This work assesses the effect of lyophilization on the cytotoxicity and residual aldehyde concentration of glutaraldehyde-treated and lyophilized bovine pericardium (group A), comparing it to conventional glutaraldehyde-treated bovine pericardium (group B). Cytotoxicity was measured by incubating a pericardium sample from each group in saline and assessing the eluant's influence on cellular growth. Residual aldehydes were measured by HPLC. Although both groups' eluants exhibited some cytotoxicity, the eluant from group A was less cytotoxic, with a cytotoxicity index (IC50(%)) of 41%. Group B eluants all had marked cytotoxic effects; cell growth was 24.15% of the negative control at the most dilute eluant concentration (6.25%). The mean residual glutaraldehyde level was less in group A than in group B (2.36 +/- 0.11 and 9.90 +/- 3.70 g/l, respectively; n=3, P < 0.05), but residual formaldehyde levels did not differ. These results demonstrate that compared with conventional glutaraldehyde-treated bovine pericardium, lyophilized pericardium is less cytotoxic, with fewer glutaraldehyde residues.

No abstract

A polyglycerol with dendritic structure (PGLD) was synthesized by ring-opening polymerization of deprotonated glycidol using a polyglycerol as core functionality in a step-growth process. Then, PGLD reacted with O-carboxymethylated chitosan to obtain PGLD-chitosan dendrimer (PGLD-Ch). After the reaction of PGLD-Ch with boric acid, there was a marked increase in the bulk viscosity evidencing physically that boron can initiate a charge transfer complex formation, (PGLD-Ch)B. Gel permeation chromatography analysis was used to characterize the molecular weight and the polydispersivity of the synthesized PGLD-Ch. A dendritic structure with a molecular mass of 16.7 kDa and a narrow polydispersity (Mw/Mn = 1.05) was obtained. 1H-NMR and 13C-NMR measurements were employed to assess the degree of branching in PGLD. The obtained value of 0.85 indicates the tendency toward a dentritic structure for PGLD. The glass transition temperature values of (PGLD-Ch)B membranes containing 10% and 30% PGLD were -19 degrees C and -26 degrees C, respectively, which favor its potential use as surface coating of several polymers. The in vitro cytotoxicity was evaluated using the minimum essential medium elution test assay. Extracts of boron-complexed PGLD exhibited lower cytotoxicity than the controls, suggesting that the material has an improved biocompatibility. Antibacterial studies of (PGLD-Ch)B against Staphylococcus aureus and Pseudomonas aeruginosa showed a significant activity. Our study confirms and supports the effectiveness of (PGLD-Ch)B as an antimicrobial coating due to its capacity in suppressing the bacterial proliferation. The best in vivo response was found for (PGLD-Ch)B-30 membranes, which exhibited higher synthesis of collagen fibers than PGLD-ChB-10.

Gyroxin is one of main serine proteases of Crotalus durissus terrificus venom, representing about 2% of the protein content in the crude venom. It is a 33 kDa glycoprotein with 3.8% by weight of sugar moiety. This toxin induces hemotoxicity in mice and a neurological condition called barrel rotation syndrome. In the present work, we report the molecular cloning of five new nucleotide sequences from a cDNA library of the venom glands of a single specimen of C. d. terrificus. These sequences have been analyzed in silico with respect to their cDNA organization and similarity with other snake venom serine proteases (SVSPs). We also describe a rapid and efficient method for screening vectors for mammalian cell expression, based on the fact that SVSPs are difficult-to-express toxins due to the presence of several disulfide bonds and glycosylation in their structures. Thus, one of the Gyroxin cDNAs was subcloned into pSectag2 HygroA and pED vectors and used to transfect COS-7 cells. Expression of the functional recombinant Gyroxin isoform was achieved with this cell line with esterase activity in the conditioned culture medium, as revealed by immunoblot of secreted protein and standard anti-crotalic serum from Butantan Institute.

No abstract