ARC Centre of Excellence in Convergent Bio-Nano Science and Technology, Monash Institute of Pharmaceutical Sciences, Monash University, 381 Royal Parade, Parkville, VIC 3052, Australia.
ARC Centre of Excellence in Convergent Bio-Nano Science and Technology, Future Industries Institute, University of South Australia, University Boulevard, Mawson Lakes, SA 5095, Australia.
Nanoscale. 2018 Jun 14;10(23):10863-10875. doi: 10.1039/c8nr00835c.
Polyethylene glycol (PEG) is a gold standard against protein fouling. However, recent studies have revealed surprising adverse effects of PEG, namely its immunogenicity and shortened bio-circulation upon repeated dosing. This highlights a crucial need to further examine 'stealth' polymers for controlling the protein 'corona', a new challenge in nanomedicine and bionanotechnology. Poly(2-ethyl-2-oxazoline) (PEtOx) is another primary form of stealth polymer that, despite its excellent hydrophilicity and biocompatibility, has found considerably less applications compared with PEG. Herein, we performed label-free proteomics to compare the associations of linear PEG- and PEtOx-grafted nano-graphene oxide (nGO) sheets with human plasma proteins, complemented by cytotoxicity and haemolysis assays to compare the cellular interactions of these polymers. Our data revealed that nGO-PEG enriched apolipoproteins, while nGO-PEtOx displayed a preferred binding with pro-angiogenic and structural proteins, despite high similarities in their respective top-10 enriched proteins. In addition, nGO-PEG and nGO-PEtOx exhibited similar levels of enrichment of complement proteins. Both PEG and PEtOx markedly reduced nGO toxicity to HEK 293 cells while mitigating nGO haemolysis. This study provides the first detailed profile of the human plasma protein corona associated with PEtOx-grafted nanomaterials and, in light of the distinctions of PEtOx in chemical adaptability, in vivo clearance and immunogenicity, validates the use of PEtOx as a viable stealth alternative to PEG for nanomedicines and bionanotechnologies.
聚乙二醇（PEG）是抗蛋白污染的金标准。然而，最近的研究揭示了 PEG 的惊人的不良反应，即其免疫原性和在重复给药时缩短的生物循环。这凸显了进一步研究“隐形”聚合物以控制蛋白质“冠”的迫切需要，这是纳米医学和生物纳米技术的一个新挑战。聚（2-乙基-2-恶唑啉）（PEtOx）是另一种主要的隐形聚合物，尽管其具有极好的亲水性和生物相容性，但与 PEG 相比，其应用要少得多。在此，我们进行了无标记蛋白质组学研究，以比较线性 PEG 和 PEtOx 接枝纳米氧化石墨烯（nGO）片与人血浆蛋白的结合情况，并通过细胞毒性和溶血试验来比较这些聚合物的细胞相互作用。我们的数据表明，nGO-PEG 富含载脂蛋白，而 nGO-PEtOx 则与促血管生成和结构蛋白优先结合，尽管它们各自的前 10 种富含蛋白非常相似。此外，nGO-PEG 和 nGO-PEtOx 均表现出补体蛋白的相似富集水平。PEG 和 PEtOx 均明显降低了 nGO 对 HEK 293 细胞的毒性，同时减轻了 nGO 的溶血作用。本研究首次详细描述了与 PEtOx 接枝纳米材料相关的人血浆蛋白冠，并鉴于 PEtOx 在化学适应性、体内清除率和免疫原性方面的差异，验证了 PEtOx 作为纳米医学和生物纳米技术中 PEG 的可行隐形替代品的使用。
