RESEARCH ARTICLE

Human erythrocyte-derived nanovesicles can readily be loaded with doxorubicin and act as anticancer agents.

 

Running title: Erythrocyte derived nanovesicles and drug delivery

Authors: Louise Dubois , Liza Löf , Anders Larsson 1, KjellHultenby3, Anders Waldenström 4, Masood Kamali-Moghaddam 2, Gunnar Ronquist 1, K Göran Ronquist 1*

 

Affiliations:

1 Department of Medical Sciences, Clinical Chemistry, Uppsala University, SE-751 85 Uppsala, Sweden

2 Department of Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala University, SE-751 08 Uppsala, Sweden

3 Department of Laboratory Medicine, Karolinska Institutet, SE-141 86 Huddinge, Sweden

4 Department of Public Health and Clinical Medicine, Umeå University, SE-901 85 Umeå, Sweden

 

* Corresponding author: K Göran Ronquist

E-mail goran.ronquist@slu.se (KGR)

These authors contributed equally to this work

  

Abstract

Purpose: In future therapeutics new formulas are needed that assure lower doses, fewer side effects, targeted administration and protection of the drug from degradation. In a first step to fulfil the requirements defined above, we carried out an in vitro study by developing a new procedure to use native vesicles first from prostasomes and then from erythrocyte membranes known to be well tolerated. The new method for production of drug delivery vehicles implicated osmotic loading of detergent resistant membranes (DRMs).

Materials and methods: DRMs of prostasomes and prepared human erythrocyte membranes were extracted and separated in a sucrose gradient at a density of 1.10 g/mL containing 1% Triton X-100. These DRMs were characterized by electron microscopy (transmission and scanning EM) and loaded with low and high molecular compounds. PC3 prostate cancer cells were treated with doxorubicin loaded DRMs in triplicate. DAPI (nuclear fluorescent stain) was included and fluorescence microscopic pictures were taken before the cells were trypsinized and counted after 48h.

Results: The content of the well separated band was observed ultrastructurally as small spherical, double layered membrane vesicles, (DRM vesicles) which harbored hyperosmolar sucrose of the gradient. Encapsulated hyperosmolar sucrose induced a transient osmotic lysis of the DRM vesicles when suspended in isotonic buffer containing loading molecules allowing vesicular inclusion. After this proof of concept, the method was finally employed for doxorubicin loading of DRM vesicles from human erythrocytes. When incubating such vesicles with PC3 cells a complete arrest of growth was observed in sharp contrast to PC3 cells incubated with plain doxorubicin in similar conditions.

Conclusion: The present results open up new possibilities for using DRM vesicles as drug delivery vehicles.

Keywords: Prostasomes, Prostate cancer, DRM vesicles, drug delivery

 

 

 

 

 

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