Lucy Ono

In vitro and in vivo antiviral properties of sulfated galactomannans against yellow fever virus (BeH111 strain) and dengue 1 virus (Hawaii strain).

Two galactomannans, one extracted from seeds of Mimosa scabrella, having a mannose to galactose ratio of 1.1, and another with a 1.4 ratio from seeds of Leucaena leucocephala, were sulfated. The products from M. scabrella (BRS) and L. leucocephala (LLS) had a degree of sulfation of 0.62 and 0.50, and an average molecular weight of 620x10(3) and 574x10(3) gmol(-1), respectively. Their activities against yellow fever virus (YFV; BeH111 strain) and dengue 1 virus (DEN-1; Hawaii strain) were evaluated. This was carried out in young mice following intraperitoneal infection with YFV. At a dose of 49 mgkg(-1), BRS and LLS gave protection against death in 87.7 and 96.5% of the mice, respectively. When challenged with 37.5 LD50 of YFV, mice previously inoculated with BRS+virus or LLS+virus, showed 93.3 and 100% resistance, respectively, with neutralization titers similar to mice injected with 25 LD50 of formaldehyde-inactivated YFV. In vitro experiments with YFV and DEN-1 in C6/36 cell culture assays in 24-well microplates showed that concentrations that produced a 100-fold decrease in virus titer of YFV were 586 and 385 mgl(-1) for BRS and LLS, respectively. For DEN-1 they were 347 and 37 mgl(-1), respectively. Sulfated galactomannans, thus demonstrate in vitro and in vivo activity against flaviviruses.

Lysozyme-Triggered Epidermal Growth Factor Release from Bacterial Cellulose Membranes Controlled by Smart Nanostructured Films

A novel wound-dressing biodevice, sensitive to lysozyme, an enzyme commonly found at infected skin wounds, was assembled by the layer-by-layer deposition of nanopolymeric chitosan and alginate films onto oxidized bacterial cellulose membranes incorporated with epidermal growth factor (EGF). Distinct EGF release profiles were obtained according to specific stimuli caused by infection. In in vitro conditions simulating noninfected wounds, the EGF rate and burst release effect were reduced by three deposited layers (Mt /M∞ of 0.25 at 3 h) in a process dependent on the porosity of the compact chitosan-alginate complex. The importance of the organized structure was revealed when an infected wound was simulated by adding lysozyme to the release medium, thus inducing the formation of a loosely polyelectrolyte architecture that caused rapid EGF diffusion (Mt /M∞ of 0.75 at 30 min). The results indicate that the nanopolymeric layers were capable of slowly releasing EGF as required for normal wound repair and rapidly undergoing architectural transitions that allow the diffusion of massive amounts of drug to enhance the process of re-epithelialization. In summary, the proposed system comprises the roles of both wound dressing and local delivery mechanism to recognize infections and respond with a burst of EGF release.

Bioactive nanocomposites of bacterial cellulose and natural hydrocolloids

The aim of this work was to develop bioactive films from bacterial cellulose and hydrocolloids (guar gum and hyaluronic acid), coated or not with collagen. After mechanical treatment, a suspension of cellulose nanofibres was obtained which, combined with the dispersions of hydrocolloids, was used to produce bionanocomposite films by wet casting. The materials were stable in physiological solution and presented better swelling capacity than that of the bacterial cellulose. The films were coated with collagen by dipping. Cell adhesion tests and surface analysis by tensiometry, X-ray photoelectron spectroscopy and atomic force microscopy showed that the surface properties of the films can be adjusted by changing the proportions of the components. The collagen coating presented a self-assembling pattern resembling that of living tissues. The materials developed in this work showed potential for applications in the medical field as bioactive wound dressings, scaffolds for cellular growth and sustained drug release systems. The films were obtained by simple production and purification methods, including the use of low toxicity solvents. Thus, in addition to potential cost saving, the development of these bionanocomposites is in accordance with green chemistry principles.

In vitro antiherpetic and antirotaviral activities of a sulfate prepared from Mimosa scabrella galactomannan.

A chemically sulfated galactomannan (BRS) from seeds of Mimosa scabrella had in vitro antiviral activity against Herpes simplex virus 1 (HSV-1), but not against Simian rotavirus A/SA11 (SiRV-A/SA11). It was examined by (13)C NMR spectroscopy, which showed the sulfate groups to be mainly at C-6 of galactose residues. BRS had a selective inhibition against HSV-1 during its attachment step, having an IC(50) lower than 2.5microg/ml, determined by plaque reduction, and a selectivity index of greater than 181, suggesting that the antiviral effect is likely due to interactions between the virus and BRS, being influenced its overall surface charge.

Hydrophilicity improvement of mercerized bacterial cellulose films by polyethylene glycol graft

In this work, polyethylene glycol (PEG), of tree distinct molar masses (200, 300 and 400 g mol(-1)), was grafted onto mercerized bacterial nanocellulose (BNCm) and applied to produce nanofilms (BNCm-PEG). The products BNCm-PEG were characterized by NMR and thermal analysis. Solid-state NMR and X-ray diffraction analyses exhibited no significant differences in index of BNCm-PEG derivatives compared to BNCm, indicating that grafting reaction did not modify the BNCm crystalline structure. The apparent contact angle of the films showed that BNCm-PEG films exhibited a pronounced increase in the polar components (BNCm: 8.1 mN m(-1) vs BNCm-PEG400: 29.4 mN m(-1)), and a decrease in dispersive components (BNCm: 41.7 mN m(-1) vs BNCm-PEG400: 35.2 mN m(-1)) of the surface free energy. The BNCm-PEG films were more hydrophilic than BNCm and retained the biocompatibility with L929 fibroblast cells culture.

Nanometric organisation in blends of gellan/xyloglucan hydrogels

Mixtures of gellan gum (GL) and a xyloglucan (XGJ) extracted from Hymenaea courbaril seeds were prepared in a solution of 0.15 mol L(-1) NaCl. Rheology measurements revealed that 2.4 g L(-1) pure GL formed a brittle hydrogel, and GL-XGJ blends showed improved pseudoplastic character with higher XGJ contents. SAXS analyses showed that the Rg dimensions ranged from 1.3 to 4.9 nm, with larger values occurring as the amount of XGJ increased, and diffusion tests indicated that better diffusion of methylene blue dye was obtained in the network with a higher XGJ content. AFM topographic images of the films deposited onto mica revealed fewer heterogeneous surfaces with increased XGJ contents. The water contact angle revealed more hydrophobic character on all of the films, and the wettability decreased with increasing amounts of XGJ. Therefore, the demonstrated benefit of using XGJ blends is the production of a soft material with improved interface properties.

Tuning Fe3O4 nanoparticle dispersion through pH in PVA/guar gum/electrospun membranes.

Polyvinyl Alcohol (PVA)/guar gum (GG) membranes with different loads of paramagnetic iron oxide Fe3O4 nanoparticles were successfully electrospun using both non-alkaline and alkaline stock solutions. The nanoparticle homogeneity distribution was clearly enhanced in fibers obtained from alkaline stock solutions. This is mainly due to the interaction between GG and the metallic ion, which also leads to further dispersion of remained uncoated nanoparticles in the mixture. It was also noticed that GG favors nanoparticle stability in the mixture and contributes to nanoparticle encapsulation. X-ray results showed that all membranes were semi-crystalline. FTIR-ATR spectra showed that Fe-O absorption band intensity improved with increasing nanoparticle load, reaching saturation at 3.5mg/ml Fe3O4 concentration under alkaline conditions. VSM analyses showed that the nanoparticles are paramagnetic and were successfully incorporated by the fibers. In vitro biocompatibility tests using L929 cells indicates adequate levels of cytotoxicity and cell adhesion/proliferation assays for both membranes obtained from non-alkaline and alkaline stock solutions. Therefore, they have potential for biomedical applications as biodegradable wound dressing.

Physicochemical and in vitro biocompatibility of films combining reconstituted bacterial cellulose with arabinogalactan and xyloglucan.

Reconstituted cellulose films were generated using residual bacterial cellulose membranes mechanically defibrillated (RBC fibrils) recycled following wound dressing production via a dry-cast process. Arabinogalactan (AG) extracted from Pereskia aculeata leaves and/or a xyloglucan (GHXG) from Guibourtia hymenifolia seeds were incorporating into the RBC at various compositions, and new films were created using the same process. Biocomposite properties were evaluated by scanning electron microscopy, contact angle (CA), and X-ray diffraction measurements. The attachment and proliferation of murine L929 fibroblasts on RBC and RBC/Hydrocolloids (HD) were also evaluated. RBC films with 20-30% GHXG replacement improved film stability and the inclusion of HD increased microfiber aggregation and reduced porous regions. Changes in the hydrophilic characteristics were also observed and owing to the adhesion effect the inclusion of HD on RBC led to a statistically significant effect of the mechanical properties of films. The RBC/AG films supported L929 adhesion similar to that observed for commercial bacterial cellulose, indicating their potential use for biomedical applications.