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At least 19 records

Fabrication of poly(ethylene glycol) hydrogel microstructures using photolithography

The fabrication of hydrogel microstructures based upon poly(ethylene glycol) diacrylates, dimethacrylates, and tetraacrylates patterned photolithographically on silicon or glass substrates is described. A silicon/silicon dioxide surface was treated with 3-(trichlorosilyl)propyl methacrylate to form a self-assembled monolayer (SAM) with pendant acrylate groups. The SAM presence on the surface was verified using ellipsometry and time-of-flight secondary ion mass spectrometry. A solution containing an acrylated or methacrylated poly(ethylene glycol) derivative and a photoinitiator (2,2-dimethoxy-2-phenylacetophenone) was spin-coated onto the treated substrate, exposed to 365 nm ultraviolet light through a photomask, and developed with either toluene, water, or supercritical CO2. As a result of this process, three-dimensional, cross-linked PEG hydrogel microstructures were immobilized on the surface. Diameters of cylindrical array members were varied from 600 to 7 micrometers by the use of different photomasks, while height varied from 3 to 12 micrometers, depending on the molecular weight of the PEG macromer. In the case of 7 micrometers diameter elements, as many as 400 elements were reproducibly generated in a 1 mm2 square pattern. The resultant hydrogel patterns were hydrated for as long as 3 weeks without delamination from the substrate. In addition, micropatterning of different molecular weights of PEG was demonstrated. Arrays of hydrogel disks containing an immobilized protein conjugated to a pH sensitive fluorophore were also prepared. The pH sensitivity of the gel-immobilized dye was similar to that in an aqueous buffer, and no leaching of the dye-labeled protein from the hydrogel microstructure was observed over a 1 week period. Changes in fluorescence were also observed for immobilized fluorophore labeled acetylcholine esterase upon the addition of acetyl acholine.

NASA Discipline Life Sciences Technologies

Poly(ethylene glycol) hydrogel microstructures encapsulating living cells

We present an easy and effective method for the encapsulation of cells inside PEG-based hydrogel microstructures fabricated using photolithography. High-density arrays of three-dimensional microstructures were created on substrates using this method. Mammalian cells were encapsulated in cylindrical hydrogel microstructures of 600 and 50 micrometers in diameter or in cubic hydrogel structures in microfluidic channels. Reducing lateral dimension of the individual hydrogel microstructure to 50 micrometers allowed us to isolate 1-3 cells per microstructure. Viability assays demonstrated that cells remained viable inside these hydrogels after encapsulation for up to 7 days.

Non-NASA Center

Hydrogels with covalent and noncovalent crosslinks

A method for targeted delivery of therapeutic compounds from hydrogels is presented. The method involves administering to a cell a hydrogel in which a therapeutic compound is noncovalently bound to heparin. The hydrogel may contain covalent and non-covalent crosslinks.

Kilck, Kristi L.

Enabling Single Cell Research in Hollow Hydrogel Microparticles

Studying biology at the single cell level enables novel discoveries through ultra-high throughput identification and isolation of rare cells and interesting subpopulations. Encapsulation of single cells in hollow hydrogel microparticles, or PicoShells, enables easy visualization, manipulation, and assaying of single cells. The PicoShell’s porous hydrogel shell allows diffusion of nutrients and wastes into and out of the hollow liquid core, allowing uninhibited cell growth. The porous shell also allows diffusion of assay reagents, enabling solution changes, and multi-step assays. Finally, single cells and their progeny can be tracked through time, for example to measure single cell growth rates. However, PicoShells are fabricated using microfluidic techniques, which require specialized expertise, equipment, and facilities. This limits PicoShell’s widespread utility for single cell biology. For Saccharomyces cerevisiae and other desiccation-tolerant microbes, desiccating PicoShells containing cells of interest offers a solution, enabling those without microfluidic capabilities to obtain dried, shelf-stable PicoShells containing their cells of interest from a specialized microfluidics facility. Researchers could then rehydrate the PicoShells and perform their single cell assays, bypassing the microfluidic steps. Desiccated PicoShells could also be flown in space, where desiccation is already a common practice for microbes (i.e. BioSentinel and BioNutrients), bringing single cell analysis capabilities to space. In this work, we develop a method to desiccate PicoShells containing S. cerevisiae while maintaining characteristics similar to fresh, non-desiccated PicoShells. We show good microparticle morphology and hydrogel pore size as well as high yeast viability in line with previous studies. We also demonstrate sorting of a fast-growing yeast population using size-based filtration, as S. cerevisiae in PicoShells are able to physically stretch the microparticle as they grow, increasing the PicoShell diameter. These efforts contribute to the feasibility of leveraging microparticle PicoShells for single cell space biology research on Earth and in space.

Simon Ng

Use of Nanofibers to Strengthen Hydrogels of Silica, Other Oxides, and Aerogels

Research has shown that including up to 5 percent w/w carbon nanofibers in a silica backbone of polymer crosslinked aerogels improves its strength, tripling compressive modulus and increasing tensile stress-at-break five-fold with no increase in density or decrease in porosity. In addition, the initial silica hydrogels, which are produced as a first step in manufacturing the aerogels, can be quite fragile and difficult to handle before cross-linking. The addition of the carbon nanofiber also improves the strength of the initial hydrogels before cross-linking, improving the manufacturing process. This can also be extended to other oxide aerogels, such as alumina or aluminosilicates, and other nanofiber types, such as silicon carbide.

Meador, Mary Ann B.

Tensile Properties of Hydrogels and of Snake Skin

Stimulus-responsive or 'smart' gels are of potential interest as sensors and actuators, in industrial separations, and as permeable delivery systems. In most applications, a certain degree of mechanical strength and toughness will be required, yet the large-strain behavior of gels has not been widely reported. Some exceptions include work on gelatin and other food gels, some characterization of soft gels applicable for in-vitro cell growth studies, and toughness determinations on commercial contact lens materials. In general, it can be anticipated that the gel stiffness will increase with increasing degree of crosslinking, but the tensile strength may go through a maximum. Gel properties can be tailored by varying not only the degree of crosslinking, but also the polymer concentration and the nature of the polymer backbone (e.g. its stiffness or solubility). Polypeptides provide an especially interesting case, where secondary structure affects trends in moduli and conformational transitions may accompany phase changes. A few papers on the tensile properties of responsive gels have begun to appear. The responsive hydrogel chosen for the present study, crosslinked hydroxypropylcellulose, shrinks over a rather narrow temperature range near 44 C. Some vertebrate skin is also subject to substantial strain. Among reptiles, the morphologies of the skin and scales show wide variations. Bauer et al. described the mechanical properties and histology of gecko skin; longitudinal tensile properties of snake skin were examined by Jayne with reference to locomotion. The present measurements focus on adaptations related to feeding, including the response of the skin to circumferential tension. Tensile properties will be related to interspecific and regional variation in skin structure and folding.

Hinkley, Jeffrey A.

Tensile Properties of a Cellulose Ether Hydrogel

Poly(hydroxycellulose) solutions were molded into dumbell-shaped specimens crosslinked with divinyl sulfone. The resulting hydrogels were tested in tension at room temperature and also at a temperature above the 40 C shrinkage transition. In contrast to behavior seen in some other responsive gels, apparent initial tangent moduli were lower in the shrunken state; breaking elongations were significantly higher. Possible molecular mechanisms are suggested, and implications for the design of temperature-responsive actuators ("artificial muscles") from this material are discussed.

Hinkley, Jeffrey A.

Conductive hydrogel containing 3-ionene

Cationic polyelectrolytes formed by the polymerization in absence of oxygen of a monomer of the general formula: dispersed ##STR1## where x is 3 or more than 6 and Z is I, Br or Cl to form high charge density linear polymers are dispered in a water-soluble polymer such as polyvinyl alcohol to form a conductive hydrogel.

Rembaum, Alan

Single-Cell Analysis of Yeast (Saccharomyces cerevisiae) Using Hydrogel Encapsulation

Space radiation poses a major health risk to astronauts. To fulfill NASA’s mission of exploration beyond Earth, the biological effects of Galactic Cosmic Radiation and gamma radiation must be investigated to elucidate cellular damage mechanisms and inform countermeasure protocols to safely bring humans beyond Earth’s magnetosphere. Budding yeast (Saccharomyces cerevisiae) are commonly used in experiments as a model organism for studying the effects of radiation on eukaryotes. Radiobiology of yeast at the single cell level is poorly understood, yet crucial for informing models to aid in the design and interpretation of experiments. We are using a novel method of microencapsulation in hydrogel particles (PicoShells) to enable analysis of the distribution of radiation-induced damage among yeast cells at the single-cell level, in high throughput. Here we describe the development of methods for culturing, visualization, and quantification of encapsulated yeast. The encapsulated yeast are cultured in Yeast extract-Peptone-Dextrose (YPD) medium, fixed in formaldehyde or ethanol, and stained with DAPI or propidium iodide, then visualized using microscopy or enumerated using flow cytometry, with the aim of developing a protocol to enumerate the distribution of viable cells in each PicoShell. This will allow us to quantify how different forms of radiation can generate different distributions of damage across a population of cells, ultimately providing insight into the biological effects of space-relevant ionizing radiation.

yeast

Preparation and characterization of methacrylate hydrogels for zeta potential control

A technique based on the measurement of streaming potentials has been developed to evaluate the effects of hydrophilic coatings on electroosmotic flow. The apparatus and procedure are described as well as some results concerning the electrokinetic potential of glass capillaries as a function of ionic strength, pH, and temperature. The effect that turbulence and entrance flow conditions have on accurate streaming potential measurements is discussed. Various silane adhesion promoters exhibited only a slight decrease in streaming potential. A coating utilizing a glycidoxy silane base upon which methylcellulose is applied affords a six-fold decrease over uncoated tubes. Hydrophilic methacrylate gels show similar streaming potential behavior, independent of the water content of the gel. By introduction of positive or negative groups into the hydrophilic methacrylate gels, a range of streaming potential values are obtained having absolute positive or negative signs.

Gregonis, D. E.

Design of intelligent mesoscale periodic array structures utilizing smart hydrogel

Mesoscale Periodic Array Structures (MPAS, also known as crystalline colloidal arrays), composed of aqueous or nonaqueous dispersions of self-assembled submicron colloidal spheres are emerging toward the development of advanced optical devices for technological applications. This is because of their unique optical diffraction properties and the ease with which these intriguing properties can be modulated experimentally. Moreover our recent advancements in this area which include 'locking' the liquid MPAS into solid or semisolid polymer matrices for greater stability with longer life span, and incorporation of CdS quantum dots and laser dyes into colloidal spheres to obtain nonlinear optical (NLO) responses further corroborate the use of MPAS in optical technology. Our long term goal is fabrication of all-optical and electro-optical devices such as spatial light modulators for optical signal processing and flat panel display devices by utilizing intelligent nonlinear periodic array structural materials. Here we show further progress in the design of novel linear MPAS which have the ability to sense and respond to an external source such as temperature. This is achieved by combining the self-assembly properties of polymer colloidal spheres and thermoshrinking properties of smart polymer gels. At selected temperatures the periodic array efficiently Bragg diffracts light and transmits most of the light at other temperatures. Hence these intelligent systems are of potential use as fixed notch filters optical switches or limiters to protect delicate optical sensors from high intensity laser radiation.

Sunkara, H. B.

Templated native silk smectic gels

One aspect of the present invention relates to a method of preparing a fibrous protein smectic hydrogel by way of a solvent templating process, comprising the steps of pouring an aqueous fibrous protein solution into a container comprising a solvent that is not miscible with water; sealing the container and allowing it to age at about room temperature; and collecting the resulting fibrous protein smectic hydrogel and allowing it to dry. Another aspect of the present invention relates to a method of obtaining predominantly one enantiomer from a racemic mixture, comprising the steps of pouring an aqueous fibrous protein solution into a container comprising a solvent that is not miscible with water; sealing the container and allowing it to age at about room temperature; allowing the enantiomers of racemic mixture to diffuse selectively into the smectic hydrogel in solution; removing the smectic hydrogel from the solution; rinsing predominantly one enantiomer from the surface of the smectic hydrogel; and extracting predominantly one enantiomer from the interior of the smectic hydrogel. The present invention also relates to a smectic hydrogel prepared according to an aforementioned method.

Jin, Hyoung-Joon

Templated Native Silk Smectic Gels

One aspect of the present invention relates to a method of preparing a fibrous protein smectic hydrogel by way of a solvent templating process, comprising the steps of pouring an aqueous fibrous protein solution into a container comprising a solvent that is not miscible with water; sealing the container and allowing it to age at about room temperature; and collecting the resulting fibrous protein smectic hydrogel and allowing it to dry. Another aspect of the present invention relates to a method of obtaining predominantly one enantiomer from a racemic mixture, comprising the steps of pouring an aqueous fibrous protein solution into a container comprising a solvent that is not miscible with water; sealing the container and allowing it to age at about room temperature; allowing the enantiomers of racemic mixture to diffuse selectively into the smectic hydrogel in solution; removing the smectic hydrogel from the solution; rinsing predominantly one enantiomer from the surface of the smectic hydrogel; and extracting predominantly one enantiomer from the interior of the smectic hydrogel. The present invention also relates to a smectic hydrogel prepared according to an aforementioned method.

Jin, Hyoung-Joon

Templated Native Silk Smectic Gels

One aspect of the present invention relates to a method of preparing a fibrous protein smectic hydrogel by way of a solvent templating process, comprising the steps of pouring an aqueous fibrous protein solution into a container comprising a solvent that is not miscible with water; sealing the container and allowing it to age at about room temperature; and collecting the resulting fibrous protein smectic hydrogel and allowing it to dry. Another aspect of the present invention relates to a method of obtaining predominantly one enantiomer from a racemic mixture, comprising the steps of pouring an aqueous fibrous protein solution into a container comprising a solvent that is not miscible with water; sealing the container and allowing it to age at about room temperature; allowing the enantiomers of racemic mixture to diffuse selectively into the smectic hydrogel in solution; removing the smectic hydrogel from the solution; rinsing predominantly one enantiomer from the surface of the smectic hydrogel; and extracting predominantly one enantiomer from the interior of the smectic hydrogel. The present invention also relates to a smectic hydrogel prepared according to an aforementioned method.

Jin, Hyoung-Joon

Optimizing the calcium content of a copolymer acrylamide gel matrix for dark-grown seedlings

A copolymer acrylamide acrylate gel was investigated as the sole root matrix for dark-grown seedlings of soybean (Glycine max Merr. 'Century 84'). Increasing Ca2+ in the hydrating solution of the hydrogel from 1 to 10 mM decreased its water-holding capacity from 97 to 46 mL g-1, yet water potential of the medium remained high, sufficient for normal plant growth at all Ca2+ concentrations tested. Elongation rate of dark-grown soybean seedlings over a 54-hour period was 0.9, 1.5, and 1.8 mm h-1 with 1.0, 2.5, or 5.0 mM Ca2+, respectively, but did not increase with further increases in Ca2+ concentration. Further study revealed that Na+ was released from the hydrogel medium and was taken up by the seedlings as Ca2+ increased in the medium. In dry hypocotyl tissue, sodium content correlated negatively with calcium content. Despite the presence of Na+ in the hydrogel, seedling growth was normal when adequate Ca2+ was added in the hydrating solution. Acrylamide hydrogels hold good potential as a sole growth matrix for short-term experiments with dark-grown seedlings without irrigation.

NASA Discipline Plant Biology

Alkali-metal silicate binders and methods of manufacture

A paint binder is described which uses a potassium or sodium silicate dispersion having a silicon dioxide to alkali-metal oxide mol ratio of from 4.8:1 to 6.0:1. The binder exhibits stability during both manufacture and storage. The process of making the binder is predictable and repeatable and the binder may be made with inexpensive components. The high mol ratio is achieved with the inclusion of a silicon dioxide hydrogel. The binder, which also employs a silicone, is in the final form of a hydrogel sol.

Schutt, J. B.

High Resolution, Single-Step Patterning of Silica Aerogels

Three-dimensional metallic structures are fabricated with high spatial resolution in silica aerogels. In our method, silica hydrogels are prepared with a standard base-catalyzed route, and exchanged with an aqueous solution typically containing Ag' ions (1 M) and 2-propanol (0.2 M). The metal ions are reduced photolytically with a table-top ultraviolet lamp, or radiolytically, with a focused X-ray beam. We fabricated dots and lines as small as 30 x 70 km, protruding for several mm into the bulk of the materials. The hydrogels are eventually supercritically dried to yield aerogels, without any measurable change in the shape and spatial resolution of the lithographed structures. Transmission electron microscopy shows that illuminated regions are composed of Ag clusters with a size of several pm, separated by thin layers of silica.

Bertino, M. F.