Dendrimers loaded into liposomes as a new dual drug delivery system for cancer treatment

Primárne karty

ISBN: 978-80-972360-6-9

Dendrimers loaded into liposomes as a new dual drug delivery system for cancer treatment

Veronika Šubjaková1 , Zuzana Garaiová , Sylwia Michlewska2 , Jakub Magiera , Šimon Šutý , Marcin Holota , Maksim Ionov , Natalia Sanz-del Olmo3 , Iveta Waczulikova , Francisco Javier de la Mata , Maria Bryszewska , Tibor Hianik ,
1 Faculty of Mathematics, Physics and Informatics Comenius University, Department of Nuclear Physics and Biophysics, Bratislava, Slovakia
2 Faculty of Biology and Environmental protection, Department of General Biophysics, University of Lodz, Laboratory of Microscopic Imaging and Specialized Biological Techniques, Lodz, Poland
3 Department of Organic and Inorganic Chemistry, and Research Institute in Chemistry ”Andrés M. Del Río” (IQAR) ) and Institute "Ramón y Cajal" for Health Research (IRYCIS), University of Alcalá, Madrid, Spain.. Networking Research Center on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Spain
subjakov@gmail.com

Studying separately, dendrimers and liposomes have shown promising results as drug delivery carriers.    In this study, we focused on their combination in order to merge their individual benefits into a new dual delivery system for more effective and safer cancer treatment [1]. We have worked with Ruthenium metallodendrimers, specifically with fluorescently labeled FITC-CRD13 dendrimer [2]. Ruthenium complexes play an important role as anticancer agents among metal drugs [3]. Liposomes were prepared by hydration method using zwitterionic 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC). We examined encapsulation of dendrimers into the DMPC liposomes by means of passive loading during vesicle formation: FITC-CRD13 (150 mM) was added either into the step of 1) dissolving lipids in an organic solvent or 2) lipid film hydration. Samples were analyzed by fluorescence confocal microscopy. The obtained microscopic images indicate the presence of dendrimers in the liposomal structure consisting of several lamellae forming vesicles with the size in micrometer range. To ensure size reduction, extrusion through 400 nm polycarbonate membranes was performed. The centrifugation was applied for removal of non-encapsulated dendrimers. The supernatant and pellet were analyzed via fluorescence intensity measurements and dynamic light scattering. The size and polydispersity index (PDI) of the samples were determined following each purification cycle. For liposomes being rehydrated with dendrimers, a more intense coloring of extrusion membrane as well as higher fluorescence intensity signal of supernatant was observed in comparison with liposomes where dendrimers were initially added into the organic solvent. This may suggest aggregating behavior of FITC-CRD13 dendrimer in the buffer solution and its weak binding/low loading into the DMPC liposomes, respectively. PDI index for liposomes being rehydrated with FITC-CRD13 was lower (~0,2) than in the case of an “organic” encapsulated sample (~0.5).The hydrodynamic diameter (Z-average size) of the liposomes increased for both types of prepared samples with each centrifugation step, probably as a result of the loss of smaller liposomes, which had a low pelletization ability, thus shifting the average size to higher values (~ 500 – 600 nm after 5th purification cycle). In conclusion, the passive loading of FITC-CRD13 dendrimer was investigated with a large concentration of non-encapsulated drug observed that needs to be removed prior further usage. To increase a capturing efficiency, active encapsulation techniques should be tested with minimizing the need for post-synthesis processing thus preventing potential damage/release of encapsulated material. The effect of various lipid composition should also be investigated.

Poďakovanie: 

This work was supported by the Slovak Research and Development Agency, APVV (Projects No. SK-PL-18-0080, APVV-14-0267, SK-BY-RD-19-0019), COST CA17140, VEGA 1/0756/20, KEGA 041UK-4/2020 and by Pl-SK 2019–2020 bilateral project of NAWA PPN/BIL/2018/1/00150; Polish National Agency for Academic Exchange, NAWA, PPI/APM/2018/1/00007/U/001.

Zdroje: 

[1] K. Gardikis, S. Hatziantoniou, M. Bucos, D. Fessas, M. Signorelli, T. Felekis, M. Zervou, C.G. Screttas, B.R. Steele, M. Ionov, M. Micha-Screttas, B. Klajnert, M. Bryszewska, C. Demetzos, J Pharm Sci. 2010, 99, 3561-71. doi: 10.1002/jps.22121.
[2] S. Michlewska, M. Kubczak, M. Maroto-Díaz, N.  Sanz del Olmo,  P. Ortega,  D. Shcharbin,  R. Gomez Ramirez, F. Javier de la Mata,  M. Ionov,  M. Bryszewska, Biomolecules 2019, 9,411. doi:10.3390/biom9090411
[3] I. Kostova, Current Medicinal Chemistry 2016, 13, 1085-1107.