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Probing the effects of dextran-coated CeO2 nanoparticles on lung fibroblasts using multivariate single-cell Raman
Mirjana Mićević1, Sonja Čalija1, Lela Korićanac2
1Institute of Physics Belgrade, University of Belgrade, Belgrade, Serbia.
Abstract:
In this study, we investigated the cytotoxic effect of highly soluble dextran-coated CeO2 nanoparticles on human fetal lung fibroblasts MRC-5. We examined individual nanoparticle-treated cells by Raman spectroscopy and analyzed Raman spectra using non-negative principal component analysis and k-means clustering. In this way, we determined dose-dependent differences between treated cells, which were reflected through the intensity change of lipid, phospholipid and RNA-related Raman modes. Performing standard biological tests for cell growth, viability and induction of apoptosis in parallel, these changes were correlated with nanoparticle-induced apoptotic processes. The cells with specific spectral characteristics, referring to non-apoptotic, but possibly autophagic cell death modality, were also detected. Additionally, Raman imaging combined with principal component and vertex component analysis was used to map the spatial distribution of biological molecules in treated and untreated cells. This work provided the description of different resulting states of the treated cells depending on the dextran-coated CeO2 nanoparticles dose, which can be later used in the design of the nanoparticles for industrial or medical applications. The wide content of information resulting from single-cell Raman spectroscopy has the potential to detect biochemical changes caused by nanoparticles that would otherwise require a series of expensive and time-consuming standard biological techniques.
Insights
Highly soluble dextran-coated cerium oxide nanoparticles (CeO2) induce dose-dependent biochemical changes in human lung fibroblasts. Single-cell Raman spectroscopy reveals distinct cellular states, including apoptosis and autophagy, offering insights for nanoparticle design.
Area of Science:
- Nanotechnology
- Cell Biology
- Spectroscopy
Background:
- Cerium oxide nanoparticles (CeO2) have emerging applications but their biological effects require detailed investigation.
- Understanding nanoparticle-cell interactions is crucial for safe and effective use in medicine and industry.
- Dextran coating enhances nanoparticle solubility and influences their biological interactions.
Purpose of the Study:
- To investigate the cytotoxic effects of dextran-coated CeO2 nanoparticles on human fetal lung fibroblasts (MRC-5).
- To correlate biochemical changes detected by Raman spectroscopy with cellular responses like apoptosis and autophagy.
- To establish dose-dependent cellular states induced by these nanoparticles.
Main Methods:
- Single-cell Raman spectroscopy was employed to analyze nanoparticle-treated MRC-5 cells.
- Non-negative principal component analysis and k-means clustering were used for spectral data analysis.
- Standard biological assays for cell growth, viability, and apoptosis were performed in parallel.
- Raman imaging combined with principal component and vertex component analysis mapped molecular distribution.
Main Results:
- Dose-dependent changes in lipid, phospholipid, and RNA-related Raman modes were observed in treated cells.
- These spectral changes correlated with nanoparticle-induced apoptosis.
- Specific spectral signatures indicated a non-apoptotic, potentially autophagic cell death modality.
- Raman imaging revealed spatial distribution differences of biomolecules in treated versus untreated cells.
Conclusions:
- Dextran-coated CeO2 nanoparticles induce distinct cellular states in a dose-dependent manner.
- Single-cell Raman spectroscopy provides a comprehensive biochemical profile of nanoparticle-affected cells.
- This approach offers a powerful, efficient alternative to conventional biological techniques for nanoparticle safety assessment and design optimization.
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