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.

Regeneration of skin tissue promoted by mesenchymal stem cells seeded in nanostructured membrane.

BACKGROUND: The mesenchymal stem cell therapy has proven to be an effective option in the treatment of skin injuries. The combination of these cells with nanostructured membranes seems to be the future for tissues recovery. The aim of this project was to use biomolecules of polysaccharides to be incorporated on regenerated cellulose membranes and to prospect the improvement as bioactive wound dressings with mesenchymal stem cells. METHODS: The biocomposites were obtained after defibrillation with the use of never-dried bacterial cellulose to form a pulp, and, after the films were regenerated, in the presence of gellan gum with or without fluconazole. Membrane atomic force microscopy was performed for comparison of their structures. RESULTS: Adipose-derived mesenchymal stem cells were obtained from human adipose tissue liposuction in accordance with Zuk et al. The flow cytometric analysis and induction tests for adipocytes and osteocytes were performed. In vitro assays were performed on different membranes to evaluate the ability of these cells to adhere at 2 hours and proliferate at 7 days; the results were obtained by use of the MTT cell counting technique. In vivo testing allowed us to observe cell migration and participation in wound-healing by fluorescence labeling of the cells with BrdU. The bioactive curative, seeded with cells, was tested in skin burned in a murine model. CONCLUSIONS: The bacterial cellulose with gelan gum membrane incorporated with fluconazole presented the best performance in adhesion and proliferation tests. The cells can be identified in burned host tissue after occurrence of the wound.

Chemically sulfated galactomannan from Dimorphandra gardneriana seed: characterization and toxicity evaluation.

Dimorphandra gardneriana galactomannan (DG) was sulfated in pyridine:formamide using chlorosulfonic acid as the sulfation agent. The degree of substitution was 0.32, determined from the sulfur percentage. Confirmation of sulfation was obtained by FTIR spectroscopy through the presence of an asymmetrical SO stretching vibration at 1,259 cm(-1). NMR data showed that the sulfation occurred on primary hydroxyl groups. NMR and GPC data indicate degradation during reaction with elimination of galactose. At the maximum tested concentration of 1,000 µg/mL, unmodified DG polysaccharide did not show a statistically significant cytotoxicity in Vero cells by the MTT method. Therefore, the CC50>1,000 µg/mL obtained for the sulfated polysaccharides from D. gardneriana in Vero cells point to its lower cytotoxicity than the sulfated galactomannan from Mimosa scabrella.

Atividade anti-HSV-1 in vitro de galactomanana quimicamente sulfatada de sementes de Leucaena leucocephala / In vitro anti-HSV-1 activity of a chemically sulfated galactomannan from Leucaena leucocephala seeds

Obteve-se uma galactomanana quimicamente sulfatada (LLS-2) a partir de polissacarídeo extraído de sementes de Leucaena leucocephala, a qual apresentou 15.2% de sulfato e grau de derivatização de 0,60, e, seu efeito antiviral sobre a replicação do vírus Herpes simplex tipo 1 (HSV-1) em células Vero foi avaliado pela metodologia de redução do número de unidades formadoras de placas. LLS-2 apresentou 93.7% de inibição da replicação viral à concentração de 2,5 ?g/ml, quando adicionado durante as etapas iniciais de replicação, com um índice de seletividade maior que 1.000, sugerindo que LLS-2 inibe a ligação de HSV-1 às células hospedeiras.

A galactomannan extracted from the seeds of Leucaena leucocephala was sulfated chemically, yielding a polymer (LLS-2) with 15.2% sulfate by weight (degree of sulfation 0.60), and its effect on the replication of Herpes simplex virus 1 (HSV-1) in Vero cells was investigated. When added during infection and early replication, LLS-2 showed 93.7% inhibition of HSV-1 replication at a concentration of 2.5 ?g/mL, according to the reduction in the number of viral plaques, and a selectivity index higher than 1,000, suggesting that it inhibits HSV-1 binding to the host cell.

Nanocomposites coated with xyloglucan for drug delivery: In vitro studies.

Enalaprilate (Enal), an active pharmaceutical component, was intercalated into a layered double hydroxide (Mg/Al-LDH) by an ion exchange reaction. The use of a layered double hydroxide (LDH) to release active drugs is limited by the low pH of the stomach (pH approximately 1.2), in whose condition it is readily dissolved. To overcome this limitation, xyloglucan (XG) extracted from Hymenaea courbaril (jatobá) seeds, Brazilian species, was used to protect the LDH and allow the drug to pass through the gastrointestinal tract. All the materials were characterized by X-ray diffraction, Fourier transform infrared spectroscopy, elemental analyses, transmission electronic microscopy, thermal analyses, and a kinetic study of the in vitro release was monitored by ultraviolet spectroscopy. The resulting hybrid system containing HDL-Enal-XG(3) slowly released the Enal. In an 8-h of test, the system protected 40% (w/v) of the drug. The kinetic profile showed that the drug release was a co-effect behavior, involving dissolution of inorganic material and ion exchange between the intercalated anions in the lamella and those of phosphate in the buffer solution. The nanocomposite coated protection with XG was therefore efficient in obtaining a slow release of Enal.

A xyloglucan from seeds of the native Brazilian species Hymenaea courbaril for micropropagation of Marubakaido and Jonagored apples.

Xyloglucan was extracted from seeds of Hymenaea courbaril and mixed with agar to prepare a solid culture medium used for micropropagation of the Marubakaido apple rootstock (Malus prunifolia Borkh) and cv. Jonagored (Malus domestica). The performance on gels created from a blend of 0.4%agar and 0.2% xyloglucan (w/v) was compared with that on media gelled with a standard concentration 0.6% (w/v) of agar. The growth of shoots and the multiplication rate were higher on the modified culture medium than on the agar-gelled medium. The occurrence of hyperhydric shoots was lower on the modified medium. In the absence of auxin, shoot rooting reached 70% (Marubakaido) and 66% (Jonagored) on the agar-xyloglucan medium and 6.7% and 10.4%, respectively, on the agar medium. When 0.25 microM indole-3-butyric acid (IBA) was added to both media, the modified medium gave better results in terms of rooting percentage and quality of roots than the agar-gelled medium.

Complexes of arabinogalactan of Pereskia aculeata and Co2+, Cu2+, Mn2+, and Ni2+.

The main interest in the biopolymer arabinogalactan is that it is edible. Complementing its high protein percentage, when complexed to essential metal ions, widens the use in food and pharmacology industries and technologies. The binding constants of Co2+, Cu2+, Mn2+ and Ni2+ with arabinogalactan, extracted from the leaves of Pereskia aculeata from Brazil were determined by potentiometric titrations and also the speciation according to pH values. The complexed species proposed by potentiometric titrations and the unique complexing ability of galacturonic acid groups towards Cu2+ and Ni2+ in the tridimensional web structure of arabinogalactan were confirmed by IR and EPR spectroscopies. The thermal stability of the complexed species also varied with the metal ion employed in the complexation when compared to the biopolymer alone. These complexes are new sources of additives for the food and pharmacology industries and carriers of essential metal ions to animal and vegetal biochemistry.

Polysaccharides from the seeds of Senna multijuga.

The seeds of Senna multijuga were extracted with water or 1% acetic acid and treated with ethanol, resulting in two insoluble fractions. After purification, the major one (FIA, 23%) was shown to be a galactomannan (Man:Gal 2.3:1; [alpha] = +54.6; [eta] = 1340 ml g-1). It consists of a main chain of (1-->4)-linked beta-D-mannopyranosyl residues substituted at O6 by single-unit alpha-D-galactopyranosyl side chains. The second fraction (FIB, 2.5%) was an O-acetyl-glucuronoarabinoxylan from the seed coats (O-acetyl 8.3 mol%; glucuronic acid 11.7%, Xyl:Ara ratio 20:1), which showed a predominance of 4-O-substituted Xylp units (84.4%), branched at O3 with non-reducing end units of Xylp, Araf and glucuronic acid. The O-acetyl positions in D-xylosyl units are at O2 (4.8%), O3 (4.4%) and O2,3 (0.9%). The ratio between O3 and O2 determined by 13C-nuclear magnetic resonance spectroscopy is 1.5:1.

Oligosaccharides derived from the xyloglucan isolated from the seeds of Hymenaea courbaril var. stilbocarpa.

On aqueous extraction, Hymenaea courbaril var. stilbocarpa, known in Brazil as jatobá, furnishes a high yield of viscous xyloglucan (45%) from its seeds. The crude polysaccharide (B1) was hydrolysed and the products, analysed as alditol acetates, were glucose, xylose, galactose and arabinose in the ratio 50:35:13:2. After further fractionation on DEAE-cellulose column (chloride form), the main fraction (70% yield, B2) was obtained. The basic structure of the xyloglucan was determined as a cellulose-type (1-->4)-linked beta-D-glucan backbone partially substituted with side chains at O6 of alpha-D-xylopyranose, some of which were themselves substituted at O2 by the units of beta-D-galactopyranose. Treatment of the xyloglucan (B2) with commercial cellulase from Trichoderma sp. yielded six oligosaccharides. These oligosaccharides were isolated by preparative paper chromatography, and their structures were determined by gas-liquid chromatography-mass spectroscopy of the derived partially O-methylated alditol acetates. These results confirm the structure proposed for jatobá seed xyloglucan.