Naderi Naghmeh, Griffin Michelle, Malins Edward, Becer Remzi, Mosahebi Afshin, Whitaker Iain S, Seifalian Alexander M
Reconstructive Surgery & Regenerative Medicine Group, Institute of Life Sciences (ILS), Swansea University Medical School, Swansea, UK Welsh Centre for Burns & Plastic Surgery, ABMU Health Board, Swansea, UK UCL Centre for Nanotechnology & Regenerative Medicine, University College London, Royal Free London NHS Foundation Trust, London, UK
UCL Centre for Nanotechnology & Regenerative Medicine, University College London, Royal Free London NHS Foundation Trust, London, UK.
J Biomater Appl. 2016 Feb;30(7):1114-24. doi: 10.1177/0885328215613666. Epub 2015 Nov 3.
Selection of the appropriate sterilisation method for biodegradable materials has been a challenging task. Many conventional sterilisation methods are not suitable for the next generation of biomaterials, mainly due to their complex composition, based on nanomaterials, often incorporating bioactive moieties. In this study, we investigate sterilisation efficacy of slow chlorine releasing compound sodium dichloroisocyanurate dihydrate (SDIC) for polyhedral oligomeric silsesquioxane (POSS)-poly(caprolactone urea-urethane) (PCL) scaffolds in comparison with conventional sterilisation methods.
POSS-PCL scaffolds were subjected to 70% ethanol, UV, and SDIC sterilisation methods. Samples were immersed in tryptone soya broth (TSB) and thioglycollate medium (THY) and after seven days visually inspected for signs of microbial growth. Bulk and surface properties and molecular weight distribution profiles of the scaffolds after sterilization were investigated using FTIR analysis, surface hydrophilicity, scanning electron microscopy analysis, tensile strength testing, and gel-permeation chromatography (GPC). Adipose-derived stem cells (ADSC) were seeded on the scaffolds and AlamarBlue® viability assay was performed to investigate cell metabolic activity. Confocal imaging of rhodamine phalloidin and Dapi stained ADSC on scaffolds was used to demonstrate cell morphology.
GPC results showed that autoclaving led to a significant decrease in the molecular weight of POSS-PCL, whereas ethanol caused visible deformation of the polymer 3D structure and UV radiation did not effectively sterilise the scaffolds. AlamarBlue® analysis showed metabolic activity close to that of tissue culture plastic for ethanol and SDIC.
SDIC sterilisation can be safely applied to biodegradable scaffolds unsuitable for the more common sterilisation methods.
为可生物降解材料选择合适的灭菌方法一直是一项具有挑战性的任务。许多传统灭菌方法不适用于下一代生物材料,主要是因为它们基于纳米材料的复杂成分,通常包含生物活性部分。在本研究中,我们将缓慢释放氯的化合物二水合二氯异氰尿酸钠(SDIC)对多面体低聚倍半硅氧烷(POSS)-聚(己内酯脲-聚氨酯)(PCL)支架的灭菌效果与传统灭菌方法进行了比较。
对POSS-PCL支架采用70%乙醇、紫外线和SDIC灭菌方法。将样品浸入胰蛋白胨大豆肉汤(TSB)和硫乙醇酸盐培养基(THY)中,7天后目视检查微生物生长迹象。使用傅里叶变换红外光谱(FTIR)分析、表面亲水性、扫描电子显微镜分析、拉伸强度测试和凝胶渗透色谱(GPC)研究灭菌后支架的整体和表面性质以及分子量分布情况。将脂肪来源干细胞(ADSC)接种在支架上,并进行AlamarBlue®活力测定以研究细胞代谢活性。使用罗丹明鬼笔环肽和Dapi染色的ADSC在支架上的共聚焦成像来展示细胞形态。
GPC结果表明,高压灭菌导致POSS-PCL的分子量显著降低,而乙醇导致聚合物3D结构出现明显变形,紫外线辐射未有效对支架进行灭菌。AlamarBlue®分析表明,乙醇和SDIC处理后的代谢活性与组织培养塑料相近。
SDIC灭菌可安全应用于不适用于更常见灭菌方法的可生物降解支架。