Dipartimento di Scienze e Tecnologie Chimiche, Università degli Studi di Roma "Tor Vergata", Rome, Italy.
Dipartimento di Fisica, Università degli Studi di Roma "Sapienza", Rome, Italy.
Sci Rep. 2019 Aug 14;9(1):11845. doi: 10.1038/s41598-019-48292-0.
Ultrasound (US) induced transient membrane permeabilisation has emerged as a hugely promising tool for the delivery of exogenous vectors through the cytoplasmic membrane, paving the way to the design of novel anticancer strategies by targeting functional nanomaterials to specific biological sites. An essential step towards this end is the detailed recognition of suitably marked nanoparticles in sonoporated cells and the investigation of the potential related biological effects. By taking advantage of Synchrotron Radiation Fourier Transform Infrared micro-spectroscopy (SR-microFTIR) in providing highly sensitive analysis at the single cell level, we studied the internalisation of a nanoprobe within fibroblasts (NIH-3T3) promoted by low-intensity US. To this aim we employed 20 nm gold nanoparticles conjugated with the IR marker 4-aminothiophenol. The significant Surface Enhanced Infrared Absorption provided by the nanoprobes, with an absorbance increase up to two orders of magnitude, allowed us to efficiently recognise their inclusion within cells. Notably, the selective and stable SR-microFTIR detection from single cells that have internalised the nanoprobe exhibited clear changes in both shape and intensity of the spectral profile, highlighting the occurrence of biological effects. Flow cytometry, immunofluorescence and murine cytokinesis-block micronucleus assays confirmed the presence of slight but significant cytotoxic and genotoxic events associated with the US-nanoprobe combined treatments. Our results can provide novel hints towards US and nanomedicine combined strategies for cell spectral imaging as well as drug delivery-based therapies.
超声（US）诱导的瞬时膜通透性已成为通过细胞质膜传递外源性载体的极具前途的工具，为通过将功能纳米材料靶向特定生物部位来设计新型抗癌策略铺平了道路。为此，至关重要的一步是在超声处理的细胞中详细识别适当标记的纳米颗粒，并研究潜在的相关生物学效应。通过利用同步辐射傅里叶变换红外微光谱（SR-microFTIR）在单细胞水平上提供高灵敏度的分析，我们研究了低强度超声促进的纳米探针在成纤维细胞（NIH-3T3）中的内化。为此，我们使用了与 IR 标记物 4-巯基苯胺偶联的 20nm 金纳米颗粒。纳米探针提供的显著表面增强红外吸收，吸收增加高达两个数量级，使我们能够有效地识别它们在细胞内的包含。值得注意的是，从已内化纳米探针的单个细胞中进行的选择性和稳定的 SR-microFTIR 检测显示出光谱轮廓的形状和强度都发生了明显变化，突出了生物学效应的发生。流式细胞术、免疫荧光和小鼠细胞有丝分裂阻断微核实验证实了与 US-纳米探针联合处理相关的轻微但显著的细胞毒性和遗传毒性事件的存在。我们的结果可以为超声和纳米医学联合策略提供新的线索，用于细胞光谱成像以及基于药物输送的治疗。
