Identification of Cholesterol in Plaques of Atherosclerotic Using Magnetic Resonance Spectroscopy and 1D U-Net Architecture

Cholesterol plays a fundamental role in the human body-it stabilizes cell membranes, modulates gene expression, and is a precursor to steroid hormones, vitamin D, and bile salts. Its correct level is crucial for homeostasis, while both excess and deficiency are associated with serious metabolic and health consequences. Excessive accumulation of cholesterol leads to the development of atherosclerosis, while its deficiency disrupts the transport of fat-soluble vitamins. Magnetic resonance spectroscopy (MRS) enables the detection of cholesterol esters and the differentiation between their liquid and crystalline phases, but the technical limitations of clinical MRI systems require the use of dedicated coils and sequence modifications. This study demonstrates the feasibility of using MRS to identify cholesterol-specific spectral signatures in atherosclerotic plaque through ex vivo analysis. Using a custom-designed experimental coil adapted for small-volume samples, we successfully detected characteristic cholesterol peaks from plaque material dissolved in chloroform, with spectral signatures corresponding to established NMR databases. To further enhance spectral quality, a deep-learning denoising framework based on a 1D U-Net architecture was implemented, enabling the recovery of low-intensity cholesterol peaks that would otherwise be obscured by noise. The trained U-Net was applied to experimental MRS data from atherosclerotic plaques, where it significantly outperformed traditional denoising methods (Gaussian, Savitzky-Golay, wavelet, median) across six quantitative metrics (SNR, PSNR, SSIM, RMSE, MAE, correlation), enhancing low-amplitude cholesteryl ester detection. This approach substantially improved signal clarity and the interpretability of cholesterol-related resonances, supporting more accurate downstream spectral assessment. The integration of MRS with NMR-based lipidomic analysis, which allows the identification of lipid signatures associated with plaque progression and destabilization, is becoming increasingly important. At the same time, the development of high-resolution techniques such as μOCT provides evidence for the presence of cholesterol crystals and their potential involvement in the destabilization of atherosclerotic lesions. In summary, nanotechnology-assisted MRI has the potential to become an advanced tool in the proof-of-concept of atherosclerosis, enabling not only the identification of cholesterol and its derivatives, but also the monitoring of treatment efficacy. However, further clinical studies are necessary to confirm the practical usefulness of these solutions and their prognostic value in assessing cardiovascular risk.