Paracoccidioidomycosis: follow up of acquired immunity parameters and of the activation state of phagocytes in susceptible isogenic mice submitted to the antifungal therapy.

Os efeitos da administração de anfotericina B a camundongos suscetíveis ao P. brasiliensis foram avaliados. A L-AmB reverte o padrão de suscetibilidade para o de resistência de forma mais eficiente do que a c-AmB, como observado na quantificação de UFC, NO e IgG2b. Porém, os níveis de TNF-a, IL-12, IFN-g, GM-CSF, IgG total, IgM, IgG1, IgG2a e IgA não são significativamente alterados. Neutrófilos e macrófagos peritoneais co-cultivados com Pb e L-AmB tendem a apresentar maior capacidade fungicida, mas não maior síntese de mediadores. O melhor desempenho de L-AmB poderia se dever a sua interação com TLR4. Em TLR4-deficientes ou não, a progressão da doença é similar. A eficácia da terapia, porém, é menor nos deficientes, como observado na quantificação de UFC; os perfis leucocitários e as concentrações de NO, TNF-a, IL-12 e GM-CSF não são significativamente alterados. Logo, a droga é capaz de reverter os parâmetros micológicos, mas não os imunológicos. A interação entre TLR4, P. brasiliensis e L-AmB não parece ser importante para o estabelecimento da imunidade.Amphotericin B effects in mice susceptible to P. brasiliensis were evaluated. L-AmB reverts the susceptibility pattern to the resistant one with more efficiency than c-AmB, as confirmed by the CFU, NO e IgG2b quantifications. However, TNF-a, IL-12, IFN-g, GM-CSF, total IgG, IgM, IgG1, IgG2a and IgA levels are not significantly altered. Neutrophils and macrophages cocultivated with Pb e L-AmB tend to present higher fungicidal ability, but not enhanced synthesis of mediators. The better performance of L-AmB could be due to its interaction with TLR4. In TLR4-deficient or sufficient mice, progression of the disease is similar. The efficiency of the therapy, however, is lower in deficient animals, as seen on CFU; leukocyte profiles and NO, TNF-a, IL-12 and GM-CSF levels are not significantly altered by L-AmB-TLR4 interaction. Therefore, the drug administration is capable of reverting mycological parameters, but not the immunological ones. Interaction between TLR4, P. brasiliensis and L-AmB does not seem to play a special role in the establishment of immunity

Photodynamic action of Tri-meso (N-methyl-pyridyl), meso (N-tetradecyl-pyridyl) porphine on Staphylococcus epidermidis biofilms grown on Ti6Al4V alloy

Staphylococcus epidermidis is a leading cause of nosocomial infections, and its virulence is attributable to formation of biofilm, especially on implanted devices. Photodynamic treatment (PDT) has been actively investigated for the eradication of bacterial biofilm growing on dental plaques and oral implants. In this study, we used Tri-meso (N-methyl-pyridyl), meso (N-tetradecyl-pyridyl) porphine (C14) for inactivation of two structurally distinct S. epidermidis biofilms grown on Ti6Al4V alloy and compared its photosensitizing efficiency with that of the parent molecule, tetra-substituted N-methyl-pyridyl-porphine (C1). A more significant reduction in bacterial survival was observed when both bacterial biofilms were exposed to a lower dose of C14, and simultaneously to visible light in comparison with C1. The different responses of both staphylococcal biofilms to C1- or C14-treatment appeared to depend on photosensitizer endocellular concentration. C14 bound to both biofilms to a greater extent than C1. Moreover, C14 penetrates deeper into the bacterial membranes, as determined by fluorescence quenching experiments with methylviologen, allowing for better bacterial killing photoefficiency. Confocal laser scanning microscope (CLSM) analysis indicated damage to bacterial cell membranes in both photodynamically treated biofilms, while disruption of PDT-treated biofilm was confirmed by scanning electron microscopy (SEM). In summary, C14 may be a potential photosensitizer for the inactivation of staphylococcal biofilms for many device-related infections which are accessible to visible light

Antimicrobial Properties and Cytocompatibility of PLGA/Ag Nanocomposites

The purpose of this study was to investigate the antimicrobial properties of multifunctional nanocomposites based on poly(dl-Lactide-co-Glycolide) (PLGA) and increasing concentration of silver (Ag) nanoparticles and their effects on cell viability for biomedical applications. PLGA nanocomposite films, produced by solvent casting with 1 wt%, 3 wt% and 7 wt% of Ag nanoparticles were investigated and surface properties were characterized by atomic force microscopy and contact angle measurements. Antibacterial tests were performed using an Escherichia coli RB and Staphylococcus aureus 8325-4 strains. The cell viability and morphology were performed with a murine fibroblast cell line (L929) and a human osteosarcoma cell line (SAOS-2) by cell viability assay and electron microscopy observations. Matrix protein secretion and deposition were also quantified by enzyme-linked immunosorbent assay (ELISA). The results suggest that the PLGA film morphology can be modified introducing a small percentage of silver nanoparticles, which induce the onset of porous round-like microstructures and also affect the wettability. The PLGA/Ag films having silver nanoparticles of more than 3 wt% showed antibacterial effects against E. coli and S. aureus. Furthermore, silver-containing PLGA films displayed also a good cytocompatibility when assayed with L929 and SAOS-2 cells; indicating the PLGA/3Ag nanocomposite film as a promising candidate for tissue engineering applications

Antimicrobial properties and cytocompatibility of PLGA/Ag nanocomposites

The purpose of this study was to investigate the antimicrobial properties of multifunctional nanocomposites based on poly(dl-Lactide-co-Glycolide) (PLGA) and increasing concentration of silver (Ag) nanoparticles and their effects on cell viability for biomedical applications. PLGA nanocomposite films, produced by solvent casting with 1 wt%, 3 wt% and 7 wt% of Ag nanoparticles were investigated and surface properties were characterized by atomic force microscopy and contact angle measurements. Antibacterial tests were performed using an Escherichia coli RB and Staphylococcus aureus 8325-4 strains. The cell viability and morphology were performed with a murine fibroblast cell line (L929) and a human osteosarcoma cell line (SAOS-2) by cell viability assay and electron microscopy observations. Matrix protein secretion and deposition were also quantified by enzyme-linked immunosorbent assay (ELISA). The results suggest that the PLGA film morphology can be modified introducing a small percentage of silver nanoparticles, which induce the onset of porous round-like microstructures and also affect the wettability. The PLGA/Ag films having silver nanoparticles of more than 3 wt% showed antibacterial effects against E. coli and S. aureus. Furthermore, silver-containing PLGA films displayed also a good cytocompatibility when assayed with L929 and SAOS-2 cells; indicating the PLGA/3Ag nanocomposite film as a promising candidate for tissue engineering applications

Nitric oxide participation in granulomatous response induced by Paracoccidioides brasiliensis infection in mice

The role of nitric oxide (NO) in granulomas of Paracoccidioides brasiliensis-infected inducible NO synthase-deficient C57BL/6 mice (iNOS KO) and their wild-type counterparts and its association with osteopontin (OPN) and matrix metalloproteinases (MMPs) was studied. At 15 days after infection (DAI), iNOS KO mice showed compact and necrotic granulomas with OPN+ macrophages and multinucleated giant cells, whereas wild-type mice developed loose granulomas with many fungi and OPN+ cells distributed throughout the tissue. In addition, high OPN levels and fungal load were observed in iNOS KO mice. Both experimental groups had MMP-9 activity. At 120 DAI, iNOS KO had smaller granulomas with OPN+ cells, lower OPN levels, lower fungal load and decreased MMP-9 activity compared with wild-type mice. These findings suggest that NO has an important role in granuloma modulation, by controlling OPN and MMP production, as well as by inducing loose granulomas formation and fungal dissemination, resulting, at later phases, in progression of paracoccidioidomycosis