Marcano Aracelys, Bou Haidar Naila, Marais Stéphane, Valleton Jean-Marc, Duncan Anthony C
Normandie University, UNIROUEN, INSA Rouen, CNRS, PBS, 76000 Rouen, France.
ACS Biomater Sci Eng. 2017 Dec 11;3(12):3654-3661. doi: 10.1021/acsbiomaterials.7b00552. Epub 2017 Nov 28.
One major factor inhibiting natural wound-healing processes is infection through bacterial biofilms, particularly in the case of chronic wounds. In this study, the micro/nanostructure of a wound dressing was optimized in order to obtain a more efficient antibiofilm protein-release profile for biofilm inhibition and/or detachment. A 3D substrate was developed with asymmetric polyhydroxyalkanoate (PHA) membranes to entrap Dispersin B (DB), the antibiofilm protein. The membranes were prepared using wet-induced phase separation (WIPS). By modulating the concentration and the molecular weight of the porogen polymer, polyvinylpyrrolidone (PVP), asymmetric membranes with controlled porosity were obtained. PVP was added at 10, 30, and 50% w/w, relative to the total polymer concentration. The physical and kinetic properties of the quaternary nonsolvent/solvent/PHA/PVP systems were studied and correlated with the membrane structures obtained. The results show that at high molecular weight ( = 360 kDa) and high PVP content (above 30%), pore size decreased and the membrane became extremely brittle with serious loss of physical integrity. This brittle effect was not observed for low molecular weight PVP ( = 40 kDa) at comparable contents. Whatever the molecular weight, porogen content up to 30% increased membrane surface porosity and consequently protein uptake. Above 30% porogen content, the pore size and the physical integrity/mechanical robustness both decreased. The PHA membranes were loaded with DB and their antibiofilm activity was evaluated against biofilms. When the bacterial biofilms were exposed to the DB-loaded PHA membrane, up to 33% of the biofilm formation was inhibited, while 26% of the biofilm already formed was destroyed. These promising results validate our approach based on the development of bioactive-protein-loaded asymmetric membranes for antibiofilm strategies in situations where traditional antibiotic therapies are ineffective.
抑制自然伤口愈合过程的一个主要因素是细菌生物膜感染，在慢性伤口的情况下尤其如此。在本研究中，对伤口敷料的微/纳米结构进行了优化，以获得更有效的抗生物膜蛋白释放曲线，用于抑制生物膜和/或使其脱离。开发了一种带有不对称聚羟基脂肪酸酯（PHA）膜的三维基质，用于包裹抗生物膜蛋白分散素B（DB）。这些膜采用湿诱导相分离（WIPS）制备。通过调节致孔聚合物聚乙烯吡咯烷酮（PVP）的浓度和分子量，获得了具有可控孔隙率的不对称膜。相对于总聚合物浓度，PVP的添加量为10%、30%和50%（w/w）。研究了四元非溶剂/溶剂/PHA/PVP体系的物理和动力学性质，并将其与所得膜结构相关联。结果表明，在高分子量（ = 360 kDa）和高PVP含量（高于30%）时，孔径减小，膜变得极其脆弱，物理完整性严重丧失。在可比含量下，低分子量PVP（ = 40 kDa）未观察到这种脆化效应。无论分子量如何，致孔剂含量高达30%时都会增加膜表面孔隙率，从而提高蛋白质摄取量。致孔剂含量超过30%时，孔径和物理完整性/机械强度都会降低。将DB负载到PHA膜上，并评估其对生物膜的抗生物膜活性。当细菌生物膜暴露于负载DB的PHA膜时，高达33%的生物膜形成受到抑制，而26%已形成的生物膜被破坏。这些有前景的结果验证了我们基于开发用于抗生物膜策略的负载生物活性蛋白的不对称膜的方法，适用于传统抗生素治疗无效的情况。
