Thrombosis Research Unit, Sree Chitra Tirunal Institute for Medical Sciences and Technology, Trivandrum, India.
Tissue Eng Part A. 2011 Feb;17(3-4):439-49. doi: 10.1089/ten.TEA.2009.0791. Epub 2010 Nov 9.
Use of silver nanoparticles (SNPs) for control of implant-associated infection is a promising strategy, if optimum antimicrobial yet nontoxic dose to mammalian cells is identified. This study was done to determine essential quantity of SNPs, which stimulate antimicrobial activity without cytotoxicity, when immobilized on poly (ɛ-caprolactone) (PCL) scaffold proposed for vascular tissue engineering. During SNP synthesis and scaffold preparation, nanoparticle aggregation was protected using poly (ethylene glycol). Transmission electron microscopy was used to characterize SNP size and to detect its mobilization from scaffold to culture medium. Antimicrobial property of the SNP and its dose response was tested using both Gram-positive and Gram-negative bacteria by zone of inhibition assay. Endothelial cells (ECs), the main cell type required for vascular tissue engineering, were grown on scaffolds to identify the nontoxic dose. After seeding EC on scaffolds, cell attachment, spreading, and viability/survival were detected using specific markers by flow cytometric/fluorescence microscopic analysis. Real-time polymerase chain reaction detected effect of SNPs on mRNA expression of selected EC-specific functional proteins. Results suggest that even devoid of antibiotics in the medium, 0.1% (w/w) SNP on PCL scaffold is antimicrobial while nontoxic to EC at cellular and molecular level once cultured on the SNP-PCL scaffold.
如果能确定对哺乳动物细胞既具有最佳抗菌效果又无毒的银纳米粒子(SNPs)剂量,那么将其用于控制植入物相关感染是一种很有前途的策略。本研究旨在确定 SNPs 的必需数量,即在固定于拟用于血管组织工程的聚(ε-己内酯)(PCL)支架上时,既能刺激抗菌活性又不会产生细胞毒性。在 SNP 合成和支架制备过程中,使用聚乙二醇(PEG)来保护纳米颗粒聚集。通过透射电子显微镜对 SNP 的大小进行了表征,并检测了其从支架向培养基中的迁移情况。通过抑菌圈试验,分别使用革兰氏阳性菌和革兰氏阴性菌测试了 SNP 的抗菌性能及其剂量反应。将内皮细胞(ECs),即血管组织工程所需的主要细胞类型,种植在支架上,以确定无毒剂量。将 EC 接种到支架上后,通过流式细胞术/荧光显微镜分析使用特定标记物检测细胞的附着、铺展和活力/存活情况。实时聚合酶链反应检测了 SNPs 对选定的 EC 特异性功能蛋白的 mRNA 表达的影响。结果表明,即使培养基中不含抗生素,在 PCL 支架上的 0.1%(w/w) SNP 也具有抗菌作用,而一旦在 SNP-PCL 支架上培养,在细胞和分子水平上对 EC 均无毒。