Cytotoxicity tests for nanostructured chitosan/PEO membranes using the agar diffusion method

Electrospinning is used to produce fibers in the nanometer range by stretching a polymeric jet using electric fields of high magnitude. Chitosan is an abundant natural polymer that can be used to obtain biocompatible nanostructured membranes. The objectives of this work were to obtain nanostructured membranes based on blends of chitosan and polyoxyethylene (PEO), and evaluate their thermal and morphological properties, as well as their in vitro biocompatibility by agar diffusion cytotoxicity tests for three different cell lines. A nanostructured fibrous membrane with fiber diameters in the order of 200 nm was obtained, which presented a rough surface and thickness ranging from one to two millimeters. The results of the cytotoxicity tests evidenced that the chitosan/PEO membranes are non-toxic to the cells studied in this work. Further, the electrospinning technique was effective in obtaining nanostructured chitosan/PEO membranes, which showed biocompatibility according to in vitro preliminary tests using the cell lines.

Internal fit of two all-ceramic systems and metal-ceramic crowns

OBJECTIVES: The aim of this study was to investigate the internal fit (IF) of glass-infiltrated alumina (ICA - In-Ceram Alumina), yttria-stabilized tetragonal zirconia polycrystals (Y-TZP - IPS e.max ZirCAD), and metal-ceramic (MC - Ni-Cr alloy) crowns. MATERIAL AND METHODS: Sixty standardized resin-tooth replicas of a maxillary first molar were produced for crown placement and divided into 3 groups (n=20 each) according to the core material used (metal, ICA or Y-TZP). The IF of the crowns was measured using the replica technique, which employs a light body polyvinyl siloxane impression material to simulate the cement layer thickness. The data were analyzed according to the surfaces obtained for the occlusal space (OS), axial space (AS) and total mean (TM) using two-way ANOVA with Tukey s multiple comparison test (p<0.05). RESULTS: No differences among the different areas were detected in the MC group. For the Y-TZP and ICA groups, AS was statistically lower than both OS and TM. No differences in AS were observed among the groups. However, OS and TM showed significantly higher values for ICA and Y-TZP groups than MC group. Comparisons of ICA and Y-TZP revealed that OS was significantly lower for Y-TZP group, whereas no differences were observed for TM. CONCLUSIONS: The total mean achieved by all groups was within the range of clinical acceptability. However, the metal-ceramic group demonstrated significantly lower values than the all-ceramic groups, especially in OS.

Statistical models for experimental model membranes.

Biological membranes are constituted from lipid bilayers and proteins. Investigation of protein-membrane interaction, essential for biological function of cells, must rest upon solid knowledge of lipid bilayer behavior. Thus, extensive studies of an experimental model for membranes, lipid bilayers in water solution, have been undertaken in the last decades. These systems present structural, thermal and electrical properties which depend on temperature, ionic strength or concentration. In this talk, we shall discuss statistical models for lipid bilayers, as well as the relation between their properties and results for properties of lipid dispersions investigated by the laboratories supervised by Teresa Lamy (IF-USP) and Amando Ito (FFCL-USP).

Probing the interaction of brain fatty acid binding protein (B-FABP) with model membranes

Brain fatty acid-binding protein (B-FABP) interacts with biological membranes and delivers polyunsaturated fatty acids (FAs) via a collisional mechanism. The binding of FAs in the protein and the interaction with membranes involve a motif called "portal region", formed by two small α-helices, A1 and A2, connected by a loop. We used a combination of site-directed mutagenesis and electron spin resonance to probe the changes in the protein and in the membrane model induced by their interaction. Spin labeled B-FABP mutants and lipidic spin probes incorporated into a membrane model confirmed that BFABP interacts with micelles through the portal region and led to structural changes in the protein as well in the micelles. These changes were greater in the presence of LPG when compared to the LPC models. ESR spectra of B-FABP labeled mutants showed the presence of two groups of residues that responded to the presence of micelles in opposite ways. In the presence of lysophospholipids, group I of residues, whose side chains point outwards from the contact region between the helices, had their mobility decreased in an environment of lower polarity when compared to the same residues in solution. The second group, composed by residues with side chains situated at the interface between the α-helices, experienced an increase in mobility in the presence of the model membranes. These modifications in the ESR spectra of B-FABP mutants are compatible with a less ordered structure of the portal region inner residues (group II) that is likely to facilitate the delivery of FAs to target membranes. On the other hand, residues in group I and micelle components have their mobilities decreased probably as a result of the formation of a collisional complex. Our results bring new insights for the understanding of the gating and delivery mechanisms of FABPs.