Design and Development of Biomimetic Nanovesicles Using a Microfluidic Approach

Abstract

The advancement of nanotechnology toward more sophisticated bioinspired approaches has highlighted the gap between the advantages of biomimetic and biohybrid platforms and the availability of manufacturing processes to scale up their production. Though the advantages of transferring biological features from cells to synthetic nanoparticles for drug delivery purposes have recently been reported, a standardizable, batch-to-batch consistent, scalable, and high-throughput assembly method is required to further develop these platforms. Microfluidics has offered a robust tool for the controlled synthesis of nanoparticles in a versatile and reproducible approach. In this study, the incorporation of membrane proteins within the bilayer of biomimetic nanovesicles (leukosomes) using a microfluidic-based platform is demonstrated. The physical, pharmaceutical, and biological properties of microfluidic-formulated leukosomes (called NA-Leuko) are characterized. NA-Leuko show extended shelf life and retention of the biological functions of donor cells (i.e., macrophage avoidance and targeting of inflamed vasculature). The NA approach represents a universal, versatile, robust, and scalable tool, which is extensively used for the assembly of lipid nanoparticles and adapted here for the manufacturing of biomimetic nanovesicles.

Biohybrid Nanoparticles to Negotiate with Biological Barriers

“…Biohybridization of synthetic and biological materials could be done via bio‐orthogonal chemistries, biointerfering chemistries, extrusion based techniques, and microfluidics …”

“…These components as well as desired synthetic NPs are then hybridized using filter extrusion (Figure I). An emerging approach for biohybridization is based on microfluidics which offers a robust tool for NPs synthesis in a reproducible fashion (Figure J) …”

Lipid Nanovesicles by Microfluidics: Manipulation, Synthesis, and Drug Delivery

“…c) Biomimetic lipid nanoparticles. Reproduced with permission . Copyright 2017, American Chemical Society.…”

“…Biomimetic lipid nanoparticles to mimic erythrocytes, leukocytes, platelets, and cancer cells are a promising therapeutic platform with intrinsic biological properties and good delivery capability . Traditionally, the synthesis requires multiple rounds of extrusion, which may cause the loss of proteins, disruption of the membrane structure, and difficulties in scaling up.…”

“…Despite the promising results, the potential disruption to membrane proteins by electroporation could be a challenge for fabricating other biomimetic nanoparticles. Molinaro et al recently described the assembly of lipid nanovesicles incorporated with leukocyte‐derived membrane proteins (leukosomes) in a Y‐shaped microchannels with herringbone structures at a flow rate of 1 mL h −1 and an aqueous‐to‐organic phase flow rate of 2, without the use of any additional process such as electroporation (Figure c) . These leukosomes exhibited intrinsic biological properties including delaying macrophage uptake and targeting of inflamed endothelium.…”