The development and function of T helper (Th) cells and regulatory T cells (Tregs) are plastic processes that are regulated by cytokines. In our project we first analyzed the effect of different cytokines on the development of induced (i) Tregs. It has been demonstrated that iTregs arise from CD4+ CD25- T cells upon stimulation with alloantigen in the presence of transforming growth factor β (TGF-β). The development of these Tregs and their proliferation were inhibited by interleukin (IL)-4 and IL-12. The aquired results also demonstrated distinct responses of naturally occuring (n) Tregs and iTregs to the regulatory action of IL-4 and an opposite role of IL-4 in maintenance of nTregs and iTregs phenotype. An important role in the induction of T cell subsets may play also mesenchymal stem cells (MSCs) which can, under specific conditions, produce TGF-β and IL-6. Depending on the current production of TGF-β or IL-6, MSCs can qualitatively regulate the ration between Tregs and Th17 cells. Anti-inflammatory Tregs and pro-inflammatory Th17 cells are induced upon stimulation in the presence of TGF-β and TGF-β and IL-6, respectively. In addition to our previous work we studied the role of IL-12 in the development of Tregs and Th17 cells. It was shown that Treg and also Th17 cell differentiation was...

1 Abstract Vitamin D3 is important for keeping the right concetration of Ca2+ in plasma. Therefore it is essential for proper bone growth and development. Nevertheless, vitamin D3 has also a number of immunomodulating effects. Our thesis has been targeted on evaluation and comparison of vitamin D3 influence on expression of chosen surface markers (CD14, CD54, HLA-DR, CD16, CD36 and CD163) with THP-1 cells and monocytes gained from human peripheral blood. Other aims have been analysing the vitamin D3 influence on longevity of THP-1 cells and measuring the soluble CD14 and IL-8 production with THP-1 cells under the vitamin D3 influence. The cells have been stimulated with five different concentrations of vitamin D3 for the time 24, 48 and 72 hours. Higher used concetrations of vitamin D3, i.e. 100 nM and 1000 nM have increased the expression of CD14 with THP-1 cells in the time 48 and 72 hours of the stimulation time. With the monocytes from peripheral human blood the increase of the CD14 expression hasn't been remarkable from the physiological point of view. Together with the vitamin D3 concentration increase the sCD14 production with THP­1 cells was considerably higher. The sCD14 was the highest in the time 72 hours after the stimulation with the highest used vitamin D3 concetration. The IL-8 quantity with...

Monocytes and macrophages are important components of the innate immune response. These mononuclear phagocytes form a heterogeneous cell population, of which phenotype and functions can be modified under the influence of different signals coming from the surrounding microenvironment. The aim of this work was to modulate the phenotype of these cells by a variety of stimulants and to compare the changes induced on the model of THP-1 monocytic cell line and on the human peripheral blood monocytes. Surface marker expression was analyzed by flow cytometry. Further on, IL-8 production was evaluated by Luminex assay and the concentration of soluble calprotectin was assessed by ELISA. The most significant changes in surface marker expression were induced by exposure to IFNγ. This cytokine increased the expression of CD54, CD14 and HLA-DR on the surface of THP-1 cell line. Higher concentrations of IFNγ promoted higher apoptotic rate and augmented calprotectin expression and production in THP-1 cell line. On the surface of monocytes, IFNγ stimulation resulted only in the upregulation of CD54 expression. IL-4 increased the expression of CD36 by THP-1 cell line and inhibited the expression of CD163 by human monocytes. LPS stimulation caused the suppression of HLA-DR activation in monocytes and enhanced IL-8...

Introdução: A rinossinusite crônica (RSC) é uma doença prevalente, multifatorial, de fisiopatologia ainda muito pouco compreendida e que tem se apresentado como grande desafio na dificuldade de tratar a doença não controlada. Embora tenha sido aceito recentemente que a RSC precisa ser diferenciada em RSC com e sem pólipos nasais, tornou-se claro que essa distinção é insuficiente para definir claramente subgrupos com fisiopatologia e padrões de citocinas uniformes. Objetivos: Estudar o perfil inflamatório TH nos pacientes da região de Ribeirão Preto-SP, Brasil, e comparar com o perfil de outras populações. Casuística e Metodos: Foi realizado estudo transversal prospectivo para identificar o perfil inflamatório (Th1/Th2/Th17) pela concentração das seguintes citocinas: IFN-?1, IFN-?, TGF-?, IL-33, IL-10, IL- 17A, IL-1?, IL-2 e IL-5, em pacientes com RSC com pólipo nasal (RSCcPN), RSC sem pólipo nasal (RSCsPN) e controles. Foram colhidas amostras de tecido nasal (concha média, seio maxilar, seio etmoidal e pólipo nasal) e analisadas por PCR em tempo real. Resultados: Foram avaliados 117 pacientes, sendo que 67 com RSCcPN e 29 com RSCsPN e 21 controles. Pacientes com RSCcPN apresentaram aumento de todas citocinas em relação ao grupo controle, e os com RSCsPN demonstraram aumento de IFN-?1, IFN-?, IL-10, IL-17A, IL1?, IL-2 e IL-5. Na comparação entre RSCcPN e RSCsPN observou-se diferença nas IFN-?, TGF-?, IL- 2, IL-1? e IL-10. Em relação ao pólipo verificou-se padrão eosinofílico em 70% dos casos e relação com IL-5 e AERD (doença respiratória exacerbada por aspirina, do inglês Aspirin-exacerbated respiratory disease). Conclusões: Os resultados mostraram que não existe um perfil inflamatório padrão de RSCcPN e RSCsPN, confirmando que há uma diversidade ampla nas diferentes populações, podendo estar associado a diferentes fatores ainda a serem estudados. / Introduction: The chronic rhinosinusitis (CRS) is a prevalent, multifactorial disease of pathophysiology that is still poorly understood and has been presented as a major challenge in the difficulty of treating uncontrolled disease. Although it has now been accepted that chronic rhinosinusitis needs to be differentiated into CRS with and without nasal polyps, it has become clear that this distinction is insufficient to clearly define subgroups with uniform cytokine pathophysiology and patterns. Objectives: The objective was to study the inflammatory TH profile in region of the Ribeirão Preto-SP, Brazil, patients and compare it with the profile of other populations. Casuistic and Methods: A prospective cross-sectional study was conducted to identify the inflammatory profile (Th1 / Th2 / Th17) by concentration of the following cytokines: IFN-?1, IFN-?, TGF-?, IL-33, IL-10, IL-17A, IL-1?, IL-2 and IL-5 in patients with chronic rhinosinusitis with nasal polyps (CRSwNP), chronic rhinosinusitis without nasal polyps (CRSsNP) and controls. Samples of nasal tissue (medial turbinate, maxillary sinus, ethmoidal sinus and nasal polyp) were collected and analyzed by real-time PCR. Results: A total of 117 patients were studied, of which 67 had CRSwNP and 29 with CRSsNP and 21 controls. Patients with CRSwNP showed an increase in all cytokines compared to the control group, patients with CRSsNP showed an increased in IFN-?1, IFN-?, IL-10, IL-17A, IL-1b, IL-2 and IL-5. In the comparison between CRSwNP and CRSsNP we found difference in IFN-?, TGF-?, IL-2, IL-1? and IL-10. In relation to the polyp we found an eosinophilic endotypes (70%) and relation with IL-5 and AERD (Aspirin-exacerbated respiratory disease). Conclusions: Our results show that there is no standard inflammatory profile of CRSwNP and CRSsNP, confirming that there is a wide diversity in the different populations, and may be associated with different factors still to be studied.

Magister Scientiae (Medical Bioscience) - MSc(MBS) / Graphene oxide (GO) is a single carbon layer, oxygen bearing graphene derivative, containing hydroxyl and carboxyl groups. Graphene oxide nanoparticles (GONPs) are promising nanomaterials for a variety of applications such as electrochemical analysis, adsorption of biomolecules, biosensors and drug and vaccine delivery systems. While these newly engineered nanoparticles hold great potential for developments in industry and medicine, the widespread use of these material will inevitably result in GO residues in the environment where they could possibly pose a risk to human and wildlife health. Interaction of the nanoparticles and biota can affect numerous biological processes. In humans they can affect any of the physiological systems such as the immune, endocrine, reproductive and cardiovascular systems. Although studies have indicated that GO exposure cause increased reactive oxygen species in cells, they mechanisms whereby GO act on the cell are still poorly understood. A few studies have investigated the effects of GONP and other graphene nanoparticle derivatives on the immune system. The aim of this study was to investigate the in vitro effects of GONPs on the immune system by the exposure of the murine macrophage cell line, RAW 264.7, to different concentrations of GONPs.

CD8 T lymphocytes are a crucial component of the adaptive immune system and mediate control of infections and malignancy, but also autoimmunity and allograft rejection. Given their central role in the immune system, CD8 T cell responses are tightly regulated by costimulatory signals and cytokines. Strategies targeting signals that are critical for T cell activation have been employed in a transplantation setting to impede alloreactive T cell responses and prevent graft rejection. The goal of my thesis is to understand how costimulatory signals and inflammation regulate alloreactive CD8 T cell responses and how to target these pathways to develop more effective tools to prevent graft rejection. Costimulation blockade is an effective approach to prolong allograft survival in murine and non-human primate models of transplantation and is an attractive alternative to immunosuppressants. I describe a novel murine anti-CD40 monoclonal antibody that prolongs skin allograft survival across major histocompatibility barriers and attenuates alloreactive CD8 T cell responses. I find that the pro-apoptotic proteins Fas and Bim function concurrently to regulate peripheral tolerance induction to allografts. Activation of the innate immune system by endogenous moIecules released during surgery or infections in transplant recipients can modulate T cell responses. However, the direct impact of inflammation on alloreactive CD8 T cell responses is not clear. Using a T cell receptor (TCR) transgenic mouse modeI, I demonstrate that inflammatory stimuli bacterial lipopolysaccharide (LPS) and the viral dsRNA mimetic poly(I:C) differentially regulate donor-reactive CD8 T cell responses by generating distinct cytokine milieus. Finally I demonstrate the role of pro-inflammatory cytokines stem cell factor (SCF), granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-3 (IL-3) in improving human B cell development in humanized NOD-scid IL2Rγnull (NSG) mice.

Neste estudo caracterizamos a expressão de Mif nas células trofoblásticas (CT), placentárias e decidua (D) na gestação em camundongos.A imunolocalização foi realizada nos dias de gestação(dg) 7,5, 10,5, 13,5 e 17,5.Com cones ectoplacentários e placentas fetais (PL) realizou-se ensaios de Western blotting e qRT-PCR nos mesmos dg.D foram submetidas a qPCR.Aos 7,5 dg as CT gigantes e D apresentaram imunomarcação.Dos 10,5 ao 17,5 dg o Mif concentrou-se nas CTG e no espongiotrofoblasto.Na D, a imunomarcação foi menor que aos 7,5 dg.A expressão protéica na PL aumentou do 7,5 dg para o 10,5 dg (p=0,005) e para o 13,5 dg (p=0.03).A maior expressão gênica foi em 10,5 dg e diferente em 13,5 dg (p=0,048) e 17,5 dg (p=0,009).Na D, a maior expressão gênica foi em 7,5 dg e diferente dos 10,5 (0,012) e 13,5 (0,032) dg.O aumento da expressão de Mif na PL coincide com a organização em quatro camadas e com o início da circulação fetal.Esta distribuição temporal e tecido-específica sugere o MIF como modulador no início da placentação ou na sua adaptação ao ambiente uterino / The goal of this study was to characterize Mif expression by trophoblast (TC),fetal placenta (FP) and decidua (D) during mouse pregnancy.Mif was immunolocalized at TC and D on gestation days (gd) 7.5, 10.5, 13.5 and 17.5.Ectoplacental cones (EC) and FP were used for Western blotting and qPCR.D were also used for qRT-PCR.On gd 7.5,DC and TC giant (TGC) showed strong reactivity.On gds 10.517.5,were concentrated in TGC and spongiotrophoblast cells. D reactivity was weaker than 7.5 gd.Protein expression at FP increased from gd 7.5 to 10.5 (p=0.005) and to 13.5 (p=0.03).Higher mRNA expression was found on gd 10.5 and was different from gds 13.5 (p=0.048) and 17.5 (p=0.009).At D on gd 7.5 was greater than those on gds 10.5 (0,012) and 13.5 (0,032).The up-regulation of Mif coincides with the stage that placenta assumes its four-layered organization and the fetal blood circulation begins,This temporal tissue-specific distribution and expression data suggests that Mif may play a modulator role in the onset of placentation or in it adaptation to the uterine environment

O ligamento periodontal e o tecido gengival são formados por tecido conjuntivo frouxo, sendo constituídos por diversas células, dentre as quais os fibroblastos são as mais numerosas. Ao serem submetidas a agressões diversas, estas células respondem com a liberação de substâncias, tais como citocinas e quimiocinas, que participam de maneira ativa no processo inflamatório, e com a produção de componentes da matriz extracelular fundamentais para o reparo, como por exemplo, o colágeno. Assim sendo, este trabalho teve como objetivo: Avaliar e comparar a expressão e a produção de prócolágeno tipo I, IL6, MIP1 e SDF1 por fibroblastos humanos, cultivados de ligamento periodontal e de tecido gengival, estimulados por lipopolissacarídeo (LPS) de Porphyromonas gingivalis. Foram coletados ligamentos periodontais de terceiros molares não irrompidos e biópsias de gengiva de um mesmo indivíduo (n=3). Estes tecidos foram picotados e mantidos em meio de cultura adequado para fibroblastos, que foram utilizados na quarta passagem. Após adesão dos fibroblastos a placas de 24 poços, o meio de cultura contendo 0,1 10 g/mL de LPS de P. gingivalis foi adicionado às placas em duplicata. O sobrenadante e as células foram coletados após 1, 6 e 24 horas e analisados por ELISA e PCR em tempo real, respectivamente. A análise estatística foi realizada por meio do programa GraphPad Prism, aplicandose o teste ANOVA a 1 critério com nível de significância de 5%. A expressão de prócolágeno tipo I mostrouse - ligeiramente diferente entre fibroblastos de ligamento periodontal e de gengiva. A produção de IL6, MIP1 e SDF1 foi significativamente maior em fibroblastos gengivais. A citocina IL6 foi produzida de maneira tempodependente com LPS de P gingivalis, principalmente por fibroblastos gengivais. Para MIP1, os fibroblastos gengivais mostraram maior produção com a menor concentração de estímulo (0,1g/ml). Para SDF1, foi detectada produção constitutiva que foi inibida com o aumento da concentração de LPS ao longo do tempo nestas mesmas células. Já para fibroblastos de ligamento periodontal, não foi observado um padrão homogêneo e linear, apesar de a produção basal de SDF1 também existir, porém em níveis bem mais discretos, como aquele observado para a produção de MIP1. A capacidade dos fibroblastos modificarem o padrão de produção dessas citocinas frente ao estímulo com LPS de P. gingivalis reforça a importância dessas células no contexto da resposta imune do indivíduo frente à doença periodontal. / The fibroblast is considered an important cell in periodontitis because it is the predominant cell type in the periodontal connective tissue. When challenged by different agents, fibroblasts respond through the release of substances, such as cytokines and chemokines that participate in an active way in the inflammatory process as well as the production of basic components of the extracellular matrix for repair, like collagen. The aim of this study was to: to evaluate and to compare the expression and production of type I procollagen, IL6, MIP1 and SDF1 by cultured human periodontal ligament and gingival fibroblasts challenged with lipopolyssacharide from Porphyromonas gingivalis. Human periodontal ligament and gingival fibroblasts were cultured from biopsies of the same donor and were used on the fourth passage. After confluence in 24well plates, the culture medium alone (control) or with 0,1 10 ug/mL of LPS from P. gingivalis were added and after 1, 6 and 24 hours, the supernatant and the cells were collected and analysed by ELISA and Real time PCR, respectively. Data were analysed by GraphPad Prism Program (1 way ANOVA test) and a significance level of 5% was adopted. Procollagen type I expression by Real Time PCR differ between periodontal ligament and gingival fibroblasts. In vitro experiments revealed that IL6, MIP 1 and SDF1 production were significantly greater in gingival fibroblasts when compared to periodontal ligament. In addition, IL6 was upregulated in a timedependent manner, mainly by the gingival fibroblasts. On one hand, MIP1 was stimulated with a low concentration (0,1ugml) of LPS by gingival fibroblasts. On the other hand, SDF1 was constitutively secreted by the same cells but its production was inhibited when challenged by a higher concentration of LPS from P gingivalis. In general, periodontal ligament fibroblasts did not show a pattern of production of these cytokines under the challenge with LPS, despite of the basal production of SDF1 in lower levels than gingival cells and the low production of MIP1 over time. The differential ability of the gingival and periodontal ligament fibroblasts to secrete these cytokines emphasizes their crucial role in the inflammatory microenvironment and in the host immune response to periodontal disease.

A alimentação não é mais vista apenas como uma fonte de calorias e nutrientes, mas sim como agente capaz de induzir benefícios na prevenção de doenças, bem como na promoção de bem estar físico e mental. Assim, uma boa alimentação da população contribui com a redução de custos em cuidados com a saúde e com o aumento da longevidade. Estratégias nutricionais podem, por exemplo, como no caso da adição de antioxidantes, reduzir o risco de doenças, melhorar a qualidade de vida e aumentar o próprio tempo de vida do indivíduo. O dano oxidativo é um fator causador do desenvolvimento de doenças e os antioxidantes têm a capacidade de prevenir ou atenuar estes processos. Sendo assim, há crescente interesse na ingestão de alimentos naturais e produtos orgânicos, na forma de alimentos funcionais e suplementos dietéticos. Entre os alimentos que, comprovadamente, exercem efeito antioxidante, está o açaí, Euterpe oleracea MARTIUS, fruta nativa da região amazônica consumida rotineiramente pela população na forma de polpa, suco, vinho e em outras variedades. Nos últimos anos o açaí tem sido objeto de muita atenção devido ao seu alto potencial antioxidante, anti inflamatório e seu papel como alimento funcional. Sendo assim, foi nosso interesse estudar os efeitos do consumo crônico da fruta, oferecida como consumida pela população em sucos. Os resultados apontam que, quando associado ao consumo de altas taxas de glicose, as propriedades do açaí podem estar reduzidas, uma vez que não foi observada ampla capacidade antioxidante e verificou-se um desvio para o perfil pró-inflamatório no tecido adiposo, induzido pelo consumo de carboidrato em alta concentração. / Feeding is no longer seen as just a source of calories and nutrients, but as an agent capable to induce benefits to prevent diseases, and also to promote physical and mental wellbeing. Therefore, a good nutritional status of the population contributes to cost reductions of health care services and increases longevity. Nutritional strategies, for instance the supplementation with antioxidants, could reduce the risk of diseases that are associated with oxidative stress. The oxidative damage is an important factor in aging and in the development of illness, such as insulin resistence, arteriosclerosis, cancer and others. Antioxidants have the ability to prevent or reduce these processes. Hence, there is a growing interest in consuming natural foods and organic products as functional foods and dietary supplements. The açai berry Euterpe oleracea MARTIUS, a native fruit of Amazonia, recently gained scientific interest due to its high antioxidant and anti inflammatory effects. Açai is frequently consumed by the population as pulp, juice, wine or other varieties that additionally qualifies this fruit as a functional food. Therefore, our interest was to investigate the chronic effects of açai consumption by the population in form of beverages. The results indicate that the açai properties do not prevail, if the consumption is associated with high levels of glucose. The açai properties may be reduced, once the antioxidant ability was not observed, also there was a deviation to pro-inflammatory profile in adipose-tissue, induced by the high levels of carbohydrate consume.

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. / 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.