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Incorporation of antimicrobial peptides on electrospun nanofibres for biomedical applications

机译:将抗微生物肽掺入生物医学应用的电纺纳米纤维中

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The aim of this work was to immobilize antimicrobial peptides onto a fibrous scaffold to create functional wound dressings. The scaffold was produced by electrospinning from a mixture of the water soluble polymers poly(acrylic acid) and poly(vinyl alcohol) and subsequently heat cured at 140 °C to produce a stable material with fibre diameter below micron size. The peptides were incorporated into the negatively charged scaffold by electrostatic interaction. The best results were obtained for lysozyme impregnated at pH 7, which rendered a loading of up to 3.0 × 10 ~(?4) mmol mg ~(?1) . The dressings were characterized using SEM, ATR-FTIR, elemental analysis, ζ-potential and confocal microscopy using fluorescamine as an amine-reactive probe. The dressings preserved their fibrous structure after impregnation and peptides were distributed homogeneously throughout the fibrous network. The antibacterial activity was assessed by solid agar diffusion tests and growth inhibition in liquid cultures using Staphylococcus aureus , a pathogenic strain generally found in infected wounds. The antibacterial activity caused clear halo inhibition zones for lysozyme-loaded dressings and a 4-fold decrease in S. aureus viable colonies after two weeks of contact of dressings with bacterial liquid cultures. The release profile in different media showed sustained release in acidic environments, and a rapid discharge at high pH values. The incorporation of lysozyme resulted in dressing surfaces essentially free of microbial growth after 14 days of contact with bacteria at pH 7.4 attributed to the peptide that remained attached to the dressing surface.
机译:这项工作的目的是将抗菌肽固定在纤维支架上,以创建功能性伤口敷料。支架是通过将水溶性聚合物聚(丙烯酸)和聚(乙烯醇)的混合物进行静电纺丝制成的,随后在140°C加热固化,以生产出纤维直径低于微米尺寸的稳定材料。通过静电相互作用将肽掺入带负电荷的支架中。 pH值为7的溶菌酶的浸渍效果最好,其负载量高达3.0×10〜(?4)mmol mg〜(?1)。使用SEM,ATR-FTIR,元素分析,ζ电位和共聚焦显微镜(使用荧光胺作为胺反应探针)对敷料进行表征。敷料在浸渍后保持其纤维结构,并且肽均匀分布在整个纤维网络中。使用金黄色葡萄球菌(一种通常在感染伤口中发现的致病菌株),通过固体琼脂扩散测试和液体培养物中的生长抑制作用来评估抗菌活性。敷料与细菌液体培养物接触两周后,抗菌活性导致溶菌酶载敷料出现明显的光晕抑制区,金黄色葡萄球菌存活菌落减少了4倍。在不同介质中的释放曲线显示出在酸性环境中的持续释放,以及在高pH值下的快速释放。溶菌酶的掺入导致敷料表面与pH 7.4的细菌接触14天后基本没有微生物的生长,这归因于肽仍然附着在敷料表面上。

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