Halamoda-Kenzaoui Blanka, Ceridono Mara, Urbán Patricia, Bogni Alessia, Ponti Jessica, Gioria Sabrina, Kinsner-Ovaskainen Agnieszka
European Commission Joint Research Centre, Directorate for Health, Consumers and Reference Materials, Via E. Fermi 2749, TP 127, 21027, Ispra, VA, Italy.
J Nanobiotechnology. 2017 Jun 26;15(1):48. doi: 10.1186/s12951-017-0281-6.
Significant progress of nanotechnology, including in particular biomedical and pharmaceutical applications, has resulted in a high number of studies describing the biological effects of nanomaterials. Moreover, a determination of so-called "critical quality attributes", that is specific physicochemical properties of nanomaterials triggering the observed biological response, has been recognised as crucial for the evaluation and design of novel safe and efficacious therapeutics. In the context of in vitro studies, a thorough physicochemical characterisation of nanoparticles (NPs), also in the biological medium, is necessary to allow a correlation with a cellular response. Following this concept, we examined whether the main and frequently reported characteristics of NPs such as size and the agglomeration state can influence the level and the mechanism of NP cellular internalization.
We employed fluorescently-labelled 30 and 80 nm silicon dioxide NPs, both in agglomerated and non-agglomerated form. Using flow cytometry, transmission electron microscopy, the inhibitors of endocytosis and gene silencing we determined the most probable routes of cellular uptake for each form of tested silica NPs. We observed differences in cellular uptake depending on the size and the agglomeration state of NPs. Caveolae-mediated endocytosis was implicated particularly in the internalisation of well dispersed silica NPs but with an increase of the agglomeration state of NPs a combination of endocytic pathways with a predominant role of macropinocytosis was noted.
We demonstrated that the agglomeration state of NPs is an important factor influencing the level of cell uptake and the mechanism of endocytosis of silica NPs.
纳米技术取得了重大进展，特别是在生物医学和制药应用方面，这导致了大量描述纳米材料生物效应的研究。此外，确定所谓的“关键质量属性”，即引发观察到的生物反应的纳米材料的特定物理化学性质，已被认为对于评估和设计新型安全有效的治疗方法至关重要。在体外研究的背景下，对纳米颗粒（NPs）进行全面的物理化学表征，包括在生物介质中的表征，对于将其与细胞反应相关联是必要的。按照这一概念，我们研究了NPs的主要且经常报道的特征，如尺寸和团聚状态，是否会影响NP细胞内化的水平和机制。
我们使用了荧光标记的30纳米和80纳米二氧化硅NPs，它们均有团聚和非团聚形式。通过流式细胞术、透射电子显微镜、内吞作用抑制剂和基因沉默技术，我们确定了每种测试二氧化硅NPs形式最可能的细胞摄取途径。我们观察到细胞摄取存在差异，这取决于NPs的尺寸和团聚状态。小窝介导的内吞作用尤其与分散良好的二氧化硅NPs的内化有关，但随着NPs团聚状态的增加，注意到内吞途径的组合，其中巨胞饮作用起主要作用。
我们证明了NPs的团聚状态是影响二氧化硅NPs细胞摄取水平和内吞作用机制的重要因素。
