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Published on: June 23, 2020
RGD-functionalized Wolffia globosa-derived Exosome-like Nanoparticles for Targeted Doxorubicin Delivery in Breast
Chatchai Nensat1,2, Worawat Songjang1,3, Manatsanan Piamano1
1Integrative Biomedical Research Unit (IBRU), Faculty of Allied Health Sciences, Naresuan University, Phitsanulok, 65000, Thailand.
Introduction:
Plant-derived exosome-like nanoparticles (ELNs) have attracted considerable attention as biocompatible drug delivery vehicles. Wolffia globosa (Kai-pum), a native Thai aquatic plant, represents a novel and underexplored source of ELNs. This study describes the isolation, characterization, and functional evaluation of W. globosa-derived exosome-like nanoparticles (WGENs) that are surface-modified with the integrin-targeting Arg-Gly-Asp (RGD) peptide and loaded with doxorubicin (Dox) for targeted breast cancer therapy.
Methods:
WGENs were isolated by differential ultracentrifugation and characterized by nanoparticle tracking analysis (NTA) and transmission electron microscopy (TEM). RGD functionalization was achieved via DSPE-PEG-RGD membrane insertion, followed by doxorubicin loading through electroporation. Cellular uptake was assessed by fluorescence microscopy and live-cell imaging in 4T1 and CL-S1 cell lines. Cytotoxicity was evaluated by LDH assay.
Results:
NTA revealed WGEN mode sizes of 94.1 and 106.9 nm. TEM confirmed cup-shaped vesicular morphology. RGD-Exo-Dox treatment resulted in 4-fold greater intracellular doxorubicin accumulation in 4T1 cells than in CL-S1 cells (4.3% vs. 1.1% red fluorescence at 16 h). No statistically significant difference in cytotoxicity between RGD-Exo-Dox and free doxorubicin was detected in 4T1 cells (all p > 0.05); the observed numerical differences did not constitute positive evidence of differential cytotoxicity.
Discussion:
Preferential intracellular doxorubicin accumulation was observed in αvβ3-overexpressing 4T1 cells compared with CL-S1 cells, which is consistent with RGD-mediated integrin targeting. This interpretation is subject to important limitations, including the absence of a scrambled-peptide control nanoparticle and unconfirmed surface RGD density; the differential uptake may therefore reflect nonspecific factors. Comparative formulation data are presented as preliminary, descriptive data pending formal statistical validation.
Conclusion:
RGD-functionalized WGENs constitute a promising plant-derived nanocarrier platform for proof-of-concept doxorubicin delivery, with enhanced uptake in αvβ3-overexpressing 4T1 cells. These preliminary findings warrant further investigation with appropriate mechanistic controls and in vivo validation.
