Submicron bioactive glass fibers 70S30C (70 mol% SiO(2), 30 mol% CaO) acting as bone tissue scaffolds were fabricated by electrospinning method. The scaffold is a hierarchical pore network that consists of interconnected fibers with macropores and mesopores. The structure, morphological characterization and mechanical properties of the submicron bioactive glass fibers were studied by XRD, EDS, FIIR, SEM, N(2) gas absorption analyses and nanoindentation. The effect of the voltage on the morphology of electrospun bioactive glass fibers was investigated. It was found that decreasing the applied voltage from 19 to 7 kV can facilitate the formation of finer fibers with fewer bead defects. The hardness and Young's modulus of submicron bioactive glass fibers were measured as 0.21 and 5.5 GPa, respectively. Comparing with other bone tissue scaffolds measured by nanoindentation, the elastic modulus of the present scaffold was relatively high and close to the bone.
Disulfide-crosslinked hollow polyelectrolyte microcapsules composed of thiolated chitosan (CS-SH) and hyaluronic acid (HA-SH) were prepared by combining the layer-by-layer (LBL) technique and horseradish peroxidase (HRP)-mediated oxidative cross-linking reaction in mild conditions. FITC-dextran-doped CaCO microspheres were used as template core and removed after LBL depositing CS-SH and HA-SH on the surface. The disulfide-crosslinked (CS/HA) microcapsules were readily fabricated by HRP-mediated oxidative coupling of the thiol groups in CS/HA shell layer in the presence of HRP (10 units/mL) and Tyramine hydrochloride (Tyr, 35 mmol/L). The kinetics of enzymatic disulfide-crosslinking reaction was investigated through the real-time monitoring of the consumption of thiol groups by UV absorption spectra. It found that the formation of disulfide linkages by the enzymatic thiol oxidation reaction showed a gradual acceleration. The disulfide-crosslinked CS/HA hydrogel were rapidly formed in gelation time between approximately 17 and 30 min, which were dependent on the concentrations of HRP and Tyr. The disulfide linkages endowed the microcapsule-enhanced physical stability and low permeability under physiological conditions and redox-responsive degradability in reducing environments. The structural stability of disulfide-crosslinked (CS/HA) microcapsules was visualized by confocal laser scanning microscopy in phosphate-buffered saline containing 5.0 mmol/L dithiothreitol (DTT) to evaluate the redox-responsive disassembly process. Redox-responsive controlled release of encapsulated FITC-dextran from the disulfide-crosslinked (CS/HA) microcapsules were obtained. The release profiles of FITC-dextran could be manipulated by controlling the shell thickness and the concentration of DTT. The conformational stability analyses and more than 94% esterase activity of released bovine serum albumin (BSA) from (CS/HA) microcapsules conformed that the structural integrity and bioactivity were well preserved during the encapsulation and release process. The microcapsules exhibited excellent cytocompatibility for HEK 293 cells up to a concentration of 1.0 mg/mL. The microcapsules efficiently delivered loaded FITC-BSA into HeLa cells and released the protein in the reducing cytosol. This study proposed a novel approach for producing disulfide-crosslinked microcarriers for intracellular delivery and redox-responsive controlled release of protein.
Some lactobacilli strains had beneficial effects on human beings due to their antioxidant activities. In this study lactobacilli strains stored in our laboratory were screened for potential antioxidant activities by investigating their 1,1-diphenyl-2-picrylhydrazyl free radical scavenging activity, oxygen radical absorbance capacity, resistance to H 2 O 2 , and hydroxyl free radical scavenging activity; then the antioxidant activities of the screened strains were evaluated by cellular antioxidant assay and protection for HT-29 cells against H 2 O 2 injury assay. The results showed that Lactobacillus plantarum Y44 could scavenge oxygen free radicals, inhibit the production of intracellular reactive oxygen species without creating obvious cytotoxic effects, and protect HT-29 cells against H 2 O 2 injury evidenced by the significant decrease of the Bcl-2-associated X protein (Bax)/B-cell lymphoma 2 (Bcl-2) ratio and heat shock protein 70 expression, increase of superoxide dismutase and glutathione peroxidase activities, and decrease of malondialdehyde level of HT-29 cells damaged by H 2 O 2 . It was speculated that L. plantarum Y44 protect HT-29 cells against oxygen radical injury through scavenging reactive oxygen species and activating intracellular antioxidant enzymes. A significant correlation was observed among the results of the hydroxyl radical scavenging assay, protection assay for HT-29 cells against H 2 O 2 injury, and the cellular antioxidant assay. The findings indicated that L. plantarum Y44 could be a probiotic candidate with antioxidant properties and combining several chemical antioxidant methods and antioxidant cellular models could be an effective procedure to screen lactobacilli strains with antioxidant activity.
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