Exploring New Horizons in Liquid Compartmentalization via Microfluidics.

Spatial organization of cellular processes is crucial to efficiently regulate life's essential reactions. Nature does this by compartmentalization, either using membranes, such as the cell and nuclear membrane, or by liquid-like droplets formed by aqueous liquid-liquid phase separation. Aqueous liquid-liquid phase separation can be divided in two different phenomena, associative and segregative phase separation, of which both are studied for their membraneless compartmentalization abilities. For centuries, segregative phase separation has been used for the extraction and purification of biomolecules. With the emergence of microfluidic techniques, further exciting possibilities were explored because of their ability to fine-tune phase separation within emulsions of various compositions and morphologies and achieve one of the simplest forms of compartmentalization. Lately, interest in aqueous liquid-liquid phase separation has been revived due to the discovery of membraneless phases within the cell. In this Perspective we focus on segregative aqueous phase separation, discuss the theory of this interesting phenomenon, and give an overview of the evolution of aqueous phase separation in microfluidics.

High-Throughput Design of Biocompatible Enzyme-Based Hydrogel Microparticles with Autonomous Movement.

Micro- and nanomotors and their use for biomedical applications have recently received increased attention. However, most designs use top-down methods to construct inorganic motors, which are labour-intensive and not suitable for biomedical use. Herein, we report a high-throughput design of an asymmetric hydrogel microparticle with autonomous movement by using a microfluidic chip to generate asymmetric, aqueous, two-phase-separating droplets consisting of poly(ethylene glycol) diacrylate (PEGDA) and dextran, with the biocatalyst placed in the PEGDA phase. The motor is propelled by enzyme-mediated decomposition of fuel. The speed of the motors is influenced by the roughness of the PEGDA surface after diffusion of dextran and was tuned by using higher molecular weight dextran. This roughness allows for easier pinning of oxygen bubbles and thus higher speeds of the motors. Pinning of bubbles occurs repeatedly at the same location, thereby resulting in constant circular or linear motion.

Anisotropic, Hydrogel Microparticles as pH-Responsive Drug Carriers for Oral Administration of 5-FU.

In the last 20 years, the development of stimuli-responsive drug delivery systems (DDS) has received great attention. Hydrogel microparticles represent one of the candidates with the most potential. However, if the role of the cross-linking method, polymer composition, and concentration on their performance as DDS has been well-studied, still, a lot needs to be explained regarding the effect caused by the morphology. To investigate this, herein, we report the fabrication of PEGDA-ALMA-based microgels with spherical and asymmetric shapes for 5-fluorouracil (5-FU) on-demand loading and in vitro pH-triggered release. Due to anisotropic properties, the asymmetric particles showed an increased drug adsorption and higher pH responsiveness, which in turn led to a higher desorption efficacy at the target pH environment, making them an ideal candidate for oral administration of 5-FU in colorectal cancer. The cytotoxicity of empty spherical microgels was higher than the cytotoxicity of empty asymmetric microgels, suggesting that the gel network's mechanical proprieties of anisotropic particles were a better three-dimensional environment for the vital functions of cells. Upon treatment with drug-loaded microgels, the HeLa cells' viability was lower after incubation with asymmetric particles, confirming a minor release of 5-FU from spherical particles.

Spatial Control over Catalyst Positioning for Increased Micromotor Efficiency.

Motion is influenced by many different aspects of a micromotor's design, such as shape, roughness and the type of materials used. When designing a motor, asymmetry is the main requirement to take into account, either in shape or in catalyst distribution. It influences both speed and directionality since it dictates the location of propulsion force. Here, we combine asymmetry in shape and asymmetry in catalyst distribution to study the motion of soft micromotors. A microfluidic method is utilized to generate aqueous double emulsions, which upon UV-exposure form asymmetric microgels. Taking advantage of the flexibility of this method, we fabricated micromotors with homogeneous catalyst distribution throughout the microbead and micromotors with different degrees of catalyst localization within the active site. Spatial control over catalyst positioning is advantageous since less enzyme is needed for the same propulsion speed as the homogeneous system and it provides further confinement and compartmentalization of the catalyst. This proof-of-concept of our new design will make the use of enzymes as driving forces for motors more accessible, as well as providing a new route for compartmentalizing enzymes at interfaces without the need for catalyst-specific functionalization.

Adaptive insertion of a hydrophobic anchor into a poly(ethylene glycol) host for programmable surface functionalization.

Covalent and non-covalent molecular binding are two strategies to tailor surface properties and functions. However, the lack of responsiveness and requirement for specific binding groups makes spatiotemporal control challenging. Here, we report the adaptive insertion of a hydrophobic anchor into a poly(ethylene glycol) (PEG) host as a non-covalent binding strategy for surface functionalization. By using polycyclic aromatic hydrocarbons as the hydrophobic anchor, hydrophilic charged and non-charged functional modules were spontaneously loaded onto PEG corona in 2 min without the assistance of any catalysts and binding groups. The thermodynamically favourable insertion of the hydrophobic anchor can be reversed by pulling the functional module, enabling programmable surface functionalization. We anticipate that the adaptive molecular recognition between the hydrophobic anchor and the PEG host will challenge the hydrophilic understanding of PEG and enhance the progress in nanomedicine, advanced materials and nanotechnology.

Reversible speed control of one-stimulus-double-response, temperature-sensitive asymmetric hydrogel micromotors.

Soft, one-stimulus-double-response, thermo-sensitive, PNIPAm-based microgels are designed for controlled autonomous motion under stimuli. At higher temperature, the motors with physically encapsulated catalase move faster, while motors in which catalase is chemically linked to PNIPAm ceased moving. The phenomenon is reversible over multiple cycles of temperature.