Natural and SyntheticLDL-Based Imaging Probes forthe Detection of Atherosclerotic Plaques

Low-density lipoprotein (LDL) is the primary natural carrier of lipids in the bloodstream and plays a central role in the development of atherosclerosis. By leveraging LDL’s natural tendency to accumulate at sites of plaque formation, LDL can be employed as a carrier to selectively deliver the imaging probes to efficiently detect atherosclerotic plaques. In our previous studies, we reported several LDL-based magnetic resonance imaging contrast agents (MRI-CAs) formed by modifying natural LDL (nLDL) or developing LDL-mimetic (synthetic LDL, sLDL) from lipid nanoparticles (LNPs) utilizing chemical reactions on the nanoparticle surface, including preliminary MRI tests. In this study, we report the <i>in vivo</i> biological functionality of these LDLs (both nLDL and sLDL)-based Gd(III)-based contrast agents (GBCAs) by conducting detailed <i>in vivo</i> studies on two types of atherosclerosis murine models, namely, <i>apoE</i>^{<i>–/–</i>} and <i>LDLr</i>^{<i>–/–</i>}. We provide more comprehensive MRI data accompanied by <i>ex vivo</i> results, including microscopic analysis of aorta segments for LDL accumulation and whole-body cryoVIZ analysis for biodistribution of the probe. We also tested <i>in vitro</i> cellular internalization of sLDL on two cell lines (RAW 264.7 and THP-1), which are derived from macrophages and monocytes, respectively, in order to observe sLDL uptake by macrophages, which are often present at the vulnerable types of atherosclerotic plaques. In conclusion, our current study demonstrates that modified LDLsboth nLDL and sLDLfacilitate MRI detection of atheroplaques by efficient uptake by macrophages. Taken together with the high loading capacity of Gd(III)-chelate molecules on LDL, especially sLDL, the LDL-based MRI contrast agents reported here hold significant potential for the early detection of atherosclerosis, including vulnerable ones, and should be useful for preventive diagnosis strategies.