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FITC-CRD13 dendrimer (un)locked in liposomal vesicles characterized by size exclusion chromatography and dynamic light scattering methods

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FITC-CRD13 dendrimer (un)locked in liposomal vesicles characterized by size exclusion chromatography and dynamic light scattering methods

Petra Kohútová 1 Zuzana Garaiová 1 Milan Zvarík 1 Veronika Šubjaková 1 Sylwia Michlewska 2,3 Maksim Ionov 2 Iveta Waczulíková 1 Javier de la Mata 4,5,6 Maria Bryszewska 2 Tibor Hianik 1

1Department of Nuclear Physics and Biophysics, Faculty of Mathematics, Physics and Informatics, Comenius University, Bratislava, Slovakia
2Department of General Biophysics, Faculty of Biology and Environmental Protection, University of Lodz, Lodz, Poland
3Laboratory of Microscopic Imaging and Specialized Biological Techniques, Faculty of Biology and Environmental Protection, University of Lodz, Poland
4Inorganic Chemistry Department, University Alcala, Alcala de Henares, Spain
5Ramón y Cajal Health Research Institute (IRYCIS), IRYCIS, Spain
6Networking Research Center on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Spain
petakohutova@gmail.com

Different types of nanoparticles were synthetized and studied either as a drug itself or as a drug delivery system aiming to combine benefits of individual particles. In this work we focused on the encapsulation of fluorescently labelled ruthenium dendrimers (FITC-CRD13) possessing anticancer properties [1] into the lipid vesicles composed of DMPC/DMPG/Cholesterol. FITC-CRD13 dendrimers (10 µM) were encapsulated into the liposomes by passive loading during the hydration step of lipid film (t = 1.5 h, T = 40°C) in sodium-phosphate buffer (10 mM, pH 7.4) upon the continuous shaking. Subsequently, the prepared vesicles were extruded through 400nm polycarbonate membrane and characterized by 1) absorption and fluorescence spectra of nanoparticles using size exclusion chromatography (SEC-HPLC) on a 30 nm size pore column; and 2) size measurements using dynamic light scattering (DLS). Absorbance at 220 nm and fluorescence at 495/520 nm was used for detection of liposomes and FITC-CRD13 dendrimers, respectively.  Firstly, liposomes were eluted at 3.35 min. Fluorescence intensity measured along the elution profile centred at this time point was monitored for dendrimers being locked in liposomes. The coelution, thus encapsulation was less extensive because the subsequent peaks in dendrimer elution profile were observed, indicating the presence of non-entrapped material sized around 70 nm at the later retention times. Hydrodynamic diameter of purified dendrimer-liposomal sample that coeluted at 3.35 min was 210±10 nm with polydispersity index 0.1 reflecting an uniform sample with respect to the particle size. The obtained results can serve for optimization of the encapsulation protocol and as a base for further studies of the dendrimer-liposomal drug delivery platform in the cancer treatment.

This work has been financially supported by Science Grant Agency VEGA, project No. 1/0756/20; by Agency for Promotion Research and Development, project No. SK-PL-18-0080 and SK-BY-RD-19-0019; by KEGA, project No. 041UK-4/2020 and by NAWA International Academic Partnership Programme EUROPARTNER.
[1] Michlewska, S. et al., 2017. Ruthenium metallodendrimers with anticancer potential in an acute promyelocytic leukemia cell line (HL60), European Polymer Journal. Elsevier Ltd, 87, pp. 39–47. doi: 10.1016/j.eurpolymj.2016.12.011.
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Size and encapsulation volume 22.05.2021 08:44
Nice piece of work within a basic research in the field of nanomedicine ...both, liposomes and dendrimers are attractive systems for localized and controlled delivery of potentially toxic agents that canno...Zobraziť celý komentár
Re: Size and encapsulation volume 28.05.2021 10:02
Thank you for your question. As you have pointed out, the process of opsonization can be overcome by the modification of the surface of liposomes, for instance with PEG (polyethylene glycol), which can prolo...Zobraziť celý komentár
Re: Re: Size and encapsulation volume 28.05.2021 19:40
Thank you for the comprehensive answer, and I wish you all the success in your future professional life.
iwa
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