The University of Sydney, Sydney Nano Institute, Faculty of Medicine and Health, Sydney Pharmacy School, Sydney , Australia.
Brigham & Women's Hospital, Harvard Medical School , Boston , MA , USA.
Nanotoxicology. 2019 Nov;13(9):1210-1226. doi: 10.1080/17435390.2019.1650970. Epub 2019 Sep 14.
Current methods for the assessment of nanoparticle safety that are based on 2D cell culture models and fluorescence-based assays show limited sensitivity and they lack biomimicry. Consequently, the health risks associated with the use of many nanoparticles have not yet been established. There is a need to develop models that mimic physiology more accurately and enable high throughput assessment. There is also a need to set up new assays that offer high sensitivity and are label-free. Here we developed 'mini-liver' models using scaffold-free bioprinting and used these models together with label-free nanoscale techniques for the assessment of toxicity of nanodiamond produced by laser-assisted technology. Results showed that NDs induced cytotoxicity in a concentration and exposure-time dependent manner. The loss of cell function was confirmed by increased cell stiffness, decreased cell membrane barrier integrity and reduced cells mobility. We further showed that NDs elevated the production of reactive oxygen species and reduced cell viability. Our approach that combined mini-liver models with label-free high-resolution techniques showed improved sensitivity in toxicity assessment. Notably, this approach allowed for label-free semi-high throughput measurements of nanoparticle-cell interactions, thus could be considered as a complementary approach to currently used methods.
目前基于 2D 细胞培养模型和荧光检测的纳米颗粒安全性评估方法灵敏度有限，且缺乏仿生特性。因此，许多纳米颗粒的使用所带来的健康风险尚未得到明确。我们需要开发更准确模拟生理机能、实现高通量评估的模型，还需要建立具有高灵敏度、无需标记的新检测方法。在这里，我们使用无支架生物打印技术开发了“迷你肝脏”模型，并将这些模型与无标记纳米技术结合，用于评估激光辅助技术生产的纳米金刚石的毒性。结果表明，NDs 以浓度和暴露时间依赖的方式诱导细胞毒性。细胞功能的丧失通过增加细胞硬度、降低细胞膜屏障完整性和降低细胞迁移率得到证实。我们进一步表明，NDs 会增加活性氧的产生并降低细胞活力。我们的方法将迷你肝脏模型与无标记高分辨率技术相结合，在毒性评估中显示出更高的灵敏度。值得注意的是，这种方法允许对纳米颗粒与细胞的相互作用进行无标记的半高通量测量，因此可以被认为是对当前使用方法的补充。
