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Detection of lipid efflux from foam cell models using a label-free infrared method

Xie, Bowen; Njoroge, Wanjiku; Dowling, Lewis M.; Sulé-Suso, Josep; Cinque, Gianfelice; Yang, Ying

Detection of lipid efflux from foam cell models using a label-free infrared method Thumbnail


Authors

Bowen Xie

Wanjiku Njoroge

Lewis M. Dowling

Gianfelice Cinque



Abstract

Cardiovascular diseases are still among the leading causes of mortality and morbidity worldwide. The build-up of fatty plaques in the arteries, leading to atherosclerosis, is the most common cause of cardiovascular diseases. The central player in atherosclerotic plaque formation is the foam cell. Foam cells are formed when monocytes infiltrate from the blood stream into the sub-endothelial space, differentiating into macrophages. With the subsequent uptake and storage of lipoprotein, especially low-density lipoprotein (LDL), they change their phenotype to lipid laden cells. Lowering circulating LDL levels, or initiating cholesterol efflux/reverse cholesterol transport in foam cells, is one of the current clinical therapies. Prescription of the pleiotropic drugs, statins, is the most successful therapy for the treatment and prevention of atherosclerosis. In this study, we used a foam cell model from the macrophage cell line, RAW 246.7, and applied the label-free Fourier Transform Infrared Spectroscopy (FTIR) method, i.e. synchrotron-based microFTIR spectroscopy, to study the lipid efflux process initiated by statins in a dose and time dependent manner. We used glass coverslips as substrates for IR analysis. The optical images (visible and fluorescent light) clearly identify the localization and lipid distribution within the foam cells, and the associated changes before and after culturing them with atorvastatin at concentrations of 0.6, 6 and 60 µg mL-1, for a culture duration between 24 to 72 hours. MicroFTIR spectroscopic spectra uniquely displayed the reduction of lipid content, with higher lipid efflux observed at higher doses of, and longer incubation time with, atorvastatin. Principal Component Analysis (PCA) and t-distributed Stochastic Neighbor Embedding (t-SNE) analysis demonstrated defined cluster separation at both lipid (3000-2800 cm-1) and fingerprint (1800-1350 cm-1) regions, with more profound discrimination for the atorvastatin dose treatment than time treatment. The data indicate that combining synchrotron-based microFTIR spectroscopy and using glass substrates for foam cells can offer an alternative tool in atherosclerosis investigation at a molecular level, and through cell morphology.

Citation

Xie, B., Njoroge, W., Dowling, L. M., Sulé-Suso, J., Cinque, G., & Yang, Y. (2022). Detection of lipid efflux from foam cell models using a label-free infrared method. Analyst, 147(23), 5372-5385. https://doi.org/10.1039/D2AN01041K

Journal Article Type Article
Acceptance Date Oct 17, 2022
Publication Date Oct 26, 2022
Publicly Available Date May 30, 2023
Journal Analyst
Print ISSN 0003-2654
Publisher Royal Society of Chemistry
Volume 147
Issue 23
Pages 5372-5385
DOI https://doi.org/10.1039/D2AN01041K
Public URL https://keele-repository.worktribe.com/output/424772
Publisher URL https://pubs.rsc.org/en/content/articlelanding/2022/AN/D2AN01041K

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