Monti Daria Maria, Guarnieri Daniela, Napolitano Giuliana, Piccoli Renata, Netti Paolo, Fusco Sabato, Arciello Angela
Department of Chemical Sciences, University of Naples Federico II, Naples, Italy; Istituto Nazionale di Biostrutture e Biosistemi (INBB), Rome, Italy.
Center for Advanced Biomaterials for Health Care@CRIB, Istituto Italiano di Tecnologia, and Interdisciplinary Research Centre on Biomaterials, Naples, Italy.
J Biotechnol. 2015 Jan 10;193:3-10. doi: 10.1016/j.jbiotec.2014.11.004. Epub 2014 Nov 18.
Recent years have witnessed an unprecedented growth in the number of applications—such as drug delivery, nutraceuticals and production of improved biocompatible materials—in the areas of nanoscience and nanotechnology. Engineered nanoparticles (NPs) are an important tool for the development of quite a few of these applications. Despite intense research activity, mechanisms regulating the uptake of NPs into cells are not completely defined, being the phenomenon dramatically influenced by physico-chemical properties of NPs and cell-specific differences. Since the cellular uptake of NPs is a prerequisite for their use in nanomedicine, the definition of their internalization pathway is crucial. For this reason, we used 44 nm polystyrene NPs as a model to analyze the uptake and endocytosis pathways in primary human renal cortical epithelial (HRCE) cells, which play a key role in the clearance of drugs. NPs were found not to affect the viability and cell cycle progression of HRCE cells. Distinct internalization pathways were analyzed by the use of drugs known to inhibit specific endocytosis routes. Analyses, performed by confocal microscopy in combination with quantitative spectrofluorimetric assays, indicated that NPs enter HRCE cells through multiple mechanisms, either energy-dependent (endocytosis) or energy-independent.
近年来,纳米科学和纳米技术领域的应用数量呈现出前所未有的增长,例如药物递送、营养保健品以及性能更优的生物相容性材料的生产等。工程化纳米颗粒(NPs)是其中不少应用开发的重要工具。尽管研究活动十分密集,但调节纳米颗粒进入细胞的机制尚未完全明确,这一现象受到纳米颗粒物理化学性质以及细胞特异性差异的显著影响。由于纳米颗粒的细胞摄取是其在纳米医学中应用的前提条件,因此明确其内化途径至关重要。出于这个原因,我们使用44纳米的聚苯乙烯纳米颗粒作为模型,来分析原代人肾皮质上皮(HRCE)细胞的摄取和内吞途径,这些细胞在药物清除过程中起关键作用。研究发现纳米颗粒不会影响HRCE细胞的活力和细胞周期进程。通过使用已知可抑制特定内吞途径的药物,分析了不同的内化途径。利用共聚焦显微镜结合定量荧光光谱分析进行的研究表明,纳米颗粒通过多种机制进入HRCE细胞,这些机制包括能量依赖型(内吞作用)和能量非依赖型。