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Abstract：
Regenerative medicine has become an extremely valuable tool offering an alternative to conventional therapies for the repair and regeneration of tissues. The re-establishment of tissue and organ functions can be carried out by tissue engineering strategies or by using medical devices such as implants. However, with any material being implanted inside the human body, one of the conundrums that remains is the ease with which these materials can get contaminated by bacteria. Bacterial adhesion leads to the formation of mature, alive and complex three-dimensional biofilm structures, further infection of surrounding tissues and consequent development of complicated chronic infections. Hence, novel tissue engineering strategies delivering biofilm-targeted therapies, while at the same time allowing tissue formation are highly relevant. In this study our aim was to develop surface modified polyhydroxyalkanoate-based fiber meshes with enhanced bacterial anti-adhesive and juvenile biofilm disrupting properties for tissue regeneration purposes. Using reactive and amphiphilic star-shaped macromolecules as an additive to a polyhydroxyalkanoate spinning solution, a synthetic antimicrobial peptide, Amhelin, with strong bactericidal and anti-biofilm properties, and Dispersin B, an enzyme promoting the disruption of exopolysaccharides found in the biofilm matrix, were covalently conjugated to the fibers by addition to the solution before the spinning process. Staphylococcus epidermidis is one of the most problematic pathogens responsible for tissue-related infections. The initial antibacterial screening showed that Amhelin proved to be strongly bactericidal at 12 μg/ml and caused >50% reductions of biofilm formation at 6 μg/ml, while Dispersin B was found to disperse >70% of pre-formed biofilms at 3 μg/ml. Regarding the cytotoxicity of the agents toward L929 murine fibroblasts, a CC50 of 140 and 115 μg/ml was measured for Amhelin and Dispersin B, respectively. Optimization of the electrospinning process resulted in aligned fibers. Surface activated fibers with Amhelin and Dispersin B resulted in 83% reduction of adhered bacteria on the surface of the fibers. Additionally, the materials developed were found to be cytocompatible toward L929 murine fibroblasts. The strategy reported in this preliminary study suggests an alternative approach to prevent bacterial adhesion and, in turn biofilm formation, in materials used in regenerative medicine applications such as tissue engineering.
摘要：
再生医学已成为一种非常有价值的工具，为组织的修复和再生提供了常规疗法的替代方法。组织和器官功能的重建可以通过组织工程策略或通过使用医疗装置如植入物来进行。然而,将任何材料植入人体内，剩下的难题之一是这些材料容易被细菌污染。细菌粘附导致成熟的形成，活的和复杂的三维生物膜结构，周围组织的进一步感染和随之而来的复杂的慢性感染的发展。因此，提供生物膜靶向治疗的新型组织工程策略，同时允许组织形成是高度相关的。在这项研究中，我们的目标是开发表面改性的基于聚羟基链烷酸酯的纤维网，具有增强的细菌抗粘附性和少年生物膜破坏特性，以用于组织再生目的。使用反应性和两亲性星形大分子作为聚羟基链烷酸酯纺丝溶液的添加剂，合成抗菌肽,Amhelin,具有很强的杀菌和抗生物膜特性，和分散素B，一种促进生物膜基质中胞外多糖破坏的酶，通过在纺丝过程之前添加到溶液中，与纤维共价共轭。表皮葡萄球菌是导致组织相关感染的最有问题的病原体之一。最初的抗菌筛选表明，Amhelin在12μg/ml时被证明具有强烈的杀菌性，在6μg/ml时导致生物膜形成减少>50％，而发现分散素B以3μg/ml分散>70%的预先形成的生物膜。关于试剂对L929鼠成纤维细胞的细胞毒性，Amhelin和DispersinB的CC50为140和115μg/ml，分别。静电纺丝工艺的优化导致纤维对齐。具有Amhelin和分散素B的表面活化纤维导致纤维表面上粘附的细菌减少83%。此外,发现开发的材料对L929鼠成纤维细胞具有细胞相容性。这项初步研究报告的策略提出了一种预防细菌粘附的替代方法，进而形成生物膜，用于再生医学应用的材料，如组织工程。
