Microfluidics Synthesis of Gene Silencing Cubosomes

Abstract

The success of gene technologies hinges on our ability to engineer superior encapsulation and delivery vectors. Cubosomes are lipid-based nanoparticles where membranes, instead of enveloping into classic liposomes, intertwine into complex arrays of pores well-ordered in a cubic lattice. These complex nanoparticles encapsulate large contents of siRNA compared to a liposomal analogue. Importantly, the membranes that form cubosomes have intrinsic fusogenic properties that promote fast endosomal escape. Despite the great potential, traditional routes of forming cubosomes lead to particle sizes too large to fulfill the state-of-the art requirements of delivery vectors. To overcome this challenge, we utilize a microfluidic nanomanufacturing device to synthesize cubosomes and siRNA-loaded cubosomes, termed cuboplexes. Utilizing cryogenic TEM and small angle X-ray scattering we elucidate the time-resolved mechanisms in which microfluidic devices allow the production of small cubosomes and cuboplexes (75 nm) that outperform commercially available delivery vectors, as well as liposome-based systems.

Microfluidics for Production of Particles: Mechanism, Methodology, and Applications

“…The other commonly used method is typically based upon smart design of the channel geometries. Various types of microfluidic chips with complex geometry, such as Tesla‐shape, herringbone‐shaped, serpentine‐shaped,, planar asymmetric split, spiral/semispiral, zigzag‐shaped channel, etc., were designed and applied for preparing NPs ( Figure a–e). For example, herringbone‐shaped microchannel containing patterned microgrooves of varying shapes and angles can induce chaotic stirring at a low Re (Re = 10 −2 –10 2 ).…”

The stabilization of primitive bicontinuous cubic phases with tunable swelling over a wide composition range

“…Lipid bicontinuous phases are found in biological systems 1,2 and have numerous biotechnology applications, including drug delivery, 3,4 nucleic acid delivery, 5–8 biosensing, 9,10 medical imaging, 4,11,12 membrane protein crystallization, 13–16 and foods. 17 Fig.…”

Microfluidic Generation of Nanomaterials for Biomedical Applications

“…Kim et al applied the similar microfluidic chip in manufacturing of colloidally stable cubosomes (a lipid‐based cubic NPs). The mixing kinetics in small volumes was significantly improved, the process of solvent extraction was precisely controlled, and the generated cubosomes were uniform with an average diameter of about 200 nm . In another experiment, Jahn et al utilized HF microfluidics to fabricate LNPs with a desired relative standard deviation (Figure d) .…”

“…The mixing kinetics in small Small 2020, 16,1901943 volumes was significantly improved, the process of solvent extraction was precisely controlled, and the generated cubosomes were uniform with an average diameter of about 200 nm. [133] In another experiment, Jahn et al utilized HF microfluidics to fabricate LNPs with a desired relative standard deviation ( Figure 3d). [134] The on-chip synthesis could also offer a platform to investigate the self-assembly process of lipid molecules under various reaction parameters.…”