Injectable Microfluidic Hydrogel Microspheres for Cell and Drug Delivery

Summary Cell-free extract derived from the eggs of the African clawed frog Xenopus laevis is a well-established model system that has been used historically in bulk aliquots. Here, we describe a microfluidic approach for isolating discrete, biologically relevant volumes of cell-free extract, with more expansive and precise control of extract shape compared with extract-oil emulsions. This approach is useful for investigating the mechanics of intracellular processes affected by cell geometry or cytoplasmic volume, including organelle scaling and positioning mechanisms. For complete details on the use and execution of this protocol, please refer to Geisterfer et al. (2020) .

Microfluidic encapsulation of Xenopus laevis cell-free extracts using hydrogel photolithography.

Facile Modification on the Oxygen Inhibited Layer of Photopolymerized Acrylates via Aza‐Michael Addition

Interfacially-mediated oxygen inhibition for precise and continuous poly(ethylene glycol) diacrylate (PEGDA) particle fabrication.

Protein therapeutics have become increasingly popular for the treatment of a variety of diseases owing to their specificity to targets of interest. However, challenges associated with them have limited their use for a range of ailments, including the limited options available for local controlled delivery. To address this challenge, degradable hydrogel microparticles, or microgels, loaded with model biocargoes were created with tunable release profiles or triggered burst release using chemistries responsive to endogenous or exogeneous stimuli, respectively. Specifically, microfluidic flow-focusing was utilized to form homogenous microgels with different spontaneous click chemistries that afforded degradation either in response to redox environments for sustained cargo release or light for on-demand cargo release. The resulting microgels were an appropriate size to remain localized within tissues upon injection and were easily passed through a needle relevant for injection, providing means for localized delivery. Release of a model biopolymer was observed over the course of several weeks for redox-responsive formulations or triggered for immediate release from the light-responsive formulation. Overall, we demonstrate the ability of microgels to be formulated with different materials chemistries to achieve various therapeutic release modalities, providing new tools for creation of more complex protein release profiles to improve therapeutic regimens.

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Microgels Formed by Spontaneous Click Chemistries Utilizing Microfluidic Flow Focusing for Cargo Release in Response to Endogenous or Exogenous Stimuli

Hydrogels have been engineered for a variety of biomedical applications, including biosensing, drug delivery, cell delivery, and tissue engineering. The fabrication of hydrogels into nanoscale and microscale particles has been a subject of intense activity and promises to extend their range of applicability. As hydrogels are reduced in size, their interfacial properties represent an increasingly significant contribution to their function and behavior. Hydrogel microparticle-based biosensing and drug delivery platforms, for instance, requires delicate spatial control over the conjugation of biofunctional groups and network architecture, which impacts their mechanical properties and molecular permeability and diffusivity. Here, we demonstrate the ability to tune, with extraordinary precision, the interfacial properties of PEGDA particles generated in a droplet microfluidic device exploiting oxygen-inhibited photopolymerization. We demonstrate the broad utility of these engineered microgels by creating spherical particles with complex but predictable radial crosslinking density gradients. Immunoassays were conducted to examine the network properties of these particles, revealing a high degree of structural tenability, which, in turn, dictates macromolecule encapsulation and release profiles, as well as the presence of radial crosslinking gradients that impact the availability of functional groups.

Structured Hydrogel Particles With Nanofabricated Interfaces via Controlled Oxygen Inhibition

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Structured Hydrogel Particles With Nanofabricated Interfaces via Controlled Oxygen Inhibition