Malatesta M, Grecchi S, Chiesa E, Cisterna B, Costanzo M, Zancanaro C
University of Verona.
Eur J Histochem. 2015 Feb 18;59(1):2492. doi: 10.4081/ejh.2015.2492.
Chitosan-based nanoparticles (chiNPs) are considered to be potentially good carriers for the sustained intracellular delivery of specific molecules. However, scarce attention has been paid to the long-lasting permanence of these NPs in the intracellular milieu, as well as to their intracellular fate (i.e., distribution, interaction with cell organelles, and degradation) in the long term. In the present study, the presence and subcellular location of FITC-labelled chiNPs were monitored in HeLa cells up to 14 days post-administration using multicolor-fluorescence confocal microscopy and diaminobenzidine photo-oxidation at transmission electron microscopy. The main result of the present study is the demonstration that internalized chiNPs persist inside the cell up to two weeks, occurring in both the cytoplasm and nucleus; accordingly, chiNPs are able to pass from mother to daughter cells through several mitotic cycles. The cells did not show increased mortality or structural damage up to 14 days after chiNP exposure.
基于壳聚糖的纳米颗粒（chiNP）被认为是特定分子持续细胞内递送的潜在良好载体。然而，这些纳米颗粒在细胞内环境中的长期持久性以及它们长期的细胞内命运（即分布、与细胞器的相互作用和降解）却很少受到关注。在本研究中，使用多色荧光共聚焦显微镜和透射电子显微镜下的二氨基联苯胺光氧化法，在给药后长达14天的时间里监测了HeLa细胞中FITC标记的chiNP的存在和亚细胞定位。本研究的主要结果表明，内化的chiNP在细胞内持续存在长达两周，存在于细胞质和细胞核中；因此，chiNP能够通过几个有丝分裂周期从母细胞传递到子细胞。在chiNP暴露后长达14天，细胞未显示出死亡率增加或结构损伤。
