Compagnin Chiara, Baù Luca, Mognato Maddalena, Celotti Lucia, Miotto Giovanni, Arduini Maria, Moret Francesca, Fede Caterina, Selvestrel Francesco, Rio Echevarria Iria M, Mancin Fabrizio, Reddi Elena
Department of Biology, University of Padova, Padova, Italy.
Nanotechnology. 2009 Aug 26;20(34):345101. doi: 10.1088/0957-4484/20/34/345101. Epub 2009 Aug 4.
Nanosized objects made of various materials are gaining increasing attention as promising vehicles for the delivery of therapeutic and diagnostic agents for cancer. Photodynamic therapy (PDT) appears to offer a very attractive opportunity to implement drug delivery systems since no release of the sensitizer is needed to obtain the therapeutic effect and the design of the nanovehicle should be much easier. The aim of our study was to investigate the use of organic-modified silica nanoparticles (NPs) for the delivery of the second-generation photosensitizer meta-tetra(hydroxyphenyl)chlorin (mTHPC) to cancer cells in vitro. mTHPC was entrapped in NPs (approximately 33 nm diameter) in a monomeric form which produced singlet oxygen with a high efficiency. In aqueous media with high salt concentrations, the NPs underwent aggregation and precipitation but their stability could be preserved in the presence of foetal bovine serum. The cellular uptake, localization and phototoxic activity of mTHPC was determined comparatively in human oesophageal cancer cells after its delivery by the NPs and the standard solvent ethanol/poly(ethylene glycol) 400/water (20:30:50, by vol). The NP formulation reduced the cellular uptake of mTHPC by about 50% in comparison to standard solvent while it did not affect the concentration-dependent photokilling activity of mTHPC and its intracellular localization. Fluorescence resonance energy transfer measurements, using NPs with mTHPC physically entrapped and a cyanine covalently linked, and ultracentrifugation experiments indicated that mTHPC is transferred from NPs to serum proteins when present in the medium. However, the coating of the NP surface with poly(ethylene glycol) largely prevented the transfer to proteins. In conclusion, mTHPC is rapidly transferred from the uncoated nanoparticles to the serum proteins and then internalized by the cells as a protein complex, irrespective of its modality of delivery.
由各种材料制成的纳米级物体作为癌症治疗和诊断药物递送的有前景载体正受到越来越多的关注。光动力疗法(PDT)似乎为实施药物递送系统提供了一个非常有吸引力的机会,因为获得治疗效果无需释放敏化剂,并且纳米载体的设计应该会容易得多。我们研究的目的是调查有机改性二氧化硅纳米颗粒(NPs)在体外将第二代光敏剂间四(羟苯基)氯卟啉(mTHPC)递送至癌细胞的用途。mTHPC以单体形式包封在NPs(直径约33 nm)中,其能高效产生活性单线态氧。在高盐浓度的水性介质中,NPs会发生聚集和沉淀,但在胎牛血清存在下其稳定性可得以保持。在通过NPs和标准溶剂乙醇/聚乙二醇400/水(体积比为20:30:50)递送mTHPC后,比较测定了其在人食管癌细胞中的细胞摄取、定位和光毒性活性。与标准溶剂相比,NP制剂使mTHPC的细胞摄取减少了约50%,同时它不影响mTHPC的浓度依赖性光杀伤活性及其细胞内定位。使用物理包封mTHPC的NPs和共价连接的花青进行荧光共振能量转移测量以及超速离心实验表明,当mTHPC存在于介质中时,它会从NPs转移至血清蛋白。然而,用聚乙二醇包被NP表面在很大程度上阻止了其向蛋白的转移。总之,无论递送方式如何,mTHPC都会从未包被的纳米颗粒迅速转移至血清蛋白,然后作为蛋白复合物被细胞内化。