RESUMO OBJETIVO: Descrever a imunofenotipagem de granulomas TB pulmonar de pacientes autopsiados com tuberculose (grupo TB) e de pacientes infectados pelo HIV com tuberculose (grupo TB/HIV) e identificar as espécies de Mycobacterium em cadeia da polimerase utilizando reação PCR. MÉTODOS: Foram selecionadas amostras pulmonares de 15 pacientes do grupo TB e 23 pacientes do grupo TB/HIV. Realizamos Histopatológia e imuno-histoquímica para descrever os granulomas e para detectar o agente infeccioso (anti-BCG). Avaliamos CD4, CD8, CD20 e CD68 para caracterizar a imunofenotipagem dos granulomas e realizamos a reação PCR para identificar as espécies de micobactéria. RESULTADOS: linfócitos T CD4 + foram as células com densidade mais elevada no grupo de TB, ao passo que as células CD68 exibiram maior densidade no grupo TB/HIV. A comparação entre os grupos mostrou que a densidade de linfócitos T CD4 + foi significativamente maior nos pacientes com TB, ao passo que a densidade CD68 foi significativamente maior nos pacientes com TB/HIV. M. tuberculosis foi identificado em 8 casos de cada grupo; M. avium só foi encontrado em um caso do grupo TB/HIV. CONCLUSÃO: Com o advento da AIDS, o perfil imunológico da tuberculose se alterou. Isto pode estar associado com a depleção de linfócitos T CD4 + em granulomas pulmonares. M. tuberculosis foi o principal agente etiológico da tuberculose em ambos os grupos.

OBJECTIVE: To describe the immunophenotype of pulmonary TB granulomas from autopsied patients with tuberculosis (TB group) and from HIV patients with tuberculosis (TB/HIV group), and to identify the Mycobacterium species using polymerase chain reaction (PCR) technique. METHODS: Lung samples of 15 TB group patients and 23 TB/HIV group patients were selected. Histopathologic analyses and immunohistochemistry tests were performed to describe the granulomas and to detect the infectious agent (anti-BCG). CD4, CD8, CD20 and CD68 were evaluated to characterize the immnophenotype of the granulomas. Polymerase Chain Reaction was performed to identify the mycobacterium species. RESULTS: CD4 + T lymphocytes were the cells with highest density in the TB group, whereas CD68 cells exhibited the highest density in the TB/HIV group. Comparison between groups showed that the CD4 + T density was significantly higher in the TB patients; whereas, CD68 density was significantly higher in the TB/HIV patients. M. tuberculosis was identified in 8 cases of each group; M. avium was only found in one case of the TB/HIV group. CONCLUSION: With the advent of AIDS, the immunological profile of TB has changed. This may be associated with the depletion of CD4 + T lymphocytes in lung granulomas. M. tuberculosis was the major etiological agent of TB in both groups.

OBJECTIVES: Goldenhar syndrome (GS) is a relatively common developmental disorder characterized by craniofacial anomalies in association with vertebral, cardiac, renal, and central nervous system defects. This paper describes GS features with special emphasis on oral characteristics. MATERIAL AND METHODS: The clinical features of 6 patients with GS aged 3 months to 12 years are described, and a brief review of the literature about this genetic disorder is presented. RESULTS: All patients demonstrated the classical triad of GS, including mandibular hypoplasia resulting in facial asymmetry, ear and/or eye malformation, and vertebral anomalies. In addition, renal and gastrointestinal abnormalities were observed in 2 patients. Regarding the oral involvement, 2 patients presented cleft lip and palate, and 1 patient had temporomandibular joint malformation. Malocclusion was found in all patients. CONCLUSION: Our orofacial findings correlate with the reported cases in the literature, and point out that after diagnosis GS patients need to be examined for systemic abnormalities.

Cyclosporin A (CyA) induces gingival overgrowth via its stimulatory effects on expression of transforming growth factor-beta1 (TGF-β1) and collagen. It is not known whether CyA has a direct effect on gingival fibroblasts or induces its effect indirectly via stimulation of myofibroblast transdifferentiation. The present study was undertaken to examine the in vivo and in vitro effect of CyA on myofibroblast transdifferentiation. Rats were treated for 60 days with a daily subcutaneous injection of CyA, and the gingival overgrowth tissue was analyzed by immunohistochemistry. In vitro, fibroblasts from normal gingiva (NG) were cultured in the presence of different concentrations of CyA, and subjected to semi-quantitative reverse transcriptase-polymerase chain reaction and western blot. Although CyA treatment stimulated TGF-β1 expression by NG fibroblasts, it lacked to induce expression and production of isoform α of smooth muscle actin (α-SMA), the specific myofibroblast marker. The expression levels of connective tissue growth factor (CTGF), which has been considered a key molecule to promote the transdifferentiation of myofibroblasts via TGF-β1 activation, were unaffected by CyA. Our results demonstrate that CyA-induced gingival overgrowth is not associated with activation of myofibroblast transdifferentiation, since CyA is not capable to increase CTGF expression.

O objetivo deste estudo foi comparar as características morfológicas, o potencial proliferativo e a produção protéica de fibroblastos do ligamento periodontal (FLP) e de fibroblastos gengivais (FG). Os fibroblastos foram cultivados pela técnica do explante a partir de fragmentos da gengiva e do ligamento periodontal de um mesmo indivíduo. As células foram isoladas e plaqueadas para análise por microscopia de contraste de fase e microscopia óptica. O índice de proliferação celular foi determinado por contagem automática de células e pelo ensaio de incorporação de bromodioxiuridina (BrdU). A produção de proteína total foi verificada por eletroforese em gel de poliacrilamida e o perfil enzimático por análise zimográfica. Os FLP são maiores e mais alongados que os FG em condições de subconfluência e confluência celular. Os FLP demonstraram um potencial proliferativo significantemente maior que os FG. Os perfis protéico e enzimático foram similares entre os FLP e FG. Os resultados demonstram que os FLP e FG são diferentes na morfologia e na capacidade proliferativa, porém são semelhantes na produção protéica.

The objective of this study was to compare fibroblasts from the periodontal ligament (PLF) and gingival fibroblasts (GF) as to morphology, proliferation rate and protein synthesis. PLF and GF were explanted from tissues of the same patient. To characterize and compare the morphology of cells, PLF and GF were plated and analyzed under phase-contrast and optical microscopies. Proliferation rates were determined by means of automated counts carried out in days 1, 4, 7, 15 and 21, and also by means of the bromodeoxyuridine labelling index (BrdU). Total protein content was analyzed by means of electrophoresis in 10% polyacrylamide gel and zimography containing gelatin as substrate. PLF were bigger and more elongated than GF in subconfluence and confluence conditions. The proliferative rate of PLF was higher than that of GF at 1, 4, and 7 days (p < 0.05). At 15 and 21 days, there was no statistically significant difference as to the number of cells. PLF presented a significantly greater proliferative potential, in relation to GF (p < 0.05). The synthesis of protein in a period of 24 hours was similar for both PLF and GF. Our results demonstrated that PLF and GF are different as to morphology and proliferative capacity, however, they do not differ as to protein synthesis.

Fibromatose gengival hereditária (FGH) é uma condição bucal rara clinicamente manifestada por um aumento gengival generalizado e fibrótico, podendo apresentar-se de forma isolada ou associada a outras alterações, como parte de síndromes. Os mecanismos biológicos envolvidos na FGH são desconhecidos, e os resultados de estudos de cultura celulares são controversos. Para elucidar as características fenotípicas dos fibroblastos de FGH, nós isolamos quatro linhagens celulares de fibroblastos de FGH de indivíduos de uma mesma família e comparamos as características morfológicas e proliferativas com fibroblastos provenientes de pacientes com gengiva clinicamente normal (GN). Fibroblastos de GN e FGH em condições de subconfluência celular apresentaram típicas características morfológicas, como formato fusiforme, núcleo central e longos prolongamentos citoplasmáticos, mas em condições de saturação da densidade celular, os fibroblastos de FGH apresentaram dimensões menores que as células controle. A relação núcleo/citoplasma foi sempre menor para todas as linhagens celulares de fibroblastos de FGH, sugerindo que a redução celular, é proveniente de uma redução ou compactação citoplasmática e não nuclear. A capacidade proliferativa de fibroblastos de FGH foi maior que a de fibroblastos de GN. Estes resultados sugerem que diferenças morfológicas e proliferativas dos fibroblastos de FGH podem estar associadas aos eventos biológicos envolvidos na etiopatogenia do aumento gengival observado em pacientes com FGH.

Hereditary gingival fibromatosis (HGF) is a rare oral condition, clinically manifested through a generalized and fibrotic enlargement of the gingiva, which may present as an isolated clinical finding or in association with other features, as part of a syndrome. The biological mechanisms involved in HGF are unknown, and the results of cell-culture studies are controversial. To elucidate the phenotypic and proliferative characteristics of HGF fibroblasts, we isolated 4 cell lines of gingival fibroblasts from members of the same family with HGF, and compared with gingival fibroblasts from 4 healthy patients (NG). HGF and NG fibroblasts, in subconfluent culture densities, showed typical morphological characteristics, such as spindle form with a central spherical nucleus and long cytoplasmatic prolongations, but in saturation density, HGF cells were shorter than control cells. The nucleus/cytoplasm relation was always smaller in all HGF cell lines, suggesting that the cellular reduction is derived from reduction or compaction of the cytoplasm and not of the nucleus. The proliferation rate was higher in fibroblasts from HGF than in the ones from NG. These results suggest that differences in the morphology and proliferation of HGF fibroblasts may be associated with the biological events involved in the pathogenesis of gingival overgrowth in HGF patients.