Department of Biochemistry and Molecular Biology, Miller School of Medicine, University of Miami, Miami, FL 33136, USA.
Department of Chemistry, University of Georgia, Athens, GA 30602, USA.
Nanoscale. 2016 Dec 1;8(47):19581-19591. doi: 10.1039/c6nr05882e.
The field of intracellular organelle targeting using nanoparticle (NP) is mushrooming rapidly. Thus, the area of nanotechnology-enabled targeting of mitochondrion, the cellular powerhouse, for diseases characterized by mitochondrial dysfunctions such as cancer, diseases of the central nervous system, and cardiovascular diseases is also growing at a rapid pace. Optimization of a NP's ability to target the mitochondria requires quantification of the particles in this subcellular organelle and isolation of mitochondria from the cells. Conventional gradient centrifugation used in currently available methods may not be appropriate for NP containing mitochondria isolation as these particles undergo Brownian motion under centrifugal forces yielding irreproducible results. There is only one method for centrifugation-free mitochondria isolation; however, this method requires immunoprecipitation. Thus, a reliable centrifugation and immunoprecipitation free method is urgently needed to support this growing field of nanotechnology-based mitochondria targeting. Here, we report a mitochondria-targeted magnetic NP, Mito-magneto, to avoid centrifugation and immunoprecipitation methods for isolation of functional, respiration active pure mitochondria, which can be used to analyze and quantify mitochondria targeting properties of various NPs as an important tool for the growing field of "mitochondrial nanomedicine".
利用纳米粒子（NP）进行细胞内细胞器靶向的领域正在迅速发展。因此，利用纳米技术靶向线粒体的领域也在迅速发展，线粒体是细胞的动力源，可用于治疗各种因线粒体功能障碍引起的疾病，如癌症、中枢神经系统疾病和心血管疾病。优化 NP 靶向线粒体的能力需要定量测定亚细胞器中的颗粒，并从细胞中分离出线粒体。目前可用方法中使用的常规梯度离心可能不适用于含有线粒体的 NP 分离，因为这些颗粒在离心力作用下会发生布朗运动，导致结果不可重现。目前只有一种无离心力的线粒体分离方法，但这种方法需要免疫沉淀。因此，迫切需要一种可靠的无需离心和免疫沉淀的方法来支持基于纳米技术的靶向线粒体这一日益发展的领域。在这里，我们报告了一种线粒体靶向磁性 NP，Mito-magneto，用于避免离心和免疫沉淀方法来分离功能、呼吸活跃的纯线粒体，可用于分析和量化各种 NP 的线粒体靶向特性，作为“线粒体纳米医学”这一日益发展领域的重要工具。
