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)

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.

Peripheral blood mononuclear phagocytes from patients with chronic periodontitis are primed for osteoclast formation.

BACKGROUND: During inflammatory periodontal disease, peripheral blood mononuclear cells (PBMCs) are attracted to bone and differentiate into active bone-resorbing osteoclasts (OCs), thus providing evidence that the impact of chronic periodontitis (CP) on the activity of circulating mononuclear cells is of central importance. The authors test the hypothesis that peripheral blood mononuclear phagocytes (PBMPs) from patients with CP are activated and more susceptible to differentiation into OCs, which in turn would lead to more intense bone resorption. METHODS: In vitro cytokine production by both unstimulated and lipopolysaccharide-stimulated PBMCs from individuals with (n = 10) or without (n = 12) periodontitis was determined by cytokine array. OC differentiation from CD14(+) PBMCs was induced by receptor activator of nuclear factor-kappa B ligand (RANKL), either alone or in the presence of macrophage colony-stimulating factor (M-CSF). PBMC differentiation to OCs was confirmed by tartrate-resistant acid phosphatase staining; bone resorbing activity was assessed by using an osteologic plate assay (bone resorption pit formation). RESULTS: PBMCs from patients with CP produced tumor necrosis factor-α and higher amounts of interferon-γ, interleukin (IL)-1α, IL-1ß, IL-1rα, CXC motif chemokine 10, macrophage migration inhibitory factor, macrophage inflammatory protein (MIP)-1α, and MIP-1ß than the control cells. OC differentiation was induced by RANKL alone in PBMCs from patients with CP, but not in PBMCs from the healthy controls, which required the addition of M-CSF. In addition, PBMC-derived OCs from patients with CP showed significantly higher resorption activity than that observed in the healthy controls. Also, the circulating concentrations of M-CSF were significantly higher in patients with CP than in the control participants. CONCLUSIONS: These data indicate that in patients with CP, circulating PBMCs are primed for increased proinflammatory activity and that M-CSF plays a central role in this process by increasing OC formation and the consequent bone resorption activity.

Kinetics of interleukin-6 and chemokine ligands 2 and 3 expression of periodontal tissues during orthodontic tooth movement.

INTRODUCTION: Mechanical loading induces remodeling of the periodontal ligament and the alveolar bone and is mediated by cytokines and chemokines. In this study, we investigated the kinetics of interleukin-6 and chemokine ligands 2 and 3 levels in periodontal ligaments subjected to orthodontic forces. METHODS: We used 64 premolars in this split-mouth design study. The experimental group consisted of premolars subjected to a force of 0.980 N in the apical direction for 3 hours, 15 hours, 3 days, 12 days, or 21 days with a 0.017 × 0.025-in beta-titanium alloy cantilever. The contralateral teeth, without orthodontic appliances, were used as controls. The premolars were extracted for orthodontic reasons, and the periodontal ligaments were scraped for analysis of cytokine levels by ELISA. RESULTS: Compared with the control group, an increase in chemokine ligand 2 was observed on days 3 and 12, and increases in interleukin-6 and chemokine ligand 3 were observed on day 12 in the experimental group. CONCLUSIONS: Our data demonstrated differential expressions of interleukin-6 and chemokine ligands 2 and 3 in periodontal ligaments after mechanical loading; this might reflect the distinct roles of these molecules in the bone remodeling process.

Salivary immunity in elderly individuals presented with Candida-related denture stomatitis.

OBJECTIVES: Elderly individuals with Candida-related denture stomatitis (DS) present with a reduced defence against Candida albicans. This study evaluated levels of antimicrobial mediators in the elderly DS saliva and salivary neutrophils' activation characteristics compared with elderly and young without DS. METHODS: Salivary peroxidases (SPO) and elastase activities (ELA), nitric oxide (NO), transforming growth factor beta (TGF-ß), IL-6 and CCL3 production were determined in saliva from elderly with or without DS, and young control individuals. TLR4, CXCR1, CD11b, CD16 and CD32 expression on salivary neutrophils were evaluated. Correlations between number and apoptosis rate of salivary neutrophils, enzymatic activities and cytokine levels were determined. RESULTS: Elderly DS individuals exhibited the lowest SPO and ELA activities. Also, the activity of both enzymes was low in elderly without DS. Although both elderly groups showed higher salivary NO and TGF-ß levels compared to young control groups, elderly DS presented the highest salivary NO, TGF-ß, IL-6 and CCL3 levels. Decreased percentages of salivary TLR4(+) and CD16(+) neutrophils were detected in both elderly groups. Although these damages could influence the establishment and persistence of DS, the highest levels of salivary IL-6 and CCL3 in elderly DS could be preventing more serious complications.

Adjuvant effect of a probiotic fermented milk in the protection against Salmonella enteritidis serovar typhimurium infection: mechanisms involved.

Probiotics may offer protection against Salmonella enteritidis serovar Typhimurium infection via different mechanisms. The aim of this study is to investigate, using mouse models, the effect of the administration of fermented milk containing the probiotic bacteria L. casei DN-114 001 in the protection against Salmonella enteritidis serovar Typhimurium when this product is administered continuously before and after infection or only post-infection. The adjuvant effect of this probiotic fermented milk (PFM) against S. Typhimurium was also evaluated in newborn mice, whose mothers received the PFM during the suckling period or their offspring after weaning. The results obtained showed that PFM administration after salmonella infection was useful to decrease the severity of the infection. The best effect was obtained with continuous PFM administration. In the newborn mice model, PFM administration to the newborn mice after weaning showed the best effect against the pathogen. PFM administration to the mother during the suckling period was beneficial against this enterophatogen when their offspring did not receive probiotics after weaning. Continuous PFM administration to adult mice (before and after infection) was important to maintain the intestinal barrier and the immune surveillance in optimal conditions to diminish the pathway of entrance of salmonella and the spread of this pathogen to deeper tissues. In the newborn mice model, it was observed that PFM administration to the offspring after weaning or their mother during the suckling period had a protective effect against salmonella infection, however, in the mice from mothers that received PFM during nursing which were fed with PFM after weaning, we found a down regulated immune maturity that was not protective against this infection.

CCR5 down-regulates osteoclast function in orthodontic tooth movement.

During orthodontic tooth movement, there is local production of chemokines and an influx of leukocytes into the periodontium. CCL5 plays an important role in osteoclast recruitment and activation. This study aimed to investigate whether the CCR5-receptor influences these events and, consequently, orthodontic tooth movement. An orthodontic appliance was placed in wild-type mice (WT) and CCR5-deficient mice (CCR5(-/-)). The expression of mediators involved in bone remodeling was evaluated in periodontal tissues by Real-time PCR. The number of TRAP-positive osteoclasts and the expression of cathepsin K, RANKL, and MMP13 were significantly higher in CCR5(-/-). Meanwhile, the expression of two osteoblastic differentiation markers, RUNX2 and osteocalcin, and that of bone resorption regulators, IL-10 and OPG, were lower in CCR5(-/-). Analysis of the data also showed that CCR5(-/-) exhibited a greater amount of tooth movement after 7 days of mechanical loading. The results suggested that CCR5 might be a down-regulator of alveolar bone resorption during orthodontic movement.