Raman imaging for monitoring deuterated squalene-gemcitabine nanomedicines in single living breast cancer cells.

We have investigated the impact of gemcitabine (Gem) and deuterated gemcitabine-squalene (GemSQ-d6) nanoparticles (NPs) on MCF7 and MDA-MB-231 breast cancer cell lines by Raman spectroscopy. Quantification of LDL expression levels in both cell lines revealed a four-fold increase in MDA-MB-231 cells compared to MCF7 cells. In in vitro antitumor assessments, Gem displayed 13.5 times more effectiveness than GemSQ NPs against MCF7 cells, whereas GemSQ NPs induced a 14-fold increase in cytotoxicity compared to Gem for MDA-MB-231 cells. Oil Red O staining revealed that the treatment with GemSQ-d6 NPs induced a higher accumulation of lipid droplets at the periphery of the nucleus in MDA-MB-231 cells compared to MCF7 cells. Raman spectroscopy was employed to assess the impact of these drugs (50 µM, 24 hrs) on these breast cancer cell lines. By using the silent region (2000-2400 cm), we demonstrated that the accumulation of the GemSQ-d6 bioconjugate was higher in the cytoplasm of MDA-MB-231 cells than in MCF7 cells. This difference in drug accumulation is likely correlated with their expression levels of low-density lipoprotein receptors (LDLR). However, no information was obtained on Gem in this spectral region. We identified Raman features of squalene (SQ) in 700-1800 cm fingerprint region that allowed us to observe almost the same distribution of GemSQ as that observed in the silent region for both cell lines treated with GemSQ-d6 or SQ-d6. Subsequently, the effects of Gem and GemSQ-d6 on cellular components such as proteins, nucleic acids, and cytochrome C were monitored within the fingerprint spectral region. Our results revealed distinct features in the subcellular accumulation of these biomolecules in response to Gem and GemSQ treatments.