microfluidic-fabrication-polymeric-biohybrid-fibers-with-predesigned

Microfluidic Fabrication of Polymeric and Biohybrid Fibers with Predesigned Size and Shape

A &#8220;sheath&#8221; fluid passing through a microfluidic channel at low Reynolds number can be directed around another &#8220;core&#8221; stream and ...

US Naval Research Laboratory

An alkyne-terminated anthracene and azide-terminated carbazole were joined through a copper-catalyzed cycloaddition to form a joined donor/acceptor pair. The photonic pair exhibited energy transfer when excited at the peak absorbance of carbazole and fluoresced with an anthracene spectral response. The fluorescent behavior was confirmed as Förster energy transfer (FRET). The lysate of Pseudomonas resinovorans CA10, a member of a predominant group of soil microorganisms that can metabolize a host of substrates, was employed to degrade the pair and alter the luminance spectral characteristics. The FRET was diminished and the corresponding, individual fluorescence of carbazole and anthracene returned. This general approach may find applications in single-cell metabolic studies and bioactivity assays.

Manipulation of Förster energy transfer of coupled fluorophores through biotransformation by Pseudomonas resinovorans CA10.

The isolation of a single type of protein from a complex mixture is vital for the characterization of the function, structure, and interactions of the protein of interest and is typically the most laborious aspect of the protein purification process. In this work, a model system is utilized to show the efficacy of synthesizing a "baited" nanoparticle to capture and recycle enzymes (proteins that catalyze chemical reactions) from crude cell lysate. Enzyme trapping and recycling is illustrated with the carbazole 1,9a-dioxygenase (CARDO) system, an enzyme important in bioremediation and natural product synthesis. The enzymes are baited with azide-modified carbazolyl moieties attached to poly(propargyl acrylate) nanoparticles through a click transformation. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF) and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis indicates the single-step procedure to immobilize the enzymes on the particles is capable of significantly concentrating the protein from raw lysate and sequestering all required components of the protein to maintain bioactivity. These results establish a universal model applicable to concentrating and extracting known substrate-protein pairs, but it can be an invaluable tool in recognizing unknown protein-ligand affinities.

Substrate-baited nanoparticles: a catch and release strategy for enzyme recognition and harvesting.

Composite nanoclusters with chemical, magnetic, and biofunctionality offer broad opportunities for targeted cellular imaging. A key challenge is to load a high degree of targeting, imaging, and therapeutic functionality onto stable metal-oxide nanoparticles. Here we report a route for producing magnetic nanoclusters (MNCs) with alkyne surface functionality that can be utilized as multimodal imaging probes. We form MNCs composed of magnetic Fe(3)O(4) nanoparticles and poly(acrylic acid-co-propargyl acrylate) by the co-precipitation of iron salts in the presence of copolymer stabilizers. The MNCs were surface-modified with near-infrared (NIR) emitting fluorophore used in photodynamic therapy, an azide-modified indocyanine green. The fluorophores engaged and complexed with bovine serum albumin, forming an extended coverage of serum proteins on the MNCs. These proteins isolated indocyanine green fluorophores from the aqueous environment and induced an effective "turn-on" of NIR emission.

Magnetic nanoclusters exhibiting protein-activated near-infrared fluorescence.

The integration of biological extracellular matrix (ECM) components and synthetic materials is a promising pathway to fabricate the next generation of hydrogel-based tissue scaffolds that more accurately emulate the microscale heterogeneity of natural ECM. We report the development of a bio/synthetic interpenetrating network (BioSINx), containing gelatin methacrylamide (GelMA) polymerized within a poly(ethylene glycol) (PEG) framework to form a mechanically robust network capable of supporting both internal cell encapsulation and surface cell adherence. The covalently crosslinked PEG network was formed by thiol-yne coupling, while the bioactive GelMA was integrated using a concurrent thiol-ene coupling reaction. The physical properties (i.e. swelling, modulus) of BioSINx were compared to both PEG networks with physically-incorporated gelatin (BioSINP) and homogenous hydrogels. BioSINx displayed superior physical properties and significantly lower gelatin dissolution. These benefits led to enhanced cytocompatibility for both cell adhesion and encapsulation; furthermore, the increased physical strength provided for the generation of a micro-engineered tissue scaffold. Endothelial cells showed extensive cytoplasmic spreading and the formation of cellular adhesion sites when cultured onto BioSINx; moreover, both encapsulated and adherent cells showed sustained viability and proliferation.

Interpenetrating networks based on gelatin methacrylamide and PEG formed using concurrent thiol click chemistries for hydrogel tissue engineering scaffolds.

Michael A. Daniele

Center for Bio/Molecular Science & Engineering

US Naval Research Laboratory

Microfluidic Fabrication of Polymeric and Biohybrid Fibers with Predesigned Size and Shape

Michael A. Daniele

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US Naval Research Laboratory

Microfluidic Fabrication of Polymeric and Biohybrid Fibers with Predesigned Size and Shape

Center for Bio/Molecular Science & Engineering