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Engineered iron oxide magnetic nanoparticles (MNPs) are one of the most promising tools in nanomedicine-based diagnostics and therapy. However, increasing evidence suggests that their specific delivery efficiency and potential long-term cytotoxicity remain a great concern. In this study, using 12 nm γ-Fe2O3 MNPs, we investigated three types of uptake pathways for MNPs into HepG2 cells: (1) a conventional incubation endocytic pathway; (2) MNPs co-administrated with microbubbles under ultrasound exposure; and (3) ultrasound delivery of MNPs covalently coated on the surface of microbubbles. The delivery efficiency and intracellular distribution of MNPs were evaluated, and the cytotoxicity induced by reactive oxygen species (ROS) was studied in detail. The results show that MNPs can be delivered into the lysosomes via classical incubation endocytic internalization; however, microbubbles and ultrasound allow the MNPs to pass through the cell membrane and enter the cytosol via a non-internalizing uptake route much more evenly and efficiently. Further, these different delivery routes result in different ROS levels and antioxidant capacities, as well as intracellular glutathione peroxidase activity for HepG2 cells. Our data indicate that the microbubble–ultrasound treatment method can serve as an efficient cytosolic delivery strategy to minimize long-term cytotoxicity of MNPs.中文摘要磁性纳米颗粒在纳米生物医学诊断和治疗研究领域是极具潜力的一种纳米材料. 如何实现纳米颗粒在特定细胞或靶器官的高效率传输以及如何降低细胞毒性是目前纳米材料研究的重点内容. 本文首先研究了12 nm的γ-Fe2O3磁性纳米颗粒进入细胞的三种不同途径: (1) 纳米颗粒与肿瘤细胞共孵育后的内吞途径; (2) 纳米颗粒与微气泡共混合后超声辐照传输途径; (3) 纳米颗粒化学偶联到微气泡膜壳表面后超声辐照传输途径. 其次, 基于上述三种不同的纳米颗粒传输途径, 对纳米颗粒引起的细胞氧化应激毒性进行了深入研究. 结果表明, 纳米颗粒与肿瘤细胞共孵育后的内吞途径使纳米颗粒通过溶酶体包裹进入细胞; 通过超声微气泡辐照, 纳米颗粒能够以更高效率通过非内吞途径直接传输进入细胞质而不被溶酶体包裹. 不同传输途径导致纳米颗粒分别进入溶酶体和细胞质, 造成对细胞内氧化应激水平、总抗氧化能力以及谷胱甘肽过氧化物酶活性的响应不同. 综上研究表明, 超声微气泡介导的磁性纳米颗粒传输能够成为一种高效无损的细胞纳米颗粒输运新方法, 同时通过控制纳米颗粒进入细胞质降低了纳米颗粒的毒性, 从而能够更广泛应用于纳米生物医学的应用研究.

Altering the response of intracellular reactive oxygen to magnetic nanoparticles using ultrasound and microbubbles