Nanomedizinische Ansätze zur Prävention und Behandlung von Atherosklerose
Zusammenfassung der Projektergebnisse
In this project, we investigated the suitability of nanotechnological approaches to atherosclerosis using endothelial cells in vitro, isolated human arteries ex vivo, and a rabbit model of atherosclerosis. Within the project duration, several new developments and challenges lead to the modifications of the original work tasks, as described in detail above. The in vitro studies covered the extensive analyses of endothelial biocompatibility and functional effects of different types of SPIONs. Furthermore, the spontaneous as well as magnetically-targeted accumulation of circulating SPIONs was investigated in the flow model of bifurcations. In the subsequent step, we established a novel ex vivo flow model based on the human umbilical artery, which constitutes a time- and cost-efficient, 3R-compliant tool to assess magnetic targeting of SPIONs under flow. Using this model, the magnetic targeting of different types of SPIONs under various external magnetic field and flow conditions was investigated by atomic emission spectroscopy and histology. Furthermore, the effect of blood components on particle margination was investigated. From the obtained results, SPION-2 with good biocompatibility and good magnetic targeting ability emerged as an appropriate candidate for magnetic drug targeting applications. SPION-3 emerged as a nanosystem with exceptional biocompatibility and very low internalization by nonphagocytic cells. These particle characteristics constitute the features of a superb candidate for magnetic resonance imaging (MRI) contrast agent and currently undergo further studies in this direction. Additionally, although we did not achieve magnetic accumulation of SPION-3 suspended in cell culture media, in the presence of RBCs, SPION-3 were successfully accumulated at the same value of magnetic field gradient. This indicates their potential use for magnetic targeting in the future. In vivo, the rabbit model of balloon injury- and Western diet-induced atherosclerosis in abdominal aorta and the magnetic targeting protocol were successfully established. For the purpose of in vivo feasibility studies on magnetic drug targeting to aortic plaques, SPION-2 have proven the most suitable candidate and were successfully targeted to abdominal aorta in experimental rabbits as confirmed by histology. Subsequently, the SPION-2 have been conjugated with dexamethasone phosphate (SPION-2-DEXA) and evaluated in vivo, in the early-treatment (acute vascular injury followed by high cholesterol diet) and late-treatment set up (advanced atherosclerotic plaque). The results demonstrated that treatment of inflammation in early vascular injury combined with hypercholesterolemia with dexamethasone-SPION-2 leads to enhanced accumulation and macrophage lipotoxicity, as recently reported also for liposomal prednisolone. To target other types of drugs, including anti-angiostatic compounds, two different types of particles were developed, namely SPION-APTES and Cit-Au-SPIONs. Due to experimental challenges, some of the studies concerning directed binding of peptides to SPIONs, as well as their biocompatibility and efficacy are still ongoing. SPION-3 particles have already been tested for liver imaging and underwent the size modification in order to achieve USPIO (ultrasmall superparamagnetic iron oxide nanoparticles) with diameters below 30 nm, which are suitable for atherosclerotic plaque imaging. The studies in a rabbit model of balloon injury- and high cholesterol diet-induced atherosclerosis confirmed their accumulation in atherosclerotic lesions by MRI and histology. Taken together, the obtained results greatly improve our understanding of the interactions of magnetic nanoparticles with endothelial cells and the biological effect of SPIONs in physiological-like settings. The newly established ex vivo artery model allows a rapid estimation of magnetic targeting capacity of different SPION types, facilitating the selection of candidate particles for drug delivery without the necessity of in vivo testing. Contrary to our expectations, corticosteroid-containing nanosystems did not reduce plaque inflammation, indicating that the efficacy of a classical anti-inflammatory compound in an inflammatory disease cannot be extrapolated to the lipid-rich environment of an atherosclerotic plaque. Finally, the successful development of diagnostic SPION-3 and their size-adapted version (USPIODex) suitable for atherosclerotic plaque can, upon the successful preclinical evaluation, contribute to the reduction of cardiovascular disease burden by earlier detection of atherosclerotic lesions and enable safe serial imaging of the plaques.
Projektbezogene Publikationen (Auswahl)
- Endothelial biocompatibility and accumulation of SPION under flow conditions. Journal of Magnetism and Magnetic Materials 2015, 380, 20
Matuszak J, Zaloga J, Friedrich RP, Lyer S, Nowak J, Odenbach S, Alexiou C, Cicha I
(Siehe online unter https://doi.org/10.1016/j.jmmm.2014.09.005) - Flow cytometry for intracellular SPION quantification: specificity and sensitivity in comparison with spectroscopic methods. Int J Nanomedicine. 2015 Jun 26;10:4185-201
Friedrich RP, Janko C, Poettler M, Tripal P, Zaloga J, Cicha I, Dürr S, Nowak J, Odenbach S, Slabu I, Liebl M, Trahms L, Stapf M, Hilger I, Lyer S, Alexiou C
(Siehe online unter https://doi.org/10.2147/ijn.s82714) - Shell matters: Magnetic targeting of SPIONs and in vitro effects on endothelial and monocytic cell function. Clin Hemorheol Microcirc. 2015;61(2):259-77
Matuszak J, Dörfler P, Zaloga J, Unterweger H, Lyer S, Dietel B, Alexiou C, Cicha I
(Siehe online unter https://doi.org/10.3233/ch-151998) - Nanoparticles for intravascular applications: physicochemical characterization and cytotoxicity testing. Nanomedicine (Lond). 2016;11(6):597-616
Matuszak J, Baumgartner J, Zaloga J, Juenet M, da Silva AE, Franke D, Almer G, Texier I, Faivre D, Metselaar JM, Navarro FP, Chauvierre C, Prassl R, Dézsi L, Urbanics R, Alexiou C, Mangge H, Szebeni J, Letourneur D, Cicha I
(Siehe online unter https://doi.org/10.2217/nnm.15.216) - A novel human artery model to assess the magnetic accumulation of SPIONs under flow conditions. Sci Rep. 2017;7:42314
Janikowska A, Matuszak J, Lyer S, Schreiber E, Unterweger H, Zaloga J, Groll J, Alexiou C, Cicha I
(Siehe online unter https://doi.org/10.1038/srep42314) - Non-immunogenic dextran-coated superparamagnetic iron oxide nanoparticles: a biocompatible, size-tunable contrast agent for magnetic resonance imaging. Int J Nanomedicine. 2017;12:5223-5238
Unterweger H, Janko C, Schwarz M, Dézsi L, Urbanics R, Matuszak J, Lyer S, Őrfi E, Fülöp T, Bäuerle T, Szebeni J, Boccaccini AR, Alexiou C, Cicha I
(Siehe online unter https://doi.org/10.2147/ijn.s138108) - Comparative analysis of nanosystems' effects on human endothelial and monocytic cell functions. Nanotoxicology 2018, 12(9):957-974
Matuszak J, Doerfler P, Lyer S, Unterweger H, Juenet M, Chauvierre C, Alaarg A, Franke D, Almer G, Texier I, Metselaar JM, Prassl R, Alexiou C, Mangge H, Letourneur D, Cicha I
(Siehe online unter https://doi.org/10.1080/17435390.2018.1502375) - Dextran-coated superparamagnetic iron oxide nanoparticles for magnetic resonance imaging: Evaluation of size-dependent imaging properties, storage stability and safety. Int J Nanomed 2018;13:1899-1915
Unterweger H, Dézsi L, Matuszak J, Janko C, Poettler M, Jordan J, Bäuerle T, Szebeni J, Fey T, Boccaccini AR, Alexiou C, Cicha I
(Siehe online unter https://doi.org/10.2147/ijn.s156528) - Drug delivery to atherosclerotic plaques using superparamagnetic iron oxide nanoparticles. Int J Nanomedicine. 2018;13:8443-8460
Matuszak J, Lutz B, Sekita A, Zaloga J, Alexiou C, Lyer S, Cicha I
(Siehe online unter https://doi.org/10.2147/ijn.s179273) - Magnetic Accumulation of SPIONs under Arterial Flow Conditions: Effect of Serum and Red Blood Cells. Molecules. 2019;24(14). pii: E2588
Hennig TL, Unterweger H, Lyer S, Alexiou C, Cicha I
(Siehe online unter https://doi.org/10.3390/molecules24142588) - Synthesis and characterization of citrate-stabilized gold-coated superparamagnetic iron oxide nanoparticles for biomedical applications. Molecules. 2020; 25: 442
Stein R, Friedrich B, Mühlberger M, Cebulla N, Schreiber E, Tietze R, Cicha I, Alexiou C, Dutz S, Boccaccini AR, Unterweger H
(Siehe online unter https://doi.org/10.3390/molecules25194425)
