Microfluidic-Assisted Coating of Nanoparticles by Membranes from Extracellular Vesicles

High Resolution Image Download MS PowerPoint Slide Extracellular vesicle (EV)-based nanotherapies represent a promising alternative in cancer nanomedicine thanks to their targeting properties. However, two major issues remain when combining nanoparticles (NPs) with EVs: (1) safety concerns of cancer-derived EVs related to their oncogenic nature and (2) lack of robust, reproducible, and efficient strategies for NPs coating with EVs membranes. To overcome these main challenges, in this study, we have combined a microfluidic platform to direct the reassembly of previously isolated EVs-derived membranes from lung cancer cells onto fluorescent silica NPs (FSNs). These FSNs leverage the versatile surface functionalization of silica NPs together with intrinsic fluorescent properties for precise intracellular tracking of our hybrids, avoiding membrane-labeling fluorescent probes. While preserving the tumor-targeting properties of natural lung cancer-derived EVs, the proposed microfluidic strategy offers superior handling and manipulation of the process to guarantee (1) homogeneous EV-like coating (6.3 nm thick), (2) high reproducibility, (3) efficient coating yields (87.8% of the NPs were coated), and (4) selective tumor-targeting properties (8.0- and 4.3-fold increases for parental A549 tumoral cells compared to HeLa and fibroblasts at 4 h, respectively). This technology enables the fabrication of biomimetic hybrid core–shell structures with high precision and stability in a continuous process that is more amenable to scaling up.