Inorganic Chemistry Laboratory, Chemistry Department, National and Kapodistrian University of Athens, Panepistimiopolis Zografou, Greece.
Institute of Nanoscience and Nanotechnology, NCSR Demokritos, Agia Paraskevi, Attikis, Greece.
Methods Mol Biol. 2021;2207:85-97. doi: 10.1007/978-1-0716-0920-0_7.
This chapter focuses on the in vitro biological evaluation of multisensitive nanocontainers as drug delivery systems for cancer treatment. Cancer tissues possess some unique characteristics such as increased temperature due to inflammation, thermal vulnerability (40-45 °C), low cellular pH, and redox instabilities. The employment of polymers bearing pH, thermo, and/or redox sensitivities in the synthesis of hollow polymeric nanostructures has led to the formulation of a variety of drug delivery vehicles that are capable of targeted delivery and trigger specific drug release. The cavity in the structure allows for the encapsulation of anticancer drugs as well as other moieties with anticancer activity, like iron oxide magnetic nanoparticles. The drug loading and release capability of the nanocontainers is evaluated prior to biological studies in order to determine the concentration of the drug in the structure. The in vitro assessment includes cytotoxicity studies, quantitatively through the colorimetric MTT assay as well as qualitatively via the scratch-wound healing assay, on both cancer and healthy cell lines. The cellular localization of the studied drug-loaded and unloaded nanocontainers is determined through confocal fluorescence microscopy.
这一章主要关注多敏感纳米容器作为癌症治疗药物传递系统的体外生物学评估。癌症组织具有一些独特的特征，如由于炎症导致的温度升高、热脆弱性（40-45°C）、低细胞 pH 值和氧化还原不稳定性。在合成中空聚合物纳米结构时使用带有 pH 值、热和/或氧化还原敏感性的聚合物，导致了多种药物传递载体的形成，这些载体能够进行靶向传递并触发特定的药物释放。结构中的空腔允许封装抗癌药物以及具有抗癌活性的其他部分，如氧化铁磁性纳米粒子。在进行生物学研究之前，纳米容器的药物负载和释放能力会进行评估，以确定结构中药物的浓度。体外评估包括细胞毒性研究，通过比色 MTT 测定法进行定量研究，以及通过划痕愈合测定法进行定性研究，针对癌细胞和健康细胞系进行研究。通过共聚焦荧光显微镜确定研究的载药和未载药纳米容器的细胞定位。
