Preparation, characterization, and evaluation of peptide-functionalized liposomes for the targeting of placenta
Loading...
Date
2021-06-23
Authors
Difonzo, Marinella
Journal Title
Journal ISSN
Volume Title
Publisher
Abstract
The lack of safe medicaments for pregnant women, not implying risk of fetal harm, is a critical issue when it comes to placental dysfunctions, for which no curative treatment is currently available. Nanomedicines, such as liposomes, can be used to modulate biodistribution and reduce off-target side effects of classical drugs.
In this project, the optimization of a functionalization procedure to produce RGD-decorated liposomes for the potential targeting of the placenta was accomplished. Effective peptide binding to liposomes surface was determined by fluorescence measurements, and liquid chromatography-tandem mass spectrometry analysis. Liposomes with different ratios of DSPE-PEG2000 and RGD were produced, and their efficacy to target placental cells tested on BeWo cells, in vitro, and human placental explants, ex vivo. Flow cytometry results showed that RGD functionalized liposomes had a higher uptake than non-functionalized ones. They also suggested that higher amounts of DSPE-PEG2000 in the lipidic composition might hinder liposomal uptake by BeWo cells. Confocal images of human placental explants confirmed these results, and allowed localization of functionalized liposomes in the external syncytiotrophoblasts layer of the placenta, suggesting the lack of transplacental passage. However, further testing on a human perfused placenta would be necessary to exclude functionalized liposomes transfer to the fetal circulation. In conclusion, this study showed the ability of RGD-decorated liposomes to target placental cells, and the impact of both PEGylation and RGD functionalization degree on liposomes internalization. Besides, it provides an easy, versatile, and effective thiol-maleimide conjugation procedure, whose use can be extended to other placenta targeting peptides and liposomes produced by different preparation methods.
Description
Keywords
Pregnancy, Liposomes