Amariei Georgiana, Kokol Vanja, Boltes Karina, Letón Pedro, Rosal Roberto
Department of Chemical Engineering, University of Alcalá E-28871 Alcalá de Henares Madrid Spain
Institute of Engineering Materials and Design, University of Maribor SI-2000 Maribor Slovenia.
RSC Adv. 2018 Aug 6;8(49):28013-28023. doi: 10.1039/c8ra03861a. eCollection 2018 Aug 2.
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 mmol mg. 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 , 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 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.
这项工作的目的是将抗菌肽固定在纤维支架上,以制造功能性伤口敷料。该支架是通过静电纺丝由水溶性聚合物聚丙烯酸和聚乙烯醇的混合物制成的,随后在140℃下热固化,以生产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。在pH值为7时浸渍溶菌酶获得了最佳结果,其负载量高达3.0×10 mmol/mg。使用扫描电子显微镜(SEM)、衰减全反射傅里叶变换红外光谱(ATR-FTIR)、元素分析、ζ电位以及以荧光胺作为胺反应性探针的共聚焦显微镜对敷料进行了表征。浸渍后敷料保留了其纤维结构,并且肽均匀地分布在整个纤维网络中。通过固体琼脂扩散试验和使用感染伤口中常见的病原菌在液体培养物中的生长抑制来评估抗菌活性。对于负载溶菌酶的敷料,抗菌活性产生了明显的晕圈抑制区,并且在敷料与细菌液体培养物接触两周后,活菌菌落减少了4倍。在不同介质中的释放曲线表明,在酸性环境中为持续释放,在高pH值下为快速释放。在pH 7.4下与细菌接触14天后,溶菌酶的掺入导致敷料表面基本没有微生物生长,这归因于仍附着在敷料表面的肽。