Biomedical Research Institute, Hasselt University, Diepenbeek, Belgium.
Biomedical Research Institute, Hasselt University, Diepenbeek, Belgium; Flemish Institute for Technological Research, Environmental Risk and Health Unit, Mol, Belgium.
J Control Release. 2015 Nov 28;218:82-93. doi: 10.1016/j.jconrel.2015.09.064. Epub 2015 Oct 3.
The study of cell-nanoparticle interactions is an important aspect for understanding drug delivery using nanocarriers. In this regard, advances in fluorescence based microscopy are useful for the investigation of temporal and spatial behavior of nanoparticles (NPs) within the intracellular environment. In this work, we focus on the delivery of the naturally-occurring hydrophobic photosensitizer Hypericin in human lung carcinoma A549 cells by using biodegradable poly L-lactic acid NPs. For the first time, Hypericin containing NPs are prepared by combining the miniemulsion technique with the solvent evaporation method. This approach yields an efficient loading of the NPs with Hypericin and allows for additional cargo molecules. To monitor the release of Hypercin from the NPs, an additional fluorescent lipophilic dye Coumarin-6 is incorporated in the NPs. Temporal and spatiotemporal image correlation spectroscopy is used to determine the fate of the NPs carrying the potential cargo. Both directed and non-directed motions are detected. By using image cross-correlation spectroscopy and specific fluorescent labeling of endosomes, lysosomes and mitochondria, the dynamics of the cargo loaded NPs in association with the organelles is studied.
细胞-纳米颗粒相互作用的研究是理解纳米载体药物递送的一个重要方面。在这方面，基于荧光的显微镜技术的进步可用于研究纳米颗粒（NPs）在细胞内环境中的时间和空间行为。在这项工作中，我们专注于通过使用可生物降解的聚 L-乳酸 NPs 来递送人肺癌 A549 细胞中天然存在的疏水性光敏剂 Hypericin。首次通过将 miniemulsion 技术与溶剂蒸发法相结合来制备含有 Hypericin 的 NPs。这种方法可以有效地将 Hypericin 负载到 NPs 中，并允许额外的货物分子。为了监测 Hypercin 从 NPs 中的释放，将另外一种荧光亲脂性染料 Coumarin-6 掺入到 NPs 中。时间和时空图像相关光谱用于确定携带潜在货物的 NPs 的命运。检测到定向和非定向运动。通过使用图像互相关光谱和内体、溶酶体和线粒体的特异性荧光标记，研究了与细胞器相关的负载货物的 NPs 的动力学。
