This study presents the development and spectroscopic characterization of an erlotinib functionalized gold nanoparticle (erlotinib:AuNP) nanosystem designed for targeted delivery to metastatic non small cell lung cancer H1299 cells. Initial MTS assays demonstrated that free erlotinib induced a concentration dependent reduction in cell viability, while 0.1 uM erlotinib exhibited negligible cytotoxicity and was therefore selected for nanosystem fabrication. AuNPs alone showed minimal toxicity toward H1299 cells over the investigated concentration range. Following conjugation of erlotinib with AuNPs, the resulting nanosystems reduced cell viability to approximately 60%, indicating enhanced biological activity of the drug after nanoparticle-assisted delivery. Fluorescence microscopy confirmed the intracellular internalization of the nanosystems in H1299 cells, with nanoparticle aggregates predominantly localized in the perinuclear and perimitochondrial regions. Three-dimensional Raman spectroscopy (3D RS) mapping further verified the intracellular localization of the conjugates through characteristic Raman signatures of erlotinib:AuNPs. Importantly, 3D RS enabled detection of nanosystems at concentrations below the sensitivity limit of fluorescence imaging, demonstrating superior analytical performance for intracellular nanosystem tracking. Atomic force microscopy infrared (AFMIR) spectroscopy coupled with principal component analysis (PCA) demonstrated substantial biochemical modifications induced by the erlotinib:AuNP nanosystems, including enhanced lipid related spectral features and significant alterations in protein secondary structure, particularly the increased contribution of unordered and antiparallel beta turn conformations. The obtained results demonstrate that combining plasmonic nanocarriers with advanced vibrational spectroscopy enables highly sensitive monitoring of intracellular drug delivery and nanosystem-induced biochemical responses in cancer cells.
Piergies, N., Ocwieja, M., Pogoda, K., Panek, A., Roman, M., Raszka, K., Kwiatek, W. M.
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