One-Step Generation of Multisomes from Lipid-Stabilized Double Emulsions

Abstract

Multisomes are multicompartmental structures formed by a lipid-stabilized network of aqueous droplets, which are contained by an outer oil phase. These biomimetic structures are emerging as a versatile platform for soft matter and synthetic biology applications. While several methods for producing multisomes have been described, including microfluidic techniques, approaches for generating biocompatible, monodisperse multisomes in a reproducible manner remain challenging to implement due to low throughput and complex device fabrication. Here, we report on a robust method for the dynamically controlled generation of multisomes with controllable sizes and high monodispersity from lipid-based double emulsions. The described microfluidic approach entails the use of three different phases forming a water/oil/water (W/O/W) double emulsion stabilized by lipid layers. We employ a gradient of glycerol concentration between the inner core and outer phase to drive the directed osmosis, allowing the swelling of lamellar lipid layers resulting in the formation of small aqueous daughter droplets at the interface of the inner aqueous core. By adding increasing concentrations of glycerol to the outer aqueous phase and subsequently varying the osmotic gradient, we show that key structural parameters, including the size of the internal droplets, can be specifically controlled. Finally, we show that this approach can be used to generate multisomes encapsulating small-molecule cargo, with potential applications in synthetic biology, drug delivery, and as carriers for active materials in the food and cosmetics industries.

A Lipid-Based Droplet Processor for Parallel Chemical Signals

“… 42 , 43 For further complexity in signaling, engineered αΗL nanopores might be used at high concentrations to modulate signal transmission with small molecules, 26 , 44 , 45 light, 46 or other stimuli, 47 , 48 enabling future structures to perform more intricate tasks. The processors in this work have been constructed manually and would benefit from high-throughput production methods such as microfluidics 24 , 25 , 49 and 3D-printing. 16 , 20 These methods would eliminate the need for assembly by manual manipulation using a Teflon-coated silver wire.…”

“…Accordingly, multicompartment structures consisting of aqueous droplet compartments inside a lipid-containing oil drop have been generated in water. 23 − 25 The compartments are connected to each other and the external environment through lipid bilayers. Multicompartment structures have been formed that degrade by design in response to an external pH or temperature change.…”

“…Networks of droplets interconnected by these droplet interface bilayers (DIBs) can be generated manually , or by 3D-printing. , Within oil, these structures can respond to external light, , or mechanical or electrical stimuli, but to handle inputs from water-soluble signaling molecules, multicompartment structures must operate in an aqueous environment. Accordingly, multicompartment structures consisting of aqueous droplet compartments inside a lipid-containing oil drop have been generated in water. − The compartments are connected to each other and the external environment through lipid bilayers. Multicompartment structures have been formed that degrade by design in response to an external pH or temperature change .…”

Advanced Microfluidic Technologies for Lipid Nano-Microsystems from Synthesis to Biological Application

“…Droplet microfluidics has also been reported for multicompartment vesicles production [156,159,161]. These multisomal systems, also known as vesosomes or vesicles-invesicles, have been reported as alternative systems with high potential as advanced drug delivery vehicles, bioreactors, and artificial cells.…”

Microfluidic production of monodisperse emulsions for cosmetics

“…If an emulsion with the same droplet size is uniformly dispersed in another liquid phase and has a sufficient size for direct observation, its applicability is very high in the cosmetic industry. Due to these advantages, many studies have been conducted for designing uniform sized emulsion drops based on a microfluidic technology, employing various surface-active substances such as low molecular surfactants, 8-10 polymeric surfactants, [11][12][13] and colloidal particles. [14][15][16][17] For visual enjoyment that comes from the appearance of emulsion products, several cosmetic companies have attempted to generate monodisperse emulsions while varying their size, shape, and texture.…”