RESUMO
We report a microfluidic approach for one-step fabrication of polyelectrolyte microcapsules in aqueous conditions. Using two immiscible aqueous polymer solutions, we generate transient water-in-water-in-water double emulsion droplets and use them as templates to fabricate polyelectrolyte microcapsules. The capsule shell is formed by the complexation of oppositely charged polyelectrolytes at the immiscible interface. We find that attractive electrostatic interactions can significantly prolong the release of charged molecules. Moreover, we demonstrate the application of these microcapsules in encapsulation and release of proteins without impairing their biological activities. Our platform should benefit a wide range of applications that require encapsulation and sustained release of molecules in aqueous environments.
Assuntos
Fluoresceína/química , Técnicas Analíticas Microfluídicas , Polieletrólitos/química , Estreptavidina/química , Cápsulas/química , Tamanho da Partícula , Eletricidade Estática , Propriedades de Superfície , Água/químicaRESUMO
Supramolecular microgel capsules based on polyethylene glycol (PEG) are a promising class of soft particulate scaffolds with tailored properties. An approach to fabricate such particles with exquisite control by droplet-based microfluidics is presented. Linear PEG precursor polymers that carry bipyridine moieties on both chain termini are gelled by complexation to iron(II) ions. To investigate the biocompatibility of the microgels, living mammalian cells are encapsulated within them. The microgel elasticity is controlled by using PEG precursors of different molecular weights at different concentrations and the influence of these parameters on the cell viabilities, which can be optimized to exceed 90% is studied. Reversion of the supramolecular polymer cross-linking allows the microcapsules to be degraded at mild conditions with no effect on the viability of the encapsulated and released cells.
Assuntos
Hidrogéis/química , Ferro/química , Teste de Materiais , Técnicas Analíticas Microfluídicas , Polietilenoglicóis/química , Animais , Cápsulas , Sobrevivência Celular , Células Imobilizadas/citologia , Células Imobilizadas/metabolismo , Humanos , Células K562 , CamundongosRESUMO
pH-Cleavable cell-laden microgels with excellent long-term viabilities were fabricated by combining bioorthogonal strain-promoted azide-alkyne cycloaddition (SPAAC) and droplet-based microfluidics. Poly(ethylene glycol)dicyclooctyne and dendritic poly(glycerol azide) served as bioinert hydrogel precursors. Azide conjugation was performed using different substituted acid-labile benzacetal linkers that allowed precise control of the microgel degradation kinetics in the interesting pH range between 4.5 and 7.4. By this means, a pH-controlled release of the encapsulated cells was achieved upon demand with no effect on cell viability and spreading. As a result, the microgel particles can be used for temporary cell encapsulation, allowing the cells to be studied and manipulated during the encapsulation and then be isolated and harvested by decomposition of the microgel scaffolds.
Assuntos
Sobrevivência Celular/fisiologia , Microfluídica/métodos , Polietilenoglicóis/química , Química Click , Microscopia ConfocalRESUMO
Micrometer-sized hydrogel particles that contain living cells can be fabricated with exquisite control through the use of droplet-based microfluidics and bioinert polymers such as polyethyleneglycol (PEG) and hyperbranched polyglycerol (hPG). However, in existing techniques, the microgel gelation is often achieved through harmful reactions with free radicals. This is detrimental for the viability of the encapsulated cells. To overcome this limitation, we present a technique that combines droplet microfluidic templating with bio-orthogonal thiol-ene click reactions to fabricate monodisperse, cell-laden microgel particles. The gelation of these microgels is achieved via the nucleophilic Michael addition of dithiolated PEG macro-cross-linkers to acrylated hPG building blocks and does not require any initiator. We systematically vary the microgel properties through the use of PEG linkers with different molecular weights along with different concentrations of macromonomers to investigate the influence of these parameters on the viability and proliferation of encapsulated yeast cells. We also demonstrate the encapsulation of mammalian cells including fibroblasts and lymphoblasts.
Assuntos
Géis , Microfluídica/métodos , Animais , Células Cultivadas , Cinética , Mamíferos , Saccharomyces cerevisiae/químicaRESUMO
Single cell-laden three-dimensional (3D) microgels that can serve to mimic stem cell niches in vitro, and are therefore termed microniches, can be efficiently fabricated by droplet-based microfluidics. In this technique an aqueous polymer solution along with a highly diluted cell solution is injected into a microfluidic device to create monodisperse pre-microgel droplets that are then solidified by a polymer crosslinking reaction to obtain monodisperse single cell-laden microniches. However, problems limiting this approach studying the fate of single cells include Poisson encapsulation statistics that result in mostly empty microniches, and cells egressing from the microniches during subsequent cell culture. Here, we present a strategy to bypass Poisson encapsulation statistics in synthetic microniches by selective crosslinking of only cell-laden pre-microgel droplets. Furthermore, we show that we can position cells in the center of the microniches, and that even in protease-sensitive microniches this greatly reduces cell egress. Collectively, we present the development of a versatile protocol that allows for unprecedented efficiency in creation of synthetic protease-sensitive microniches for probing single stem cell fate in 3D.
Assuntos
Técnicas de Cultura de Células/métodos , Microambiente Celular/fisiologia , Técnicas Analíticas Microfluídicas/métodos , Análise de Célula Única/métodos , Animais , Linhagem Celular , Camundongos , Peptídeo HidrolasesRESUMO
Hybrid polymer-network gels built by both physical and covalent polymer crosslinking combine the advantages of both these crosslinking types: they exhibit high mechanical strength along with excellent fracture toughness and extensibility. If these materials are extensively deformed, their physical crosslinks can break such that strain energy is dissipated and irreversible fracturing is restricted to high strain only. This mechanism of energy dissipation is determined by the kinetics and thermodynamics of the physical crosslinking contribution. In this paper, we present a poly(ethylene glycol) (PEG) based material toolkit to control these contributions in a rational and custom fashion. We form well-defined covalent polymer-network gels with regularly distributed additional supramolecular mechanical fuse links, whose strength of connectivity can be tuned without affecting the primary polymer-network composition. This is possible because the supramolecular fuse links are based on terpyridineâ»metal complexation, such that the mere choice of the fuse-linking metal ion adjusts their kinetics and thermodynamics of complexationâ»decomplexation, which directly affects the mechanical properties of the hybrid gels. We use oscillatory shear rheology to demonstrate this rational control and enhancement of the mechanical properties of the hybrid gels. In addition, static light scattering reveals their highly regular and well-defined polymer-network structures. As a result of both, the present approach provides an easy and reliable concept for preparing hybrid polymer-network gels with rationally designed properties.
RESUMO
Stimuli-responsive hydrogels are able to change their physical properties such as their elastic moduli in response to changes in their environment. If biocompatible polymers are used to prepare such materials and if living cells are encapsulated within these networks, their switchability allows the cell-matrix interactions to be investigated with unprecedented consistency. In this paper, thermo-responsive macro- and microscopic hydrogels are presented based on azide-functionalized copolymers of poly(N-(2-hydroxypropyl)-methacrylamide) and poly(hydroxyethyl methacrylate) grafted with poly(N-isopropylacrylamide) side chains. Crosslinking of these comb polymers is realized by bio-orthogonal strain-promoted azide-alkyne cycloaddition with cyclooctyne-functionalized poly(ethylene glycol). The resulting hybrid hydrogels exhibit thermo-tunable elasticity tailored by the polymer chain length and grafting density. This bio-orthogonal polymer crosslinking strategy is combined with droplet-based microfluidics to encapsulate living cells into stimuli-responsive microgels, proving them to be a suitable platform for future systematic stem-cell research.
Assuntos
Hidrogéis/química , Resinas Acrílicas/química , Animais , Azidas/química , Materiais Biocompatíveis/química , Elasticidade , Concentração de Íons de Hidrogênio , Camundongos , Microfluídica , Células NIH 3T3 , Polietilenoglicóis/química , Polímeros/química , Ácidos Polimetacrílicos/químicaRESUMO
Biohybrid hydrogels that change their mechanical properties in response to pharmacological cues hold high promises as externally controlled drug depots for biomedical applications. In this study, we devise a generically applicable method for the synthesis of micrometer-scale, injection-ready biohybrid materials. We use droplet-based microfluidics to generate monodisperse pre-microgel fluid droplets, wherein which we react fluorescein-modified 8-arm poly(ethylene glycol) with a thiol-functionalized humanized anti-fluorescein single chain antibody fragment and vinylsulfone-functionalized 8-arm poly(ethylene glycol), resulting in the formation of stable, narrowly dispersed supramolecular microgels (30 and 150 µm diameter). We demonstrate that the addition of free fluorescein to these microgels results in a weakening of their hydrogel structure, eventually leading to its disintegration. This method of formation of pharmacologically responsive biohybrid hydrogels in an injection-ready formulation is a pioneering example of a general approach for the synthesis of biohybrid hydrogel-based drug depots for biomedical applications.
Assuntos
Portadores de Fármacos/química , Portadores de Fármacos/síntese química , Hidrogéis/química , Hidrogéis/síntese química , Técnicas Analíticas Microfluídicas/métodos , Polietilenoglicóis/química , Anticorpos de Cadeia Única/químicaRESUMO
Tin dioxide coatings are widely applied in glasses and ceramics to improve not only optical, but also mechanical properties. In this work, we report a new method to prepare SnO(2) coatings from aqueous dispersions of polymer/organotin hybrid nanoparticles. Various liquid organotin compounds were encapsulated in polymeric nanoparticles synthesized by miniemulsion polymerization. Large amounts of tetrabutyltin and bis(tributyltin) could be successfully incorporated in cross-linked and noncross-linked polystyrene nanoparticles that served as sacrificial templates for the formation of tin oxide coatings after etching with oxygen plasma or calcination. Cross-linked polystyrene particles containing bis(tributyltin)--selected for having a high boiling point--were found to be especially suited for the oxide coating formation. The content of metal in the particles was up to 12 wt %, and estimations by thermogravimetrical indicated that at least 96% of the total organotin compound was converted to SnO(2). The resulting coatings were mainly identified as tetragonal SnO(2) (cassiterite) by X-ray diffraction, although a coexistence of this phase with orthorhombic SnO(2) was observed for samples prepared with bis(tributyltin).