbiological, lipid and immunological profiles in children with gingivitis and type 1 diabetes mellitus

Abstract Objective The aim of this study was to compare the prevalence of periodontal pathogens, systemic inflammatory mediators and lipid profiles in type 1 diabetes children (DM) with those observed in children without diabetes (NDM), both with gingivitis. Material and methods Twenty-four DM children and twenty-seven NDM controls were evaluated. The periodontal status, glycemic and lipid profiles were determined for both groups. Subgingival samples of periodontal sites were collected to determine the prevalence of periodontal microorganisms by PCR. Blood samples were collected for IL-1-β, TNF-α and IL-6 analysis using ELISA kits. Results Periodontal conditions of DM and NDM patients were similar, without statistical differences in periodontal indices. When considering patients with gingivitis, all lipid parameters evaluated were highest in the DM group; Capnocytophaga sputigena and Capnocytophaga ochracea were more prevalent in the periodontal sites of DM children. “Red complex” bacteria were detected in few sites of DM and NDM groups. Fusobacterium nucleatum and Campylobacter rectus were frequently found in both groups. Similar levels of IL-1-β, TNF-α and IL-6 were detected in DM and NDM children. Conclusion Clinical and immunological profiles are similar between DM and NDM children. The presence of Capnocytophaga sputigena and Capnocytophaga ochracea were associated with gingivitis in DM children

Periodontitis causes abnormalities in the liver of rats

Periodontitis not only causes injury to the periodontium, but also damages other tissues such as: articulate, renal, cardiac, and hepatic. The objective of this study was to investigate periodontitis induced alterations in liver function and structure using an experimental model.Twenty female rats (Rattus norvegicus) were allocated into two groups: control and periodontitis. Gingival bleeding index and oxidative stress parameters and specific circulating biomarkers were measured. Immunohistochemistry was carried out using alkaline phosphatase (AlkP) staining of the liver. Hepatic tissues, cytokines, and lipid contents were measured. Histopathologic evaluation of the liver was carried out using light and electron microscopy.Liver histopathologic and immunohistochemistry assessment showed increase in steatosis score, and presence of binucleate hepatocytes and positive cells for AlkP in periodontitis versus control group. Ultrastructural evaluation showed significant increase in size and number of lipid droplets (LD), distance between the cisterns of rough endoplasmic reticulum (RER), mitochondria size, foamy cytoplasm, and glycogen accumulation in the liver of the periodontitis group compared with the control group. In addition, plasma levels of AlkP, high‐density lipoprotein (HDL), triglycerides, and total cholesterol were also changed.Experimental periodontitis caused immunohistochemistry, histopathologic, ultrastructural, oxidative, and biochemical changes in the liver of rats.903295305CAPES - Coordenação de Aperfeiçoamento de Pessoal e Nível SuperiorCNPQ - Conselho Nacional de Desenvolvimento Científico e Tecnológico455104/2014-0Não te