Análise da expressão imuno-histoquímica da ciclofilina A, EMMPRIN e MMP-7 na doença periodontal

Periodontal disease is an infectious disease resulting from the immunoinflammatory response of the host to microorganisms present in the dental biofilm which causes tissue destruction. The objective of this study was to evaluate the immunohistochemical expression of cyclophilin A (CYPA), extracellular matrix metalloproteinase inducer (EMMPRIN) and matrix metalloproteinase 7 (MMP-7) in human specimens of clinically healthy gingiva (n=32), biofilm-induced gingivitis (n=28), and chronic periodontitis (n=30). Immunopositivity for CYPA, EMMPRIN and MMP-7 differed significantly between the three groups, with higher percentages of staining in chronic periodontitis specimens, followed by chronic gingivitis and healthy gingiva specimens (p < 0.001). Immunoexpression of CYPA and MMP-7 was higher in the intense inflammatory infiltrate observed mainly in cases of periodontitis. Analysis of possible correlations between the immunoexpression of EMMPRIN, MMP-7 and CYPA and between the expression of these proteins and clinical parameters (probing depth and clinical attachment loss) showed a positive correlation of CYPA expression with MMP-7 (r = 0.831; p < 0.001) and EMMPRIN (r = 0.289; p = 0.006). In addition, there was a significant positive correlation between probing depth and expression of MMP-7 (r = 0.726; p < 0.001), EMMPRIN (r = 0.345; p = 0.001), and CYPA (r = 0.803; p < 0.001). These results suggest that CYPA, EMMPRIN and MMP-7 are associated with the pathogenesis and progression of periodontal disease

Avaliação clínica da resposta do tratamento periodontal não-cirúrgico (Full mouth desinfection) de pacientes diabéticos e não diabéticos

Tem sido demonstrado que o diabetes influencia no desenvolvimento e progressão da doença periodontal. Acredita-se ainda que há uma relação bi-direcional entre o Diabetes Mellito e a Doença Periodontal. Por isso, o tratamento periodontal pode responder de forma diferente em pacientes com e sem o quadro de diabetes. O objetivo desse estudo foi avaliar clinicamente o efeito da terapia periodontal não cirúrgica em pacientes com periodontite e diabetes mellito (grupo teste) e sem o quadro de diabetes (grupo controle). Para isso, realizou-se o tratamento periodontal não-cirúrgico FMSRP (Full mouth Scalling and Root Planing) e verificou-se os parâmetro clínicos periodontais (profundidade de sondagem, nível clínico de inserção, mobilidade, índice gengival e índice de placa) no início do estudo e após 3 meses. Na análise estatística, o paciente foi considerado como unidade de análise (p<0,05). Para as variáveis categóricas utilizou-se o teste de Fisher. Nas comparações inter-grupo, foi utilizado o Mann-Whitney Test e para comparações intra-grupo (baseline e três meses) utilizou-se o Teste de Wilcoxon. Participaram 26 pacientes no grupo controle e 14 no grupo teste. O índice de placa era 71,20% no início do estudo para o grupo teste e ficou 47,12% no fim; já no controle, os valores eram de 48,52% passando para 37,50%. E o índice gengival no grupo teste no baseline era 42,67 diminuindo para 26,81 e no grupo controle diminuiu de 41,36 para 30,62. A profundidade de sondagem foi no grupo teste 2,71mm passando para 2,40mm; já no controle, os valores foram de 2,84mm diminuindo para 2,55mm. O grupo controle ganhou 0,34 mm de inserção e o grupo teste perdeu 0,44mm de inserção. A recessão gengival aumentou 0,33mm no grupo teste e 0,04mm no grupo controle após os 3 meses. Houve diferença significativa inter-grupo para o índice de placa e gengival no baseline, também foi encontrado na recessão após 3 meses; já intra-grupo verificou-se diferença significativa para todas as variáveis, exceto para o nível clinico de inserção e mobilidade. Quando se categorizou a profundidade de sondagem em ≤3mm, >3 e≤6mm e >6mm, não foi encontrado diferença entre os grupos, mas verificou-se diferença significativa entre os períodos tanto para o grupo teste como para o controle. A hemoglobina glicada no grupo controle foi de 5,90% e no teste aumentou de 7,79% para 8,10%. Portanto, verifica-se que há uma melhora dos parâmetros clínicos periodontais em ambos os grupos, contudo não se verificou uma diferença significativa entre eles. Não obstante, o FMSRP promove um efeito benéfico em relação à condição de saúde periodontal, melhora dos parâmetros clínicos periodontais, em curto prazo (3 meses) tanto em indivíduos diabéticos como em não diabéticos, não sendo possível observar um melhor quadro glicêmico nos diabéticos

Influência da laserterapia na proliferação de células-tronco do ligamento periodontal humano

Low level laser irradiation (LLLI) has been used in Dentistry to promote wound healing and tissue regeneration. The literature shows a positive effect of LLLI on cell proliferation, but little is known about their effectiveness in promoting stem cells proliferation. The aim of this study was to evaluate the effect of LLLI on the proliferative rate of human periodontal ligament stem cells. Extracts of periodontal ligament were isolated from two third molars removed by surgical and/or orthodontic indication. After enzymatic digestion, the cells were grown in α-MEM culture medium supplemented with antibiotics and 15% fetal bovine serum. On the third subculture, the cells were irradiated with a InGaAlP-diode laser, using two different energy densities (0,5J/cm 2 - 16 seconds and 1,0J/cm² - 33 seconds), with wavelength of 660nm and output power of 30mW. A new irradiation, using the same parameters, was performed 48h after the first. A control group (non irradiated) was kept under the same experimental culture conditions. The Trypan blue exclusion test and the mitochondrial activity of the cells measured by MTT [3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide] essay were performed to assess the cell proliferation in the intervals of 0, 24, 48 e 72 h after irradiation. The data of cell counts were submitted to nonparametrical statistical tests (Kruskal-Wallis and Mann-Whitney), considering a confidence interval of 95%. DAPI (4 -6-Diamidino-2-phenylindole) staining of the cells was performed at 72h interval to evaluate possible nuclear morphological changes induced by LLLI. The results of this study show that the energy density of 1,0 J/cm² promoted greater cell proliferation compared to the other groups (control and 0,5 J/cm²) at intervals of 48 and 72h. The mitochondrial activity measured by MTT essay showed similar results to the Trypan blue cell counting test. The group irradiated with 1,0J/cm² exhibited a significantly higher MTT activity in the intervals of 48 and 72h, when compared to the group irradiated with 0,5J/cm². No nuclear morphological change was observed in the cells from the three groups studied. It is concluded that LLLI has stimulatory effects on the proliferation of human periodontal ligament stem cells. Therefore, the use of laser irradiation in this cell type may be important to promote future advances in periodontal regeneration

Análise imuno-histoquímica de proteínas relacionadas às respostas Th1, Th2 e Th17 na doença periodontal

Periodontal disease is an inflammatory condition of infectious nature characterized by destruction of protecting and supporting dental tissues. It happens as a response produced by the host when attacked by microorganisms. Several factors are involved in this process. Among them, cytokines are key regulatory molecules in this immune response, playing a role either protective and/or destructive in lesion progression. Thus, this study investigated the immunohistochemical expression of IFN- , GATA-3, IL-17, IL-23, IL-6 and TGF- in gingival tissues of humans, in an attempt to gain a better understanding of the participation of Th1, Th2 and Th17 immune responses in the development of periodontal disease processes. To this end, eighty-two samples of gingival tissues were divided into three groups: Group 1 = 15 (samples of healthy gum tissue as controls), Group 2 = 36 (samples with chronic gingivitis) and Group 3 = 31 (samples with chronic periodontitis). All cases were submitted to morphological analysis from sections stained with hematoxylin and eosin and then subjected to staining by immunohistochemistry using the streptavidin-biotin method. Results showed positive labeling for all proteins. Nonetheless, we observed a greater expression of Th1 cytokines and Th17 cells in group 3. We found statistically significant difference between TGF- expression and the clinical condition of the samples (p=0.02). We conclude that Th1 and Th17 responses may act synergistically in the destructive process of periodontal tissue, overlapping the Th2 response that was also present in these tissues