Altın nanoparçacıkların yüzeyde zenginleştirilmiş raman saçılmasına dayalı endositozunun takibi

Understanding the interaction of nanomaterials with living systems is critical to use these novel materials in various fields including nanomedicine. Due to being the first step at the interaction interface, endocytosis has a major role governing the intracellular destinations and behaviors of molecular and non-molecular species including nanoparticles. In this thesis, origin of the intracellular surface-enhanced Raman spectroscopy (SERS) spectra was investigated based on the protein corona structure on the AuNPs surface, endocytosis pathway development and pathway choice of AuNPs. For this purpose, firstly protein corona formation on AuNPs were examined with different cell culture conditions by SERS and effect of the protein corona on the obtained intracellular SERS spectra was investigated. Then, endocytosis pathway choice of AuNPs was investigated with the help of specific inhibitors for each endocytosis pathway. It is aimed to understand whether SERS spectra can tell us endocytosis route into a living cell. For this, endocytosis pathways are inhibited and the obtained SERS spectra are evaluated. Further, the source of the obtained SERS spectra is examined by isolating endosomes and comparing obtained SERS spectra with cellular spectra as time dependent. Finally, the effect of the endocytosis on the macrophage activation and AuNPs phagocytosis is investigated. From the results, it is concluded that protein corona alterations based on different cell culture conditions can be tracked by SERS, and it can affect the intracellular SERS spectra directly. It is also found that SERS can significantly contribute to the investigation of different endosomal pathways, and macrophage activation from single living cells. Furthermore, it is also seen that source of the SERS cannot be thought as only from AuNPs in the endosomes but also from escaped AuNPs, which are evidenced to be found in both cytosol and nucleus accumulated in a time dependent manner. Thus, the origin of the intracellular SERS spectra cannot be considered only from endosomes. Consequently, it is revealed that SERS can distinguish significantly not only protein corona alterations and interference with the intracellular spectra, but also pathways used for nanoparticle uptake and different metabolisms including macrophage activation and phagocytosis.